53
Hindawi Publishing Corporation Evidence-Based Complementary and Alternative Medicine Volume 2013, Article ID 340215, 51 pages http://dx.doi.org/10.1155/2013/340215 Review Article Bidens pilosa L. (Asteraceae): Botanical Properties, Traditional Uses, Phytochemistry, and Pharmacology Arlene P. Bartolome, 1,2 Irene M. Villaseñor, 1 and Wen-Chin Yang 2,3,4,5,6 1 Institute of Chemistry, University of the Philippines, Diliman, Quezon City, Philippines 2 Agricultural Biotechnology Research Center, Academia Sinica, Taipei 115, Taiwan 3 Institute of Pharmacology, Yang-Ming University, Taipei 112, Taiwan 4 Department of Life Sciences, National Chung-Hsing University, Taichung 402, Taiwan 5 Institute of Zoology, National Taiwan University, Taipei 106, Taiwan 6 Agricultural Biotechnology Research Center, Academia Sinica, No. 128, Academia Sinica Road, Section 2, Nankang, Taipei 11529, Taiwan Correspondence should be addressed to Wen-Chin Yang; [email protected] Received 31 January 2013; Accepted 29 April 2013 Academic Editor: Gail B. Mahady Copyright © 2013 Arlene P. Bartolome et al. is is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. ere are 230 to 240 known Bidens species. Among them, Bidens pilosa is a representative perennial herb, globally distributed across temperate and tropical regions. B. pilosa has been traditionally used in foods and medicines without obvious adverse effects. Despite significant progress in phytochemical and biological analyses of B. pilosa over the past few years, comprehensive and critical reviews of this plant are anachronistic or relatively limited in scope. e present review aims to summarize up-to-date information on the phytochemistry, pharmacology, and toxicology of B. pilosa from the literature. In addition to botanical studies and records of the traditional use of B. pilosa in over 40 diseases, scientific studies investigating the potential medicinal uses of this species and its constituent phytochemicals for a variety of disorders are presented and discussed. e structure, bioactivity, and likely mechanisms of action of B. pilosa and its phytochemicals are emphasized. Although some progress has been made, further rigorous efforts are required to investigate the individual compounds isolated from B. pilosa to understand and validate its traditional uses and develop clinical applications. e present review provides preliminary information and gives guidance for further basic and clinical research into this plant. 1. Introduction e United Nations World Health Organization estimates that as many as 5.6 billion people, 80% of the world popu- lation, utilize herbal medicine for primary health care [1]. 1 Plants have formed the foundation of complicated traditional medicine systems for thousands of years. Medicinal herbs are applied to treat a wide range of disease categories. e first written documentation of the use of medicinal herbs dates from the 26th century BCE in Mesopotamia, and the first record of the use of medicinal herbs by the Egyptians and Greeks dates from 18th century BCE and the 5th century BCE, respectively. Starting around the 11th century BCE, the Chinese and Indians started to develop herbal medicine systems, Chinese herbal medicine, and Ayurvedic medicine, respectively, that continue to be widely practiced today [2]. erefore, since antiquity, medicinal herbs have played a prominent role in human health. B. pilosa is an easy-to-grow herb that is widely distributed all over the world. It is considered to be a rich source of food and medicine for humans and animals [3, 4]. ere is increasing global interest in the use of B. pilosa as shown by the many studies conducted on the plant in recent years. e folkloric use of B. pilosa has been recorded in America, Africa, Asia, and Oceania [5]. To explore the potential clinical application of B. pilosa, it is important to link its traditional use with rigorous evidence-based scientific study. e present review focuses on recent studies on the botany, traditional

ReviewArticle Bidens pilosa L. (Asteraceae): Botanical

  • Upload
    others

  • View
    3

  • Download
    0

Embed Size (px)

Citation preview

Hindawi Publishing CorporationEvidence-Based Complementary and Alternative MedicineVolume 2013 Article ID 340215 51 pageshttpdxdoiorg1011552013340215

Review ArticleBidens pilosa L (Asteraceae) Botanical Properties TraditionalUses Phytochemistry and Pharmacology

Arlene P Bartolome12 Irene M Villasentildeor1 and Wen-Chin Yang23456

1 Institute of Chemistry University of the Philippines Diliman Quezon City Philippines2 Agricultural Biotechnology Research Center Academia Sinica Taipei 115 Taiwan3 Institute of Pharmacology Yang-Ming University Taipei 112 Taiwan4Department of Life Sciences National Chung-Hsing University Taichung 402 Taiwan5 Institute of Zoology National Taiwan University Taipei 106 Taiwan6Agricultural Biotechnology Research Center Academia Sinica No 128 Academia Sinica Road Section 2 Nankang Taipei 11529Taiwan

Correspondence should be addressed to Wen-Chin Yang wcyanggatesinicaedutw

Received 31 January 2013 Accepted 29 April 2013

Academic Editor Gail B Mahady

Copyright copy 2013 Arlene P Bartolome et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

There are 230 to 240 known Bidens species Among them Bidens pilosa is a representative perennial herb globally distributedacross temperate and tropical regions B pilosa has been traditionally used in foods and medicines without obvious adverse effectsDespite significant progress in phytochemical and biological analyses of B pilosa over the past few years comprehensive and criticalreviews of this plant are anachronistic or relatively limited in scopeThe present review aims to summarize up-to-date informationon the phytochemistry pharmacology and toxicology of B pilosa from the literature In addition to botanical studies and records ofthe traditional use of B pilosa in over 40 diseases scientific studies investigating the potential medicinal uses of this species and itsconstituent phytochemicals for a variety of disorders are presented and discussedThe structure bioactivity and likely mechanismsof action of B pilosa and its phytochemicals are emphasized Although some progress has been made further rigorous efforts arerequired to investigate the individual compounds isolated from B pilosa to understand and validate its traditional uses and developclinical applicationsThe present review provides preliminary information and gives guidance for further basic and clinical researchinto this plant

1 Introduction

The United Nations World Health Organization estimatesthat as many as 56 billion people 80 of the world popu-lation utilize herbal medicine for primary health care [1]1Plants have formed the foundation of complicated traditionalmedicine systems for thousands of years Medicinal herbs areapplied to treat a wide range of disease categories The firstwritten documentation of the use of medicinal herbs datesfrom the 26th century BCE in Mesopotamia and the firstrecord of the use of medicinal herbs by the Egyptians andGreeks dates from 18th century BCE and the 5th centuryBCE respectively Starting around the 11th century BCEthe Chinese and Indians started to develop herbal medicine

systems Chinese herbal medicine and Ayurvedic medicinerespectively that continue to be widely practiced today [2]Therefore since antiquity medicinal herbs have played aprominent role in human health

B pilosa is an easy-to-grow herb that is widely distributedall over the world It is considered to be a rich source offood and medicine for humans and animals [3 4] There isincreasing global interest in the use of B pilosa as shownby the many studies conducted on the plant in recent yearsThe folkloric use of B pilosa has been recorded in AmericaAfrica Asia andOceania [5] To explore the potential clinicalapplication of B pilosa it is important to link its traditionaluse with rigorous evidence-based scientific studyThe presentreview focuses on recent studies on the botany traditional

2 Evidence-Based Complementary and Alternative Medicine

Table 1 Taxonomy of B pilosa [6]

Kingdom PlantaeSubkingdom TracheobiontaSuperdivision SpermatophytaDivision MagnoliophytaClass MagnoliopsidaSubclass AsteridaeOrder AsteralesFamily AsteraceaeGenus BidensSpecies Bidens Pilosa L

usage phytochemistry pharmacology and toxicology of Bpilosa The information provided here highlights the pos-sible usefulness of B pilosa and its isolated compoundsand offers insights into possible future research directionsStudies of B pilosa are divided into three groups (1) thebotany ethnomedical uses plant chemistry pharmacologyand biosafety of B pilosa (2) scientific studies that validatethe ethnomedical uses ofB pilosa and (3) the therapeutic andfuture research potential of B pilosa

11 Botany B pilosa was first collected and named by CarlLinnaeus in 1753 [3] Taxonomically it is assigned to theBidens genus (Asteraceae) as shown in Table 1 This genus isestimated to include 230 to 240 species worldwide [3 4] Bpilosa has several varieties such as B pilosa var radiata varminor var pilosa and var bisetosa Alongside examination ofmorphological traits authentication of B pilosa can be aidedby chemotaxonomy and molecular characterization [8]

B pilosa is an erect perennial herb widely distributedacross temperate and tropical regions B pilosa is eitherglabrous or hairy with green opposite leaves that are serratelobed or dissected It has white or yellow flowers and longnarrow ribbed black achenes (seeds) It grows to an averageheight of 60 cm and a maximum of 150 cm in favorableenvironments [9] (Figure 1) B pilosa prefers full sun andmoderately dry soil However it can grow in arid and barrenland from low to high elevations With the advantage ofbeing fast-growing in the 1970s the Food and AgriculturalOrganization actively promoted the cultivation of B pilosain Africa [10] B pilosa propagates via seeds A single plantcan produce 3000ndash6000 seeds Dry mature seeds from Bpilosa can be germinated in 3 to 4 days in moist soil orafter being soaking in water Seeds are viable for at least 3years [11] Minimal agricultural techniques are required forB pilosa cultivation Due to its invasive tendencies B pilosais generally considered to be a weed [12]

B pilosa is thought to have originated in South Americaand subsequently spread all over the world [13] Bidensspecies and their varieties bear vernacular names based ontheir characteristics For example Bidens species are knownby such names as Spanish needles beggarrsquos ticks devilrsquosneedles cobblerrsquos pegs broom stick pitchforks and farmersrsquofriends in English and some other languages because of theirsticky achenes [9] and are sometimes known as xian feng cao

(ldquoall bountiful grassrdquo) in Chinese because of their prosperousgrowth

12 Traditional Uses B pilosa is used as an herb and asan ingredient in teas or herbal medicines Its shoots andleaves dried or fresh are utilized in sauces and teas [1415] In the 1970s the United Nations Food and AgricultureOrganization (FAO) promoted the cultivation of B pilosa inAfrica because it is easy to grow edible palatable and safe[7] The nutritional value of B pilosa is shown in Table 2

All parts of B pilosa plant the whole plant the aerialparts (leaves flowers seeds and stems) andor the rootsfresh or dried are used as ingredients in folk medicines It isfrequently prepared as a dry powder decoction macerationor tincture [16] Generally this plant is applied as drypowder or tincture when used externally and as a powdermaceration or decoction when used as an internal remedy[15]

As summarized in Table 3 B pilosa either as a wholeplant or different parts has been reported to be useful inthe treatment of more than 40 disorders such as inflamma-tion immunological disorders digestive disorders infectiousdiseases cancers metabolic syndrome wounds and manyothers [26ndash29] B pilosa is usually ingested however it canalso be utilized externally For instance fresh B pilosa is usedto treat snake bites and wounds [23] and in Trinidad andTobago the aqueous solution of the leaves of B pilosa is usedto bathe babies and children [22]

B pilosa is sometimes used alone it is also used asan ingredient in medicinal mixtures together with othermedicinal plants such as Aloe vera Plectranthus mollisValeriana officinalis andCissus sicyoides among others [17 1820 25] Whether the commonly used mixtures of B pilosaor the mixtures of the compounds found in B pilosa affordsynergistic effects is not yet clear and needs to be verifiedby further studies However one study has suggested thatB pilosa varieties share similar phytochemical compositionsand may be substituted for each other [8]

2 Phytochemicals

Interest in basic research and application of B pilosa hasincreased since its first identification in 1753 This is mainlydue to its wide application in medicines foods and drinksAround 116 publications have documented the exploitationand medical use of B pilosa To date 201 compoundscomprising 70 aliphatics 60 flavonoids 25 terpenoids 19phenylpropanoids 13 aromatics 8 porphyrins and 6 othercompounds have been identified from this plant as compiledpreviously [30] The structures of these compounds are pre-sented in Tables 4 5 6 7 8 9 and 10 respectively Howeverthe association between B pilosa phytochemicals and theirbioactivities is not yet fully established and should becomea future research focus In the present review we explorepossible associations (Table 11) describe the importance ofthe known compounds in relation to their biological activity

Evidence-Based Complementary and Alternative Medicine 3

Table 2 Nutritional facts about B pilosa courtesy of the United Nations Food and Agriculture Organization [7]

Plant(100 g)

Energy(kcal)

Moisture()

Protein(g)

Fat(g)

Carbohydrate(g)

Fiber(g)

Ash(g)

Calcium(mg)

Phosphorus(mg)

Iron(120583g)

Caroteneequivalent

(120583g)

Thiamine(mg)

Raw 43 851 38 05 84 39 22 340 67 mdash 1 800 mdashDried 33 886 28 06 6 13 2 111 39 23 mdash mdashldquomdashrdquo denotes not detectable

(a) (b)

(c)

Figure 1 B pilosa (a) and its flowers (b) and achenes (c)

and discuss their likelymechanisms of action (Table 3) Com-pelling evidence suggests that the various diverse bioactivitiesreported for B pilosa reflect its phytochemical complexity

B pilosa is an extraordinary source of phytochemicalsparticularly flavonoids and polyynes Plant flavonoids arecommonly reported to possess anticancer antiinflammatoryantioxidant and other bioactivities However the bioactivi-ties of only seven of the 60 flavonoids present in B pilosa havebeen studiedThe bioactivities of the remaining 53 flavonoidsare poorly understood and deserve further investigationOther classes of compounds found inB pilosa are also in needof further examination as shown in Table 11

3 Pharmacological Properties

As outlined in Table 3 B pilosa is traditionally used totreat a wide variety of ailments Different preparations ofits whole plant andor parts have been purported to treatover 40 categories of illnesses Scientific studies although notextensive have demonstrated that B pilosa extracts andorcompounds have antitumor [43 69 77ndash83] antiinflamma-tory [27 75 79 84ndash88] antidiabetic and antihyperglycemic[8 24 49 51 53] antioxidant [70 89 90] immunomodu-latory [50 69] antimalarial [47 77] antibacterial [47 9192] antifungal [92 93] antihypertensive vasodilatory [2894] and antiulcerative [26] activities In this section the

4 Evidence-Based Complementary and Alternative Medicine

Table 3 Ethnomedical information about B pilosa

Disorder Plant partb Mode of use Regioncountry References

Stomachache LE Decoction IndiaAfrica [17ndash19]

Colics WP Decoction AfricaChina [18]

Catarrh WP Juice or decoctiontaken orally Cuba [20]

Diarrhea LE andWP

Fresh leaves orDecoction

AfricaUganda [18 21]

Constipation WP Decoction Africa [18]DysenteryBacillarydysentery WP Decoction Africa

China [18]

Choleretic WP Decoction Middle America [18]

Anti-inflammatory WP Not stated ChinaCuba [20 22]

Asthma WPDecoction ormaceration takenorally

CubaChina [20 23]

Antirheumatic RT andWP Juice and decoction Hong Kong

Zulu Africa [18 22]

Acute appendicitis WP Decoction Hong Kong [23]

Enteritis WP Decoction AfricaChina [18]

Pruritus WP Decoction Hong Kong [23]

Conjunctivitis WP Decoction AfricaChina [18]

Otitis WP Decoction AfricaChina [18]

Pharyngitis WP Decoction AfricaChina [18]

Gastritis WP Juice taken orally Cuba [20]

Diabetes WP Decoction takenorally

CubaTaiwan [20 24]

Headache WP Decoction Bafia Cameroon [25]Diuretic WP Decoction Middle America [18]

Hypotensive WP Decoction takenorally Bafia Cameroon [25]

Colds LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21 23]

Yellow Fever LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21ndash23]

Influenza LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21 23]

Acute infectioushepatitis WP Decoction Hong Kong [23]

Intestinal worms WP Decoction Africa [18]

Malaria RT andWP Juice Africa

China [18 23]

Evidence-Based Complementary and Alternative Medicine 5

Table 3 Continued

Disorder Plant partb Mode of use Regioncountry References

Eye Infection LE andWP Fresh or juice Uganda

Middle America [18 21]

Antimicrobial APDecoction fordrinkingbathingexternal use

Trinidad andTobago [22]

Pulmonarytuberculosis WP

Decoction ormaceration takenorally

CubaChina [20 23]

Bacterial infectionsingastrointestinal tracts

WP Decoction Trinidad andTobago [22]

Renal infection LE Decoction takenorally Cuba [20]

Sore throat LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21 23]

Cough WP Decoction takenorally

CubaChina [20 23]

Coolness of theuterus WP Decoction taken

orally Cuba [20]

Menstrualirregularities WP Decoction taken

orally Cuba [20]

Dysmenorrhea WP Decoction Bafia Cameroon [25]Hyperemesisgravidarum(morning sickness)

WP Decoction Africa [18]

Hemorrhoids WP Decoction Hong Kong [23]

Nose bleeds LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21 23]

Stomach ulcers LE andWP

Maceration or juicetaken orally

CubaMiddle AmericaUganda

[18 20 21 23]

Cuts burns and skinproblems

LE andWP

Fresh plant ordecoction topicalapplicationbathing

Trinidad andTobagoAfricaChinaCameroonBrazilVenezuela

[18 22 26]

Wounds WP Crushed herb

ChinaAfricaCentral AmericaHawaii

[23]

Snake bites WP Pulverized herb China [23]bLE leaves ST stem FW flower WP whole plant RT roots AP aerial parts

primary pharmacological properties of B pilosa extracts andphytochemicals are presented and discussed

31 AntiCancer Activity Folkloric reports revealed the pos-sible antitumor efficacy of B pilosa and several scientific in2vitro studies have supported the claim that B pilosa extractsand isolated compounds possess anti-cancer activities againsta variety of cancer cells Several studies have used bioassay

guided isolation and fractionation methods to discover newcompounds from B pilosa For example Kviecinski andcolleagues tested hydroalcoholic crude extracts chloroformethyl acetate and methanol fractions for anti-tumor activity[82] The cytotoxicity of the extracts was assessed usingbrine shrimp hemolytic MTT and neutral red uptake(NRU) assays In vivo studies were performed using Ehrlichascites carcinoma in isogenic BALBc mice Among them

6 Evidence-Based Complementary and Alternative Medicine

Table 4 Aliphatic natural products isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

1 Heneicosane CH3(CH2)19CH3 Aerial (Tanzania) [18]2 Docosane CH3(CH2)20CH3 Aerial (Tanzania) [18]3 Tricosane CH3(CH2)21CH3 Aerial (Tanzania) [18]4 Tetracosane CH3(CH2)22CH3 Aerial (Tanzania) [18]5 Pentacosane CH3(CH2)23CH3 Aerial (Tanzania) [18]6 Hexacosane CH3(CH2)24CH3 Aerial (Tanzania) [18]7 Heptacosane CH3(CH2)25CH3 Aerial (Tanzania) [18]

8 Octacosone CH3(CH2)26CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

9 Nonacosane CH3(CH2)27CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

10 Triacontane CH3(CH2)28CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

11 Hentriacontane CH3(CH2)29CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

12 Dotriacontane CH3(CH2)30CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

13 Tritriacontane CH3(CH2)31CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

14 2-Butoxy-ethanol CH3(CH2)3OCH2CH2OH Whole (Taiwan) [32]15 Tetracosan-1-ol CH3(CH2)22CH2OH Aerial (Tanzania) [18]16 Hexacosan-1-ol CH3(CH2)24CH2OH Aerial (Tanzania) [18]17 1-Octacosanol CH3(CH2)26CH2OH Aerial (Tanzania) [18]

18 1-Hentriacontanol CH3(CH2)29CH2OHNot found(Taiwan) [31]

19 Tetradecanoic acid Myristic acid CH3(CH2)12CO2H Aerial (Tanzania) [18]20 Hexadecanoic acid Palmitic acid CH3(CH2)14CO2H Aerial (Tanzania) [18]21 Octadecanoic acid Stearic acid CH3(CH2)16CO2H Aerial (Tanzania) [18]22 Eicosanoic acid Arachidic acid CH3(CH2)18CO2H Aerial (Tanzania) [18]

23 Docosanoic acid Behenic acid CH3(CH2)20CO2HLeaves(Philippines) [33]

24 2-butenedioic acidO

OHO

HO Aerial (China) [34 35]

25 (Z)-9-Octadecenoicacid Oleic acid

O

OH Aerial (Tanzania) [18]

Evidence-Based Complementary and Alternative Medicine 7

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

26 (E)-9-Octadecenoicacid Elaidic acid

O

HOLeaves(Philippines) [33]

27 (ZZ)-912-Octadecadienoic acid

Linolic acid or Linoleicacid

O

OHAerial(Tanzania)whole (Taiwan)

[18 36]

28(ZZZ)-91215-Octadecatrienoicacid

120572-Linoleic acid

O

OH Whole (Taiwan) [32]

29(ZZ)-912-Octadecadienoic acidethyl ester

Ethyl linoleate O

O

Whole (Taiwan) [32]

30(ZZZ)-91215-Octadecatrienoicacid methyl ester

Methyl linolenate O

O

Whole (Taiwan) [32]

31(ZZZ)-91215-Octadecatrienoicacid ethyl ester

Ethyl linolenate O

O

Whole (Taiwan) [32]

32(Z)-9-Octadecenoicacid 2-butoxyethylester

2-Butoxyethyl oleate O

OO

Whole (Taiwan) [32]

33 2-Butoxyethyllinoleate O

OO Whole (Taiwan) [32]

34(ZZZ)-91215-Octadecatrienoicacid butoxyethyl ester

2-Butoxyethyl linoleate O

OO

Whole (Taiwan) [32]

3517E9E15E-Heptadecatetraene-1113-diyne

Heptadeca-2E8E10E16-tetraen-46-diyne

Not found(China) [37]

8 Evidence-Based Complementary and Alternative Medicine

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

36 111-Tridecadiene-3579-tetrayne

Roots (notstated) [38]

37 1-Tridecene-357911-pentayne Pentayneene

Leaves (notstated) andnot found(Egypt)

[38 39]

38 5-Tridecene-7911-triyne-3-ol

HO

not found(Egypt) [39]

39 21012-Tridecatriene-468-triyn-1-ol

OHPart not specified(Not stated) [40]

40 212-Tridecadiene-46810-tetrayn-1-ol

111-Tridecadiene-3579-

tetrayn-13-ol

OH Roots (Notstated) notfound(Egypt)

[38 39]

41 212-Tridecadiene-46810-tetraynal

111-Tridecadiene-3579-

tetrayn-13-al

O

Roots (Germany) [41]

42212-Tridecadiene-46810-tetrayn-1-ol1-acetate

111-Tridecadiene-3579-

tetrayn-13-acetateO

O

Roots (notstated) [38]

43(5E)-15-Tridecadiene-79-diyn-3413-triol OH

OH

HO

Aerial (China) [42]

44(6E12E)-3-oxo-tetradeca-612-dien-810-diyn-1-ol

OH

O

Aerial (China) [42]

45 (E)-5-Tridecene-7911-triyne-12-diol

12-Dihydroxy-5(E)-tridecene-7911-triyne

HO

OHWhole (Taiwan) [43]

46 (E)-6-Tetradecene-81012-triyne-13-diol

13-Dihydroxy-6(E)-tetradecene-81012-triyne

OH

OHWhole (Taiwan) [43ndash45]

47(2R3E11E)-311-Tridecadiene-579-triyne-12-diol

Safynol

OH

HO

Not found (Egyptand China) [37 39]

Evidence-Based Complementary and Alternative Medicine 9

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

4857911-Tridecatetrayne-12-diol

12-Dihydroxytrideca-57911-tetrayne

HOOH

Whole (Taiwan) [43 44 46]

49(R)-357911-Tridecapentayne-12-diol

(R)-12-Dihydroxytrideca-357911-pentayne

HO

OH

Aerial (Japan) [47]

50

(4E)-1-(Hydroxymethyl)-4-dodecene-6810-triyn-1-yl-120573-D-glucopyranoside

2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-

tridecene-7911-triyne

O

HO

H

OH

H

H

HOH

O

OH

HO

H Aerial (USA)whole (Taiwan)leaves (Taiwan)

[8 45 48ndash51]

51

(4E)-1-(2-Hydroxyethyl)-4-dodecene-6810-triyn-1-yl-120573-D-glucopyranoside

3-120573-D-Glucopyranosyloxy-1-hydroxy-6(E)-

tridecene-81012-triyne

OHO

H

OH

H

H

HOH

O

OH

OH

HAerial (USA andChina) whole(Taiwan) leaves(Taiwan)

[8 35 4245 48ndash51]

52

3-Hydroxy-6-tetradecene-81012-triynyl-120573-D-glucopyranoside

120573-D-Glucopyranosyloxy-

3-hydroxy-6E-tetradecence-81012-triyne

OH

O

OHO

H

OH

H

H

HOH

OH

HWhole (Mexico) [52]

53

1-(Hydroxymethyl)-46810-dodecatetrayne-1-yl-120573-D-glucopyranoside

Cytopiloyne2-120573-D-

Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne

O

HO

OHO

H

OH

H

H

HOH

OH

HWhole (Taiwan)leaves (Taiwan) [8 50 53]

54

2-O-D-Glucosyltrideca-11E-en-3579-tetrayn-12-diol

OH

O

O

OHH

HO

H

H

HHOH

OH

Leaves (Brazil) [27]

10 Evidence-Based Complementary and Alternative Medicine

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

55

(R)-1-(Hydroxymethyl)-246810-dodecapentayn-1-yl-a-D-glucopyranoside

2- 120573 -D-Glucopyranosyloxy-1-hydroxytrideca-357911-pentayne

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Chinaand Japan) [35 47]

56

1-[[(Carboxyacetyl)oxy]methyl]46810-dodecatetraynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Japan) [54]

57

(4E)-1-[[(Carboxyacetyl)oxy]-methyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHO

H

OH

Aerial (Japan) [54]

58

(4E)-1-[[(Carboxyacetyl)oxy]-ethyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

O

O

O

OHO

O

OHOH

H

HO

H

H

HHO

HOH

Aerial (Japan) [54]

59 (5E)-5-Heptene-13-diyn-1-yl-benzene

1-Phenylhepta-13-diyn-5en Whole (Taiwan) [32]

60 7-Phenyl-2(E)-heptene-46-diyn-1-ol

OHRoots (notstated) Aerial(China)

[38 42]

617-Phenyl-2(E)-heptene-46-diyn-1-ol-acetate

OO

Roots (notstated Brazil) [38 55 56]

62 7-Phenyl-46-heptadiyn-2-ol Pilosol A

OH

Whole (Taiwan)aerial (China) [42 57]

Evidence-Based Complementary and Alternative Medicine 11

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

63 7-Phenylhepta-46-diyn-12-diol HO

OH

Aerial (China) [42]

64 135-Heptatriyn-1-yl-benzene

1-Phenylhepta-135-triyne

Leaves (notstated) leaves oftissue culture(not stated)aerial (TanzaniaChina)Whole (Taiwan)roots (Brazil)

[18 34 3842 56ndash58]

65 7-Phenyl-246-heptatriyn-1-ol HO

Leaves (notstated) aerial(China)

[38 42]

667-Phenyl-246-heptatriyn-1-ol-acetate O

O Leaves (notstated) [38]

675-(2-Phenylethynyl)-2-thiophenemethanol

S OHAerial (China) [42]

685-(2-Phenylethynyl)-2120573-glucosylmethyl-thiophene

O

O

OH

H

OH

H

H

HOH H

O

HO

S

Aerial (China) [42]

69

3- 120573 -D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne

OH

O

OH

HOHO

H

H

HHOH

OH

Leaves(Cameroon) [28]

701-Phenyl-13-diyn-5-en-7-ol-acetate O

O

Leaves (Brazil) [27]

SN denotes serial number

the chloroform fraction was the most toxic with a halfmaximal inhibitory concentration (IC

50) of 97 plusmn 72 and 83 plusmn

52 120583gmL in NRU and MTT respectively [82] Kumari andcolleagues also reported the anti-cancer and anti-malarialactivities of B pilosa leaves [77] Based on a cytotoxicity-directed fractionation strategy they identified phenyl-135-heptatriene with IC

50values of 8plusmn001 049plusmn045 07plusmn001

and 10 plusmn 001 120583gmL against human oral liver colon andbreast cancer cell lines respectively However phenyl-135-heptatriyne showed lower activity against breast cancer celllines than the chloroform leaf extract which had an IC

50

value of 65 plusmn 001 120583gmL Moreover the positive controltaxol showed higher activity than phenyl-135-heptatriyne[77] Furthermore in vitro comet assays were performed toevaluate the toxicity of n-hexane chloroform and methanolextracts of B pilosa and its ethyl acetate acetone and waterfractions onHela andKB cellsThe ethyl acetate fraction fromthe methanol extract exhibited the highest activity with halfmaximal cytotoxic concentrations (CTC

50) of 9652120583gmL

and 5862120583gmL against Hela and KB cells respectivelyDespite the moderate toxicity these findings suggest thatthese B pilosa extractsfractions could be useful for future

12 Evidence-Based Complementary and Alternative Medicine

Table5Flavon

oids

isolated

from

Bpilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

71

2-[(34-Dihydroxyph

enyl)-

methylene]-

6-hydroxy-3(2H

)-benzofuranon

e

Sulfu

retin

O

OHO

OH

OH

Aeria

l(Ch

ina)

[35]

72

2-[(34-Dihydroxyph

enyl)-

methylene]-

67-dihydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-673101584041015840-

tetrahydroxy

maritimetin

O

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

73

2-[(34-Dihydroxyph

enyl)-

methylene]-6-

(120573-D

-glucopyrano

syloxy)-7-

hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-120573-D

-glucop

yranosyl-

673101584041015840-te

trahydroxy

maritimein

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japanand

China)aerial(Ch

ina)

[355960]

74

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

(120573-D

-glucopyrano

syloxy)-

6-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-7-O-120573-D

-glucop

yranosyl-

673101584041015840-

tetrahydroxy

O

OHO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japan)

[59]

75

6-[(6-O-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-acetyl-

120573-D

-glucopyrano

syl)

-673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OHO

H HO

H

H

HOH

HO

O

O

Leaves

(Not

stated)

aeria

l(Ch

ina)

[3559]

Evidence-Based Complementary and Alternative Medicine 13

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

76

6-[(36-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-

2-[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone(Z)-6-O

-(36-di-O

-acetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

bideno

sideA

O

O

HO

OH

OH

OHOH O

H

H

HOH

HO

O

O

O

Leaves

(China)

[60]

77

6-[(46-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(410158401015840610158401015840-

diacetyl-120573-D

-glucropyrano

syl)-

673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

HO

O

OO

Leaves

(not

stated)

aeria

l(Ch

ina)

[343561]

78

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

hydroxy-6-

[(246-tri-O

-acetyl-120573

-D-

glucop

yranosyl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(210158401015840410158401015840610158401015840-

triacetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

O

OOO

O

Leaves

(not

stated)

aeria

l(Ch

ina)

[3461]

79

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-6-

[(346-tri-O

-acetyl-120573

-D-glucopyrano

syl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(310158401015840410158401015840610158401015840-

triacetyl-120573

-D-glucrop

yranosyl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H OH

H

HOH

HO

OOO O

Aeria

l(Ch

ina)

[3435]

80

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-

6-[[6-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propenyl]-120573-

D-glucopyrano

syl]o

xy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-

p-coum

aroyl-

120573-D

-glucropyrano

syl)-

673101584041015840-te

trahydroxy

OH

O

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OLeaves

(Japan)

[59]

14 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

81

1-[2-(120573-D

-Glucopyrano

syloxy)-

4-hydroxyphenyl]-2-

hydroxy-3-(3-

hydroxyphenyl)-2-

prop

en-1-on

e

Chalcone120572

32101584041015840-

tetrahydroxy-

21015840-O

-120573-D

-glucop

yranosyl

HO

O

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[35]

821-(24-Dihydroxyph

enyl)-

3-(34-dihydroxyph

enyl)-

2-prop

en-1-on

eBu

tein

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

833-(34-D

ihydroxyph

enyl)-

1-(234-trihydroxyph

enyl)-

2-prop

en-1-on

eOkanin

O

HO

HO

OH

OH

OH

Leaves

(China)

[60]

84

3-(34-D

ihydroxyph

enyl)-

1-[3-(120573-D

-glucop

yranosyloxy)-

24-dihydroxyphenyl]-2-

prop

en-1-on

e

Okanin

31015840-O

-120573-D

-glucosid

e

O

HO

OH

OH

OH

OO

HO

H

HO

H

H

HOH

H

OH

Leaves

(Germany)

flowers(Germany)

[61ndash63]

85

3-(34-D

ihydroxyph

enyl)-

1-[4-(120573-D

-glucop

yranosyloxy)-

23-dihydroxyphenyl]-

2-prop

en-1-on

e

Okanin

41015840-O

-120573-D

-glucosid

e

OOH

OH

OH

HOO

OHO

H

HO

H

H

HOH

H

OH

Flow

ers(Germany)

leaves

(Japan)

[5963]

Evidence-Based Complementary and Alternative Medicine 15

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

86Okanin41015840-O

-120573-D

-(610158401015840-

O-acetylglucosid

e)

O

O

OH

OH

OH

OHO

H

HO

H

H

HOH

H

HO

O

O

Flow

ers(Germany)

[63]

87

1-[4-[(46-di-O

-Acetyl-

120573-D

-glucopyrano

syl)-oxy]-

23-dihydroxyphenyl]-3-

(34-dihydroxyph

enyl)-2-

prop

en-1-on

e

Okanin

41015840-O

-120573-D

-(410158401015840610158401015840-diacetyl)-

glucop

yranoside

O

O

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Aeria

l(Ch

ina)

[34]

88Okanin41015840-O

-120573-D

-(210158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[64]

89Okanin41015840-O

-120573-D

-(310158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

O

OAe

rial(Ch

ina)

[34]

16 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

90

1-[23-Dihydroxy-4-

[[6-O-[3-(4-hydroxyph

enyl)-

1-oxo-2-prop

enyl]-120573-D

-glucop

yranosyl]oxy]-ph

enyl]-

3-(34-dihydroxyph

enyl)-

2-prop

en-1on

e

Okanin

41015840-O

-120573-D

-(610158401015840-tr

ans-

p-coum

aroyl)-

glucoside

O

O

OH

OH

OH

OHOH

HO

H

H

HOH

H

HO

OO

OH

Leaves

(Germany)

[64]

91Okanin41015840-O

-120573-D

-(410158401015840-acetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Leaves

(Germany)

[65]

92

Okanin

41015840-O

-120573-D

-(210158401015840410158401015840-diacetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[65]

Evidence-Based Complementary and Alternative Medicine 17

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

93

Okanin

41015840-O

-120573-D

-(310158401015840410158401015840-diacetyl-610158401015840-

trans-p-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

OH

O O

Leaves

(Germany)

[65]

94

Okanin

41015840-O

-[120573-D

-glucopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

OOH

OH

OH

HO

OH

HO

HHO

H

HOH

HO

OH

OH

HO

HHO

H

HOH

HO

Flow

er(G

ermany)

[63]

95Okanin

3101584041015840-di-O

-120573-D

-glucosid

e

OOH

OH

OH

O OO

H HO

HHOH

HOH

HHO

OHO

H

HO

H

H

HOH

H

OH

Flow

er(G

ermany)

[63]

18 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

96

1-[3-(120573-D

-Glucopyrano

syloxy)-

24-dihydroxyphenyl]-

3-(3-hydroxy)-

4-metho

xyph

enyl)-

2-prop

en-1-on

e

Okanin

4-methylether-

31015840-O

-120573-D

-glucop

yranoside

OOH

O

OH

HO

O

H HO

HHOH

HOH

H

OHO

Leaves

(Germany)aerial

(China)

[3562]

97

Okanin4-methylether-

3101584041015840-di-O

-120573-(41015840101584061015840101584041015840101584010158406101584010158401015840-

tetraacetyl)-

glucop

yranoside

OOH

OOH

O

HH

OH

HO

HOH

H

OO

O

Ac

Ac

AcAc

OO

H

HO

H

H

HOH

H

O

Aeria

l(Ch

ina)

[42]

98Ch

alcone210158404101584061015840-tr

imetho

xy-

4-O-D

-glucopyrano

syl-

dihydro

Not

foun

dLeaves

(China)

[60]

992-(34-D

ihydroxyph

enyl)-23-

dihydro-78

-dihydroxy-

4H-1-

benzop

yran-4-one

Okanin

isoOO

OH

HO

OH

OH

Leaves

(China)

[60]

100

2-(34-D

ihydroxyph

enyl)-

23-dihydro-8-hydroxy-

7[(246-tr

i-O-acetyl-120573

-D-

glucop

yranosyl)oxy]-4H

-1-b

enzopyran-4-on

eOO

OH

OH

OH

OH

O

HHO

H

HO

HO

OAc

Ac

Ac

Aeria

l(Ch

ina)

[34]

Evidence-Based Complementary and Alternative Medicine 19

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

101

57-Dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

OO

OH

HO

OH

Aeria

l(TanzaniaC

hina)

[4266]

102

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

7-O-glucopyrano

side

OO

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Tanzania)

[66]

103

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-4H

-1-benzop

yran-4-one

Luteolin

OO

HO

OH

OH

OH

Aeria

l(TanzaniaC

hina

Vietnam)

[343542

6667]

104

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5hydroxy-4H-1-

benzop

yran-4-one

Luteolin

7-O-120573-D

-glucop

yranoside

OOOH

OH

OH

OO

HO

H

HO

H

H

HOH

OH H

Aeria

l(Tanzania)

[66]

105

57-Dim

etho

xy-6-

(5-m

etho

xy-6-m

ethyl-

4-oxo-4H

-pyran-3-yl)-

2-ph

enyl-4H-1-

benzop

yran-4-one

5-O-M

ethylhosludin

O OOO

O

O

O

Aeria

l(Ugand

a)[68]

20 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

106

3-(120573-D

-Glucopyrano

syloxy)-

57-dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Astragalin

Kaem

pferol-3-O

-120573-D

-glucopyrano

side

HO

OH

O O

OH

O

O

HO

HHO

H

H

HOH

OH

H

Aeria

l(Ch

ina)

[35]

107

Kaem

pferol3-

(23-di-E

-p-cou

maroyl-

120572-L-rhamno

pyrano

side)

Not

foun

dAe

rial(Vietnam)

[67]

108

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5-hydroxy-36-dimetho

xy-

4H-1-

benzop

yran-4-one

Axillarosid

eO O

OH

OH

OO

OH

OO

HO

H

HO

H

H

HOH

H

OH

Aeria

l(Ch

ina)

[42]

109

57-Dihydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaureidin

O

O

O

O

O

HO

OH

OH

Who

le(Taiwan)

[69]

110

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaurein

O O

OO

O OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan)w

hole

(Taiwan)

[69ndash

71]

Evidence-Based Complementary and Alternative Medicine 21

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

111

5-Hydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-

67-dimetho

xy-4H-

chromen-4-one

Eupatorin

iso

OO O

OH

O

O OH

Not

foun

d(C

hina)

[3772]

112

2-(34-D

imetho

xyph

enyl)-7-

(120573-D

-glucopyrano

syloxy)-35-

dihydroxy-

8-metho

xy-4H-1-

benzop

yran-4-one

O O

OH

O O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

113

7-(120573-D

-Glucopyrano

syloxy)-

5hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-38-

dimehtoxy-4H-1-

benzop

yran-4-one

O O

O

OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

114

Isorhamnetin

3-[O

-120572-

L-rham

nopyrano

syl-

(1-2)-120573-D

-glucop

yranoside]

Not

foun

dAe

rial(Vietnam)

[67]

115

7-[(6-deoxy-120572-L-

Manno

pyrano

syl)o

xy]-

3-120573-D

-glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-4H

-1-benzop

yran-4-one

Luteoside

O O

OH

O

OH

OH

OH

HOH

H

H HO

HO

HO

OH

HO

H

HO

OH

OH

HH

Aeria

l(Ch

ina)

[42]

22 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

116

Luteolin

3-O-120573-D

-Glucopyrano

side

O O

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

HO

Aeria

l(Tanzania)

[66]

117

57-Dihydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Quercetagetin

3631015840-tr

imethylether

OOO

O

O OH

HO

OH

Aeria

l(Ch

ina)

[42]

118

6-((2S3S4S5S)-

Tetrahydro-345-tr

ihydroxy-

6-(hydroxymethyl)-2H

-pyran-2-yloxy)-5-hydroxy-

2-(4-hydroxy-3-

metho

xyph

enyl)-

37-dimetho

xy-4H-

chromen-4-one

Quercetagetin

3731015840-

trim

ethylether-6-O

-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[42]

119

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

JaceinQ

uercetin

3631015840-

trim

ethyl

ether-7-O-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan

China)

who

le(Taiwan)

[425470]

120

2-(34-D

ihydroxyph

enyl)-

357-trihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

OO

HO

OH

OH

OH

OH

Aeria

l(Ch

ina)leaves

(China)who

le(C

hina)

[356074]

Evidence-Based Complementary and Alternative Medicine 23

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

121

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-galactopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-4-one

Quercetin

3-O-120573-D

-galactosidehyperin

hyperosid

e

O O

HO

OH

OH

OH

OOHH

HOH

H

H HO

HO

OH

Aeria

l(TanzaniaJapan)

Leaves

(China)Who

le(C

hina)

[376066

717475]

122

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-glucopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

3-O-120573-

D-glucopyrano

side

O O

HO

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

Aeria

l(TanzaniaC

hina

Japan)L

eaves(Japan)

[355966

75]

123

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-

4-oxo-4H

-1-benzop

yran-3-yl-

120573-D

-glucopyrano

siduron

icacid

Quercetin

3-O-120573-D

-glucuron

opyranoside

O O

HO

OH

OH

OH

OHOH

HOH

H

H HO

HO

OHO

Aeria

l(TanzaniaJapan)

[6671]

124

3-[[6-O-(6-

Deoxy-120572-L-m

anno

pyrano

syl)-

120573-D

-galactopyrano

syl]o

xy]-

2-(34-dihydroxyph

enyl)-57-

dihydroxy-4H

-1-benzop

yran-4-one

Quercetin

3-O-rob

inob

iosid

e

O O

HO

OH

OH

OH

O

HOHH OH

HO

H HH

OO

OHH

HOH

H

H HO

HO

Aeria

l(Japan)W

hole

(Taiwan)

[7071]

24 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

125

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-

3-metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

OH

Roots(Brazil)

[55676]

126

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-

120573-D

-glucopyrano

syl]o

xy]-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120572-

L-rham

nopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

O

HHO HHO

HO H

HH

ORo

ots(Brazil)

[5676]

127

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-120573-D

-glucop

yranosyl]oxy]-5-

hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-3-metho

xy-

4H-1-

benzop

yran-4-one

Quercetin

341015840-

dimethyl

ether-7-O-rutinoside

O OOH

OH

O

O

OH O

HOH

H

HHO

H

O

HHO HHO

HO H

HH

O

HO

Aeria

l(Ch

ina)

[3435]

128

2-(34-D

ihydroxyph

enyl)-3-

(120573-D

-glucofurano

syloxy)-57-

dihydroxy-4H

-1-benzop

yran-

4-on

e

Isoq

uercitrin

O OOH

OH

OHO

H

OHH

OH

HO

H O HOH

OH

Aeria

l(Japan

China)

[3571]

Evidence-Based Complementary and Alternative Medicine 25

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

129

Quercetin

3-O-120573-D

-galactop

yranoside

O

OH

HO

OH OH

O

O

HO

OH

OHOH

OAe

rial(no

tstated)

[18]

130

Quercetin

3-O-rutinoside

O

O

R

OO

OH

OHO

H

O

OH

OH

H3C

H3C

H3C

OH

HO

Who

le(Taiwan)

[70]

26 Evidence-Based Complementary and Alternative Medicine

Table 6 Terpenoids isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(Country) References

131371111-Tetramethylbicyclo[810]undeca-

26-dieneBicyclogermacrene Leaves

(Brazil) [95]

13241111-Trimethyl-8-

methylenebicyclo[720]undec-4-ene

E-Caryophyllene Leaves(Brazil) [95]

1331-Methyl-5-methylene-8-

(1-methylethyl)-16-cyclodecadiene

Germacrene D Leaves(Brazil) [95]

1344-(15-Dimethyl-4-hexen-1-ylidene)-1-methyl-cyclohexene

Z-120574-Bisabolene Leaves(Brazil) [95]

135Decahydro-114-trimethyl-7-methylene-1H-cycloprop[e]-

azulene120573-Gurjunene Leaves

(Brazil) [95]

136 2669-Tetramethyl-148-cycloundecatriene

120572-Humulene120572-caryophyllene

Leaves(Brazil) [95]

13712344a568a-Octahydro-1-isopropyl-7-methyl-4-methylenenaphthalene

120575-Muurolene Leaves(Brazil) [95]

138

12344a568a-Octahydro-4a8-dimethyl-2-(1-methylethylidene)-

naphthalene

Selina-37(11)-diene

Leaves(Brazil) [95]

139(2E7R11R)-371115-

Tetramethyl-2-hexadecen-1-ol

Phytol

HO

Whole(Taiwan) [57]

Evidence-Based Complementary and Alternative Medicine 27

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

140 371115-Tetramethyl-2-hexadecenoic acid Phytanic acid

O

OH

Whole(Taiwan) [57]

141371115-Tetramethyl-2-

hexadecenyl ester-heptanoicacid

Phythylheptanoate O O

Leaves(Not stated) [33]

142

(3S10R13R)-17-((2R5R)-5-Ethyl-6-methylheptan-2-yl)-

2347891011121314151617-tetradecahydro-

1013-dimethyl-1H-cyclopenta[a]phenanthren-

3-ol

Campestrol

HO

Aerial(Tanzania) [18]

143 Not found Phytosterin-B Not foundNot found(TaiwanEgypt)

[39 96]

144 Stigmast-5-en-3-ol 120573-sitosterol

HOH

H

H

Aerial(Tanzania)

whole(Taiwan)

[18 31 57]

145

1314151617-Tetradecahydro-1013-

dimethyl-1H-cyclopenta[a]phenanthren-

3-yloxy)-tetrahydro-6-(hydroxymethyl)-2H-pyran-345-triol

120573-Sitosterolglucoside

H

H

HOH

HO

HHO

H

HOHH O

OH Not found(Egypt) [39]

146 5120572-Stigmasta-7-en-3120573-ol

HO

Whole(Taiwan) [57]

147 5120572-Stigmasta-722t-dien-3120573-ol

HO

Whole(Taiwan) [57]

148 Stigmasta-522-dien-3-ol Stigmasterol

HOH

H

H

Not found(Taiwan)aerial

(Tanzania)leaves (notstated)whole

(Taiwan)

[18 31 3357]

28 Evidence-Based Complementary and Alternative Medicine

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

149 Lup-20(29)-en-3-ol Lupeol

H

HH

HOH

Not found(Egypt) [39]

150 Lup-20(29)-en-3-ol acetate Lupeol acetate

H

HH

O

OH

Not found(Egypt) [39]

151 Olean-12-en-3-ol 120573-amyrin

H

HOH

Not found(Egypt) [39]

152 5913-Trimethyl-242526-trinoroleanan-3-ol Friedelan-3120573-ol

HO

Aerial(Tanzania) [18]

153 5913-Trimethyl-242526-trinoroleanan-3-one

Friedelinfriedelan-3-one

O

Aerial(Tanzania) [18]

1542610151923-

Hexamethyl-2610141822-tetracosahexaene

Squalene

Aerial(Tanzania)Leaves (Notstated)Whole(Taiwan)

[18 33 57]

155 120573120573-Carotene 120573-Carotene Leaves (Notstated) [97]

Evidence-Based Complementary and Alternative Medicine 29

studies [83] Hot water extracts of B pilosa varminor Sheriffwere also assessed for its antileukemic effects on leukemiccell lines L1210 U937 K562 Raji and P3HR1 using XTT-based colorimetric assays The extract inhibited the five celllines with IC

50values ranging from 145120583gmL to 586120583gmL

L1210 K562 Raji and P3HR1 were more sensitive to B pilosaextract with IC

50values below 200120583gmL [98]

Consistent with the antitumor activities of B pilosaextracts and fractions some of its phytochemicals alsoshowed anticancer activity as outlined in Table 11 Amongthem luteolin (103) a well-studied flavonoid with multiplebioactivities was more effective against tumor cell prolifera-tion than its derivatives with IC

50values ranging from 3 120583M

to 50120583M in cells and 5 to 10mgkg in animals Luteolin wasalso found to fight cancer as a food additive at concentrationsof 50 to 200 ppm [78] and prevent skin cancer [78] and cancer3invasion [103] Lee and colleagues reported that luteolinprevents cancer by inhibiting cell adhesion and invasion[103] Significant inhibition concentration was reported to be5 120583M and complete inhibition concentration was reported tobe 40120583M Moreover luteolin was reported to inhibit hepa-tocyte growth factor (HGF)-induced cell scattering as wellas cytoskeleton changes such as filopodia and lamellipodiawhichwas determined using phase-contrast and fluorescencemicroscopy Furthermore luteolin also inhibited the HGF-induced phosphorylation of c-Met ERK 12 and Akt aswell as the MAPKERK and P13K-Akt pathways [78 103]Other mechanisms underlying the anticancer activities ofluteolin are the inhibition of topoisomerase I and II whichinhibits cell replication and DNA repair thus promotingapoptosis regulation of PI-3-KinaseAktMAPKERKJNKactivation of apoptosis in the mitochondrial pathway byactivating caspase 9 and caspase 3 which were found inmalignant cells but not in normal human peripheral bloodmononuclear cells death receptor-induced apoptosis and acell-cycle arrest mechanism inhibition of fatty acid synthasewhich is upregulated in many cancer cells and sensitizationto chemotherapywhereby luteolin increases the susceptibilityof cancer cells to chemotherapy [78] Butein (82) is anotherflavonoid that showed a cytotoxic effect on human colonadenocarcinoma cell proliferation with a reported IC

50value

of 175 120583M Butein at 2 120583M affected the incorporation of[14C]-labeled leucine thymidine and uridine which cancause the inhibition of DNA RNA and protein synthesis ofhuman colon cancer cells Moreover butein also exhibitednoncompetitive inhibition of 1-chloro-24-dinitrobenzene(CDNB) in glutathione S-transferase (GST) activity Tumorresistance was correlated with high levels of GST thus buteininhibited proliferation of cancer cells [80] Another flavonoidpresent in B pilosa centaureidin (109) also showed anti-cancer activity in B lymphoma cells Centaureidin isolatedfrom Polymnia fruticosa inhibited tubulin polymerization invitro and induced mitotic figure formation in CA46 Burkittlymphoma cells Using turbidimetric assay the IC

50value of

centaureidin in inhibition of mitosis was 3120583M [104] Cyto-toxicity of centaureidin was further analyzed using AmericanNational Cancer Institute (NCI) 60 human tumor cell linesThe cytotoxicity potency of centaureidin expressed as GI

50

(50 growth inhibition in the NCI tumor line panel) was024120583M [81] These data mark centaureidin as a promisingantimitotic agent for tumor therapy

In addition to anti-tumor flavones polyynes found in B 4pilosa have also been shown to possess anti-tumor proper-ties Based on a bioactivity-directed isolation approach Wuand colleagues identified two polyyne aglycones from theethyl acetate fraction of B pilosa [44] 12-Dihydroxytrideca-57911-tetrayne (48) and 13-Dihydroxy-6(E)- tetradecene-81012-triyne (46) exhibited significant anticell prolifera-tion activity in primary human umbilical vein endotheliumcells (HUVEC) with IC

50values of 125 120583M and 173 120583M

respectively They also decreased angiogenesis and pro-moted apoptosis in human endothelial cells Their anti-angiogenic and cytotoxic effects correlated with activationof the CDK inhibitors and caspase-7 [44] In addition 12-Dihyroxy-5(E)-tridecene-7911-triyne (45) showed antian-giogenic effects in HUVECs with an IC

50value of 124 120583M as

evidenced by a decrease in the tube formation and migrationof HUVECs [43] The IC

50value of compound 45 in the

inhibition of basic fibroblast growth factor-induced HUVECgrowth was 282120583M However it had higher IC

50values

than those for lung carcinoma cells and keratinocytes Thiscompound could also inhibit cell proliferation of HUVECslung carcinoma A549 cells and HACAT keratinocytes Themechanism by which compound 45 inhibits HUVEC growthand angiogenesis is complicated and includes decreasing theexpression of cell cycle regulators (CDK4 cyclins D1 and Aretinoblastoma (Rb) and vascular endothelial growth factorreceptor 1) caspase-mediated activation of CDK inhibitorsp21 (Cip1) and p27 (Kip) upregulation of Fas ligand expres-sion downregulation of Bcl-2 expression and activation ofcaspase-7 and poly (ADP-ribose) polymerase [43]

32 Anti-Inflammatory Activity B pilosa is commonly usedto treat inflammatory disorders The anti-inflammatory phy-tochemicals present in B pilosa are listed in Table 11 5Cyclooxygenase-2 (COX-2) is a physiologically importantenzyme that converts arachidonic acid to prostaglandin(PGE2) Its expression is induced by a wide variety of external

stimuli indicating its involvement in inflammatory diseasesand it is used as an inflammatory marker [88] Yoshida andcolleagues studied the effects of the aqueous extracts of Bpilosa aerial parts in the production of COX-2 and PGE

2as

well as on the activation of mitogen activated protein kinases(MAPKs) in normal human dermal fibroblasts (HDFs) inresponse to inflammatory cytokine IL-1120573 This work showedthat IL-1120573 activated MAPKs such as ERK12 p38 and JNKto different extents and induced COX-2 expression TheCOX-2 expression in HDFs was regulated mainly by p38following IL-1120573 stimulation Consistently the p38 inhibitorSB203580 blocked this expression Using this cell platformB pilosa extracts were tested for inhibition of inflammationThe extract dose-dependently suppressed the activation ofp38 and JNK and moderately suppressed ERK12 as wellas suppressing COX-2 expression and PGE

2production

30 Evidence-Based Complementary and Alternative Medicine

Table7Ph

enylprop

anoids

isolatedfro

mB

pilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

156

3-(4-H

ydroxyph

enyl)-2-prop

enoica

cid

p-Cou

maricacid

OHO

OH

Who

le(Ja

pan)

[99]

157

2-Metho

xy-4-(2-prop

en-1-yl)-ph

enol

Eugeno

lOOH

Leaves

androots(Japan)

[99]

158

3-(4-H

ydroxy-3-m

etho

xyph

enyl)-

2-prop

enoica

cid

Ferulic

acid

O

OH

O

HO

Who

le(Ja

pan)

[99]

159

3-(34-D

ihydroxyph

enyl)-2-prop

enoic

acid

Caffeicacid

O

HO

OH

OH

Who

leandaeria

l(Japan)

[7599]

160

3-Prop

yl-3[(245-trim

etho

xyph

enyl)-

metho

xy]-24-pentanedione

3-Prop

yl-3-(245-

trim

etho

xy)benzyloxy-

pentan-24-dione

O

O

O

O

O

O

Leaves

(India)

[100]

161

3-(34-D

ihydroxyph

enyl)-2-

prop

enoica

cidethyleste

rCa

ffeateethyl

O

O

OH

OH

Not

foun

d(Taiwan)

who

le(Taiwan)

[434584]

Evidence-Based Complementary and Alternative Medicine 31Ta

ble7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

162

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

d-Erythron

icacid

2-O-caffeoyl-2-C

-methyl

O

HO

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

163

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

2-O-caffeoyl-2-C

-methyl

O

O

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

164

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-24-

dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

3-O-caffeoyl-2-C

-methyl

O

O

HO

HO

O

O

OH

OH

Leaves

(Japan)

[101]

165

4-(A

cetyloxy)-3-[[3-(34-

dihydroxyphenyl)-

1-oxo-2-propen-1-y

l-]oxy]-2-hydroxy-

2-methyl-b

utanoica

cid

O

HO

OH

O OHO

HO

OO

Not

foun

d(Ja

pan)

[73]

166

3-(34-D

ihydroxyph

enyl)-tetrahydro-4-

hydroxy-4-

methyl-5

-oxo-3-fu

ranyl

ester-2-prop

enoica

cid

3-O-C

affeoyl-2-C

-methyl-

D-erythrono

-14-la

cton

eHO

HO

O

OO

OOH

Leaves

(Japan)

[101]

167

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]-oxy]-145-

trihydroxy-cyclohexanecarboxylicacid

Chlorogenica

cid

O

HO

HO

OH

O

OH

O

OH

HO

Aeria

l(Japan)w

hole

(Taiwan)

[707175]

32 Evidence-Based Complementary and Alternative Medicine

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

168

4-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]oxy]-

135-tr

ihydroxy-cyclohexanecarboxylic

acid

4-O-C

affeoylqu

inicacid

OHO

HO

OH

OH

O

O

OH

OH

Aeria

l(Japan)

[71]

169

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-15-

dihydroxy-cyclo

hexanecarboxylicacid

34-Di-O

-caffeoylqu

inic

acid

HO

HO

O

O

O

O

OH

OH

HO

OH O

H

O

Aeria

l(Japan)W

hole

(Taiwan)

[45497071]

170

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-14-dihydroxy-

cyclo

hexanecarboxylicacid

35-Di-O

-caffeoylqu

inic

acid

HO

HO

O O

HO

O

HO

OH OO

OHOH

Aeria

l(Japan)w

hole

(Taiwan)

[45497071]

Evidence-Based Complementary and Alternative Medicine 33

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

171

34-bis[[(2E

)-3-(34-D

ihydroxyph

enyl)-1-

oxo-2-prop

en-1-

yl]oxo]-15

-dihydroxy-

cyclo

hexanecarboxylicacid

45-Di-O

-caffeoylqu

inic

acid

OHO

OH

HO

O

O

OH

OH

O

OH

HO

OWho

le(Taiwan)

[454970]

172

3-[4-[[-6-O-[3-(4-H

ydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]-phenyl]-2-prop

enoica

cid

120573-D

-p-C

oumaricacid4-O

-(6-O

-p-caoum

aroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HOH

HO

O

O

Leaves

(Japan)

[59]

173

3-[4-[[2-O

-Acetyl-6

-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]ph

enyl]2-propeno

icacid

120573-D

-p-C

oumaricacid4-O

-(2-O

-acetyl-6

-O-p-

caou

maroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HO

HO

O

O

Leaves

(Japan)aerial

(China)

[3459]

174

67-Dihydroxy-2-chrom

enon

eEsculetin

cicho

rigenin

OHO HO

O

Not

foun

d(Egypt)

[39]

34 Evidence-Based Complementary and Alternative Medicine

Table 8 Aromatic compounds isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

175 12-Benzenediol PyrocatechinHO

HOWhole(Japan) [99]

176 4-Ethyl-12-benzenediol Pyrocatechol

OH

OH

Whole(Japan) [99]

177 Dimethoxyphenol26-dimethoxyphenol

O

HO

O

Roots (Japan) [99]

178 4-Ethenyl-2-methoxy-phenol p-Vinylguaiacol

O

OHWhole(Japan) [99]

179 2-Hydroxy-6-methylbenzaldehyde

6-Methyl-salicylaldehyde O

OH

Whole(Japan) [99]

180 Benzene-ethanol 2-Phenyl-ethanolHO

Whole(Japan) [57]

181 4-Hydroxy-3-methoxy-benzaldehyde Vanillin O

O

HOAerial(Japan) [99]

182 3-Hydroxy-4-methoxy-benzaldehyde Vanillin iso

OO

OH

Leaves(Japan) [99]

183 4-Hydroxybenzoic acid p-Hydroxybenzoicacid

O

HOOH Whole

(Japan) [99]

184 2-Hydroxybenzoic acid Salicylic acidO

HO

HO

Stem androots (Japan) [99]

185 34-Dihydroxybenzoicacid Protocatechuic acid

O

HOOH

OH

Whole(Japan) [99]

1864-Hydroxy-

3methoxybenzoicacid

Vanillic acid

OOH

O

OH

Aerial(Uganda)

Roots (Japan)[68 99]

Evidence-Based Complementary and Alternative Medicine 35

Table 8 Continued

S N IUPAC names Common names Structure Plant part(country) References

187 345-Trihydroxybenzoicacid Gallic acid

O

OHHO

HO

HO

Whole(China) [102]

[88] This work supports the use of B pilosa as an anti-inflammatory agent however no compounds responsible forthe anti-inflammatory activity of B pilosa were identified

A further study also reported the anti-inflammatoryactivity as well as the antiallergic activity of B pilosa [75] Inthis study dried powder of the aerial part of B pilosa whichhad been pretreated with the enzyme cellulosine was usedfor further tests The results showed that oral administrationof the cellulosine-treated B pilosa lowered the level of serumIgE in mice 10 days after immunization with DNP (24-dintrophenyl)-Ascaris as an antigen This treatment alsoreduced dye exudation in skin induced by passive cutaneousanaphylaxis and production of inflammatory mediators his-tamine and substance P in rats [75] Phytochemical analysisshowed that cellulosine treatment increased the percentage ofcaffeic acid and flavonoids This study suggests that B pilosaand its phenolics have anti-inflammatory functions

Phenolics and polyynes are major anti-inflammatoryphytochemicals present in B pilosa (Table 11) Unsurpris-6ingly phenolics such as luteolin (103) and ethyl caffeate(161) that are major constituents of B pilosa have alsobeen reported to possess anti-inflammatory activity Lute-olin was reported to exhibit anti-inflammatory activity inmacrophages Xagorari and colleagues showed that luteolininhibited the release of inflammatory cytokines TNF-120572 andinterleukin-6 in RAW 2647 cells following LPS stimulation[87] It inhibited TNF-120572 production with an IC

50value

of 1120583M The underlying anti-inflammatory mechanism ofluteolin was reported to be the inactivation of Akt andNF-120581B activation [87] In addition luteolin was reportedto confer anti-inflammatory activity through inhibition ofLPS-stimulated iNOS expression in BV-2 microglial cellsIt inhibited LPS-activated microglia in a dose-dependentmanner with an IC

50value of 69 120583MMoreover immunoblot

and RT-PCR data proved that luteolin suppressed I120581B-120572degradation and iNOS expression in LPS-activated microglia[85] Kim and colleagues stated that luteolin may havebeneficial effects on inflammatory neural diseases throughinhibition of iNOS expression [85] A related study revealedthat luteolin decreased the transcriptional activity of NF-120581BRelA via partial inhibition of TNF-mediated NF-120581B DNAbinding activity Luteolin also inhibited Akt phosphorylationand induced degradation of a transcription factor interferonregulatory factor (IRF) [86]

Chiang and colleagues showed that ethyl caffeate(161) significantly inhibited NO production in mousemacrophages RAW 2647 cells [84] Based on MTT assays

they concluded that this inhibition was not due to thecytotoxicity of ethyl caffeate The IC

50value of ethyl caffeate

in the inhibition of NO production was 55 120583gmL slightlylower than curcumin (positive control) which has anIC50

value of 65120583gmL They demonstrated that ethylcaffeate exerted anti-inflammatory activity via the reducedtranscription and translation of iNOS (inducible nitric oxidesynthase) in RAW 2467 cells In addition this compoundalso suppressed COX-2 expression in RAW 2467 cellsand MCF-7 cells The in vivo anti-inflammatory effectof ethyl caffeate was verified by testing in TPA-treatedmouse skin Like celecoxib the positive control ethylcaffeate significantly abolished COX-2 expression in adose-dependent manner Ethyl caffeate at 1mg200120583Lsite(24mM) inhibited COX-2 expression at a level comparableto celecoxib at 1mg200120583Lsite (13mM) Remarkably ethylcaffeate at 48mM (2mg200120583Lsite) was more effectivethan celecoxib at 131mM (10mg200120583Lsite) Moreoverthis compound inhibited the activation of nuclear factor-120581B(NF-120581B) by LPS via the prevention of NF-120581B binding toDNA [84] In addition 3 ethyl caffeate analogs (ethyl 34-dihydroxyhydrocinnamate ethyl cinnamate and catechol)also showed different degrees of NF-120581B binding to DNA aslisted in Table 12 Ethyl cinnamate lacks a catechol moietywhich results in ineffective inhibition of NF-120581B binding toDNA [84]

Pereira and colleagues assessed the anti- 7inflammatory and immunomodulatory activities ofB pilosa methanol extract as well as one polyyne2-O-120573-glucosyltrideca-11(E)-en-3 579-tetrayn-12-diol(54) in T lymphocytes and a zymosan-induced arthritismouse model [27] They first examined the in vitro effect of 8the B pilosa extract and compound 54 on cell proliferation ofhuman T cells stimulated with 5120583gmL phytohemagglutinin(PHA) or 100 nM 12-O-tetradecanoyl phorbol-13-acetate(TPA) plus 15 120583M ionomycin and on cell proliferation ofmouse T cells stimulated with 5 120583gmL concanavalin A(Con A) The data demonstrated that both methanol extractand compound 54 suppressed T-cell proliferation in adose-dependent manner The estimated IC

50values of the B

pilosa extract against human T cells stimulated with 5120583gmLPHA and 100 nM TPA plus 15120583M ionomycin were 125 and25 120583gmL respectively In comparison with the methanolextract compound 54 showed 10-fold more inhibition ofhuman T-cell proliferation with an estimated IC

50value of

15 120583gmL Accordingly the B pilosa extract and compound54 dose-dependently suppressed mouse T-cell proliferation

36 Evidence-Based Complementary and Alternative Medicine

Table 9 Porphyrins isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

188

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(15S16S)-10-ethenyl-5-ethyl-1161820-tetrahydro-6111522-tetramethyl-1820-dioxo-15H-912-imino-212-

metheno-471714-dinitrilopyrano[43-

b]azacyclononadecine-16-propanoic

acid

AristophyllC

NH N

HNN

O

O OOOPhytyl

O

Leaves(Taiwan) [100]

189

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -20b-(methoxycarbonyl)-9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-dinitrilo-12-

dioxeto-[310158404101584034]-cyclo-pent[1 2]azacyclononadecine-

19-propanoic acid

Bidenphytin A

NH N

HNN

OPhytylO O O

O O

Leaves(Taiwan) [100]

190

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -2a-

hydroxy-20b-(methoxycarbonyl)-

9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-

dinitrilo-12-dioxeto[310158404101584034]-cyclo-pent[12-b]-

azacyclononadecine-19-propanoic acid

Bidenphytin B

NH N

HNN

OPhytylO O O

O O

OH

Leaves(Taiwan) [100]

191

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO O

HOOO

Leaves(Taiwan) [100]

192

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

Evidence-Based Complementary and Alternative Medicine 37

Table 9 Continued

S N IUPAC names Common names Structure Plant part(country) References

193

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

Leaves(Taiwan) [100]

194

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

195

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3S4S21R)-9-ethenyl-14-ethyl-21-(methoxycarbonyl)-

481318-tetramethyl-20-oxo-3-phorbinepropanoic acid

Pheophytin A

NH N

HNN

PhytylO

O

OO O

Leaves(Taiwan) [100]

with estimated IC50values of 30 and 25 120583gmL respectively

Taken together the data indicate that the B pilosa extractand compound 54 act on human and mouse T cells Totest the in vivo effect of the B pilosa extract and compound54 a zymosan-induced arthritis mouse model was usedThis model was established from B10ASgSnJ mice with aninjection of zymosan (015mg) The zymosan-injected mice9received an intraperitoneal injection of the B pilosa extract(1 5 or 10mg) at one dose a day for 5 days Popliteal lymphnode (PLN) weight was monitored to check the developmentof arthritis The results revealed that 10mg of the methanolextract of B pilosa extract could significantly diminishinflammation as evidenced by PLN weight [27] This worksuggests that B pilosa (and compound 54) can suppressimmune response and inflammation

33 Antidiabetic Activity Anti-diabetic agents are primarilydeveloped from plants and other natural resources [8 24 4951 53] B pilosa is one of 1200 plant species that have beeninvestigated for antidiabetic activity [120 121]B pilosa is usedas an anti-diabetic herb in America Africa and Asia [51 120

122] Many studies have indicated that B pilosa could treattype 1 diabetes (T1D) and type 2 diabetes (T2D) in animals

Etiologically speaking T1D is caused by autoimmune-mediated destruction of pancreatic 120573 cells leading to insulindeficiency hyperglycemia and complications Currentlythere is no cure for T1D Polarization ofTh cell differentiationcontrols the development of T1D Suppression of Th1 celldifferentiation and promotion of Th2 cell differentiationameliorate T1D [123] One study showed that the butanolfraction of B pilosa inhibited T-cell proliferation decreasedTh1 cells and cytokines and increased Th2 cells andcytokines leading to prevention of T1D in nonobesediabetic (NOD) mice [49] Based on a bioactivity-directedisolation strategy 3 polyynes 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne (53) also known ascytopiloyne 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D- Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne (50) were identifiedfrom B pilosa [49 53] The IC

50value of the butanol fraction

was 200120583gmL This inhibition was reported to be partiallyattributed to cytotoxicity because the butanol fraction at

38 Evidence-Based Complementary and Alternative Medicine

Table 10 Other compounds isolated from B pilosa [30]

SN IUPAC names Common names Structure Plant part(country) References

19637-Dihydro-137-

trimethyl-1H-purine-26-dione

CaffeineN

NN

NO

O

Aerial(Uganda) [68]

197

1-((2R4S5R)-Tetrahydro-4-hydroxy-5-(hydroxymethyl)furan-2-yl)-5-

methylpyrimidine-24(1H3H)-dione

ThymidineHO

HO

ON

O

NHO Not found

(China) [37]

198 1-(2-Thienyl)-ethanone 2-Acetyl-thiophene

S

O

Roots(Germany) [41]

199

(2R3S4S5S)-2-(Heptan-2-yloxy)-tetrahydro-6-

((tetrahydro-34-dihydroxy-5-

(hydroxymethyl)furan-2-yloxy)methyl)-

2H-pyran-345-triol

Heptanyl2-O-120573-xylofuranosyl-

(1rarr 6)-120573-glucopyranoside

O

OOH

OH

HO

OHO

H

HO

H

H

HOHH

O

Whole(Taiwan) [70]

200

2-[(3R7R11R)-3-Hydroxy-371115-

tetramethylhexadecyl]-356-trimethyl-

25-cyclohexadiene-14-dione

120572-Tocopherylquinone

OO

HO

Whole(Taiwan) [32]

2017-O-(410158401015840610158401015840-Diacetyl)-

120573-D-glucopyranoside

Not found Leaves(China) [60]

180 120583gmL could cause 50 death of Th1 cells Moreoverthis study suggested that the butanol fraction may preventdiabetes in NOD mice in vivo via downregulation of Th1cells or upregulation Th2 cells that have effects which areantagonistic of those of Th1 cells [49] This was proven byintraperitoneal injection of the butanol fraction at a doseof 3mgkg BW 3 times a week to NOD mice from 4 to 27weeks This dosage resulted in lower incidence of diabetes(33) At a dose of 10mgkg the butanol fraction of Bpilosa totally eliminated (0) the initiation of the diseaseTo further support this result assessment of IgG2a and IgEproduction was performed in the serum of NOD mice Asin vivo results obtained from intracellular cytokine stainingexperiments were not very conclusive levels of IgG2a and IgE

were measured since Th1 cytokine IFN120574 and Th2 cytokineIL-4 favor the production of IgG2a and IgE respectively Asexpected high levels of IgE and some decline in the levels ofIgG2a were observed in the serum Profiling of the butanolextract revealed five compounds 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne(50) 45-Di-O-caffeoyl- quinic acid 35-Di-O-caffeoylquinicacid and 34-Di-O-caffeoylquinic acid Only the first twocompounds showed similar effects on the prevention ofdiabetes in NOD mice as the B pilosa butanol fractionMoreover compound 50 showed greater activity thancompound 69 in terms of enhancement (by 34 comparedto 8) of differentiation of Th0 to Th2 at 15 120583gmL (both

Evidence-Based Complementary and Alternative Medicine 39

Table 11 Chemical constituents of B pilosa and their biological activities

SN Name Classification Molecularformula Biological activities

109 Centaureidin [91] Flavonoid C18H16O8Anti-listerial [69 91]

Cytotoxic [10]

110 Centaurein [91] Flavonoid C24H26O13

Anti-listerial [69 91]Cytotoxic [10]Anti-viral [105]

103 Luteolin [106] Flavonoid C15H10O6

Anti-viral [107 108]Cytotoxic [77]

Anti-inflammatory [109]Anti-allergic [109]

82 Butein [110] Flavonoid C15H12O5Anti-leishmanial [111]

Cytotoxic [78]48 12-Dihydroxytrideca-57911-tetrayne [112] Polyyne C13H12O2 Anti-angiogeneic [112]46 13-Dihyroxy-6(E)-tetradecene-81012-triyne [112] Polyyne C14H16O2 Anti-angiogeneic [112]

45 12-Dihyroxy-5(E)-tridecene-7911-triyne [113] Polyyne C13H14O2Anti-angiogeneic [113]Anti-proliferative [113]

64 1-Phenylhepta-135-triyne [114] Polyyne C13H8

Anti-microbial [115]Anti-malarial [3]Cytotoxic [3]Antifungal [15]

27 Linoleic acid [79] Fatty acid C18H32O2Anti-viral (100)Cytotoxic [17]

161 Ethyl caffeate [84] Phenylpropanoid C11H12O4 Anti-inflammatory [84]

54 2-O-120573-Glucosyltrideca-11(E)-en-3579-tetrayn-12-diol [26] Polyyne C19H20O7Immunosuppressive andAnti-inflammatory [26]

53 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne [50] Polyyne C19H22O7Anti-diabetic [116]

Anti-inflammatory [50]

69 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne [28] Polyyne C20H26O7Anti-diabetic [28]

Anti-inflammatory [94]

50 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne [28] Polyyne C19H24O7

Anti-diabetic [28]Anti-inflammatory [94]

Anti-malarial andantibacterial [47]

129 Quercetin 3-O-120573-D-galactopyranoside [18] Flavonoid C21H20O12 Anti-inflammatory [117]

170 35-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

171 45-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

169 34-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

126 Quercetin 331015840-dimethyl ether7-O-120572-L-rhamnopyranosyl-(1rarr 6)-120573-D-glucopyranoside [118] Flavonoid C21H20O11 Anti-malarial [119]

125 Quercetin 331015840-dimethyl ether-7-O-120573-D-glucopyranoside [118] Flavonoid C21H20O12 Anti-malarial [119]70 1-Phenyl-13-diyn-5-en-7-ol-acetate [27] Polyyne C15H12O2 Anti-malarial [27]199 Heptanyl 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside [70] Miscellaneous C18H44O10 Antioxidant [70]124 3-O-Rabinobioside [70] Saccharide C27H38O15 Antioxidant [70]130 Quercetin 3-O-rutinoside [70] Flavonoid C27H38O15 Antioxidant [70]167 Chlorogenic acid [70] Phenolic C16H26O9 Antioxidant [70]119 Jacein [70] Flavonoid C24H26O13 Antioxidant [70]

49 (R)-12-dihydroxytrideca-357911-pentayne [47] Polyyne C13H8O2Anti-malarial andAntibacterial [47]

40 Evidence-Based Complementary and Alternative Medicine

Table 12 Structure and activity relationship studies of ethyl caffeateusing in vitro NF-120581BDNA binding assays [84]

Compound Concentration(120583M)

NF-120581BDNA binding

Ethyl caffeate 50 100inhibition

Ethyl 34-dihydroxyhydrocinnamate 100 100inhibition

Catechol 400 100inhibition

Ethyl cinnamate 400 Noinhibition

Table 13 Apoptosis in cocultures of T cells and pancreatic 120573 cells[53]

Cellmedium apoptosis andnecrosis

CD4+ T cellscontrol medium lt4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 2

CD4+ T cellswith cytopiloyne-treated 120573 cells 18CD4+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 7

CD8+ T cellscontrol medium 4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 4

CD8+ T cellswith cytopiloyne-treated 120573 cells 4CD8+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 4

Table 14 Radical scavenging activities of B pilosa extracts [70]

Extractscontrol DPPH assayIC50 (120583gmL)

NBThypoxanthinesuperoxide assayIC50 (120583gmL)

Quercetin 198 15Ascorbic acid 634 Not determined120572-tocopherol 897 Not determinedEthyl acetateextract 1383 597

Butanol extract 1669 114Water extract gt100 gt100

compounds) and inhibition (by 40 compared to 10) ofdifferentiation toTh1 at the same concentration [49]

Among the three polyynes found in B pilosa cytopiloyne(53) had the most potent anti-T1D activity [53] To test the invivo effect of cytopiloyne NODmice received intraperitonealor intramuscular injection of cytopiloyne at 25120583gkg BW3 times per week Twelve-week-old NOD mice started todevelop T1D and 70 of NOD mice aged 23 weeks andover developed T1D Remarkably 12- to 30-week-old NODmice treated with cytopiloyne showed normal levels of bloodglucose (lt200mgdL) and insulin (1-2 ngmL) Consistent

Table 15 Radical scavenging activity of secondarymetabolites fromB pilosa [70]

Metabolite (Table 11)control DPPH assay IC50 (120583gmL)199 Not determined124 53130 68167 105169 33171 38119 Not determined110 Not determinedQuercetin 256Caffeic acid 890

Table 16 Antioxidant activity of the essential oils and water extractsfrom B pilosa [92]

Extract IC50 (120583gmL)Leaf essential oils 47Flower essential oils 50Leaf extract 61Flower extract 172

with T1D incidence cytopiloyne delayed and reduced theinvasion of CD4+ T cells into the pancreatic islets [53]

In vitro study showed that cytopiloyne (53) inhibited thedifferentiation of naıveTh (Th0) cells (ie CD4+ T cells) intoTh1 cells and promoted differentiation of Th0 cells into Th2cells [50] The in vitro data are consistent with the in vivoresults indicating that cytopiloyne reducedTh1differentiationand increased Th2 differentiation as shown by intracellularcytokine staining and FACS analysis [53] In line with theskewing of Th differentiation the level of serum IFN-120574 andIgG2c decreased while that of serum IL-4 and serum IgEincreased compared to the negative controls (PBS-treatedmice) Cytopiloyne also enhanced the expression of GATA-3 a master gene for Th2 cell differentiation but not theexpression of T-bet a master gene forTh1 cell differentiationfurther supporting its role in skewingTh differentiation [53]

Also importantly cytopiloyne partially depleted CD4+rather than CD8+ T cells in NOD mice [53] As shown inTable 13 coculture assays showed that the depletion of CD4+T cells was mediated through the induction of Fas ligandexpression on pancreatic islet cells by cytopiloyne leading toapoptosis of infiltrating CD4+ T cells in the pancreas via theFas and Fas ligand pathways However cytopiloyne did notinduce the expression of TNF-120572 in pancreatic islet cells andthus had no effect on CD8+ T cells [53]

In addition Chang and colleagues showed that cytopi-loyne dose-dependently inhibited T-cell proliferation stimu-lated by IL-2 plus Con A or anti-CD3 antibody using [3H]thymidine incorporation assay [53]

Overall the mechanism of action of cytopiloyne andprobably its derivatives in T1D includes inhibition of T-cellproliferation skewing of Th cell differentiation and partialdepletion of Th cells Due to the anti-diabetic mechanisms

Evidence-Based Complementary and Alternative Medicine 41

Table 17 Antibacterial activity of essential oils and flower extracts from B pilosa [92]

Strain Mean zone of inhibition (mm)Leaf essential oil Flower essential oil Leaf extract Flower extract

Micrococcus flavus 127 plusmn 03 87 plusmn 03 102 plusmn 02 108 plusmn 03Bacillus subtilis 173 plusmn 19 117 plusmn 02 109 plusmn 02 103 plusmn 02Bacillus cereus 190 plusmn 14 112 plusmn 03 118 plusmn 04 185 plusmn 10Bacillus pumilus 123 plusmn 07 108 plusmn 02 105 plusmn 04 77 plusmn 02Escherichia coli 137 plusmn 04 203 plusmn 07 102 plusmn 11 140 plusmn 13Pseudomonas ovalis 125 plusmn 08 137 plusmn 15 102 plusmn 06 125 plusmn 06

Table 18 Antibacterial activity of root extracts from B pilosa [93]

Strain MIC50 (mgmL)Methanol extract Acetone extract

Bacillus cereus 10 mdashEscherichia coli 5 5Klebsilla pneumonia 5 10Micrococcus kristinae 10 mdashPseudomonas aeruginosa 10 10Staphylococcus aureus 5 10Sraphylococcus epidermidis 5 5Serratia marcescens 10 mdashShigelea flexneri 10 mdashStreptococcus faecalis 10 mdash

Table 19 Antibacterial activity of B pilosa of compound 29 [47]

Strain MIC50 (120583gmL)Escherichia coli NIHJ 1Escherichia coli ATCC25922 1Klebsiella pneumoniae ATCC700603 128Serratia marcescens ATCC13880 16Pseudomonas aeruginosa ATCC27853 8Staphylococcus aureus FDA209P 05Staphylococcus aureus ATCC29213 025Staphylococcus aureus N315 (MRSA) 05Enterococcus faecalis ATCC29212 2Enterococcus faecalis NCTC12201 (VRE) 1Bacillus subtilis ATCC6633 05Candida albicans ATCC10231 025

of action it was hypothesized that cytopiloyne protectsNOD mice from diabetes by a generalized suppression ofadaptive immunity To evaluate this hypothesis ovalbumin(Ova) was used as a T-cell dependent antigen to primeNOD mice which had already received cytopiloyne orPBS vehicle Ova priming boosted similar anti-Ova titersin cytopiloyne-treated mice and PBS-treated mice but adifference in immunoglobulin isotype was observed in thetwo groups Thus it was concluded that cytopiloyne is animmunomodulatory compound rather than an immunosup-pressive compound [50 53]

T2D is a chronic metabolic disease with serious com-plications resulting from defects in either insulin secretion

insulin action or both [124] A study by Ubillas et alshowed that the aqueous ethanol extract of the aerial partof B pilosa at 1 gkg body weight (BW) lowered bloodglucose in dbdb mice a T2D mouse model [51] Basedon a bioactivity-guided identification compounds 69 and50 were identified Further the mixture of the compounds(69 50) in a 2 3 ratio significantly decreased blood glucoseconcentration and reduced food intake on the second dayof treatment when administered at doses of 250mgkg twicea day to C5BLKs-dbdb mice When tested at 500mgkga more substantial drop in blood glucose level as wellas the stronger anorexic effect (food intake reduced from58 gmouseday to 25 gmouseday) was observed [51] Inthis study it was suggested that the blood glucose lower-ing effect of B pilosa was caused in part by the hungersuppressing effect of its polyynes [51] However the hungersuppressing effect of the ethanol extract of B pilosa wasnot found in the studies described below In another study[24] water extracts of B pilosa (BPWE) were used indiabetic dbdb mice aged 6ndash8 weeks with postprandialblood glucose levels of 350 to 400mgdL Like oral anti-diabetic glimepiride which stimulates insulin release onesingle dose of BPWE reduced blood glucose levels from374 to 144mgdL The antihyperglycemic effect of BPWEwas inversely correlated to an increase in serum insulinlevels suggesting that BPWE acts to lower blood glucose viaincreased insulin production However BPWE had differentinsulin secretion kinetics to glimepiride [24] One flaw incurrent anti-diabetics is their decreasing efficacy over timeThe authors investigated the long term anti-diabetic effectof BPWE in dbdb mice BPWE reduced blood glucoseincreased blood insulin improved glucose tolerance andreduced the percentage of glycosylated hemoglobin (HbA1c)Both long-term and one-time experiments strongly supportthe anti-diabetic action of BPWE [24] In sharp contrast toglimepiride BPWE protected against islet atrophy in mousepancreas The investigators further evaluated anti-diabeticproperties of 3 B pilosa varieties B pilosa L var radiate(BPR) Bpilosa L var pilosa (BPP) and B pilosa L varminor(BPM) in dbdb mice [8] One single oral dose (10 50 and250mgkg body weight) of BPR BPP or BPM crude extractsdecreased postprandial blood glucose levels in dbdb micefor up to four hours and the reduction of glucose levels inthe blood appeared to be dose-dependent Comparing thethree variants BPR extract resulted in a higher reduction inblood glucose levels when administered at the same dose as

42 Evidence-Based Complementary and Alternative Medicine

Table 20 Antifungal activity of B pilosa [92]

Partextract Concentration (ppm) Strain InhibitionCortiicum rolfsii Fusarium solani Fusarium oxysporum

Leaves

Essentialoils

100 857 plusmn 09 682 plusmn 0 745 plusmn 17250 960 plusmn 08 779 plusmn 18 879 plusmn 04

AqueousExtracts

100 446 plusmn 17 605 plusmn 21 716 plusmn 07250 942 plusmn 03 689 plusmn 07 824 plusmn 19

Flowers

Essentialoils

100 604 plusmn 09 892 plusmn 04 869 plusmn 05250 894 plusmn 12 980 plusmn 03 949 plusmn 06

AqueousExtracts

100 331 plusmn 11 714 plusmn 07 573 plusmn 22250 661 plusmn 14 912 plusmn 0 900 plusmn 07

Table 21 Antifungal activity of B pilosa root extracts [93]

Strain LC50 (mgmL)Acetoneextracts

Methanolextracts

Waterextracts

Aspergillus niger 014 006 007Aspergillus flavus 1091 658 0Penicillium notatum 005 005 005

the other two varieties In terms of serum insulin levels a doseof 50mgkg of each extract was used and the BPR extracttogether with the three polyynes significantly increased theserum insulin level in dbdb mice Long-term experiments(28-day treatment) were then conducted using diabetic micewith postprandial glucose levels from 370 to 420mgdL andglimepiride was used as positive controlThe range of dosagesapplied was from 10mgkg BW to 250mgkg BW Resultsshowed that the positive control as well as the crude extractsof the three varieties lowered the blood glucose levels indbdb mice However only BPR extract containing a higherpercentage of cytopiloyne (53) reduced blood glucose levelsand augmented blood insulin levelsmore than BPP andBPMThe percentage of glycosylated hemoglobin A1c (HbA1c) wasalso measured and found to be 79 plusmn 05 in mice aged 10ndash12 weeks and 66 plusmn 02 61 plusmn 03 and 62 plusmn 03 inthe blood of age-matchedmice following treatment with BPRcrude extract (50mgkg) glimepiride (1mgkg) and com-pound 53 (05mgkg) respectively [8] Among the polyynesfound in B pilosa cytopiloyne was the most effective againstT2D Hence cytopiloyne was used for further study on anti-diabetic action and mechanism [125] The data confirmedthat cytopiloyne reduced postprandial blood glucose lev-els increased blood insulin improved glucose tolerancesuppressed HbA1c level and protected pancreatic islets indbdb mice Nevertheless cytopiloyne failed to decreaseblood glucose in streptoztocin (STZ)-treated mice whose bcells were already destroyed Additionally cytopiloyne dose-dependently increased insulin secretion and expression in bcells as well as calcium influx diacylglycerol and activationof protein kinase C120572 Collectively the mechanistic studiessuggest that cytopiloyne treats T2D via regulation of insulin

production involving the calciumDAGPKC120572 cascade in bcells

The studies detailed above point to the conclusion thatcytopiloyne and related polyynes (compounds 69 and 49)are anti-diabetics in animal models The data uncover a newbiological action of polyynes It should be noted that likeall anti-diabetic drugs cytopiloyne failed to prevent or curediabetes completely but reduced diabetic complications [125]Intriguingly 34 polyynes have been found in B pilosa so farIt remains to be seen whether all the polyynes present in thisplant have anti-diabetic activities

34 Antioxidant Activity Free radicals can damage cellularcomponents via a series of chemical reactions [89] leadingto development and progression of cardiovascular diseasecancer neurodegenerative diseases and ageing [70] Freeradicals nitric oxide (NO) and superoxide anions can beproduced in macrophages to kill microbes However anexcessive generation of the free radicals under pathologicalconditions is associated with a wide range of illnessesPlants are known to be rich in antioxidant phytochemicalsChiang and colleagues evaluated the free radical scaveng-ing activity of crude extract fractions and compounds ofB pilosa using 11-diphenyl-2-picrylhydrazyl (DPPH) andhypoxanthinexanthine oxidase assays [70] Using DPPHand hypoxanthinexanthine oxidase assays they found thatthe B pilosa crude extract and the ethyl acetate butanoland water fractions had free radical scavenging activityNine compounds Heptyl- 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside (199) 3-O-Rabinobioside (124) Quercetin3-O-rutinoside (130) Chlorogenic acid (167) 34-Di-O-caffeoylquinic acid (169) 35-Di-O-caffeoylquinic acid (170)45-Di-O-caffeoylquinic acid (171) Jacein (119) and Cen-taurein (110) had DPPH radical scavenging activity [70]The IC

50values of the B pilosa crude extractfractions and

compounds are summarized in Tables 14 and 15 respectivelyMeasurement of free radical scavenging activities is one

way of assessing the antioxidant activities of B pilosa andits fractions and compounds It is interesting that the ethylacetate and butanol fractions are more active than the waterfraction and B pilosa crude extract [70] Of the secondarymetabolites only phenolic compounds 124 130 167 169 and171 showed significant DPPH-radical scavenging activities

Evidence-Based Complementary and Alternative Medicine 43

Further analysis of the structure-activity relationship of thecompounds suggested that substitution of the C3 hydroxylgroup with glycosides increased the activity approximately2-fold (for example in compounds 124 and 130) relative toquercetin (-OH in its C3) [70] Further modification of thestructures of the active compounds needs to be performed totest the effects of various substituents on activity The reasonwhy most of the antioxidant compounds contain phenolmoieties in their structure could be that the reduction-oxidation (redox) properties of phenols allow them to actas reducing agents singlet oxygen quenchers and hydrogendonors

A complementary study by Muchuweti and colleaguesdetermined phenolic content antioxidant activity and thephenolic profile of B pilosa methanol extract [89] Theyestimated that the phenolic content of the methanol extractof B pilosawas 11028plusmn22mgg [89] Vanillin hydroxyben-zaldehyde caffeic acid coumaric acid and ferulic acid werefound in this extractThe B pilosa extract also showed DPPHradical scavenging activity Furthermore the antioxidantactivity of the flavonoids found in B pilosa was correlatedwith its hepatoprotective effects through their inhibitionof NF-120581B activation which may lessen the oxidative stresscaused by the production of free radicals during liver injury[60]This activity might also be due to the anti-inflammatoryeffects of the aqueous extracts of B pilosa aerial parts on theinhibition of COX-2 and PGE

2production [88]

Essential oils from B pilosa flowers and leaves are alsoreported to possess antioxidant activity With the aim ofreplacing chemically synthesized additives Deba and col-leagues [92] worked on the antioxidant antibacterial andantifungal activities of essential oils and water extracts of Bpilosarsquos leaves and flowers Table 16 summarizes the resultsobtained from DPPH free radical scavenging assay

It can be inferred fromTable 16 that essential oils from theleaves possessed the highest activity It is reported elsewherethat monoterpenes present in essential oils such that of Bpilosa have protective effects and antioxidant properties [92]Beta-carotene bleaching method was also performed Leavesessential oils and aqueous extracts of the leaves and flowersshowed higher activity than the flower essential oils This isdue to the volatility of the flower essential oils The activityexhibited by the aqueous extracts was accounted to thepresence of phenolic compounds that are reported to donatea hydrogen atom to free radicals such that the propagation ofthe chain reaction during lipid oxidation is terminated [92]Overall essential oils and phenolics present in B pilosa canbe thought of as major antioxidant compounds

35 Immunomodulatory Activity B pilosa is thought to bean immunomodulatory plant and is reported to be effectivein the treatment of immune disorders such as allergy [75]arthritis [75] and T1D [48 50 53]

As pointed out in the discussion of its anti-diabeticactivities (Section 33) a combination of phytochemicalsmethod and T-cell activation assays was used to studyimmunomodulatory properties of B pilosa IFN-120574 is a keycytokine released by T and NK cells that mediates immune

cells and sustains immunity against pathogens Defects inIFN-120574 expression regulation and activation result in vulner-ability to diseases caused by bacteria and viruses [69] Anelegant study performed byChang and colleagues using IFN-120574 promoter-driven luciferase reporter construct in JurkatT cells showed that hot water crude extracts of B pilosaincreased IFN-120574 promoter activity two-fold [69] Out of thesubfractions of this extract the butanol fraction but not thewater or ethyl acetate fractions increased IFN-120574 promoteractivity six-fold Centaurein (110) and centaureidin (109)were identified from the butanol fraction and were stated tocause a four-fold increase in IFN-120574 promoter activity withEC50

values of 75 120583gmL and 09120583gmL respectively Themechanism of action of centaurein was determined usingtranscription factors such as AP-1 NFAT and NF120581B whichare reported to bind to IFN-120574 promoter and regulate IFN-120574transcription Unlike with the activity of the positive controlPHA centaurein caused a four-fold increase in NFAT a3-fold increase in NF120581B and had little if any effect onAP-1 enhancer activities (23-fold three-fold and ten-foldincreases respectively were seenwith PHA) [69]The authorsconcluded that centaureinmodulates IFN-120574 expression by theNFAT and NF120581B pathways The article only determined themechanism of action of centaurein Its aglycone centaureidinmay act through the samemechanism though this conclusionneeds to be further verified

B pilosa extract and its compounds are reported to inhibitdifferentiation of naıve CD4+ helper T (Th0) cells into Th1cells [49] Using theThcell differentiation assay as a screeningplatform 3 polyynes 2-120573-D-glucopyranosyloxy-1 -hydroxytrideca-57911-tetrayne (53) 3-120573-D-glucopyranosyl-1-hydroxy-6(E)- tetradecene-81012-triyne (69) and 2-120573-D-glucopyranosyloxy-1-hydroxy-5(E)- tridecene-7911-triyne(49) were discovered from B pilosa [49 53] The data showsthat cytopiloyne and other two polyynes suppressed thedifferentiation of type 1 helper T (Th1) cells and productionof Th1 cytokines and promoted that of type 2 helper T (Th2)cells and production of Th2 cytokines thus explaining theimmunomodulatory and anti-inflammatory effects of Bpilosa and its polyynes

Chang and colleagues were the first to report the effect ofthe butanol extract of B pilosa on the autoimmune diabetesand airway inflammation inmice [48] Imbalance in the levelsofTh1 andTh2 and of various cytokines leads to autoimmunediseases T1D and other autoimmune diseases (rheumatoidarthritis Crohnrsquos disease among others) are exacerbated byan increase Th1 levels (specifically CD4+ Th1 cells) whileTh2 cells antagonize this effect [19] Moreover Th2 cellsmediate asthma in ovalbumin-induced hypersensitivity inBALBc mice [48] In their work [48] Chang and colleaguesshowed that 10mgkg butanol extracts with 15 (ww)compound 69 and 11 (ww) compound 49 (Table 11)ameliorated the development of Th1-mediated diabetes inNOD mice through inhibition of 120573 cell death and leukocyteinfiltration At the same dosage the butanol extracts alsoexacerbated ovalbumin-induced pulmonary inflammationin BALBc mice with an increase in the infiltration ofeosinophils and mast cells into the airway of the mice [48]Despite the different outcomes both mouse models proved

44 Evidence-Based Complementary and Alternative Medicine

the concept that control over the Th1Th2 shift is associatedwith autoimmune diseases and that the B pilosa butanolextract can shift the differentiation of Th0 cells to Th2 cells[48 49]

An extended study presented by Chiang and colleaguesshowed that compound 53 modulates T-cell functions [50]Using CD4+ T cells from BALBc mice they demonstratedthat cytopiloyne decreased levels of IFN-120574 producing cells(Th1) by 122 (from 72 to 598) Since Th1 and Th2 celldifferentiation is antagonistic it was expected that compound53 increased the percentage of mouse IL-4 producing cells(Th2) by 72 (from 237 to 309) Subsequent assess-ment of effect of compound 53 on the modulation of thetranscription of IL-4 and IFN-120574 showed that cytopiloyne asexpected decreased the splenocyte levels of IFN-120574 mRNAand increasing that of IL-4 in a dose-dependent manner Inthe range of 01 to 3 120583gmL of compound 53 the effects werenot attributed to its cytotoxicity Consequently using 3120583gmLcytopiloyne for 72 and 96 hours the protein concentration ofIFN-120574 decreased to 186 and 444 respectively Under thesame conditions cytopiloyne increased IL-4 concentrationsto 1985 and 2470 (for 72 and 96 hours resp) Thismodulation of T-cell differentiation exhibited by compound53 was used to explain its anti-diabetic activity [50]

The anti-diabetic role of cytopiloyne was extensivelydiscussed above (Section 33 anti-diabetic activity) Themolecular basis of the regulation of cytokine expression bycytopiloyne has been described Cytopiloyne directly elevatedthe expression level of IL-4 via GATA-3 upregulation inT cells [53] However reduction of IFN-120574 expression in Tcells seemed to come from the indirect opposing effect IL-4cytokine because the expression level of T-bet was unaltered[53] In this way cytopiloyne skewedTh1 polarization into theTh2 state conferring protection against T1D in NOD miceAside frompolarization ofThcell differentiation cytopiloynealso activated the expression of Fas ligand in pancreaticb cells this increase leading to the partial depletion of Tcells and reduction of immune response in local areas suchas the pancreas Of note cytopiloyne also inhibited T-cellproliferation and activation By targeting T cells from threeimmunomodulatory actions cytopiloyne protects againstT1D and probably other Th1-mediated autoimmune diseases[19]

The phytochemical constituents of B pilosa exert theirfunctions on different immune cells to modulate immuneresponse It is possible that some of the compounds mayhave agonistic or antagonistic effects on immune responseImmune function of B pilosa may depend on its compo-sition and amount of compounds which could explain theapparently conflicting report of B pilosa butanol extractaggravating allergy in mice [48] while cellulosine-treatedextract ameliorated allergy [48 75] IFN-120574 promoter reporterassays and T-cell differentiation assays were used to isolate2 flavonoids [69] and 3 polyynes [49 50] as immunomod-ulatory compounds from the butanol fraction of B pilosaInterestingly the flavonoids promote IFN-120574 expression inNK and T cells In marked contrast the polyynes promoteIL-4 expression and indirectly inhibit IFN-120574 expression indifferentiating T cells Sensitivity appears to be the key to

identifying structure- and bioactivity-related phytochemicalsfrom medicinal plants

36 Antimalarial Activity The use of chemical drugs againstpathogens has resulted in drug-resistant mutants Examplesof drug resistance can be found in the species of thePlasmodium that cause malaria It is important to search fornew compounds to combat Plasmodium parasites [47] Astudy of the anti-malarial activity of the leaf extracts of Bpilosa using a combination of phytochemistry and bioassaysshowed that compound 49 (Table 11) showed activity againsta malaria parasite (P falciparum NF54 strain) with an IC

50

value of 60 120583gmL [77] In addition compound 49 isolatedfrom the aerial parts of B pilosa inhibited growth of the Pfalciparum FCR-3 strain with an IC

50value of 035 120583gmL

This compound was tested for its in vivo effect in miceinfected with P berghei NK-65 strain Results showed thatcompound 49 decreased the average parasitemia in the redblood cells by 207 (from 328 of that of the control to121) after an intravenous injection of 08mgkg BWday forfour days [47] Further studies addressing the anti-malarialmechanism underlying both polyynes and clinical studies areneeded

37 Antibacterial Activity Emergence of multiple antibiotic-resistantmicrobes is becoming a global threat to public healthand a challenge to disease treatment For instance penicillinis commonly used to combat a food-borne intracellularbacteriumListeria however penicillin-resistant bacteria havebeen discovered recently [69] Chang and coworkers iso-lated centaurein (110) and centaureidin (109) from B pilosaextract [69 70] Centaurein enhances expression of IFN-120574a key cytokine for macrophage activation and consequentlyenhances bactericidal activity in macrophages [69 70] Inagreement with observed in vitro effects centaurein wasreported to prevent and treat Listeria infection in C57BL6Jmice [91] Mechanistic studies confirmed that centaureinexerted antilisterial action via IFN-120574 expression in wild-typemice but not IFN-120574 knockout mice [91] Further in vitrostudies on centaurein showed that this compound increasedIFN-120574 expression by 13 (from 17 to 20) 20 (from 21to 41) and 11 (from 6 to 17) in CD4+ T cells CD8+Tcells and NK cells respectively That is to say there wasan increase in IFN-120574 producing immune cells As expectedcentaurein also enhanced the expression level of T-bet a keynuclear factor for IFN-120574 expression Consistently centaureinaugmented the serum IFN-120574 levels inC57BL6Jmice and thisaugmentation peaked 24 hours after compound injectionThe quantity of mouse serum IFN-120574 was sufficient to activatemacrophages in vitro and eradicated GFP-producing Listeriainside macrophages However the entry of Listeria intomacrophages was not affected by centaurein-treated mousesera Centaurein treatment at 20120583g per mouse rescued 30of the mice infected with a lethal dose of Listeria (2 times106 CFU) It is noteworthy that in the presence of ampicillin(5 120583gmouse) centaurein (20 120583gmouse) rescued 70 of themice suggesting an additive effect between ampicillin andcentaurein [91] Despite lower abundance centaureidin was

Evidence-Based Complementary and Alternative Medicine 45

30 times more active than centaurein in terms of IFN-120574production [91]

Aside from the indirect antibacterial action mentionedabove extract andor compounds of B pilosa also showeddirect bacteriostatic andor bactericidal action One studyreported that essential oils and leafflower extracts of Bpilosa could suppress the growth of gram positive and gramnegative bacteria as evidenced by zone of inhibition assaysIn this study antibacterial activity of the essential oils fromthe leaves and flowers of B pilosa was determined in anattempt to identify natural products as food preservatives forprevention of microbial multiplication and food oxidationThe essential oils and extracts of B pilosa leaves and flow-ers showed moderate but different extents of antibacterialactivity (Table 17) In general essential oils had higherantibacterial activity than crude extracts One explanation forthis could be that monoterpenes in the essential oils destroycellular integrity and subsequently inhibit the respirationand ion transport processes The presence of antibacterial120573-caryophyllene could be another explanation as reportedelsewhere [92] Another study reported that the methanoland acetone extract of B pilosa roots displayed antibacterialactivities against the bacteria listed in Table 18 [93] andmethanol extracts from the roots seemed to be the mosteffective

Another study indicated that the polyyne (R)-12-dihydroxytrideca-357911-pentayne (49) from this plantalso suppressed bacterial growth as shown by the minimuminhibitory concentration required to inhibit 50 bacterialgrowth (MIC

50) in Table 19 This compound was highly

effective against several Gram positive and Gram negativebacteria including the drug-resistant bacteria Staphylococcusaureus N315 (MRSA) and Enterococcus faecalis NCTC12201(VRE) [47] Strikingly compound 49 had a similar MIC

50

value to antibiotics (ampicillin tetracycline norfloxacin andamphotericin B) in most of the bacteria tested

Antibacterial activity of B pilosa extracts and compo-nents expressed as MIC

50and the mean zone of inhibition

is tabulated in Tables 17 18 and 19The zone of inhibition forAmpicilline (positive control) ranges from 153 plusmn 03mm to443 plusmn 02mm [92]

38 Antifungal Activity B pilosa has traditionally been usedto treat microbial infection Recently different parts of Bpilosa have been tested for antifungal activities Deba andcolleagues first evaluated the antifungal effect of the hotwater extracts of the B pilosa roots stems and leaves againstCorticium rolfsii Fusarium solani and Fusarium oxysporumThey discovered that C rolfsii was most suppressed bytreatment with B pilosa as its growth was reduced at almostall the tested doses followed by F oxysporum and F solani[99] However the fungicidal activities of the stems androots were greater than the leaves [99] Moreover the samegroup assessed the antifungal activity of the essential oilsand aqueous extracts from B pilosa flowers and leaves [92]They showed that the extracts and oils had antifungal activityagainst C rolfsii F solani and F oxysporum Essential oils

appeared to have better fungicidal activity thanwater extractsas summarized in Table 20

Another study by Ashafa and colleagues showed thatacetone methanol and water extracts of the B pilosa rootsshowed antifungal activities against Aspergillus niger Aflavus and Penicillium notatum using the agar dilutionmethod The results are tabulated in Table 21 [93] Negativecontrols showed 0 growth inhibitionThe methanol extractof the B pilosa roots at 10mgmL was also effective againstCandida albicans [93]Of note B pilosa obtained fromPapuaNew Guinea had no activity against A niger and C albicans[126] but the SouthAfrican (EasternCape) ecotype exhibitedmoderate activity against C albicans [93] This discrepancymay depend on extraction solvents extraction procedureassay techniques different plant parts and abundance ofactive compounds

B pilosa produces a variety of secondary metabolitessuch as flavonoids phenylacetylenes alkaloids sterols ter-penoids and tannis [92 93] However none of them havebeen confirmed as active compounds against fungi Furtherinvestigation of active compounds from B pilosa is necessaryto further understand the antifungal efficacy of this plant

39 Hypotensive and Vasodilatory Activities In early studiesDimo and colleagues used three rat models normotensiveWistar rats (NTR) salt-loading hypertensive rats (SLHR)and spontaneous hypertensive rats (SHR) to investigate thehypotensive effect of the methanol crude extract of B pilosaleaves [28 127] The extract lowered systolic blood pressurein hypertensive rats (SLHR and SHR) to a greater degreethan NTR [28 127] In addition a decrease in urinarysodium ions and an increase in urinary potassium ions wereobserved after treatment with the methanol extract of Bpilosa leaves although neither differences were statisticallysignificant [28 127] Taking the data together the studyproposed that B pilosa leaf extract reduced blood pressurevia vasodilation [28 127] The same group continued totest the antihypertensive effect of aqueous and methylenechloride extracts of B pilosa leaves in a hypertensive ratmodel [28 127] To establish a fructose-induced hypertensionmodel male Wistar rats were given 10 fructose solutionto drink ad libitum for three weeks In addition to freeaccess to 10 fructose the rats were treated with the aqueous(150mgkg) or methyl chloride (350mgkg) extracts of Bpilosa for additional three weeks [28 127] Both extracts ofB pilosa leaves had a hypotensive effect on rats Howeverneither extracts reversed the elevation of serum insulin infructose-fed rats Therefore B pilosa lowered blood pressureirrespective of insulin [28 127] To better understand thehypotensive mechanism the authors investigated the effectof a neutral extract of B pilosa (NBP) a mixture of methanolandmethylene chloride (1 1) extract after neutralization withNaOH and HCl on the heart and the blood pressure ofNTR and SHR [110] This study showed that an intravenousinjection of the NBP resulted in a biphasic reduction insystolic blood pressure In addition one intravenous dose ofthe extract at 10 20 and 30mgkg BW decreased systolicblood pressure in normal rats by 183 425 and 30

46 Evidence-Based Complementary and Alternative Medicine

respectively and the same doses reduced the blood pressurein hypertensive rats by 258 389 and 286 respectivelyOnly the highest dose (30mgkg) affected the force of thecontraction of the heart Atropine and propranolol were usedto interfere with the hypotensive action of the NBP Atropinereduced the initial phase of the hypotensive response in NBPand completely abolished the second phase of hypotensiveresponse in NBP In contrast propranolol increased thefirst hypotensive response but partially abolished the secondhypotensive response provoked by the NBP [110]This mech-anistic study suggested that B pilosa invokes the biphasichypotensive responses via targeting cardiac pump efficiencyduring the first phase and vasodilation at the second phase[110]

A further study was performed to investigate the relaxingeffect of a neutral extract of B pilosa (NBP) on rat aorta con-tractedwithKCl (60mM) andnorepinephrine (01mM) [94]Cumulative addition of NBP relaxed the rat aorta previouslycontracted by KCl in a dose-dependent manner The EC

50

value of theNBP for vasorelaxationwas 032mgmLThedataalso showed that the NBP reduced the contraction of aortapreviously contracted by KCl irrespective of the presence ofaortic endothelium [94]

Pretreatment with glibenclamide an ATP-dependent K+channel blocker did not considerably affect the relaxant effectof the NBP on KCl-induced contraction suggesting thatthe vasodilatory effect of B pilosa was not related to theopening of this ATP-dependent K+ channel [94] On theother hand in the presence of indomethacin or pyrilaminemaleate the relaxant response induced by the plant extractwas significantly inhibited at the lower concentrations Theplant extract was able to reduce the aorta resting tone inhibitthe KCl-induced contractions by 90 at 15mgmL and theCaCl2-induced contractions by 95 at 075mgmL These

results demonstrate that B pilosa can act as a vasodilatorprobably via acting as a calcium antagonist [94]

However no specific compound for the above activityhas been identified from B pilosa to date A bioactivity-guided identification approach may be adopted to identifythe active compounds in B pilosa that possess hypotensiveand vasodilatory effects and understand their mechanism ofaction

310 Wound Healing Activity B pilosa has been traditionallyused to treat tissue injury inCameroon Brazil andVenezuela[26] Hassan and colleagues investigated the wound healingpotential of B pilosa in Wistar rats [128] Mirroring thepositive control neomycin sulfate the ethanol extract of Bpilosa had faster wound closure than control rats 3 6 and 9days after topical application Histological examination alsorevealed better collagenation angiogenesis and organizationof wound tissue seven days after application Epithelializationand total healing time in B pilosa-treated rats were compara-ble to those of neomycin sulfate Together these data suggestthat B pilosa may be a viable alternative to neomycin lotionfor the treatment of wounds

In addition to studying the wound healing effect of Bpilosa on external ulcers Tan and colleagues also examined

the effect of methanol cyclohexane and methyl chlorideextracts of B pilosa on gastric ulcers in Wistar rats fedwith 1mL HClethanol gastric necrotizing solution (150mMHCl in 60 ethanol) and macroscopically visible lesionswere scored [26] Among the three extracts methylene chlo-ride extracts exhibited the highest activity showing 464inhibition of lesion formation at a dose of 500mgkg BWand complete inhibition at 750mgkg [26] The efficacy ofthe ethylene chloride extract was followed by that of themethanol extracts which had inhibition ranging from 304to 822 at concentrations of 500mgkg and 1000mgkgBW respectively [26] The cyclohexane extracts showed thelowest activity against gastric ulcers in rats with 13340 and 797 inhibition at 500 750 and 1000mgkg BWrespectively [26] To better understand the mode of actionof the methylene chloride extract of B pilosa rats werepretreated with indomethacin a COX-2 inhibitor involved inprostaglandin synthesis Pretreatment significantly reducedthe protection against HClethanol-induced ulcers to 313inhibition at 750mgkg BW suggesting a link between theantiulcerative activity of B pilosa and prostaglandin syn-thesis Unexpectedly the methylene chloride extract of Bpilosa showed little gastric mucosal protection against gastriclesions induced by 95 ethanol (1mL) [26] Absolute alcoholis known to cause mucosalsubmucosal tissue destruction viacellular necrosis and the release of tissue-derived mediators(histamine and leukotrieneC4)Thus these data imply thatBpilosa did not prevent the generation or the necrotic action ofthese mediators on the gastric microvasculature In additionpylorus ligation can increase gastric acid secretion withoutan alteration of mucosal histamine content The methylenechloride extract of B pilosa did not possess antisecretoryactivity On the contrary it was observed that increases in thedose of the extract led to elevated gastric juice acidity [26]Results with both absolute ethanol and pylorus ligation ratmodels suggested the possibility that the ineffectiveness of Bpilosa against gastric ulcers was due to lack of antihistaminicactivity in the plant In summary overall the data suggest thatB pilosa protects against HCLethanol-mediated ulcers viainhibition of prostaglandin biosynthesis

Previous phytochemical studies showed that a groupof flavonoids acyclichalcones are present in B pilosa[129 130] and the chalcones were proposed to have anti-ulcerative activity [131] Moreover nine hydroxychalcones 10were reported to possess gastric cytoprotective effects with24-dihydroxychalcone being the most active [132] Sincemethylene chloride extracts appear to be the most active Bpilosa extracts next the specific anti-ulcerative phytochemi-cals in the methylene chloride extracts of B pilosa and theirmodes of action needs to be probed

Despite the claims listed inTable 3 relatively few scientificstudies have been conducted in vitro and in vivo to address thetraditional ethnomedical uses of B pilosa Information aboutthe use of B pilosa as a botanical therapy recorded so far isfar from complete Studies conducted thus far only serve asa starting point for further investigation of B pilosa and theultimate efficacious use of the herb in clinical applications

Evidence-Based Complementary and Alternative Medicine 47

4 Toxicology

Despite its use as an ingredient in food for human con-sumption studies on systemic toxicity (eg acute subacutechronic and subchronic toxicities) of B pilosa in humansand animals are still inadequate and insufficient So faracute andor subchronic toxicities have been evaluated inrats and mice Oral acute and 28-day toxicities of waterextract of B pilosa leaves were evaluated in Wistar rats[133] An oral dose of water extract of B pilosa leaves at10 gkg BW showed no obvious mortality or changes inthe appearance in rats [134] The same extract at 08 gkgBWday once a day showed no obvious sub-chronic toxicityin rats over 28 days as measured by survival rate bodyweight and gross examination of organs [134] These dataare consistent with our data indicating that oral deliveryof the water extract of the B pilosa whole plant at 1 gkgBWday once a day is safe in rats over 28 days (unpublisheddata) Taken together these studies suggest that ingestion ofB pilosa aqueous extract at up to at 1 gkg BWday oncea day is highly safe in rats In addition the acute toxicityof aqueous and ethanol extracts of B pilosa in mice havebeen reported [134] Five- to six-week-old mice with weightsbetween 28 and 35 g received a peritoneal injection of bothextracts at the different doses The LD

50 the dose that causes

50 lethality of the aqueous and ethanol extracts in micewas 1230 gkg BW and 615 gkg BW respectively [134] Acomplete toxicological study has not been completed forhumans Furthermore the drug interactions of B pilosa withother drugs are unknown Further safety verification andclinical trials should be performed before B pilosa can beconsidered for medicinal use

5 Conclusions

B pilosa is an erect perennial plant with green leaves whiteor yellow flowers and tiny black seeds As it is distributedworldwide and is widely used as a folk remedy B pilosacan be thought of as an extraordinary source of food andmedicine However a comprehensive up-to-date review ofresearch on B pilosa has hitherto been unavailable In thisarticle scientific studies on B pilosa have been summarizedand critically discussed from the perspectives of botanyethnomedicine phytochemistry pharmacology and toxicol-ogy B pilosa is claimed to treat more than 40 disordersand 201 compounds have been identified from this plantThe medicinal utility of B pilosa and its modes of actionin relation to its known phytochemicals were discussedherein Polyynes flavonoids phenylpropanoids fatty acidsand phenolics are the primary bioactive compounds of Bpilosa and they have been reported to be effective in thetreatment of tumors inflammationimmune modulationdiabetes viruses microbes protozoans gastrointestinal dis-eases hypertension and cardiovascular diseases Cautionshould be exercised in the therapeutic use of B pilosa forhypoglycemia hypotension bleeding and allergy

Acknowledgments

The authors thank their laboratory members for construc-tive suggestions and the authors whose publications theycited for their contributions The authors also thank MsMiranda Loney of Academia Sinica for English editing of thispaper This work was supported by Grants 99-CDA-L11 and101S0010056 from Academia Sinica Taiwan Arlene P Bar-tolome is a recipient of MECO-TECO Sandwich ScholarshipProgram between Taiwan and the Philippines

References

[1] T Shen G H Li X N Wang and H X Lou ldquoThe genusCommiphora a review of its traditional uses phytochemistryand pharmacologyrdquo Journal of Ethnopharmacology vol 142 no2 pp 319ndash330

[2] C Long P Sauleau B David et al ldquoBioactive flavonoids ofTanacetum parthenium revisitedrdquo Phytochemistry vol 64 no2 pp 567ndash569 2003

[3] P O Karis and O Ryding Asteraceae Cladistics and Classifica-tion Bremer K Eds pp 559ndash569 Timber press Portland OreUSA 1994

[4] O N Pozharitskaya A N Shikov M N Makarova et alldquoAnti-inflammatory activity of a HPLC-fingerprinted aqueousinfusion of aerial part of Bidens tripartita LrdquoPhytomedicine vol17 no 6 pp 463ndash468 2010

[5] F Q Oliveira V Andrade-Neto A U Krettli and MG L Brandao ldquoNew evidences of antimalarial activity ofBidens pilosa roots extract correlated with polyacetylene andflavonoidsrdquo Journal of Ethnopharmacology vol 93 no 1 pp 39ndash42 2004

[6] Agriculture USDA ldquoPlants databaserdquo 2013in Natural Resources Conservation Servicehttpwwwnrcsusdagovwpsportalnrcssitenationalhome

[7] Y Zhou and X Z Yan ldquoExperimental study of qi deficiencysyndrome and Codonopsis pillosulae and Astragalus injectionon the immune response in micerdquo Zhong Xi Yi Jie He Za Zhivol 9 no 5 pp 286ndash262 1989

[8] S C Chien P H Young Y J Hsu et al ldquoAnti-diabeticproperties of three common Bidens pilosa variants in TaiwanrdquoPhytochemistry vol 70 no 10 pp 1246ndash1254 2009

[9] M J Alcaraz and M J Jimenez ldquoFlavonoids as anti-inflammatory agentsrdquo Fitoterapia vol 59 no 1 pp 25ndash38 1988

[10] FAO Agriculture Food and Nutrition for AfricamdashA ResourceBook for Teachers of Agriculture Publishing ManagementGroup FAO Information Division Rome Italy 1997

[11] M B Rokaya Z Munzbergova B Timsina and K R BhattaraildquoRheum australe D Don a review of its botany ethnobotanyphytochemistry and pharmacologyrdquo Journal of Ethnopharma-cology vol 141 no 3 pp 761ndash774 2012

[12] P H Young Y J Hsu and W C Yang ldquoBidens pilosa L andits medicinal userdquo in Recent Progress in Medicinal Plants DrugPlant II A S AwaadVK Singh and JNGovil Eds StandiumPress Houston Tex USA 2010

[13] C Ge ldquoCytologic study of Bidens bipinnata LrdquoZhongguo ZhongYao Za Zhi vol 15 no 2 pp 72ndash125 1990

[14] L C Chiang J S Chang C C Chen L T Ng and C CLin ldquoAnti-herpes simplex virus activity of Bidens pilosa and

48 Evidence-Based Complementary and Alternative Medicine

Houttuynia cordatardquoAmerican Journal of ChineseMedicine vol31 no 3 pp 355ndash362 2003

[15] N P Rybalchenko V A Prykhodko S S Nagorna et alldquoIn vitro antifungal activity of phenylheptatriyne from Bidenscernua L against yeastsrdquo Fitoterapia vol 81 no 5 pp 336ndash3382010

[16] K Redl W Breu B Davis and R Bauer ldquoAnti-inflammatoryactive polyacetylenes from Bidens campylothecardquo Planta Med-ica vol 60 no 1 pp 58ndash62 1994

[17] M Ayyanar and S Ignacimuthu ldquoTraditional knowledge ofKani tribals inKouthalai of Tirunelveli hills TamilNadu IndiardquoJournal of Ethnopharmacology vol 102 no 2 pp 246ndash255 2005

[18] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant intraditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[19] AKAbbas KMMurphy andA Sher ldquoFunctional diversity ofhelper T lymphocytesrdquo Nature vol 383 no 6603 pp 787ndash7931996

[20] J H Cano and G Volpato ldquoHerbal mixtures in the traditionalmedicine of Eastern Cubardquo Journal of Ethnopharmacology vol90 no 2-3 pp 293ndash316 2004

[21] J Namukobe J M Kasenene B T Kiremire et al ldquoTraditionalplants used for medicinal purposes by local communitiesaround the Northern sector of Kibale National Park UgandardquoJournal of Ethnopharmacology vol 136 no 1 pp 236ndash245 2011

[22] C Lans ldquoComparison of plants used for skin and stomachproblems in Trinidad and Tobago with Asian ethnomedicinerdquoJournal of Ethnobiology andEthnomedicine vol 3 article 3 2007

[23] S Dharmananda ldquoA Popular Remedy Ecapes Notice ofWesternPractitionersrdquo 2013 httpwwwitmonlineorgartsbidenshtm

[24] Y J Hsu T H Lee C L T Chang Y T Huang andW C YangldquoAnti-hyperglycemic effects and mechanism of Bidens pilosawater extractrdquo Journal of Ethnopharmacology vol 122 no 2 pp379ndash383 2009

[25] E Noumi F Houngue and D Lontsi ldquoTraditional medicinesin primary health care plants used for the treatment ofhypertension in Bafia Cameroonrdquo Fitoterapia vol 70 no 2 pp134ndash139 1999

[26] P V Tan T Dimo and E Dongo ldquoEffects of methanolcyclohexane and methylene chlo ride extracts of Bidens pilosaon various gastric ulcer models in ratsrdquo Journal of Ethnophar-macology vol 73 no 3 pp 415ndash421 2000

[27] R L C Pereira T Ibrahim L Lucchetti A J R Da Silva and VL G De Moraes ldquoImmunosuppressive and anti-inflammatoryeffects of methanolic extract and the polyacetylene isolatedfrom Bidens pilosa Lrdquo Immunopharmacology vol 43 no 1 pp31ndash37 1999

[28] T Dimo J Azay P V Tan et al ldquoEffects of the aqueous andmethylene chloride extracts of Bidens pilosa leaf on fructose-hypertensive ratsrdquo Journal of Ethnopharmacology vol 76 no 3pp 215ndash221 2001

[29] CWiartMedicinal Plants of Southeast Asia Pelanduk SelabgorDarul Ehsan Malaysia 2000

[30] F L Silva D C H Fischer J F Tavares M S Silva P FDe Athayde-Filho and J M Barbosa-Filho ldquoCompilation ofsecondary metabolites from Bidens pilosa LrdquoMolecules vol 16no 2 pp 1070ndash1102 2011

[31] A H Chen S R Lin and C H Hong ldquoPhytochemical studyon Bidens pilosa L varminorrdquo Huaxue pp 38ndash42 1975

[32] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[33] M C Zulueta M Tada and C Y Ragasa ldquoA diterpene fromBidens pilosardquo Phytochemistry vol 38 no 6 pp 1449ndash14501995

[34] J Wang H Yang Z W Lin and H D Sun ldquoFlavonoids fromBidens pilosa var radiatardquo Phytochemistry vol 46 no 7 pp1275ndash1278 1997

[35] A Zhao Q Zhao L Peng et al ldquoA new chalcone glycoside fromBidens pilosardquo Acta Botanica Yunnanica vol 26 no 1 pp 121ndash126

[36] P Gao Y Wu S Cui and Y Zou ldquoSynthesis and absoluteconfiguration of the 7-phenylhepta-4 6-diyne-1 2-diol isolatedfrom Bidens pilosardquo Synthesis no 13 pp 2131ndash2135 2011

[37] S Wang B Yang D Zhu D He and L Wang ldquoActivecomponents of Bidens pilosa Lrdquo Zhongcaoyao vol 36 pp 20ndash21

[38] F Bohlmann T Burkhardt and C Zdero Naturally OccuringAcetylenes Academic Press New York NY USA 1973

[39] T M Sarg A M Ateya N M Farrag and F A AbbasldquoConstituents and biological activity of Bidens pilosa L grownin EgyptrdquoActa Pharmaceutica Hungarica vol 61 no 6 pp 317ndash323 1991

[40] H A L Valdes and H P Rego ldquoBidens pilosa Linnerdquo RevistaCubana de Plantas Medicinales vol 6 pp 28ndash33

[41] F Bohlmann H Bornowski and K M Kleine ldquoNew polyynesfrom the tribe Heliantheaerdquo Chemische Berichte vol 97 pp2135ndash2138

[42] RWang Q XWu and Y P Shi ldquoPolyacetylenes and flavonoidsfrom the aerial parts ofBidens pilosardquo PlantaMedica vol 76 no9 pp 893ndash896 2010

[43] L W Wu Y M Chiang H C Chuang et al ldquoA novelpolyacetylene significantly inhibits angiogenesis and promotesapoptosis in human endothelial cells through activation of theCDK inhibitors and caspase-7rdquo Planta Medica vol 73 no 7 pp655ndash661 2007

[44] L W Wu Y M Chiang H C Chuang et al ldquoPolyacetylenesfunction as anti-angiogenic agentsrdquo Pharmaceutical Researchvol 21 no 11 pp 2112ndash2119 2004

[45] H L Yang S C Chen N W Chang et al ldquoProtection fromoxidative damage using Bidens pilosa extracts in normal humanerythrocytesrdquo Food and Chemical Toxicology vol 44 no 9 pp1513ndash1521 2006

[46] H Q Wang S J Lu H Li and Z H Yao ldquoEDTA-enhancedphytoremediation of lead contaminated soil by Bidens maxi-mowiczianardquo Journal of Environmental Sciences vol 19 no 12pp 1496ndash1499 2007

[47] S TobinagaMK SharmaWG L Aalbersberg et al ldquoIsolationand identification of a potent antimalarial and antibacterialpolyacetylene from Bidens pilosardquo Planta Medica vol 75 no 6pp 624ndash628 2009

[48] C L T Chang H K Kuo S L Chang et al ldquoThe distincteffects of a butanol fraction of Bidens pilosa plant extract onthe development of Th1-mediated diabetes and Th2-mediatedairway inflammation in micerdquo Journal of Biomedical Sciencevol 12 no 1 pp 79ndash89 2005

[49] S L Chang C L T Chang Y M Chiang et al ldquoPolyacetyleniccompounds and butanol fraction from Bidens pilosa can mod-ulate the differentiation of helper T cells and prevent autoim-mune diabetes in non-obese diabetic micerdquo Planta Medica vol70 no 11 pp 1045ndash1051 2004

Evidence-Based Complementary and Alternative Medicine 49

[50] Y M Chiang C L T Chang S L Chang W C Yang and LF Shyur ldquoCytopiloyne a novel polyacetylenic glucoside fromBidens pilosa functions as a T helper cell modulatorrdquo Journal ofEthnopharmacology vol 110 no 3 pp 532ndash538 2007

[51] R PUbillas CDMendez S D Jolad et al ldquoAntihyperglycemicacetylenic glucosides from Bidens pilosardquo Planta Medica vol66 no 1 pp 82ndash83 2000

[52] L Alvarez S Marquina M L Villarreal D Alonso E Arandaand G Delgado ldquoBioactive polyacetylenes from Bidens pilosardquoPlanta Medica vol 62 no 4 pp 355ndash357 1996

[53] C L T Chang S L Chang Y M Lee et al ldquoCytopiloyne apolyacetylenic glucoside prevents type 1 diabetes in nonobesediabetic micerdquo Journal of Immunology vol 178 no 11 pp 6984ndash6993 2007

[54] G Kusano A Kusano and Y Seyama Novel Hypoglycemicand Anti-Inflammatory Polyacetylenic Compounds Their Com-positions Bidens Plant Extract Fractions and CompositionsContaining the Plant or Fraction JPO Tokyo Japan 2004

[55] M G L Brandao A U Krettli L S R Soares C G CNery and H C Marinuzzi ldquoAntimalarial activity of extractsand fractions from Bidens pilosa and other Bidens species(Asteraceae) correlated with the presence of acetylene andflavonoid compoundsrdquo Journal of Ethnopharmacology vol 57no 2 pp 131ndash138 1997

[56] A U Krettli V F Andrade-Neto M D G L Brandao andW M S Ferrari ldquoThe search for new anti-malarial drugs fromplants used to treat fever and malaria or plants ramdomlyselected a reviewrdquoMemorias do Instituto Oswaldo Cruz vol 96no 8 pp 1033ndash1042 2001

[57] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[58] C K Wat R K Biswas E A Graham L Bohm G H NTowers and E R Waygood ldquoUltraviolet-mediated cytotoxicactivity of phenylheptatriyne from Bidens pilosa Lrdquo Journal ofNatural Products vol 42 no 1 pp 103ndash111 1979

[59] Y Sashida K Ogawa M Kitada H Karikome Y Mimakiand H Shimomura ldquoNew aurone glucosides and new phenyl-propanoid glucosides from Bidens pilosardquo Chemical and Phar-maceutical Bulletin vol 39 no 3 pp 709ndash711 1991

[60] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens pilosa L (TFB) on animal liver injury andliver fibrosisrdquo Journal of Ethnopharmacology vol 116 no 3 pp539ndash546 2008

[61] B Hoffmann and J Holzl ldquoAcylated compounds from Bidenspilosardquo Planta Medica vol 55 pp 108ndash109

[62] B Hoffmann and J Holzl ldquoA methylated chalcone glucosidefrom Bidens pilosardquo Phytochemistry vol 27 no 11 pp 3700ndash3701 1988

[63] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[64] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[65] B Hoffmann and J Holzl ldquoWeitere acylierte chalkone ausBidens pilosardquo Planta Medica vol 54 pp 450ndash451 1988

[66] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant in11traditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[67] V V Pham V V T K P V L Hoang and V K Phan ldquoFlavonoidcompounds from the plant Bidens pilosa L (Asteraceae)rdquo TapChi Duoc Hoc vol 50 pp 48ndash53

[68] S D Sarker B Bartholomew R J Nash and N Robinson ldquo5-O-methylhoslundin an unusual flavonoid from Bidens pilosa(Asteraceae)rdquo Biochemical Systematics and Ecology vol 28 no6 pp 591ndash593 2000

[69] S L Chang Y M Chiang C L T Chang et al ldquoFlavonoidscentaurein and centaureidin from Bidens pilosa stimulate IFN-120574 expressionrdquo Journal of Ethnopharmacology vol 112 no 2 pp232ndash236 2007

[70] Y M Chiang D Y Chuang S Y Wang Y H Kuo P WTsai and L F Shyur ldquoMetabolite profiling and chemopreventivebioactivity of plant extracts from Bidens pilosardquo Journal ofEthnopharmacology vol 95 no 2-3 pp 409ndash419 2004

[71] A Kusano Y Seyama E Usami et al ldquoStudies on the antioxi-dant active constituents of the dried powder from Bidens pilosaL var radiata SchrdquoNaturalMedicines vol 57 no 3 pp 100ndash1042003

[72] I Doleckova L Rarova J Gruz et al ldquoAnti-proliferative andanti-angiogenic effects of flavone eupatorin an active con-stituent of chloroform extract ofOrthosiphon stamineus leavesrdquoFitoterapia vol 83 no 6 pp 1000ndash1007

[73] K Bairwa R Kumar R J Sharma and R K Roy ldquoAn updatedreview on Bidens pilosa Lrdquo Der Pharma Chemica vol 2 no 3pp 325ndash337

[74] Q Xia Y Liu and Y Li ldquoDetermination of hyperoside indifferent parts and different species of Herba Bidens by RP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol 50pp 48ndash53

[75] M Horiuchi and Y Seyama ldquoImprovement of the antiinflam-matory and antiallergic activity of Bidens pilosa L var radiataSCHERFF treated with enzyme (Cellulosine)rdquo Journal of HealthScience vol 54 no 3 pp 294ndash301 2008

[76] M G L Brandao C G C Nery M A S Mamao and AU Krettli ldquoTwo methoxylated flavone glycosides from Bidenspilosardquo Phytochemistry vol 48 no 2 pp 397ndash400 1998

[77] P Kumari K Misra B S Sisodia et al ldquoA promising anticancerand antimalarial component from the leaves of Bidens pilosardquoPlanta Medica vol 75 no 1 pp 59ndash61 2009

[78] G Seelinger I Merfort U Wolfle and C M Schempp ldquoAnti-carcinogenic effects of the flavonoid luteolinrdquoMolecules vol 13no 10 pp 2628ndash2651 2008

[79] G Seelinger I Merfort and C M Schempp ldquoAnti-oxidantanti-inflammatory and anti-allergic activities of luteolinrdquoPlanta Medica vol 74 no 14 pp 1667ndash1677 2008

[80] C C Yit and N P Das ldquoCytotoxic effect of butein on humancolon adenocarcinoma cell proliferationrdquo Cancer Letters vol82 no 1 pp 65ndash72 1994

[81] J A Beutler E Hamel A J Vlietinck et al ldquoStructure-activityrequirements for flavone cytotoxicity and binding to tubulinrdquoJournal of Medicinal Chemistry vol 41 no 13 pp 2333ndash23381998

[82] M R Kviecinski K B Felipe T Schoenfelder et al ldquoStudy ofthe antitumor potential of Bidens pilosa (Asteraceae) used inBrazilian folkmedicinerdquo Journal of Ethnopharmacology vol 117no 1 pp 69ndash75 2008

[83] P Sundararajan A Dey A Smith A G Doss M Rajappanand S Natarajan ldquoStudies of anticancer and antipyretic activityof Bidens pilosawhole plantrdquo African Health Sciences vol 6 no1 pp 27ndash30 2006

[84] YMChiang C P Lo Y P Chen et al ldquoEthyl caffeate suppressesNF-120581B activation and its downstream inflammatory mediatorsiNOS COX-2 and PGE2 in vitro or in mouse skinrdquo BritishJournal of Pharmacology vol 146 no 3 pp 352ndash363 2005

50 Evidence-Based Complementary and Alternative Medicine

[85] J S Kim H J Lee M H Lee J Kim C Jin and J HRyu ldquoLuteolin inhibits LPS-stimulated inducible nitric oxidesynthase expression in BV-2 microglial cellsrdquo Planta Medicavol 72 no 1 pp 65ndash68 2006

[86] P A Ruiz and D Haller ldquoFunctional diversity of flavonoidsin the inhibition of the proinflammatory NF-120581B IRF and Aktsignaling pathways in murine intestinal epithelial cellsrdquo Journalof Nutrition vol 136 no 3 pp 664ndash671 2006

[87] A Xagorari A Papapetropoulos A Mauromatis MEconomou T Fotsis and C Roussos ldquoLuteolin inhibitsan endotoxin-stimulated phosphorylation cascade andproinflammatory cytokine production in macrophagesrdquoJournal of Pharmacology and Experimental Therapeutics vol296 no 1 pp 181ndash187 2001

[88] N Yoshida T Kanekura Y Higashi and T Kanzaki ldquoBidenspilosa suppresses interleukin-1120573-induced cyclooxygenase-2expression through the inhibition of mitogen activated proteinkinases phosphorylation in normal human dermal fibroblastsrdquoJournal of Dermatology vol 33 no 10 pp 676ndash683 2006

[89] M Muchuweti C Mupure A Ndhlala T Murenje and M AN Benhura ldquoScreening of antioxidant and radical scavengingactivity of Vigna ungiculata Bidens pilosa and Cleome gynan-drardquo American Journal of Food Technology vol 2 no 3 pp 161ndash168 2007

[90] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens bipinnata L against carbon tetrachloride-induced liver fibrosis in ratsrdquo Journal of Pharmacy and Pharma-cology vol 60 no 10 pp 1393ndash1402 2008

[91] S L Chang H H Yeh Y S Lin Y M Chiang T K Wuand W C Yang ldquoThe effect of centaurein on interferon-120574 expression and Listeria infection in micerdquo Toxicology andApplied Pharmacology vol 219 no 1 pp 54ndash61 2007

[92] F Deba T D Xuan M Yasuda and S Tawata ldquoChemical com-position and antioxidant antibacterial and antifungal activitiesof the essential oils from Bidens pilosa Linn var Radiatardquo FoodControl vol 19 no 4 pp 346ndash352 2008

[93] A O T Ashafa and A J Afolayan ldquoScreening the root extractsfrom Biden pilosa L var radiata (Asteraceae) for antimicrobialpotentialsrdquo Journal of Medicinal Plant Research vol 3 no 8 pp568ndash572 2009

[94] T B Nguelefack T Dimo E P Nguelefack Mbuyo P V TanS V Rakotonirina and A Kamanyi ldquoRelaxant effects of theneutral extract of the leaves of Bidens pilosa linn on isolated ratvascular smooth musclerdquo Phytotherapy Research vol 19 no 3pp 207ndash210 2005

[95] M T Grombone-Guaratini K L Silva-Brandao V N SolferiniJ Semir and J R Trigo ldquoSesquiterpene and polyacetyleneprofile of the Bidens pilosa complex (Asteraceae Heliantheae)from Southeast of Brazilrdquo Biochemical Systematics and Ecologyvol 33 no 5 pp 479ndash486 2005

[96] L L Lin C Y Wu H C Hsiu M T Wang and H ChuangldquoStudies on diabetes mellitus I The hypoglycemic activity ofphytosterin on alloxan diabetic ratsrdquo Taiwan Yi Xue Hui Za Zhivol 66 no 2 pp 58ndash65 1967

[97] M A N Benhura and I C Chitsiku ldquoThe extractable 120573-carotene content of Guku (Bidens pilosa) leaves after cookingdrying and storagerdquo International Journal of Food Science andTechnology vol 32 no 6 pp 495ndash500 1997

[98] J S Chang L C Chiang C C Chen L T Liu K C Wangand C C Lin ldquoAtileukemic activity of Bidens pilosa l varminor (blume) sherff andHouttuynia cordata thunbrdquo AmericanJournal of Chinese Medicine vol 29 no 2 pp 303ndash312 2001

[99] F Deba T D Xuan M Yasuda and S Tawata ldquoHerbicidal andfungicidal activities and identification of potential phytotoxinsfrom Bidens pilosa L var radiata Scherff research paperrdquoWeedBiology and Management vol 7 no 2 pp 77ndash83 2007

[100] J K Kumar and A K Sinha ldquoA new disubstituted acetylacetonefrom the leaves ofBidens pilosa LinnrdquoNatural Product Researchvol 17 no 1 pp 71ndash74 2003

[101] K Ogawa and Y Sashida ldquoCaffeoyl derivatives of a sugarlactone and its hydroxy acid from the leaves of Bidens pilosardquoPhytochemistry vol 31 no 10 pp 3657ndash3658 1992

[102] Q Xia Y Liu and Y Li ldquoDetermination of gallic acid fromdifferent species and different medical parts of Herba Bidens byRP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol24 no 03 pp 308ndash310

[103] W J Lee L F Wu W K Chen C J Wang and T HTseng ldquoInhibitory effect of luteolin on hepatocyte growth fac-torscatter factor-induced HepG2 cell invasion involving bothMAPKERKs and PI3K-Akt pathwaysrdquo Chemico-BiologicalInteractions vol 160 no 2 pp 123ndash133 2006

[104] J A Beutler J H Cardellina C M Lin E Hamel G MCragg andM R Boyd ldquoCentaureidin a cytotoxic flavone fromPolymnia fruticosa inhibits tubulin polymerizationrdquo BioorganicandMedicinal Chemistry Letters vol 3 no 4 pp 581ndash584 1993

[105] A Verma A Su A M Golin B OrsquoMarrah and J K AmorosaldquoThe lateral view a screening method for knee traumardquo Aca-demic Radiology vol 8 no 5 pp 392ndash397 2001

[106] J Corren M Lemay Y Lin L Rozga and R K RandolphldquoClinical and biochemical effects of a combination botanicalproduct (ClearGuard) for allergy a pilot randomized double-blind placebo-controlled trialrdquo Nutrition Journal vol 7 no 1article 20 2008

[107] M S Gachet J S Lecaro M Kaiser et al ldquoAssessment of anti-protozoal activity of plants traditionally used in Ecuador in thetreatment of leishmaniasisrdquo Journal of Ethnopharmacology vol128 no 1 pp 184ndash197 2010

[108] W C Yang M Ghiotto B Barbarat and D Olive ldquoThe role ofTec protein-tyrosine kinase in T cell signalingrdquo The Journal ofBiological Chemistry vol 274 no 2 pp 607ndash617 1999

[109] S Tewtrakul H Miyashiro N Nakamura et al ldquoHIV-1 inte-grase inhibitory substances from Coleus parvifoliusrdquo Phytother-apy Research vol 17 no 3 pp 232ndash239 2003

[110] T Dimo T B Nguelefack P V Tan et al ldquoPossible mechanismsof action of the neutral extract from Bidens pilosa L leaves onthe cardiovascular system of anaesthetized ratsrdquo PhytotherapyResearch vol 17 no 10 pp 1135ndash1139 2003

[111] T S C Li Chinese and Related North American Herbs CRCPress New York NY USA 2002

[112] H Wu H Chen X Hua Z Shi L Zhang and J ChenldquoClinical therapeutic effect of drug-separated moxibustion onchronic diarrhea and its immunologic mechanismsrdquo Journal ofTraditional Chinese Medicine vol 17 no 4 pp 253ndash258 1997

[113] S W Wright R R Harris J S Kerr et al ldquoSynthesis chemicaland biological properties of vinylogous hydroxamic acids dualinhibitors of 5-lipoxygenase and IL-1 biosynthesisrdquo Journal ofMedicinal Chemistry vol 35 no 22 pp 4061ndash4068 1992

[114] W R Almiron and M E Brewer ldquoClassification of immaturestage habitats of Culicidae (Diptera) collected in CordobaArgentinardquo Memorias do Instituto Oswaldo Cruz vol 91 no 1pp 1ndash9 1996

[115] N L Wang J Wang X S Yao and S Kitanaka ldquoTwo newmonoterpene glycosides and a new (+)-jasmololone glucoside

Evidence-Based Complementary and Alternative Medicine 51

from Bidens parvifloraWilldrdquo Journal of Asian Natural ProductsResearch vol 9 no 5 pp 473ndash479 2007

[116] P Champagnat ldquoRole of the terminal bud in the actionexercised by the cotyledon of Bidens pilosus L var radiatus onits axillary budrdquo Comptes Rendus des Seances et Memoires de laSociete de Biologie vol 145 no 17-18 pp 1374ndash1376 1951

[117] S F Nielsen S B Christensen G Cruciani A Kharazmiand T Liljefors ldquoAntileishmaniai chalcones statistical designsynthesis and three-dimensional quantitative structure-activityrelationship analysisrdquo Journal of Medicinal Chemistry vol 41no 24 pp 4819ndash4832 1998

[118] Y C Hwang J J H Chu P L Yang W Chen and M V YatesldquoRapid identification of inhibitors that interfere with poliovirusreplication using a cell-based assayrdquo Antiviral Research vol 77no 3 pp 232ndash236 2008

[119] V F Andrade-Neto M G L Brandao F Q Oliveira et alldquoAntimalarial activity of Bidens pilosa L (Asteraceae) ethanolextracts fromwild plants collected in various localities or plantscultivated in humus soilrdquo Phytotherapy Research vol 18 no 8pp 634ndash639 2004

[120] R J Marles and N R Farnsworth ldquoAnti-diabetic plants andtheir active constituentsrdquo Phytomedicine vol 2 no 2 pp 137ndash189

[121] M Habeck ldquoDiabetes treatments get sweet help from naturerdquoNature Medicine vol 9 no 10 p 1228 2003

[122] H W Lin G Y Han and S X Liao ldquoStudies on the activeconstituents of the Chinese traditional medicine Polygonatumodoratum (Mill) DrucerdquoActa Pharmaceutica Sinica vol 29 no3 pp 215ndash222 1994

[123] C L T Chang Y C Chen H M Chen N S Yang andW C Yang ldquoNatural cures for type 1 diabetes a reviewof phytochemicals biological actions and clinical potentialrdquoCurrent Medicinal Chemistry vol 20 no 7 pp 899ndash907 2013

[124] L Dey A S Attele and C S Yuan ldquoAlternative therapies fortype 2 diabetesrdquo Alternative Medicine Review vol 7 no 1 pp45ndash58 2002

[125] C L T ChangH Y Liu T F Kuo et al ldquoAnti-diabetic effect andmode of action of cytopiloynerdquo Evidence-Based Complementaryand Alternative Medicine vol 2013 Article ID 685642 13 pages2013

[126] M R Khan M Kihara and A D Omoloso ldquoAnti-microbialactivity of Bidens pilosa Bischofia javanica Elmerillia papuanaand Sigesbekia orientalisrdquo Fitoterapia vol 72 no 6 pp 662ndash6652001

[127] T Dimo T B Nguelefack P Kamtchouing E Dongo ARakotonirina and S V Rakotonirina ldquoHypotensive effects ofmethanol extract from Bidens pilosa Linn on hypertensive ratsrdquoComptes Rendus de lrsquoAcademie des Sciences Serie III vol 322no 4 pp 323ndash329 1999

[128] A K Hassan O Deogratius J F Nyafuono O Francis and OP Engeu ldquoWound healing potential of the ethanolic extracts ofBidens pilosa and Ocimum suaverdquo African Journal of Pharmacyand Pharmacology vol 5 no 2 pp 132ndash136 2011

[129] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[130] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[131] C Kandaswami and EMiddleton ldquoFree radical scavenging andantioxidant activity of plant flavonoidsrdquo Advances in Experi-mental Medicine and Biology vol 366 pp 351ndash376 1994

[132] K Yamamoto H Kakegawa H Ueda et al ldquoGastric cytopro-tective anti-ulcerogenic actions of hydroxychalcones in ratsrdquoPlanta Medica vol 58 no 5 pp 389ndash393 1992

[133] J O C Ezeonwumelu A K Julius C N Muhoho et alldquoBiochemical and histological studies of aqueous extract ofBidens pilosa leaves from Ugandan Rift valley in ratsrdquo BritishJournal of Pharmacology and Toxicology vol 2 no 6 pp 302ndash309

[134] L Frida S Rakotonirina A Rakotonirina and J P SavineauldquoIn vivo and in vitro effects of Bidens pilosa L (Asteraceae)leaf aqueous and ethanol extracts on primed-oestrogenized ratuterine musclerdquo African Journal of Traditional Complementaryand Alternative Medicines vol 5 no 1 pp 79ndash91 2008

52 Evidence-Based Complementary and Alternative Medicine

Composition Comments

1 Please provide valid postal code to the address if

possible

2 We made the highlighted change in Table 4 for the sake

of correctness Please check

3 Please check the correctness of the highlighted part in

Table 5

4 We considered the highlighted part in Table 11 as

reference Please check similar cases throughout

5 Please check the format of Table 18

6 We reformatted Table 20 Please check

7 We made the highlighted change in Table 4 for the sake

of consistency Please check similar cases throughout the

paper

8 We made the highlighted change(s) for the sake

of consistency Please check simila highlighted cases

throughout

9 We made the highlighted change in Table 9 for the sake

of clarity Please check similar cases through out

10 We made the highlighted change(s) for the sake of

correctness Please check

11 Comment on ref [66] This reference is a repetition

of [18] Please check similar cases throughout

Author(s) Name(s)

Author 1

Author 2

Author 3

2 Evidence-Based Complementary and Alternative Medicine

Table 1 Taxonomy of B pilosa [6]

Kingdom PlantaeSubkingdom TracheobiontaSuperdivision SpermatophytaDivision MagnoliophytaClass MagnoliopsidaSubclass AsteridaeOrder AsteralesFamily AsteraceaeGenus BidensSpecies Bidens Pilosa L

usage phytochemistry pharmacology and toxicology of Bpilosa The information provided here highlights the pos-sible usefulness of B pilosa and its isolated compoundsand offers insights into possible future research directionsStudies of B pilosa are divided into three groups (1) thebotany ethnomedical uses plant chemistry pharmacologyand biosafety of B pilosa (2) scientific studies that validatethe ethnomedical uses ofB pilosa and (3) the therapeutic andfuture research potential of B pilosa

11 Botany B pilosa was first collected and named by CarlLinnaeus in 1753 [3] Taxonomically it is assigned to theBidens genus (Asteraceae) as shown in Table 1 This genus isestimated to include 230 to 240 species worldwide [3 4] Bpilosa has several varieties such as B pilosa var radiata varminor var pilosa and var bisetosa Alongside examination ofmorphological traits authentication of B pilosa can be aidedby chemotaxonomy and molecular characterization [8]

B pilosa is an erect perennial herb widely distributedacross temperate and tropical regions B pilosa is eitherglabrous or hairy with green opposite leaves that are serratelobed or dissected It has white or yellow flowers and longnarrow ribbed black achenes (seeds) It grows to an averageheight of 60 cm and a maximum of 150 cm in favorableenvironments [9] (Figure 1) B pilosa prefers full sun andmoderately dry soil However it can grow in arid and barrenland from low to high elevations With the advantage ofbeing fast-growing in the 1970s the Food and AgriculturalOrganization actively promoted the cultivation of B pilosain Africa [10] B pilosa propagates via seeds A single plantcan produce 3000ndash6000 seeds Dry mature seeds from Bpilosa can be germinated in 3 to 4 days in moist soil orafter being soaking in water Seeds are viable for at least 3years [11] Minimal agricultural techniques are required forB pilosa cultivation Due to its invasive tendencies B pilosais generally considered to be a weed [12]

B pilosa is thought to have originated in South Americaand subsequently spread all over the world [13] Bidensspecies and their varieties bear vernacular names based ontheir characteristics For example Bidens species are knownby such names as Spanish needles beggarrsquos ticks devilrsquosneedles cobblerrsquos pegs broom stick pitchforks and farmersrsquofriends in English and some other languages because of theirsticky achenes [9] and are sometimes known as xian feng cao

(ldquoall bountiful grassrdquo) in Chinese because of their prosperousgrowth

12 Traditional Uses B pilosa is used as an herb and asan ingredient in teas or herbal medicines Its shoots andleaves dried or fresh are utilized in sauces and teas [1415] In the 1970s the United Nations Food and AgricultureOrganization (FAO) promoted the cultivation of B pilosa inAfrica because it is easy to grow edible palatable and safe[7] The nutritional value of B pilosa is shown in Table 2

All parts of B pilosa plant the whole plant the aerialparts (leaves flowers seeds and stems) andor the rootsfresh or dried are used as ingredients in folk medicines It isfrequently prepared as a dry powder decoction macerationor tincture [16] Generally this plant is applied as drypowder or tincture when used externally and as a powdermaceration or decoction when used as an internal remedy[15]

As summarized in Table 3 B pilosa either as a wholeplant or different parts has been reported to be useful inthe treatment of more than 40 disorders such as inflamma-tion immunological disorders digestive disorders infectiousdiseases cancers metabolic syndrome wounds and manyothers [26ndash29] B pilosa is usually ingested however it canalso be utilized externally For instance fresh B pilosa is usedto treat snake bites and wounds [23] and in Trinidad andTobago the aqueous solution of the leaves of B pilosa is usedto bathe babies and children [22]

B pilosa is sometimes used alone it is also used asan ingredient in medicinal mixtures together with othermedicinal plants such as Aloe vera Plectranthus mollisValeriana officinalis andCissus sicyoides among others [17 1820 25] Whether the commonly used mixtures of B pilosaor the mixtures of the compounds found in B pilosa affordsynergistic effects is not yet clear and needs to be verifiedby further studies However one study has suggested thatB pilosa varieties share similar phytochemical compositionsand may be substituted for each other [8]

2 Phytochemicals

Interest in basic research and application of B pilosa hasincreased since its first identification in 1753 This is mainlydue to its wide application in medicines foods and drinksAround 116 publications have documented the exploitationand medical use of B pilosa To date 201 compoundscomprising 70 aliphatics 60 flavonoids 25 terpenoids 19phenylpropanoids 13 aromatics 8 porphyrins and 6 othercompounds have been identified from this plant as compiledpreviously [30] The structures of these compounds are pre-sented in Tables 4 5 6 7 8 9 and 10 respectively Howeverthe association between B pilosa phytochemicals and theirbioactivities is not yet fully established and should becomea future research focus In the present review we explorepossible associations (Table 11) describe the importance ofthe known compounds in relation to their biological activity

Evidence-Based Complementary and Alternative Medicine 3

Table 2 Nutritional facts about B pilosa courtesy of the United Nations Food and Agriculture Organization [7]

Plant(100 g)

Energy(kcal)

Moisture()

Protein(g)

Fat(g)

Carbohydrate(g)

Fiber(g)

Ash(g)

Calcium(mg)

Phosphorus(mg)

Iron(120583g)

Caroteneequivalent

(120583g)

Thiamine(mg)

Raw 43 851 38 05 84 39 22 340 67 mdash 1 800 mdashDried 33 886 28 06 6 13 2 111 39 23 mdash mdashldquomdashrdquo denotes not detectable

(a) (b)

(c)

Figure 1 B pilosa (a) and its flowers (b) and achenes (c)

and discuss their likelymechanisms of action (Table 3) Com-pelling evidence suggests that the various diverse bioactivitiesreported for B pilosa reflect its phytochemical complexity

B pilosa is an extraordinary source of phytochemicalsparticularly flavonoids and polyynes Plant flavonoids arecommonly reported to possess anticancer antiinflammatoryantioxidant and other bioactivities However the bioactivi-ties of only seven of the 60 flavonoids present in B pilosa havebeen studiedThe bioactivities of the remaining 53 flavonoidsare poorly understood and deserve further investigationOther classes of compounds found inB pilosa are also in needof further examination as shown in Table 11

3 Pharmacological Properties

As outlined in Table 3 B pilosa is traditionally used totreat a wide variety of ailments Different preparations ofits whole plant andor parts have been purported to treatover 40 categories of illnesses Scientific studies although notextensive have demonstrated that B pilosa extracts andorcompounds have antitumor [43 69 77ndash83] antiinflamma-tory [27 75 79 84ndash88] antidiabetic and antihyperglycemic[8 24 49 51 53] antioxidant [70 89 90] immunomodu-latory [50 69] antimalarial [47 77] antibacterial [47 9192] antifungal [92 93] antihypertensive vasodilatory [2894] and antiulcerative [26] activities In this section the

4 Evidence-Based Complementary and Alternative Medicine

Table 3 Ethnomedical information about B pilosa

Disorder Plant partb Mode of use Regioncountry References

Stomachache LE Decoction IndiaAfrica [17ndash19]

Colics WP Decoction AfricaChina [18]

Catarrh WP Juice or decoctiontaken orally Cuba [20]

Diarrhea LE andWP

Fresh leaves orDecoction

AfricaUganda [18 21]

Constipation WP Decoction Africa [18]DysenteryBacillarydysentery WP Decoction Africa

China [18]

Choleretic WP Decoction Middle America [18]

Anti-inflammatory WP Not stated ChinaCuba [20 22]

Asthma WPDecoction ormaceration takenorally

CubaChina [20 23]

Antirheumatic RT andWP Juice and decoction Hong Kong

Zulu Africa [18 22]

Acute appendicitis WP Decoction Hong Kong [23]

Enteritis WP Decoction AfricaChina [18]

Pruritus WP Decoction Hong Kong [23]

Conjunctivitis WP Decoction AfricaChina [18]

Otitis WP Decoction AfricaChina [18]

Pharyngitis WP Decoction AfricaChina [18]

Gastritis WP Juice taken orally Cuba [20]

Diabetes WP Decoction takenorally

CubaTaiwan [20 24]

Headache WP Decoction Bafia Cameroon [25]Diuretic WP Decoction Middle America [18]

Hypotensive WP Decoction takenorally Bafia Cameroon [25]

Colds LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21 23]

Yellow Fever LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21ndash23]

Influenza LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21 23]

Acute infectioushepatitis WP Decoction Hong Kong [23]

Intestinal worms WP Decoction Africa [18]

Malaria RT andWP Juice Africa

China [18 23]

Evidence-Based Complementary and Alternative Medicine 5

Table 3 Continued

Disorder Plant partb Mode of use Regioncountry References

Eye Infection LE andWP Fresh or juice Uganda

Middle America [18 21]

Antimicrobial APDecoction fordrinkingbathingexternal use

Trinidad andTobago [22]

Pulmonarytuberculosis WP

Decoction ormaceration takenorally

CubaChina [20 23]

Bacterial infectionsingastrointestinal tracts

WP Decoction Trinidad andTobago [22]

Renal infection LE Decoction takenorally Cuba [20]

Sore throat LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21 23]

Cough WP Decoction takenorally

CubaChina [20 23]

Coolness of theuterus WP Decoction taken

orally Cuba [20]

Menstrualirregularities WP Decoction taken

orally Cuba [20]

Dysmenorrhea WP Decoction Bafia Cameroon [25]Hyperemesisgravidarum(morning sickness)

WP Decoction Africa [18]

Hemorrhoids WP Decoction Hong Kong [23]

Nose bleeds LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21 23]

Stomach ulcers LE andWP

Maceration or juicetaken orally

CubaMiddle AmericaUganda

[18 20 21 23]

Cuts burns and skinproblems

LE andWP

Fresh plant ordecoction topicalapplicationbathing

Trinidad andTobagoAfricaChinaCameroonBrazilVenezuela

[18 22 26]

Wounds WP Crushed herb

ChinaAfricaCentral AmericaHawaii

[23]

Snake bites WP Pulverized herb China [23]bLE leaves ST stem FW flower WP whole plant RT roots AP aerial parts

primary pharmacological properties of B pilosa extracts andphytochemicals are presented and discussed

31 AntiCancer Activity Folkloric reports revealed the pos-sible antitumor efficacy of B pilosa and several scientific in2vitro studies have supported the claim that B pilosa extractsand isolated compounds possess anti-cancer activities againsta variety of cancer cells Several studies have used bioassay

guided isolation and fractionation methods to discover newcompounds from B pilosa For example Kviecinski andcolleagues tested hydroalcoholic crude extracts chloroformethyl acetate and methanol fractions for anti-tumor activity[82] The cytotoxicity of the extracts was assessed usingbrine shrimp hemolytic MTT and neutral red uptake(NRU) assays In vivo studies were performed using Ehrlichascites carcinoma in isogenic BALBc mice Among them

6 Evidence-Based Complementary and Alternative Medicine

Table 4 Aliphatic natural products isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

1 Heneicosane CH3(CH2)19CH3 Aerial (Tanzania) [18]2 Docosane CH3(CH2)20CH3 Aerial (Tanzania) [18]3 Tricosane CH3(CH2)21CH3 Aerial (Tanzania) [18]4 Tetracosane CH3(CH2)22CH3 Aerial (Tanzania) [18]5 Pentacosane CH3(CH2)23CH3 Aerial (Tanzania) [18]6 Hexacosane CH3(CH2)24CH3 Aerial (Tanzania) [18]7 Heptacosane CH3(CH2)25CH3 Aerial (Tanzania) [18]

8 Octacosone CH3(CH2)26CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

9 Nonacosane CH3(CH2)27CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

10 Triacontane CH3(CH2)28CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

11 Hentriacontane CH3(CH2)29CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

12 Dotriacontane CH3(CH2)30CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

13 Tritriacontane CH3(CH2)31CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

14 2-Butoxy-ethanol CH3(CH2)3OCH2CH2OH Whole (Taiwan) [32]15 Tetracosan-1-ol CH3(CH2)22CH2OH Aerial (Tanzania) [18]16 Hexacosan-1-ol CH3(CH2)24CH2OH Aerial (Tanzania) [18]17 1-Octacosanol CH3(CH2)26CH2OH Aerial (Tanzania) [18]

18 1-Hentriacontanol CH3(CH2)29CH2OHNot found(Taiwan) [31]

19 Tetradecanoic acid Myristic acid CH3(CH2)12CO2H Aerial (Tanzania) [18]20 Hexadecanoic acid Palmitic acid CH3(CH2)14CO2H Aerial (Tanzania) [18]21 Octadecanoic acid Stearic acid CH3(CH2)16CO2H Aerial (Tanzania) [18]22 Eicosanoic acid Arachidic acid CH3(CH2)18CO2H Aerial (Tanzania) [18]

23 Docosanoic acid Behenic acid CH3(CH2)20CO2HLeaves(Philippines) [33]

24 2-butenedioic acidO

OHO

HO Aerial (China) [34 35]

25 (Z)-9-Octadecenoicacid Oleic acid

O

OH Aerial (Tanzania) [18]

Evidence-Based Complementary and Alternative Medicine 7

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

26 (E)-9-Octadecenoicacid Elaidic acid

O

HOLeaves(Philippines) [33]

27 (ZZ)-912-Octadecadienoic acid

Linolic acid or Linoleicacid

O

OHAerial(Tanzania)whole (Taiwan)

[18 36]

28(ZZZ)-91215-Octadecatrienoicacid

120572-Linoleic acid

O

OH Whole (Taiwan) [32]

29(ZZ)-912-Octadecadienoic acidethyl ester

Ethyl linoleate O

O

Whole (Taiwan) [32]

30(ZZZ)-91215-Octadecatrienoicacid methyl ester

Methyl linolenate O

O

Whole (Taiwan) [32]

31(ZZZ)-91215-Octadecatrienoicacid ethyl ester

Ethyl linolenate O

O

Whole (Taiwan) [32]

32(Z)-9-Octadecenoicacid 2-butoxyethylester

2-Butoxyethyl oleate O

OO

Whole (Taiwan) [32]

33 2-Butoxyethyllinoleate O

OO Whole (Taiwan) [32]

34(ZZZ)-91215-Octadecatrienoicacid butoxyethyl ester

2-Butoxyethyl linoleate O

OO

Whole (Taiwan) [32]

3517E9E15E-Heptadecatetraene-1113-diyne

Heptadeca-2E8E10E16-tetraen-46-diyne

Not found(China) [37]

8 Evidence-Based Complementary and Alternative Medicine

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

36 111-Tridecadiene-3579-tetrayne

Roots (notstated) [38]

37 1-Tridecene-357911-pentayne Pentayneene

Leaves (notstated) andnot found(Egypt)

[38 39]

38 5-Tridecene-7911-triyne-3-ol

HO

not found(Egypt) [39]

39 21012-Tridecatriene-468-triyn-1-ol

OHPart not specified(Not stated) [40]

40 212-Tridecadiene-46810-tetrayn-1-ol

111-Tridecadiene-3579-

tetrayn-13-ol

OH Roots (Notstated) notfound(Egypt)

[38 39]

41 212-Tridecadiene-46810-tetraynal

111-Tridecadiene-3579-

tetrayn-13-al

O

Roots (Germany) [41]

42212-Tridecadiene-46810-tetrayn-1-ol1-acetate

111-Tridecadiene-3579-

tetrayn-13-acetateO

O

Roots (notstated) [38]

43(5E)-15-Tridecadiene-79-diyn-3413-triol OH

OH

HO

Aerial (China) [42]

44(6E12E)-3-oxo-tetradeca-612-dien-810-diyn-1-ol

OH

O

Aerial (China) [42]

45 (E)-5-Tridecene-7911-triyne-12-diol

12-Dihydroxy-5(E)-tridecene-7911-triyne

HO

OHWhole (Taiwan) [43]

46 (E)-6-Tetradecene-81012-triyne-13-diol

13-Dihydroxy-6(E)-tetradecene-81012-triyne

OH

OHWhole (Taiwan) [43ndash45]

47(2R3E11E)-311-Tridecadiene-579-triyne-12-diol

Safynol

OH

HO

Not found (Egyptand China) [37 39]

Evidence-Based Complementary and Alternative Medicine 9

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

4857911-Tridecatetrayne-12-diol

12-Dihydroxytrideca-57911-tetrayne

HOOH

Whole (Taiwan) [43 44 46]

49(R)-357911-Tridecapentayne-12-diol

(R)-12-Dihydroxytrideca-357911-pentayne

HO

OH

Aerial (Japan) [47]

50

(4E)-1-(Hydroxymethyl)-4-dodecene-6810-triyn-1-yl-120573-D-glucopyranoside

2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-

tridecene-7911-triyne

O

HO

H

OH

H

H

HOH

O

OH

HO

H Aerial (USA)whole (Taiwan)leaves (Taiwan)

[8 45 48ndash51]

51

(4E)-1-(2-Hydroxyethyl)-4-dodecene-6810-triyn-1-yl-120573-D-glucopyranoside

3-120573-D-Glucopyranosyloxy-1-hydroxy-6(E)-

tridecene-81012-triyne

OHO

H

OH

H

H

HOH

O

OH

OH

HAerial (USA andChina) whole(Taiwan) leaves(Taiwan)

[8 35 4245 48ndash51]

52

3-Hydroxy-6-tetradecene-81012-triynyl-120573-D-glucopyranoside

120573-D-Glucopyranosyloxy-

3-hydroxy-6E-tetradecence-81012-triyne

OH

O

OHO

H

OH

H

H

HOH

OH

HWhole (Mexico) [52]

53

1-(Hydroxymethyl)-46810-dodecatetrayne-1-yl-120573-D-glucopyranoside

Cytopiloyne2-120573-D-

Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne

O

HO

OHO

H

OH

H

H

HOH

OH

HWhole (Taiwan)leaves (Taiwan) [8 50 53]

54

2-O-D-Glucosyltrideca-11E-en-3579-tetrayn-12-diol

OH

O

O

OHH

HO

H

H

HHOH

OH

Leaves (Brazil) [27]

10 Evidence-Based Complementary and Alternative Medicine

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

55

(R)-1-(Hydroxymethyl)-246810-dodecapentayn-1-yl-a-D-glucopyranoside

2- 120573 -D-Glucopyranosyloxy-1-hydroxytrideca-357911-pentayne

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Chinaand Japan) [35 47]

56

1-[[(Carboxyacetyl)oxy]methyl]46810-dodecatetraynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Japan) [54]

57

(4E)-1-[[(Carboxyacetyl)oxy]-methyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHO

H

OH

Aerial (Japan) [54]

58

(4E)-1-[[(Carboxyacetyl)oxy]-ethyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

O

O

O

OHO

O

OHOH

H

HO

H

H

HHO

HOH

Aerial (Japan) [54]

59 (5E)-5-Heptene-13-diyn-1-yl-benzene

1-Phenylhepta-13-diyn-5en Whole (Taiwan) [32]

60 7-Phenyl-2(E)-heptene-46-diyn-1-ol

OHRoots (notstated) Aerial(China)

[38 42]

617-Phenyl-2(E)-heptene-46-diyn-1-ol-acetate

OO

Roots (notstated Brazil) [38 55 56]

62 7-Phenyl-46-heptadiyn-2-ol Pilosol A

OH

Whole (Taiwan)aerial (China) [42 57]

Evidence-Based Complementary and Alternative Medicine 11

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

63 7-Phenylhepta-46-diyn-12-diol HO

OH

Aerial (China) [42]

64 135-Heptatriyn-1-yl-benzene

1-Phenylhepta-135-triyne

Leaves (notstated) leaves oftissue culture(not stated)aerial (TanzaniaChina)Whole (Taiwan)roots (Brazil)

[18 34 3842 56ndash58]

65 7-Phenyl-246-heptatriyn-1-ol HO

Leaves (notstated) aerial(China)

[38 42]

667-Phenyl-246-heptatriyn-1-ol-acetate O

O Leaves (notstated) [38]

675-(2-Phenylethynyl)-2-thiophenemethanol

S OHAerial (China) [42]

685-(2-Phenylethynyl)-2120573-glucosylmethyl-thiophene

O

O

OH

H

OH

H

H

HOH H

O

HO

S

Aerial (China) [42]

69

3- 120573 -D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne

OH

O

OH

HOHO

H

H

HHOH

OH

Leaves(Cameroon) [28]

701-Phenyl-13-diyn-5-en-7-ol-acetate O

O

Leaves (Brazil) [27]

SN denotes serial number

the chloroform fraction was the most toxic with a halfmaximal inhibitory concentration (IC

50) of 97 plusmn 72 and 83 plusmn

52 120583gmL in NRU and MTT respectively [82] Kumari andcolleagues also reported the anti-cancer and anti-malarialactivities of B pilosa leaves [77] Based on a cytotoxicity-directed fractionation strategy they identified phenyl-135-heptatriene with IC

50values of 8plusmn001 049plusmn045 07plusmn001

and 10 plusmn 001 120583gmL against human oral liver colon andbreast cancer cell lines respectively However phenyl-135-heptatriyne showed lower activity against breast cancer celllines than the chloroform leaf extract which had an IC

50

value of 65 plusmn 001 120583gmL Moreover the positive controltaxol showed higher activity than phenyl-135-heptatriyne[77] Furthermore in vitro comet assays were performed toevaluate the toxicity of n-hexane chloroform and methanolextracts of B pilosa and its ethyl acetate acetone and waterfractions onHela andKB cellsThe ethyl acetate fraction fromthe methanol extract exhibited the highest activity with halfmaximal cytotoxic concentrations (CTC

50) of 9652120583gmL

and 5862120583gmL against Hela and KB cells respectivelyDespite the moderate toxicity these findings suggest thatthese B pilosa extractsfractions could be useful for future

12 Evidence-Based Complementary and Alternative Medicine

Table5Flavon

oids

isolated

from

Bpilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

71

2-[(34-Dihydroxyph

enyl)-

methylene]-

6-hydroxy-3(2H

)-benzofuranon

e

Sulfu

retin

O

OHO

OH

OH

Aeria

l(Ch

ina)

[35]

72

2-[(34-Dihydroxyph

enyl)-

methylene]-

67-dihydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-673101584041015840-

tetrahydroxy

maritimetin

O

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

73

2-[(34-Dihydroxyph

enyl)-

methylene]-6-

(120573-D

-glucopyrano

syloxy)-7-

hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-120573-D

-glucop

yranosyl-

673101584041015840-te

trahydroxy

maritimein

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japanand

China)aerial(Ch

ina)

[355960]

74

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

(120573-D

-glucopyrano

syloxy)-

6-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-7-O-120573-D

-glucop

yranosyl-

673101584041015840-

tetrahydroxy

O

OHO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japan)

[59]

75

6-[(6-O-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-acetyl-

120573-D

-glucopyrano

syl)

-673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OHO

H HO

H

H

HOH

HO

O

O

Leaves

(Not

stated)

aeria

l(Ch

ina)

[3559]

Evidence-Based Complementary and Alternative Medicine 13

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

76

6-[(36-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-

2-[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone(Z)-6-O

-(36-di-O

-acetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

bideno

sideA

O

O

HO

OH

OH

OHOH O

H

H

HOH

HO

O

O

O

Leaves

(China)

[60]

77

6-[(46-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(410158401015840610158401015840-

diacetyl-120573-D

-glucropyrano

syl)-

673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

HO

O

OO

Leaves

(not

stated)

aeria

l(Ch

ina)

[343561]

78

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

hydroxy-6-

[(246-tri-O

-acetyl-120573

-D-

glucop

yranosyl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(210158401015840410158401015840610158401015840-

triacetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

O

OOO

O

Leaves

(not

stated)

aeria

l(Ch

ina)

[3461]

79

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-6-

[(346-tri-O

-acetyl-120573

-D-glucopyrano

syl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(310158401015840410158401015840610158401015840-

triacetyl-120573

-D-glucrop

yranosyl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H OH

H

HOH

HO

OOO O

Aeria

l(Ch

ina)

[3435]

80

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-

6-[[6-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propenyl]-120573-

D-glucopyrano

syl]o

xy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-

p-coum

aroyl-

120573-D

-glucropyrano

syl)-

673101584041015840-te

trahydroxy

OH

O

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OLeaves

(Japan)

[59]

14 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

81

1-[2-(120573-D

-Glucopyrano

syloxy)-

4-hydroxyphenyl]-2-

hydroxy-3-(3-

hydroxyphenyl)-2-

prop

en-1-on

e

Chalcone120572

32101584041015840-

tetrahydroxy-

21015840-O

-120573-D

-glucop

yranosyl

HO

O

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[35]

821-(24-Dihydroxyph

enyl)-

3-(34-dihydroxyph

enyl)-

2-prop

en-1-on

eBu

tein

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

833-(34-D

ihydroxyph

enyl)-

1-(234-trihydroxyph

enyl)-

2-prop

en-1-on

eOkanin

O

HO

HO

OH

OH

OH

Leaves

(China)

[60]

84

3-(34-D

ihydroxyph

enyl)-

1-[3-(120573-D

-glucop

yranosyloxy)-

24-dihydroxyphenyl]-2-

prop

en-1-on

e

Okanin

31015840-O

-120573-D

-glucosid

e

O

HO

OH

OH

OH

OO

HO

H

HO

H

H

HOH

H

OH

Leaves

(Germany)

flowers(Germany)

[61ndash63]

85

3-(34-D

ihydroxyph

enyl)-

1-[4-(120573-D

-glucop

yranosyloxy)-

23-dihydroxyphenyl]-

2-prop

en-1-on

e

Okanin

41015840-O

-120573-D

-glucosid

e

OOH

OH

OH

HOO

OHO

H

HO

H

H

HOH

H

OH

Flow

ers(Germany)

leaves

(Japan)

[5963]

Evidence-Based Complementary and Alternative Medicine 15

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

86Okanin41015840-O

-120573-D

-(610158401015840-

O-acetylglucosid

e)

O

O

OH

OH

OH

OHO

H

HO

H

H

HOH

H

HO

O

O

Flow

ers(Germany)

[63]

87

1-[4-[(46-di-O

-Acetyl-

120573-D

-glucopyrano

syl)-oxy]-

23-dihydroxyphenyl]-3-

(34-dihydroxyph

enyl)-2-

prop

en-1-on

e

Okanin

41015840-O

-120573-D

-(410158401015840610158401015840-diacetyl)-

glucop

yranoside

O

O

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Aeria

l(Ch

ina)

[34]

88Okanin41015840-O

-120573-D

-(210158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[64]

89Okanin41015840-O

-120573-D

-(310158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

O

OAe

rial(Ch

ina)

[34]

16 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

90

1-[23-Dihydroxy-4-

[[6-O-[3-(4-hydroxyph

enyl)-

1-oxo-2-prop

enyl]-120573-D

-glucop

yranosyl]oxy]-ph

enyl]-

3-(34-dihydroxyph

enyl)-

2-prop

en-1on

e

Okanin

41015840-O

-120573-D

-(610158401015840-tr

ans-

p-coum

aroyl)-

glucoside

O

O

OH

OH

OH

OHOH

HO

H

H

HOH

H

HO

OO

OH

Leaves

(Germany)

[64]

91Okanin41015840-O

-120573-D

-(410158401015840-acetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Leaves

(Germany)

[65]

92

Okanin

41015840-O

-120573-D

-(210158401015840410158401015840-diacetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[65]

Evidence-Based Complementary and Alternative Medicine 17

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

93

Okanin

41015840-O

-120573-D

-(310158401015840410158401015840-diacetyl-610158401015840-

trans-p-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

OH

O O

Leaves

(Germany)

[65]

94

Okanin

41015840-O

-[120573-D

-glucopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

OOH

OH

OH

HO

OH

HO

HHO

H

HOH

HO

OH

OH

HO

HHO

H

HOH

HO

Flow

er(G

ermany)

[63]

95Okanin

3101584041015840-di-O

-120573-D

-glucosid

e

OOH

OH

OH

O OO

H HO

HHOH

HOH

HHO

OHO

H

HO

H

H

HOH

H

OH

Flow

er(G

ermany)

[63]

18 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

96

1-[3-(120573-D

-Glucopyrano

syloxy)-

24-dihydroxyphenyl]-

3-(3-hydroxy)-

4-metho

xyph

enyl)-

2-prop

en-1-on

e

Okanin

4-methylether-

31015840-O

-120573-D

-glucop

yranoside

OOH

O

OH

HO

O

H HO

HHOH

HOH

H

OHO

Leaves

(Germany)aerial

(China)

[3562]

97

Okanin4-methylether-

3101584041015840-di-O

-120573-(41015840101584061015840101584041015840101584010158406101584010158401015840-

tetraacetyl)-

glucop

yranoside

OOH

OOH

O

HH

OH

HO

HOH

H

OO

O

Ac

Ac

AcAc

OO

H

HO

H

H

HOH

H

O

Aeria

l(Ch

ina)

[42]

98Ch

alcone210158404101584061015840-tr

imetho

xy-

4-O-D

-glucopyrano

syl-

dihydro

Not

foun

dLeaves

(China)

[60]

992-(34-D

ihydroxyph

enyl)-23-

dihydro-78

-dihydroxy-

4H-1-

benzop

yran-4-one

Okanin

isoOO

OH

HO

OH

OH

Leaves

(China)

[60]

100

2-(34-D

ihydroxyph

enyl)-

23-dihydro-8-hydroxy-

7[(246-tr

i-O-acetyl-120573

-D-

glucop

yranosyl)oxy]-4H

-1-b

enzopyran-4-on

eOO

OH

OH

OH

OH

O

HHO

H

HO

HO

OAc

Ac

Ac

Aeria

l(Ch

ina)

[34]

Evidence-Based Complementary and Alternative Medicine 19

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

101

57-Dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

OO

OH

HO

OH

Aeria

l(TanzaniaC

hina)

[4266]

102

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

7-O-glucopyrano

side

OO

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Tanzania)

[66]

103

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-4H

-1-benzop

yran-4-one

Luteolin

OO

HO

OH

OH

OH

Aeria

l(TanzaniaC

hina

Vietnam)

[343542

6667]

104

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5hydroxy-4H-1-

benzop

yran-4-one

Luteolin

7-O-120573-D

-glucop

yranoside

OOOH

OH

OH

OO

HO

H

HO

H

H

HOH

OH H

Aeria

l(Tanzania)

[66]

105

57-Dim

etho

xy-6-

(5-m

etho

xy-6-m

ethyl-

4-oxo-4H

-pyran-3-yl)-

2-ph

enyl-4H-1-

benzop

yran-4-one

5-O-M

ethylhosludin

O OOO

O

O

O

Aeria

l(Ugand

a)[68]

20 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

106

3-(120573-D

-Glucopyrano

syloxy)-

57-dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Astragalin

Kaem

pferol-3-O

-120573-D

-glucopyrano

side

HO

OH

O O

OH

O

O

HO

HHO

H

H

HOH

OH

H

Aeria

l(Ch

ina)

[35]

107

Kaem

pferol3-

(23-di-E

-p-cou

maroyl-

120572-L-rhamno

pyrano

side)

Not

foun

dAe

rial(Vietnam)

[67]

108

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5-hydroxy-36-dimetho

xy-

4H-1-

benzop

yran-4-one

Axillarosid

eO O

OH

OH

OO

OH

OO

HO

H

HO

H

H

HOH

H

OH

Aeria

l(Ch

ina)

[42]

109

57-Dihydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaureidin

O

O

O

O

O

HO

OH

OH

Who

le(Taiwan)

[69]

110

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaurein

O O

OO

O OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan)w

hole

(Taiwan)

[69ndash

71]

Evidence-Based Complementary and Alternative Medicine 21

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

111

5-Hydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-

67-dimetho

xy-4H-

chromen-4-one

Eupatorin

iso

OO O

OH

O

O OH

Not

foun

d(C

hina)

[3772]

112

2-(34-D

imetho

xyph

enyl)-7-

(120573-D

-glucopyrano

syloxy)-35-

dihydroxy-

8-metho

xy-4H-1-

benzop

yran-4-one

O O

OH

O O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

113

7-(120573-D

-Glucopyrano

syloxy)-

5hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-38-

dimehtoxy-4H-1-

benzop

yran-4-one

O O

O

OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

114

Isorhamnetin

3-[O

-120572-

L-rham

nopyrano

syl-

(1-2)-120573-D

-glucop

yranoside]

Not

foun

dAe

rial(Vietnam)

[67]

115

7-[(6-deoxy-120572-L-

Manno

pyrano

syl)o

xy]-

3-120573-D

-glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-4H

-1-benzop

yran-4-one

Luteoside

O O

OH

O

OH

OH

OH

HOH

H

H HO

HO

HO

OH

HO

H

HO

OH

OH

HH

Aeria

l(Ch

ina)

[42]

22 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

116

Luteolin

3-O-120573-D

-Glucopyrano

side

O O

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

HO

Aeria

l(Tanzania)

[66]

117

57-Dihydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Quercetagetin

3631015840-tr

imethylether

OOO

O

O OH

HO

OH

Aeria

l(Ch

ina)

[42]

118

6-((2S3S4S5S)-

Tetrahydro-345-tr

ihydroxy-

6-(hydroxymethyl)-2H

-pyran-2-yloxy)-5-hydroxy-

2-(4-hydroxy-3-

metho

xyph

enyl)-

37-dimetho

xy-4H-

chromen-4-one

Quercetagetin

3731015840-

trim

ethylether-6-O

-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[42]

119

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

JaceinQ

uercetin

3631015840-

trim

ethyl

ether-7-O-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan

China)

who

le(Taiwan)

[425470]

120

2-(34-D

ihydroxyph

enyl)-

357-trihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

OO

HO

OH

OH

OH

OH

Aeria

l(Ch

ina)leaves

(China)who

le(C

hina)

[356074]

Evidence-Based Complementary and Alternative Medicine 23

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

121

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-galactopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-4-one

Quercetin

3-O-120573-D

-galactosidehyperin

hyperosid

e

O O

HO

OH

OH

OH

OOHH

HOH

H

H HO

HO

OH

Aeria

l(TanzaniaJapan)

Leaves

(China)Who

le(C

hina)

[376066

717475]

122

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-glucopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

3-O-120573-

D-glucopyrano

side

O O

HO

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

Aeria

l(TanzaniaC

hina

Japan)L

eaves(Japan)

[355966

75]

123

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-

4-oxo-4H

-1-benzop

yran-3-yl-

120573-D

-glucopyrano

siduron

icacid

Quercetin

3-O-120573-D

-glucuron

opyranoside

O O

HO

OH

OH

OH

OHOH

HOH

H

H HO

HO

OHO

Aeria

l(TanzaniaJapan)

[6671]

124

3-[[6-O-(6-

Deoxy-120572-L-m

anno

pyrano

syl)-

120573-D

-galactopyrano

syl]o

xy]-

2-(34-dihydroxyph

enyl)-57-

dihydroxy-4H

-1-benzop

yran-4-one

Quercetin

3-O-rob

inob

iosid

e

O O

HO

OH

OH

OH

O

HOHH OH

HO

H HH

OO

OHH

HOH

H

H HO

HO

Aeria

l(Japan)W

hole

(Taiwan)

[7071]

24 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

125

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-

3-metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

OH

Roots(Brazil)

[55676]

126

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-

120573-D

-glucopyrano

syl]o

xy]-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120572-

L-rham

nopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

O

HHO HHO

HO H

HH

ORo

ots(Brazil)

[5676]

127

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-120573-D

-glucop

yranosyl]oxy]-5-

hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-3-metho

xy-

4H-1-

benzop

yran-4-one

Quercetin

341015840-

dimethyl

ether-7-O-rutinoside

O OOH

OH

O

O

OH O

HOH

H

HHO

H

O

HHO HHO

HO H

HH

O

HO

Aeria

l(Ch

ina)

[3435]

128

2-(34-D

ihydroxyph

enyl)-3-

(120573-D

-glucofurano

syloxy)-57-

dihydroxy-4H

-1-benzop

yran-

4-on

e

Isoq

uercitrin

O OOH

OH

OHO

H

OHH

OH

HO

H O HOH

OH

Aeria

l(Japan

China)

[3571]

Evidence-Based Complementary and Alternative Medicine 25

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

129

Quercetin

3-O-120573-D

-galactop

yranoside

O

OH

HO

OH OH

O

O

HO

OH

OHOH

OAe

rial(no

tstated)

[18]

130

Quercetin

3-O-rutinoside

O

O

R

OO

OH

OHO

H

O

OH

OH

H3C

H3C

H3C

OH

HO

Who

le(Taiwan)

[70]

26 Evidence-Based Complementary and Alternative Medicine

Table 6 Terpenoids isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(Country) References

131371111-Tetramethylbicyclo[810]undeca-

26-dieneBicyclogermacrene Leaves

(Brazil) [95]

13241111-Trimethyl-8-

methylenebicyclo[720]undec-4-ene

E-Caryophyllene Leaves(Brazil) [95]

1331-Methyl-5-methylene-8-

(1-methylethyl)-16-cyclodecadiene

Germacrene D Leaves(Brazil) [95]

1344-(15-Dimethyl-4-hexen-1-ylidene)-1-methyl-cyclohexene

Z-120574-Bisabolene Leaves(Brazil) [95]

135Decahydro-114-trimethyl-7-methylene-1H-cycloprop[e]-

azulene120573-Gurjunene Leaves

(Brazil) [95]

136 2669-Tetramethyl-148-cycloundecatriene

120572-Humulene120572-caryophyllene

Leaves(Brazil) [95]

13712344a568a-Octahydro-1-isopropyl-7-methyl-4-methylenenaphthalene

120575-Muurolene Leaves(Brazil) [95]

138

12344a568a-Octahydro-4a8-dimethyl-2-(1-methylethylidene)-

naphthalene

Selina-37(11)-diene

Leaves(Brazil) [95]

139(2E7R11R)-371115-

Tetramethyl-2-hexadecen-1-ol

Phytol

HO

Whole(Taiwan) [57]

Evidence-Based Complementary and Alternative Medicine 27

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

140 371115-Tetramethyl-2-hexadecenoic acid Phytanic acid

O

OH

Whole(Taiwan) [57]

141371115-Tetramethyl-2-

hexadecenyl ester-heptanoicacid

Phythylheptanoate O O

Leaves(Not stated) [33]

142

(3S10R13R)-17-((2R5R)-5-Ethyl-6-methylheptan-2-yl)-

2347891011121314151617-tetradecahydro-

1013-dimethyl-1H-cyclopenta[a]phenanthren-

3-ol

Campestrol

HO

Aerial(Tanzania) [18]

143 Not found Phytosterin-B Not foundNot found(TaiwanEgypt)

[39 96]

144 Stigmast-5-en-3-ol 120573-sitosterol

HOH

H

H

Aerial(Tanzania)

whole(Taiwan)

[18 31 57]

145

1314151617-Tetradecahydro-1013-

dimethyl-1H-cyclopenta[a]phenanthren-

3-yloxy)-tetrahydro-6-(hydroxymethyl)-2H-pyran-345-triol

120573-Sitosterolglucoside

H

H

HOH

HO

HHO

H

HOHH O

OH Not found(Egypt) [39]

146 5120572-Stigmasta-7-en-3120573-ol

HO

Whole(Taiwan) [57]

147 5120572-Stigmasta-722t-dien-3120573-ol

HO

Whole(Taiwan) [57]

148 Stigmasta-522-dien-3-ol Stigmasterol

HOH

H

H

Not found(Taiwan)aerial

(Tanzania)leaves (notstated)whole

(Taiwan)

[18 31 3357]

28 Evidence-Based Complementary and Alternative Medicine

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

149 Lup-20(29)-en-3-ol Lupeol

H

HH

HOH

Not found(Egypt) [39]

150 Lup-20(29)-en-3-ol acetate Lupeol acetate

H

HH

O

OH

Not found(Egypt) [39]

151 Olean-12-en-3-ol 120573-amyrin

H

HOH

Not found(Egypt) [39]

152 5913-Trimethyl-242526-trinoroleanan-3-ol Friedelan-3120573-ol

HO

Aerial(Tanzania) [18]

153 5913-Trimethyl-242526-trinoroleanan-3-one

Friedelinfriedelan-3-one

O

Aerial(Tanzania) [18]

1542610151923-

Hexamethyl-2610141822-tetracosahexaene

Squalene

Aerial(Tanzania)Leaves (Notstated)Whole(Taiwan)

[18 33 57]

155 120573120573-Carotene 120573-Carotene Leaves (Notstated) [97]

Evidence-Based Complementary and Alternative Medicine 29

studies [83] Hot water extracts of B pilosa varminor Sheriffwere also assessed for its antileukemic effects on leukemiccell lines L1210 U937 K562 Raji and P3HR1 using XTT-based colorimetric assays The extract inhibited the five celllines with IC

50values ranging from 145120583gmL to 586120583gmL

L1210 K562 Raji and P3HR1 were more sensitive to B pilosaextract with IC

50values below 200120583gmL [98]

Consistent with the antitumor activities of B pilosaextracts and fractions some of its phytochemicals alsoshowed anticancer activity as outlined in Table 11 Amongthem luteolin (103) a well-studied flavonoid with multiplebioactivities was more effective against tumor cell prolifera-tion than its derivatives with IC

50values ranging from 3 120583M

to 50120583M in cells and 5 to 10mgkg in animals Luteolin wasalso found to fight cancer as a food additive at concentrationsof 50 to 200 ppm [78] and prevent skin cancer [78] and cancer3invasion [103] Lee and colleagues reported that luteolinprevents cancer by inhibiting cell adhesion and invasion[103] Significant inhibition concentration was reported to be5 120583M and complete inhibition concentration was reported tobe 40120583M Moreover luteolin was reported to inhibit hepa-tocyte growth factor (HGF)-induced cell scattering as wellas cytoskeleton changes such as filopodia and lamellipodiawhichwas determined using phase-contrast and fluorescencemicroscopy Furthermore luteolin also inhibited the HGF-induced phosphorylation of c-Met ERK 12 and Akt aswell as the MAPKERK and P13K-Akt pathways [78 103]Other mechanisms underlying the anticancer activities ofluteolin are the inhibition of topoisomerase I and II whichinhibits cell replication and DNA repair thus promotingapoptosis regulation of PI-3-KinaseAktMAPKERKJNKactivation of apoptosis in the mitochondrial pathway byactivating caspase 9 and caspase 3 which were found inmalignant cells but not in normal human peripheral bloodmononuclear cells death receptor-induced apoptosis and acell-cycle arrest mechanism inhibition of fatty acid synthasewhich is upregulated in many cancer cells and sensitizationto chemotherapywhereby luteolin increases the susceptibilityof cancer cells to chemotherapy [78] Butein (82) is anotherflavonoid that showed a cytotoxic effect on human colonadenocarcinoma cell proliferation with a reported IC

50value

of 175 120583M Butein at 2 120583M affected the incorporation of[14C]-labeled leucine thymidine and uridine which cancause the inhibition of DNA RNA and protein synthesis ofhuman colon cancer cells Moreover butein also exhibitednoncompetitive inhibition of 1-chloro-24-dinitrobenzene(CDNB) in glutathione S-transferase (GST) activity Tumorresistance was correlated with high levels of GST thus buteininhibited proliferation of cancer cells [80] Another flavonoidpresent in B pilosa centaureidin (109) also showed anti-cancer activity in B lymphoma cells Centaureidin isolatedfrom Polymnia fruticosa inhibited tubulin polymerization invitro and induced mitotic figure formation in CA46 Burkittlymphoma cells Using turbidimetric assay the IC

50value of

centaureidin in inhibition of mitosis was 3120583M [104] Cyto-toxicity of centaureidin was further analyzed using AmericanNational Cancer Institute (NCI) 60 human tumor cell linesThe cytotoxicity potency of centaureidin expressed as GI

50

(50 growth inhibition in the NCI tumor line panel) was024120583M [81] These data mark centaureidin as a promisingantimitotic agent for tumor therapy

In addition to anti-tumor flavones polyynes found in B 4pilosa have also been shown to possess anti-tumor proper-ties Based on a bioactivity-directed isolation approach Wuand colleagues identified two polyyne aglycones from theethyl acetate fraction of B pilosa [44] 12-Dihydroxytrideca-57911-tetrayne (48) and 13-Dihydroxy-6(E)- tetradecene-81012-triyne (46) exhibited significant anticell prolifera-tion activity in primary human umbilical vein endotheliumcells (HUVEC) with IC

50values of 125 120583M and 173 120583M

respectively They also decreased angiogenesis and pro-moted apoptosis in human endothelial cells Their anti-angiogenic and cytotoxic effects correlated with activationof the CDK inhibitors and caspase-7 [44] In addition 12-Dihyroxy-5(E)-tridecene-7911-triyne (45) showed antian-giogenic effects in HUVECs with an IC

50value of 124 120583M as

evidenced by a decrease in the tube formation and migrationof HUVECs [43] The IC

50value of compound 45 in the

inhibition of basic fibroblast growth factor-induced HUVECgrowth was 282120583M However it had higher IC

50values

than those for lung carcinoma cells and keratinocytes Thiscompound could also inhibit cell proliferation of HUVECslung carcinoma A549 cells and HACAT keratinocytes Themechanism by which compound 45 inhibits HUVEC growthand angiogenesis is complicated and includes decreasing theexpression of cell cycle regulators (CDK4 cyclins D1 and Aretinoblastoma (Rb) and vascular endothelial growth factorreceptor 1) caspase-mediated activation of CDK inhibitorsp21 (Cip1) and p27 (Kip) upregulation of Fas ligand expres-sion downregulation of Bcl-2 expression and activation ofcaspase-7 and poly (ADP-ribose) polymerase [43]

32 Anti-Inflammatory Activity B pilosa is commonly usedto treat inflammatory disorders The anti-inflammatory phy-tochemicals present in B pilosa are listed in Table 11 5Cyclooxygenase-2 (COX-2) is a physiologically importantenzyme that converts arachidonic acid to prostaglandin(PGE2) Its expression is induced by a wide variety of external

stimuli indicating its involvement in inflammatory diseasesand it is used as an inflammatory marker [88] Yoshida andcolleagues studied the effects of the aqueous extracts of Bpilosa aerial parts in the production of COX-2 and PGE

2as

well as on the activation of mitogen activated protein kinases(MAPKs) in normal human dermal fibroblasts (HDFs) inresponse to inflammatory cytokine IL-1120573 This work showedthat IL-1120573 activated MAPKs such as ERK12 p38 and JNKto different extents and induced COX-2 expression TheCOX-2 expression in HDFs was regulated mainly by p38following IL-1120573 stimulation Consistently the p38 inhibitorSB203580 blocked this expression Using this cell platformB pilosa extracts were tested for inhibition of inflammationThe extract dose-dependently suppressed the activation ofp38 and JNK and moderately suppressed ERK12 as wellas suppressing COX-2 expression and PGE

2production

30 Evidence-Based Complementary and Alternative Medicine

Table7Ph

enylprop

anoids

isolatedfro

mB

pilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

156

3-(4-H

ydroxyph

enyl)-2-prop

enoica

cid

p-Cou

maricacid

OHO

OH

Who

le(Ja

pan)

[99]

157

2-Metho

xy-4-(2-prop

en-1-yl)-ph

enol

Eugeno

lOOH

Leaves

androots(Japan)

[99]

158

3-(4-H

ydroxy-3-m

etho

xyph

enyl)-

2-prop

enoica

cid

Ferulic

acid

O

OH

O

HO

Who

le(Ja

pan)

[99]

159

3-(34-D

ihydroxyph

enyl)-2-prop

enoic

acid

Caffeicacid

O

HO

OH

OH

Who

leandaeria

l(Japan)

[7599]

160

3-Prop

yl-3[(245-trim

etho

xyph

enyl)-

metho

xy]-24-pentanedione

3-Prop

yl-3-(245-

trim

etho

xy)benzyloxy-

pentan-24-dione

O

O

O

O

O

O

Leaves

(India)

[100]

161

3-(34-D

ihydroxyph

enyl)-2-

prop

enoica

cidethyleste

rCa

ffeateethyl

O

O

OH

OH

Not

foun

d(Taiwan)

who

le(Taiwan)

[434584]

Evidence-Based Complementary and Alternative Medicine 31Ta

ble7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

162

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

d-Erythron

icacid

2-O-caffeoyl-2-C

-methyl

O

HO

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

163

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

2-O-caffeoyl-2-C

-methyl

O

O

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

164

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-24-

dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

3-O-caffeoyl-2-C

-methyl

O

O

HO

HO

O

O

OH

OH

Leaves

(Japan)

[101]

165

4-(A

cetyloxy)-3-[[3-(34-

dihydroxyphenyl)-

1-oxo-2-propen-1-y

l-]oxy]-2-hydroxy-

2-methyl-b

utanoica

cid

O

HO

OH

O OHO

HO

OO

Not

foun

d(Ja

pan)

[73]

166

3-(34-D

ihydroxyph

enyl)-tetrahydro-4-

hydroxy-4-

methyl-5

-oxo-3-fu

ranyl

ester-2-prop

enoica

cid

3-O-C

affeoyl-2-C

-methyl-

D-erythrono

-14-la

cton

eHO

HO

O

OO

OOH

Leaves

(Japan)

[101]

167

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]-oxy]-145-

trihydroxy-cyclohexanecarboxylicacid

Chlorogenica

cid

O

HO

HO

OH

O

OH

O

OH

HO

Aeria

l(Japan)w

hole

(Taiwan)

[707175]

32 Evidence-Based Complementary and Alternative Medicine

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

168

4-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]oxy]-

135-tr

ihydroxy-cyclohexanecarboxylic

acid

4-O-C

affeoylqu

inicacid

OHO

HO

OH

OH

O

O

OH

OH

Aeria

l(Japan)

[71]

169

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-15-

dihydroxy-cyclo

hexanecarboxylicacid

34-Di-O

-caffeoylqu

inic

acid

HO

HO

O

O

O

O

OH

OH

HO

OH O

H

O

Aeria

l(Japan)W

hole

(Taiwan)

[45497071]

170

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-14-dihydroxy-

cyclo

hexanecarboxylicacid

35-Di-O

-caffeoylqu

inic

acid

HO

HO

O O

HO

O

HO

OH OO

OHOH

Aeria

l(Japan)w

hole

(Taiwan)

[45497071]

Evidence-Based Complementary and Alternative Medicine 33

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

171

34-bis[[(2E

)-3-(34-D

ihydroxyph

enyl)-1-

oxo-2-prop

en-1-

yl]oxo]-15

-dihydroxy-

cyclo

hexanecarboxylicacid

45-Di-O

-caffeoylqu

inic

acid

OHO

OH

HO

O

O

OH

OH

O

OH

HO

OWho

le(Taiwan)

[454970]

172

3-[4-[[-6-O-[3-(4-H

ydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]-phenyl]-2-prop

enoica

cid

120573-D

-p-C

oumaricacid4-O

-(6-O

-p-caoum

aroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HOH

HO

O

O

Leaves

(Japan)

[59]

173

3-[4-[[2-O

-Acetyl-6

-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]ph

enyl]2-propeno

icacid

120573-D

-p-C

oumaricacid4-O

-(2-O

-acetyl-6

-O-p-

caou

maroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HO

HO

O

O

Leaves

(Japan)aerial

(China)

[3459]

174

67-Dihydroxy-2-chrom

enon

eEsculetin

cicho

rigenin

OHO HO

O

Not

foun

d(Egypt)

[39]

34 Evidence-Based Complementary and Alternative Medicine

Table 8 Aromatic compounds isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

175 12-Benzenediol PyrocatechinHO

HOWhole(Japan) [99]

176 4-Ethyl-12-benzenediol Pyrocatechol

OH

OH

Whole(Japan) [99]

177 Dimethoxyphenol26-dimethoxyphenol

O

HO

O

Roots (Japan) [99]

178 4-Ethenyl-2-methoxy-phenol p-Vinylguaiacol

O

OHWhole(Japan) [99]

179 2-Hydroxy-6-methylbenzaldehyde

6-Methyl-salicylaldehyde O

OH

Whole(Japan) [99]

180 Benzene-ethanol 2-Phenyl-ethanolHO

Whole(Japan) [57]

181 4-Hydroxy-3-methoxy-benzaldehyde Vanillin O

O

HOAerial(Japan) [99]

182 3-Hydroxy-4-methoxy-benzaldehyde Vanillin iso

OO

OH

Leaves(Japan) [99]

183 4-Hydroxybenzoic acid p-Hydroxybenzoicacid

O

HOOH Whole

(Japan) [99]

184 2-Hydroxybenzoic acid Salicylic acidO

HO

HO

Stem androots (Japan) [99]

185 34-Dihydroxybenzoicacid Protocatechuic acid

O

HOOH

OH

Whole(Japan) [99]

1864-Hydroxy-

3methoxybenzoicacid

Vanillic acid

OOH

O

OH

Aerial(Uganda)

Roots (Japan)[68 99]

Evidence-Based Complementary and Alternative Medicine 35

Table 8 Continued

S N IUPAC names Common names Structure Plant part(country) References

187 345-Trihydroxybenzoicacid Gallic acid

O

OHHO

HO

HO

Whole(China) [102]

[88] This work supports the use of B pilosa as an anti-inflammatory agent however no compounds responsible forthe anti-inflammatory activity of B pilosa were identified

A further study also reported the anti-inflammatoryactivity as well as the antiallergic activity of B pilosa [75] Inthis study dried powder of the aerial part of B pilosa whichhad been pretreated with the enzyme cellulosine was usedfor further tests The results showed that oral administrationof the cellulosine-treated B pilosa lowered the level of serumIgE in mice 10 days after immunization with DNP (24-dintrophenyl)-Ascaris as an antigen This treatment alsoreduced dye exudation in skin induced by passive cutaneousanaphylaxis and production of inflammatory mediators his-tamine and substance P in rats [75] Phytochemical analysisshowed that cellulosine treatment increased the percentage ofcaffeic acid and flavonoids This study suggests that B pilosaand its phenolics have anti-inflammatory functions

Phenolics and polyynes are major anti-inflammatoryphytochemicals present in B pilosa (Table 11) Unsurpris-6ingly phenolics such as luteolin (103) and ethyl caffeate(161) that are major constituents of B pilosa have alsobeen reported to possess anti-inflammatory activity Lute-olin was reported to exhibit anti-inflammatory activity inmacrophages Xagorari and colleagues showed that luteolininhibited the release of inflammatory cytokines TNF-120572 andinterleukin-6 in RAW 2647 cells following LPS stimulation[87] It inhibited TNF-120572 production with an IC

50value

of 1120583M The underlying anti-inflammatory mechanism ofluteolin was reported to be the inactivation of Akt andNF-120581B activation [87] In addition luteolin was reportedto confer anti-inflammatory activity through inhibition ofLPS-stimulated iNOS expression in BV-2 microglial cellsIt inhibited LPS-activated microglia in a dose-dependentmanner with an IC

50value of 69 120583MMoreover immunoblot

and RT-PCR data proved that luteolin suppressed I120581B-120572degradation and iNOS expression in LPS-activated microglia[85] Kim and colleagues stated that luteolin may havebeneficial effects on inflammatory neural diseases throughinhibition of iNOS expression [85] A related study revealedthat luteolin decreased the transcriptional activity of NF-120581BRelA via partial inhibition of TNF-mediated NF-120581B DNAbinding activity Luteolin also inhibited Akt phosphorylationand induced degradation of a transcription factor interferonregulatory factor (IRF) [86]

Chiang and colleagues showed that ethyl caffeate(161) significantly inhibited NO production in mousemacrophages RAW 2647 cells [84] Based on MTT assays

they concluded that this inhibition was not due to thecytotoxicity of ethyl caffeate The IC

50value of ethyl caffeate

in the inhibition of NO production was 55 120583gmL slightlylower than curcumin (positive control) which has anIC50

value of 65120583gmL They demonstrated that ethylcaffeate exerted anti-inflammatory activity via the reducedtranscription and translation of iNOS (inducible nitric oxidesynthase) in RAW 2467 cells In addition this compoundalso suppressed COX-2 expression in RAW 2467 cellsand MCF-7 cells The in vivo anti-inflammatory effectof ethyl caffeate was verified by testing in TPA-treatedmouse skin Like celecoxib the positive control ethylcaffeate significantly abolished COX-2 expression in adose-dependent manner Ethyl caffeate at 1mg200120583Lsite(24mM) inhibited COX-2 expression at a level comparableto celecoxib at 1mg200120583Lsite (13mM) Remarkably ethylcaffeate at 48mM (2mg200120583Lsite) was more effectivethan celecoxib at 131mM (10mg200120583Lsite) Moreoverthis compound inhibited the activation of nuclear factor-120581B(NF-120581B) by LPS via the prevention of NF-120581B binding toDNA [84] In addition 3 ethyl caffeate analogs (ethyl 34-dihydroxyhydrocinnamate ethyl cinnamate and catechol)also showed different degrees of NF-120581B binding to DNA aslisted in Table 12 Ethyl cinnamate lacks a catechol moietywhich results in ineffective inhibition of NF-120581B binding toDNA [84]

Pereira and colleagues assessed the anti- 7inflammatory and immunomodulatory activities ofB pilosa methanol extract as well as one polyyne2-O-120573-glucosyltrideca-11(E)-en-3 579-tetrayn-12-diol(54) in T lymphocytes and a zymosan-induced arthritismouse model [27] They first examined the in vitro effect of 8the B pilosa extract and compound 54 on cell proliferation ofhuman T cells stimulated with 5120583gmL phytohemagglutinin(PHA) or 100 nM 12-O-tetradecanoyl phorbol-13-acetate(TPA) plus 15 120583M ionomycin and on cell proliferation ofmouse T cells stimulated with 5 120583gmL concanavalin A(Con A) The data demonstrated that both methanol extractand compound 54 suppressed T-cell proliferation in adose-dependent manner The estimated IC

50values of the B

pilosa extract against human T cells stimulated with 5120583gmLPHA and 100 nM TPA plus 15120583M ionomycin were 125 and25 120583gmL respectively In comparison with the methanolextract compound 54 showed 10-fold more inhibition ofhuman T-cell proliferation with an estimated IC

50value of

15 120583gmL Accordingly the B pilosa extract and compound54 dose-dependently suppressed mouse T-cell proliferation

36 Evidence-Based Complementary and Alternative Medicine

Table 9 Porphyrins isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

188

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(15S16S)-10-ethenyl-5-ethyl-1161820-tetrahydro-6111522-tetramethyl-1820-dioxo-15H-912-imino-212-

metheno-471714-dinitrilopyrano[43-

b]azacyclononadecine-16-propanoic

acid

AristophyllC

NH N

HNN

O

O OOOPhytyl

O

Leaves(Taiwan) [100]

189

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -20b-(methoxycarbonyl)-9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-dinitrilo-12-

dioxeto-[310158404101584034]-cyclo-pent[1 2]azacyclononadecine-

19-propanoic acid

Bidenphytin A

NH N

HNN

OPhytylO O O

O O

Leaves(Taiwan) [100]

190

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -2a-

hydroxy-20b-(methoxycarbonyl)-

9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-

dinitrilo-12-dioxeto[310158404101584034]-cyclo-pent[12-b]-

azacyclononadecine-19-propanoic acid

Bidenphytin B

NH N

HNN

OPhytylO O O

O O

OH

Leaves(Taiwan) [100]

191

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO O

HOOO

Leaves(Taiwan) [100]

192

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

Evidence-Based Complementary and Alternative Medicine 37

Table 9 Continued

S N IUPAC names Common names Structure Plant part(country) References

193

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

Leaves(Taiwan) [100]

194

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

195

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3S4S21R)-9-ethenyl-14-ethyl-21-(methoxycarbonyl)-

481318-tetramethyl-20-oxo-3-phorbinepropanoic acid

Pheophytin A

NH N

HNN

PhytylO

O

OO O

Leaves(Taiwan) [100]

with estimated IC50values of 30 and 25 120583gmL respectively

Taken together the data indicate that the B pilosa extractand compound 54 act on human and mouse T cells Totest the in vivo effect of the B pilosa extract and compound54 a zymosan-induced arthritis mouse model was usedThis model was established from B10ASgSnJ mice with aninjection of zymosan (015mg) The zymosan-injected mice9received an intraperitoneal injection of the B pilosa extract(1 5 or 10mg) at one dose a day for 5 days Popliteal lymphnode (PLN) weight was monitored to check the developmentof arthritis The results revealed that 10mg of the methanolextract of B pilosa extract could significantly diminishinflammation as evidenced by PLN weight [27] This worksuggests that B pilosa (and compound 54) can suppressimmune response and inflammation

33 Antidiabetic Activity Anti-diabetic agents are primarilydeveloped from plants and other natural resources [8 24 4951 53] B pilosa is one of 1200 plant species that have beeninvestigated for antidiabetic activity [120 121]B pilosa is usedas an anti-diabetic herb in America Africa and Asia [51 120

122] Many studies have indicated that B pilosa could treattype 1 diabetes (T1D) and type 2 diabetes (T2D) in animals

Etiologically speaking T1D is caused by autoimmune-mediated destruction of pancreatic 120573 cells leading to insulindeficiency hyperglycemia and complications Currentlythere is no cure for T1D Polarization ofTh cell differentiationcontrols the development of T1D Suppression of Th1 celldifferentiation and promotion of Th2 cell differentiationameliorate T1D [123] One study showed that the butanolfraction of B pilosa inhibited T-cell proliferation decreasedTh1 cells and cytokines and increased Th2 cells andcytokines leading to prevention of T1D in nonobesediabetic (NOD) mice [49] Based on a bioactivity-directedisolation strategy 3 polyynes 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne (53) also known ascytopiloyne 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D- Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne (50) were identifiedfrom B pilosa [49 53] The IC

50value of the butanol fraction

was 200120583gmL This inhibition was reported to be partiallyattributed to cytotoxicity because the butanol fraction at

38 Evidence-Based Complementary and Alternative Medicine

Table 10 Other compounds isolated from B pilosa [30]

SN IUPAC names Common names Structure Plant part(country) References

19637-Dihydro-137-

trimethyl-1H-purine-26-dione

CaffeineN

NN

NO

O

Aerial(Uganda) [68]

197

1-((2R4S5R)-Tetrahydro-4-hydroxy-5-(hydroxymethyl)furan-2-yl)-5-

methylpyrimidine-24(1H3H)-dione

ThymidineHO

HO

ON

O

NHO Not found

(China) [37]

198 1-(2-Thienyl)-ethanone 2-Acetyl-thiophene

S

O

Roots(Germany) [41]

199

(2R3S4S5S)-2-(Heptan-2-yloxy)-tetrahydro-6-

((tetrahydro-34-dihydroxy-5-

(hydroxymethyl)furan-2-yloxy)methyl)-

2H-pyran-345-triol

Heptanyl2-O-120573-xylofuranosyl-

(1rarr 6)-120573-glucopyranoside

O

OOH

OH

HO

OHO

H

HO

H

H

HOHH

O

Whole(Taiwan) [70]

200

2-[(3R7R11R)-3-Hydroxy-371115-

tetramethylhexadecyl]-356-trimethyl-

25-cyclohexadiene-14-dione

120572-Tocopherylquinone

OO

HO

Whole(Taiwan) [32]

2017-O-(410158401015840610158401015840-Diacetyl)-

120573-D-glucopyranoside

Not found Leaves(China) [60]

180 120583gmL could cause 50 death of Th1 cells Moreoverthis study suggested that the butanol fraction may preventdiabetes in NOD mice in vivo via downregulation of Th1cells or upregulation Th2 cells that have effects which areantagonistic of those of Th1 cells [49] This was proven byintraperitoneal injection of the butanol fraction at a doseof 3mgkg BW 3 times a week to NOD mice from 4 to 27weeks This dosage resulted in lower incidence of diabetes(33) At a dose of 10mgkg the butanol fraction of Bpilosa totally eliminated (0) the initiation of the diseaseTo further support this result assessment of IgG2a and IgEproduction was performed in the serum of NOD mice Asin vivo results obtained from intracellular cytokine stainingexperiments were not very conclusive levels of IgG2a and IgE

were measured since Th1 cytokine IFN120574 and Th2 cytokineIL-4 favor the production of IgG2a and IgE respectively Asexpected high levels of IgE and some decline in the levels ofIgG2a were observed in the serum Profiling of the butanolextract revealed five compounds 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne(50) 45-Di-O-caffeoyl- quinic acid 35-Di-O-caffeoylquinicacid and 34-Di-O-caffeoylquinic acid Only the first twocompounds showed similar effects on the prevention ofdiabetes in NOD mice as the B pilosa butanol fractionMoreover compound 50 showed greater activity thancompound 69 in terms of enhancement (by 34 comparedto 8) of differentiation of Th0 to Th2 at 15 120583gmL (both

Evidence-Based Complementary and Alternative Medicine 39

Table 11 Chemical constituents of B pilosa and their biological activities

SN Name Classification Molecularformula Biological activities

109 Centaureidin [91] Flavonoid C18H16O8Anti-listerial [69 91]

Cytotoxic [10]

110 Centaurein [91] Flavonoid C24H26O13

Anti-listerial [69 91]Cytotoxic [10]Anti-viral [105]

103 Luteolin [106] Flavonoid C15H10O6

Anti-viral [107 108]Cytotoxic [77]

Anti-inflammatory [109]Anti-allergic [109]

82 Butein [110] Flavonoid C15H12O5Anti-leishmanial [111]

Cytotoxic [78]48 12-Dihydroxytrideca-57911-tetrayne [112] Polyyne C13H12O2 Anti-angiogeneic [112]46 13-Dihyroxy-6(E)-tetradecene-81012-triyne [112] Polyyne C14H16O2 Anti-angiogeneic [112]

45 12-Dihyroxy-5(E)-tridecene-7911-triyne [113] Polyyne C13H14O2Anti-angiogeneic [113]Anti-proliferative [113]

64 1-Phenylhepta-135-triyne [114] Polyyne C13H8

Anti-microbial [115]Anti-malarial [3]Cytotoxic [3]Antifungal [15]

27 Linoleic acid [79] Fatty acid C18H32O2Anti-viral (100)Cytotoxic [17]

161 Ethyl caffeate [84] Phenylpropanoid C11H12O4 Anti-inflammatory [84]

54 2-O-120573-Glucosyltrideca-11(E)-en-3579-tetrayn-12-diol [26] Polyyne C19H20O7Immunosuppressive andAnti-inflammatory [26]

53 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne [50] Polyyne C19H22O7Anti-diabetic [116]

Anti-inflammatory [50]

69 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne [28] Polyyne C20H26O7Anti-diabetic [28]

Anti-inflammatory [94]

50 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne [28] Polyyne C19H24O7

Anti-diabetic [28]Anti-inflammatory [94]

Anti-malarial andantibacterial [47]

129 Quercetin 3-O-120573-D-galactopyranoside [18] Flavonoid C21H20O12 Anti-inflammatory [117]

170 35-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

171 45-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

169 34-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

126 Quercetin 331015840-dimethyl ether7-O-120572-L-rhamnopyranosyl-(1rarr 6)-120573-D-glucopyranoside [118] Flavonoid C21H20O11 Anti-malarial [119]

125 Quercetin 331015840-dimethyl ether-7-O-120573-D-glucopyranoside [118] Flavonoid C21H20O12 Anti-malarial [119]70 1-Phenyl-13-diyn-5-en-7-ol-acetate [27] Polyyne C15H12O2 Anti-malarial [27]199 Heptanyl 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside [70] Miscellaneous C18H44O10 Antioxidant [70]124 3-O-Rabinobioside [70] Saccharide C27H38O15 Antioxidant [70]130 Quercetin 3-O-rutinoside [70] Flavonoid C27H38O15 Antioxidant [70]167 Chlorogenic acid [70] Phenolic C16H26O9 Antioxidant [70]119 Jacein [70] Flavonoid C24H26O13 Antioxidant [70]

49 (R)-12-dihydroxytrideca-357911-pentayne [47] Polyyne C13H8O2Anti-malarial andAntibacterial [47]

40 Evidence-Based Complementary and Alternative Medicine

Table 12 Structure and activity relationship studies of ethyl caffeateusing in vitro NF-120581BDNA binding assays [84]

Compound Concentration(120583M)

NF-120581BDNA binding

Ethyl caffeate 50 100inhibition

Ethyl 34-dihydroxyhydrocinnamate 100 100inhibition

Catechol 400 100inhibition

Ethyl cinnamate 400 Noinhibition

Table 13 Apoptosis in cocultures of T cells and pancreatic 120573 cells[53]

Cellmedium apoptosis andnecrosis

CD4+ T cellscontrol medium lt4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 2

CD4+ T cellswith cytopiloyne-treated 120573 cells 18CD4+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 7

CD8+ T cellscontrol medium 4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 4

CD8+ T cellswith cytopiloyne-treated 120573 cells 4CD8+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 4

Table 14 Radical scavenging activities of B pilosa extracts [70]

Extractscontrol DPPH assayIC50 (120583gmL)

NBThypoxanthinesuperoxide assayIC50 (120583gmL)

Quercetin 198 15Ascorbic acid 634 Not determined120572-tocopherol 897 Not determinedEthyl acetateextract 1383 597

Butanol extract 1669 114Water extract gt100 gt100

compounds) and inhibition (by 40 compared to 10) ofdifferentiation toTh1 at the same concentration [49]

Among the three polyynes found in B pilosa cytopiloyne(53) had the most potent anti-T1D activity [53] To test the invivo effect of cytopiloyne NODmice received intraperitonealor intramuscular injection of cytopiloyne at 25120583gkg BW3 times per week Twelve-week-old NOD mice started todevelop T1D and 70 of NOD mice aged 23 weeks andover developed T1D Remarkably 12- to 30-week-old NODmice treated with cytopiloyne showed normal levels of bloodglucose (lt200mgdL) and insulin (1-2 ngmL) Consistent

Table 15 Radical scavenging activity of secondarymetabolites fromB pilosa [70]

Metabolite (Table 11)control DPPH assay IC50 (120583gmL)199 Not determined124 53130 68167 105169 33171 38119 Not determined110 Not determinedQuercetin 256Caffeic acid 890

Table 16 Antioxidant activity of the essential oils and water extractsfrom B pilosa [92]

Extract IC50 (120583gmL)Leaf essential oils 47Flower essential oils 50Leaf extract 61Flower extract 172

with T1D incidence cytopiloyne delayed and reduced theinvasion of CD4+ T cells into the pancreatic islets [53]

In vitro study showed that cytopiloyne (53) inhibited thedifferentiation of naıveTh (Th0) cells (ie CD4+ T cells) intoTh1 cells and promoted differentiation of Th0 cells into Th2cells [50] The in vitro data are consistent with the in vivoresults indicating that cytopiloyne reducedTh1differentiationand increased Th2 differentiation as shown by intracellularcytokine staining and FACS analysis [53] In line with theskewing of Th differentiation the level of serum IFN-120574 andIgG2c decreased while that of serum IL-4 and serum IgEincreased compared to the negative controls (PBS-treatedmice) Cytopiloyne also enhanced the expression of GATA-3 a master gene for Th2 cell differentiation but not theexpression of T-bet a master gene forTh1 cell differentiationfurther supporting its role in skewingTh differentiation [53]

Also importantly cytopiloyne partially depleted CD4+rather than CD8+ T cells in NOD mice [53] As shown inTable 13 coculture assays showed that the depletion of CD4+T cells was mediated through the induction of Fas ligandexpression on pancreatic islet cells by cytopiloyne leading toapoptosis of infiltrating CD4+ T cells in the pancreas via theFas and Fas ligand pathways However cytopiloyne did notinduce the expression of TNF-120572 in pancreatic islet cells andthus had no effect on CD8+ T cells [53]

In addition Chang and colleagues showed that cytopi-loyne dose-dependently inhibited T-cell proliferation stimu-lated by IL-2 plus Con A or anti-CD3 antibody using [3H]thymidine incorporation assay [53]

Overall the mechanism of action of cytopiloyne andprobably its derivatives in T1D includes inhibition of T-cellproliferation skewing of Th cell differentiation and partialdepletion of Th cells Due to the anti-diabetic mechanisms

Evidence-Based Complementary and Alternative Medicine 41

Table 17 Antibacterial activity of essential oils and flower extracts from B pilosa [92]

Strain Mean zone of inhibition (mm)Leaf essential oil Flower essential oil Leaf extract Flower extract

Micrococcus flavus 127 plusmn 03 87 plusmn 03 102 plusmn 02 108 plusmn 03Bacillus subtilis 173 plusmn 19 117 plusmn 02 109 plusmn 02 103 plusmn 02Bacillus cereus 190 plusmn 14 112 plusmn 03 118 plusmn 04 185 plusmn 10Bacillus pumilus 123 plusmn 07 108 plusmn 02 105 plusmn 04 77 plusmn 02Escherichia coli 137 plusmn 04 203 plusmn 07 102 plusmn 11 140 plusmn 13Pseudomonas ovalis 125 plusmn 08 137 plusmn 15 102 plusmn 06 125 plusmn 06

Table 18 Antibacterial activity of root extracts from B pilosa [93]

Strain MIC50 (mgmL)Methanol extract Acetone extract

Bacillus cereus 10 mdashEscherichia coli 5 5Klebsilla pneumonia 5 10Micrococcus kristinae 10 mdashPseudomonas aeruginosa 10 10Staphylococcus aureus 5 10Sraphylococcus epidermidis 5 5Serratia marcescens 10 mdashShigelea flexneri 10 mdashStreptococcus faecalis 10 mdash

Table 19 Antibacterial activity of B pilosa of compound 29 [47]

Strain MIC50 (120583gmL)Escherichia coli NIHJ 1Escherichia coli ATCC25922 1Klebsiella pneumoniae ATCC700603 128Serratia marcescens ATCC13880 16Pseudomonas aeruginosa ATCC27853 8Staphylococcus aureus FDA209P 05Staphylococcus aureus ATCC29213 025Staphylococcus aureus N315 (MRSA) 05Enterococcus faecalis ATCC29212 2Enterococcus faecalis NCTC12201 (VRE) 1Bacillus subtilis ATCC6633 05Candida albicans ATCC10231 025

of action it was hypothesized that cytopiloyne protectsNOD mice from diabetes by a generalized suppression ofadaptive immunity To evaluate this hypothesis ovalbumin(Ova) was used as a T-cell dependent antigen to primeNOD mice which had already received cytopiloyne orPBS vehicle Ova priming boosted similar anti-Ova titersin cytopiloyne-treated mice and PBS-treated mice but adifference in immunoglobulin isotype was observed in thetwo groups Thus it was concluded that cytopiloyne is animmunomodulatory compound rather than an immunosup-pressive compound [50 53]

T2D is a chronic metabolic disease with serious com-plications resulting from defects in either insulin secretion

insulin action or both [124] A study by Ubillas et alshowed that the aqueous ethanol extract of the aerial partof B pilosa at 1 gkg body weight (BW) lowered bloodglucose in dbdb mice a T2D mouse model [51] Basedon a bioactivity-guided identification compounds 69 and50 were identified Further the mixture of the compounds(69 50) in a 2 3 ratio significantly decreased blood glucoseconcentration and reduced food intake on the second dayof treatment when administered at doses of 250mgkg twicea day to C5BLKs-dbdb mice When tested at 500mgkga more substantial drop in blood glucose level as wellas the stronger anorexic effect (food intake reduced from58 gmouseday to 25 gmouseday) was observed [51] Inthis study it was suggested that the blood glucose lower-ing effect of B pilosa was caused in part by the hungersuppressing effect of its polyynes [51] However the hungersuppressing effect of the ethanol extract of B pilosa wasnot found in the studies described below In another study[24] water extracts of B pilosa (BPWE) were used indiabetic dbdb mice aged 6ndash8 weeks with postprandialblood glucose levels of 350 to 400mgdL Like oral anti-diabetic glimepiride which stimulates insulin release onesingle dose of BPWE reduced blood glucose levels from374 to 144mgdL The antihyperglycemic effect of BPWEwas inversely correlated to an increase in serum insulinlevels suggesting that BPWE acts to lower blood glucose viaincreased insulin production However BPWE had differentinsulin secretion kinetics to glimepiride [24] One flaw incurrent anti-diabetics is their decreasing efficacy over timeThe authors investigated the long term anti-diabetic effectof BPWE in dbdb mice BPWE reduced blood glucoseincreased blood insulin improved glucose tolerance andreduced the percentage of glycosylated hemoglobin (HbA1c)Both long-term and one-time experiments strongly supportthe anti-diabetic action of BPWE [24] In sharp contrast toglimepiride BPWE protected against islet atrophy in mousepancreas The investigators further evaluated anti-diabeticproperties of 3 B pilosa varieties B pilosa L var radiate(BPR) Bpilosa L var pilosa (BPP) and B pilosa L varminor(BPM) in dbdb mice [8] One single oral dose (10 50 and250mgkg body weight) of BPR BPP or BPM crude extractsdecreased postprandial blood glucose levels in dbdb micefor up to four hours and the reduction of glucose levels inthe blood appeared to be dose-dependent Comparing thethree variants BPR extract resulted in a higher reduction inblood glucose levels when administered at the same dose as

42 Evidence-Based Complementary and Alternative Medicine

Table 20 Antifungal activity of B pilosa [92]

Partextract Concentration (ppm) Strain InhibitionCortiicum rolfsii Fusarium solani Fusarium oxysporum

Leaves

Essentialoils

100 857 plusmn 09 682 plusmn 0 745 plusmn 17250 960 plusmn 08 779 plusmn 18 879 plusmn 04

AqueousExtracts

100 446 plusmn 17 605 plusmn 21 716 plusmn 07250 942 plusmn 03 689 plusmn 07 824 plusmn 19

Flowers

Essentialoils

100 604 plusmn 09 892 plusmn 04 869 plusmn 05250 894 plusmn 12 980 plusmn 03 949 plusmn 06

AqueousExtracts

100 331 plusmn 11 714 plusmn 07 573 plusmn 22250 661 plusmn 14 912 plusmn 0 900 plusmn 07

Table 21 Antifungal activity of B pilosa root extracts [93]

Strain LC50 (mgmL)Acetoneextracts

Methanolextracts

Waterextracts

Aspergillus niger 014 006 007Aspergillus flavus 1091 658 0Penicillium notatum 005 005 005

the other two varieties In terms of serum insulin levels a doseof 50mgkg of each extract was used and the BPR extracttogether with the three polyynes significantly increased theserum insulin level in dbdb mice Long-term experiments(28-day treatment) were then conducted using diabetic micewith postprandial glucose levels from 370 to 420mgdL andglimepiride was used as positive controlThe range of dosagesapplied was from 10mgkg BW to 250mgkg BW Resultsshowed that the positive control as well as the crude extractsof the three varieties lowered the blood glucose levels indbdb mice However only BPR extract containing a higherpercentage of cytopiloyne (53) reduced blood glucose levelsand augmented blood insulin levelsmore than BPP andBPMThe percentage of glycosylated hemoglobin A1c (HbA1c) wasalso measured and found to be 79 plusmn 05 in mice aged 10ndash12 weeks and 66 plusmn 02 61 plusmn 03 and 62 plusmn 03 inthe blood of age-matchedmice following treatment with BPRcrude extract (50mgkg) glimepiride (1mgkg) and com-pound 53 (05mgkg) respectively [8] Among the polyynesfound in B pilosa cytopiloyne was the most effective againstT2D Hence cytopiloyne was used for further study on anti-diabetic action and mechanism [125] The data confirmedthat cytopiloyne reduced postprandial blood glucose lev-els increased blood insulin improved glucose tolerancesuppressed HbA1c level and protected pancreatic islets indbdb mice Nevertheless cytopiloyne failed to decreaseblood glucose in streptoztocin (STZ)-treated mice whose bcells were already destroyed Additionally cytopiloyne dose-dependently increased insulin secretion and expression in bcells as well as calcium influx diacylglycerol and activationof protein kinase C120572 Collectively the mechanistic studiessuggest that cytopiloyne treats T2D via regulation of insulin

production involving the calciumDAGPKC120572 cascade in bcells

The studies detailed above point to the conclusion thatcytopiloyne and related polyynes (compounds 69 and 49)are anti-diabetics in animal models The data uncover a newbiological action of polyynes It should be noted that likeall anti-diabetic drugs cytopiloyne failed to prevent or curediabetes completely but reduced diabetic complications [125]Intriguingly 34 polyynes have been found in B pilosa so farIt remains to be seen whether all the polyynes present in thisplant have anti-diabetic activities

34 Antioxidant Activity Free radicals can damage cellularcomponents via a series of chemical reactions [89] leadingto development and progression of cardiovascular diseasecancer neurodegenerative diseases and ageing [70] Freeradicals nitric oxide (NO) and superoxide anions can beproduced in macrophages to kill microbes However anexcessive generation of the free radicals under pathologicalconditions is associated with a wide range of illnessesPlants are known to be rich in antioxidant phytochemicalsChiang and colleagues evaluated the free radical scaveng-ing activity of crude extract fractions and compounds ofB pilosa using 11-diphenyl-2-picrylhydrazyl (DPPH) andhypoxanthinexanthine oxidase assays [70] Using DPPHand hypoxanthinexanthine oxidase assays they found thatthe B pilosa crude extract and the ethyl acetate butanoland water fractions had free radical scavenging activityNine compounds Heptyl- 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside (199) 3-O-Rabinobioside (124) Quercetin3-O-rutinoside (130) Chlorogenic acid (167) 34-Di-O-caffeoylquinic acid (169) 35-Di-O-caffeoylquinic acid (170)45-Di-O-caffeoylquinic acid (171) Jacein (119) and Cen-taurein (110) had DPPH radical scavenging activity [70]The IC

50values of the B pilosa crude extractfractions and

compounds are summarized in Tables 14 and 15 respectivelyMeasurement of free radical scavenging activities is one

way of assessing the antioxidant activities of B pilosa andits fractions and compounds It is interesting that the ethylacetate and butanol fractions are more active than the waterfraction and B pilosa crude extract [70] Of the secondarymetabolites only phenolic compounds 124 130 167 169 and171 showed significant DPPH-radical scavenging activities

Evidence-Based Complementary and Alternative Medicine 43

Further analysis of the structure-activity relationship of thecompounds suggested that substitution of the C3 hydroxylgroup with glycosides increased the activity approximately2-fold (for example in compounds 124 and 130) relative toquercetin (-OH in its C3) [70] Further modification of thestructures of the active compounds needs to be performed totest the effects of various substituents on activity The reasonwhy most of the antioxidant compounds contain phenolmoieties in their structure could be that the reduction-oxidation (redox) properties of phenols allow them to actas reducing agents singlet oxygen quenchers and hydrogendonors

A complementary study by Muchuweti and colleaguesdetermined phenolic content antioxidant activity and thephenolic profile of B pilosa methanol extract [89] Theyestimated that the phenolic content of the methanol extractof B pilosawas 11028plusmn22mgg [89] Vanillin hydroxyben-zaldehyde caffeic acid coumaric acid and ferulic acid werefound in this extractThe B pilosa extract also showed DPPHradical scavenging activity Furthermore the antioxidantactivity of the flavonoids found in B pilosa was correlatedwith its hepatoprotective effects through their inhibitionof NF-120581B activation which may lessen the oxidative stresscaused by the production of free radicals during liver injury[60]This activity might also be due to the anti-inflammatoryeffects of the aqueous extracts of B pilosa aerial parts on theinhibition of COX-2 and PGE

2production [88]

Essential oils from B pilosa flowers and leaves are alsoreported to possess antioxidant activity With the aim ofreplacing chemically synthesized additives Deba and col-leagues [92] worked on the antioxidant antibacterial andantifungal activities of essential oils and water extracts of Bpilosarsquos leaves and flowers Table 16 summarizes the resultsobtained from DPPH free radical scavenging assay

It can be inferred fromTable 16 that essential oils from theleaves possessed the highest activity It is reported elsewherethat monoterpenes present in essential oils such that of Bpilosa have protective effects and antioxidant properties [92]Beta-carotene bleaching method was also performed Leavesessential oils and aqueous extracts of the leaves and flowersshowed higher activity than the flower essential oils This isdue to the volatility of the flower essential oils The activityexhibited by the aqueous extracts was accounted to thepresence of phenolic compounds that are reported to donatea hydrogen atom to free radicals such that the propagation ofthe chain reaction during lipid oxidation is terminated [92]Overall essential oils and phenolics present in B pilosa canbe thought of as major antioxidant compounds

35 Immunomodulatory Activity B pilosa is thought to bean immunomodulatory plant and is reported to be effectivein the treatment of immune disorders such as allergy [75]arthritis [75] and T1D [48 50 53]

As pointed out in the discussion of its anti-diabeticactivities (Section 33) a combination of phytochemicalsmethod and T-cell activation assays was used to studyimmunomodulatory properties of B pilosa IFN-120574 is a keycytokine released by T and NK cells that mediates immune

cells and sustains immunity against pathogens Defects inIFN-120574 expression regulation and activation result in vulner-ability to diseases caused by bacteria and viruses [69] Anelegant study performed byChang and colleagues using IFN-120574 promoter-driven luciferase reporter construct in JurkatT cells showed that hot water crude extracts of B pilosaincreased IFN-120574 promoter activity two-fold [69] Out of thesubfractions of this extract the butanol fraction but not thewater or ethyl acetate fractions increased IFN-120574 promoteractivity six-fold Centaurein (110) and centaureidin (109)were identified from the butanol fraction and were stated tocause a four-fold increase in IFN-120574 promoter activity withEC50

values of 75 120583gmL and 09120583gmL respectively Themechanism of action of centaurein was determined usingtranscription factors such as AP-1 NFAT and NF120581B whichare reported to bind to IFN-120574 promoter and regulate IFN-120574transcription Unlike with the activity of the positive controlPHA centaurein caused a four-fold increase in NFAT a3-fold increase in NF120581B and had little if any effect onAP-1 enhancer activities (23-fold three-fold and ten-foldincreases respectively were seenwith PHA) [69]The authorsconcluded that centaureinmodulates IFN-120574 expression by theNFAT and NF120581B pathways The article only determined themechanism of action of centaurein Its aglycone centaureidinmay act through the samemechanism though this conclusionneeds to be further verified

B pilosa extract and its compounds are reported to inhibitdifferentiation of naıve CD4+ helper T (Th0) cells into Th1cells [49] Using theThcell differentiation assay as a screeningplatform 3 polyynes 2-120573-D-glucopyranosyloxy-1 -hydroxytrideca-57911-tetrayne (53) 3-120573-D-glucopyranosyl-1-hydroxy-6(E)- tetradecene-81012-triyne (69) and 2-120573-D-glucopyranosyloxy-1-hydroxy-5(E)- tridecene-7911-triyne(49) were discovered from B pilosa [49 53] The data showsthat cytopiloyne and other two polyynes suppressed thedifferentiation of type 1 helper T (Th1) cells and productionof Th1 cytokines and promoted that of type 2 helper T (Th2)cells and production of Th2 cytokines thus explaining theimmunomodulatory and anti-inflammatory effects of Bpilosa and its polyynes

Chang and colleagues were the first to report the effect ofthe butanol extract of B pilosa on the autoimmune diabetesand airway inflammation inmice [48] Imbalance in the levelsofTh1 andTh2 and of various cytokines leads to autoimmunediseases T1D and other autoimmune diseases (rheumatoidarthritis Crohnrsquos disease among others) are exacerbated byan increase Th1 levels (specifically CD4+ Th1 cells) whileTh2 cells antagonize this effect [19] Moreover Th2 cellsmediate asthma in ovalbumin-induced hypersensitivity inBALBc mice [48] In their work [48] Chang and colleaguesshowed that 10mgkg butanol extracts with 15 (ww)compound 69 and 11 (ww) compound 49 (Table 11)ameliorated the development of Th1-mediated diabetes inNOD mice through inhibition of 120573 cell death and leukocyteinfiltration At the same dosage the butanol extracts alsoexacerbated ovalbumin-induced pulmonary inflammationin BALBc mice with an increase in the infiltration ofeosinophils and mast cells into the airway of the mice [48]Despite the different outcomes both mouse models proved

44 Evidence-Based Complementary and Alternative Medicine

the concept that control over the Th1Th2 shift is associatedwith autoimmune diseases and that the B pilosa butanolextract can shift the differentiation of Th0 cells to Th2 cells[48 49]

An extended study presented by Chiang and colleaguesshowed that compound 53 modulates T-cell functions [50]Using CD4+ T cells from BALBc mice they demonstratedthat cytopiloyne decreased levels of IFN-120574 producing cells(Th1) by 122 (from 72 to 598) Since Th1 and Th2 celldifferentiation is antagonistic it was expected that compound53 increased the percentage of mouse IL-4 producing cells(Th2) by 72 (from 237 to 309) Subsequent assess-ment of effect of compound 53 on the modulation of thetranscription of IL-4 and IFN-120574 showed that cytopiloyne asexpected decreased the splenocyte levels of IFN-120574 mRNAand increasing that of IL-4 in a dose-dependent manner Inthe range of 01 to 3 120583gmL of compound 53 the effects werenot attributed to its cytotoxicity Consequently using 3120583gmLcytopiloyne for 72 and 96 hours the protein concentration ofIFN-120574 decreased to 186 and 444 respectively Under thesame conditions cytopiloyne increased IL-4 concentrationsto 1985 and 2470 (for 72 and 96 hours resp) Thismodulation of T-cell differentiation exhibited by compound53 was used to explain its anti-diabetic activity [50]

The anti-diabetic role of cytopiloyne was extensivelydiscussed above (Section 33 anti-diabetic activity) Themolecular basis of the regulation of cytokine expression bycytopiloyne has been described Cytopiloyne directly elevatedthe expression level of IL-4 via GATA-3 upregulation inT cells [53] However reduction of IFN-120574 expression in Tcells seemed to come from the indirect opposing effect IL-4cytokine because the expression level of T-bet was unaltered[53] In this way cytopiloyne skewedTh1 polarization into theTh2 state conferring protection against T1D in NOD miceAside frompolarization ofThcell differentiation cytopiloynealso activated the expression of Fas ligand in pancreaticb cells this increase leading to the partial depletion of Tcells and reduction of immune response in local areas suchas the pancreas Of note cytopiloyne also inhibited T-cellproliferation and activation By targeting T cells from threeimmunomodulatory actions cytopiloyne protects againstT1D and probably other Th1-mediated autoimmune diseases[19]

The phytochemical constituents of B pilosa exert theirfunctions on different immune cells to modulate immuneresponse It is possible that some of the compounds mayhave agonistic or antagonistic effects on immune responseImmune function of B pilosa may depend on its compo-sition and amount of compounds which could explain theapparently conflicting report of B pilosa butanol extractaggravating allergy in mice [48] while cellulosine-treatedextract ameliorated allergy [48 75] IFN-120574 promoter reporterassays and T-cell differentiation assays were used to isolate2 flavonoids [69] and 3 polyynes [49 50] as immunomod-ulatory compounds from the butanol fraction of B pilosaInterestingly the flavonoids promote IFN-120574 expression inNK and T cells In marked contrast the polyynes promoteIL-4 expression and indirectly inhibit IFN-120574 expression indifferentiating T cells Sensitivity appears to be the key to

identifying structure- and bioactivity-related phytochemicalsfrom medicinal plants

36 Antimalarial Activity The use of chemical drugs againstpathogens has resulted in drug-resistant mutants Examplesof drug resistance can be found in the species of thePlasmodium that cause malaria It is important to search fornew compounds to combat Plasmodium parasites [47] Astudy of the anti-malarial activity of the leaf extracts of Bpilosa using a combination of phytochemistry and bioassaysshowed that compound 49 (Table 11) showed activity againsta malaria parasite (P falciparum NF54 strain) with an IC

50

value of 60 120583gmL [77] In addition compound 49 isolatedfrom the aerial parts of B pilosa inhibited growth of the Pfalciparum FCR-3 strain with an IC

50value of 035 120583gmL

This compound was tested for its in vivo effect in miceinfected with P berghei NK-65 strain Results showed thatcompound 49 decreased the average parasitemia in the redblood cells by 207 (from 328 of that of the control to121) after an intravenous injection of 08mgkg BWday forfour days [47] Further studies addressing the anti-malarialmechanism underlying both polyynes and clinical studies areneeded

37 Antibacterial Activity Emergence of multiple antibiotic-resistantmicrobes is becoming a global threat to public healthand a challenge to disease treatment For instance penicillinis commonly used to combat a food-borne intracellularbacteriumListeria however penicillin-resistant bacteria havebeen discovered recently [69] Chang and coworkers iso-lated centaurein (110) and centaureidin (109) from B pilosaextract [69 70] Centaurein enhances expression of IFN-120574a key cytokine for macrophage activation and consequentlyenhances bactericidal activity in macrophages [69 70] Inagreement with observed in vitro effects centaurein wasreported to prevent and treat Listeria infection in C57BL6Jmice [91] Mechanistic studies confirmed that centaureinexerted antilisterial action via IFN-120574 expression in wild-typemice but not IFN-120574 knockout mice [91] Further in vitrostudies on centaurein showed that this compound increasedIFN-120574 expression by 13 (from 17 to 20) 20 (from 21to 41) and 11 (from 6 to 17) in CD4+ T cells CD8+Tcells and NK cells respectively That is to say there wasan increase in IFN-120574 producing immune cells As expectedcentaurein also enhanced the expression level of T-bet a keynuclear factor for IFN-120574 expression Consistently centaureinaugmented the serum IFN-120574 levels inC57BL6Jmice and thisaugmentation peaked 24 hours after compound injectionThe quantity of mouse serum IFN-120574 was sufficient to activatemacrophages in vitro and eradicated GFP-producing Listeriainside macrophages However the entry of Listeria intomacrophages was not affected by centaurein-treated mousesera Centaurein treatment at 20120583g per mouse rescued 30of the mice infected with a lethal dose of Listeria (2 times106 CFU) It is noteworthy that in the presence of ampicillin(5 120583gmouse) centaurein (20 120583gmouse) rescued 70 of themice suggesting an additive effect between ampicillin andcentaurein [91] Despite lower abundance centaureidin was

Evidence-Based Complementary and Alternative Medicine 45

30 times more active than centaurein in terms of IFN-120574production [91]

Aside from the indirect antibacterial action mentionedabove extract andor compounds of B pilosa also showeddirect bacteriostatic andor bactericidal action One studyreported that essential oils and leafflower extracts of Bpilosa could suppress the growth of gram positive and gramnegative bacteria as evidenced by zone of inhibition assaysIn this study antibacterial activity of the essential oils fromthe leaves and flowers of B pilosa was determined in anattempt to identify natural products as food preservatives forprevention of microbial multiplication and food oxidationThe essential oils and extracts of B pilosa leaves and flow-ers showed moderate but different extents of antibacterialactivity (Table 17) In general essential oils had higherantibacterial activity than crude extracts One explanation forthis could be that monoterpenes in the essential oils destroycellular integrity and subsequently inhibit the respirationand ion transport processes The presence of antibacterial120573-caryophyllene could be another explanation as reportedelsewhere [92] Another study reported that the methanoland acetone extract of B pilosa roots displayed antibacterialactivities against the bacteria listed in Table 18 [93] andmethanol extracts from the roots seemed to be the mosteffective

Another study indicated that the polyyne (R)-12-dihydroxytrideca-357911-pentayne (49) from this plantalso suppressed bacterial growth as shown by the minimuminhibitory concentration required to inhibit 50 bacterialgrowth (MIC

50) in Table 19 This compound was highly

effective against several Gram positive and Gram negativebacteria including the drug-resistant bacteria Staphylococcusaureus N315 (MRSA) and Enterococcus faecalis NCTC12201(VRE) [47] Strikingly compound 49 had a similar MIC

50

value to antibiotics (ampicillin tetracycline norfloxacin andamphotericin B) in most of the bacteria tested

Antibacterial activity of B pilosa extracts and compo-nents expressed as MIC

50and the mean zone of inhibition

is tabulated in Tables 17 18 and 19The zone of inhibition forAmpicilline (positive control) ranges from 153 plusmn 03mm to443 plusmn 02mm [92]

38 Antifungal Activity B pilosa has traditionally been usedto treat microbial infection Recently different parts of Bpilosa have been tested for antifungal activities Deba andcolleagues first evaluated the antifungal effect of the hotwater extracts of the B pilosa roots stems and leaves againstCorticium rolfsii Fusarium solani and Fusarium oxysporumThey discovered that C rolfsii was most suppressed bytreatment with B pilosa as its growth was reduced at almostall the tested doses followed by F oxysporum and F solani[99] However the fungicidal activities of the stems androots were greater than the leaves [99] Moreover the samegroup assessed the antifungal activity of the essential oilsand aqueous extracts from B pilosa flowers and leaves [92]They showed that the extracts and oils had antifungal activityagainst C rolfsii F solani and F oxysporum Essential oils

appeared to have better fungicidal activity thanwater extractsas summarized in Table 20

Another study by Ashafa and colleagues showed thatacetone methanol and water extracts of the B pilosa rootsshowed antifungal activities against Aspergillus niger Aflavus and Penicillium notatum using the agar dilutionmethod The results are tabulated in Table 21 [93] Negativecontrols showed 0 growth inhibitionThe methanol extractof the B pilosa roots at 10mgmL was also effective againstCandida albicans [93]Of note B pilosa obtained fromPapuaNew Guinea had no activity against A niger and C albicans[126] but the SouthAfrican (EasternCape) ecotype exhibitedmoderate activity against C albicans [93] This discrepancymay depend on extraction solvents extraction procedureassay techniques different plant parts and abundance ofactive compounds

B pilosa produces a variety of secondary metabolitessuch as flavonoids phenylacetylenes alkaloids sterols ter-penoids and tannis [92 93] However none of them havebeen confirmed as active compounds against fungi Furtherinvestigation of active compounds from B pilosa is necessaryto further understand the antifungal efficacy of this plant

39 Hypotensive and Vasodilatory Activities In early studiesDimo and colleagues used three rat models normotensiveWistar rats (NTR) salt-loading hypertensive rats (SLHR)and spontaneous hypertensive rats (SHR) to investigate thehypotensive effect of the methanol crude extract of B pilosaleaves [28 127] The extract lowered systolic blood pressurein hypertensive rats (SLHR and SHR) to a greater degreethan NTR [28 127] In addition a decrease in urinarysodium ions and an increase in urinary potassium ions wereobserved after treatment with the methanol extract of Bpilosa leaves although neither differences were statisticallysignificant [28 127] Taking the data together the studyproposed that B pilosa leaf extract reduced blood pressurevia vasodilation [28 127] The same group continued totest the antihypertensive effect of aqueous and methylenechloride extracts of B pilosa leaves in a hypertensive ratmodel [28 127] To establish a fructose-induced hypertensionmodel male Wistar rats were given 10 fructose solutionto drink ad libitum for three weeks In addition to freeaccess to 10 fructose the rats were treated with the aqueous(150mgkg) or methyl chloride (350mgkg) extracts of Bpilosa for additional three weeks [28 127] Both extracts ofB pilosa leaves had a hypotensive effect on rats Howeverneither extracts reversed the elevation of serum insulin infructose-fed rats Therefore B pilosa lowered blood pressureirrespective of insulin [28 127] To better understand thehypotensive mechanism the authors investigated the effectof a neutral extract of B pilosa (NBP) a mixture of methanolandmethylene chloride (1 1) extract after neutralization withNaOH and HCl on the heart and the blood pressure ofNTR and SHR [110] This study showed that an intravenousinjection of the NBP resulted in a biphasic reduction insystolic blood pressure In addition one intravenous dose ofthe extract at 10 20 and 30mgkg BW decreased systolicblood pressure in normal rats by 183 425 and 30

46 Evidence-Based Complementary and Alternative Medicine

respectively and the same doses reduced the blood pressurein hypertensive rats by 258 389 and 286 respectivelyOnly the highest dose (30mgkg) affected the force of thecontraction of the heart Atropine and propranolol were usedto interfere with the hypotensive action of the NBP Atropinereduced the initial phase of the hypotensive response in NBPand completely abolished the second phase of hypotensiveresponse in NBP In contrast propranolol increased thefirst hypotensive response but partially abolished the secondhypotensive response provoked by the NBP [110]This mech-anistic study suggested that B pilosa invokes the biphasichypotensive responses via targeting cardiac pump efficiencyduring the first phase and vasodilation at the second phase[110]

A further study was performed to investigate the relaxingeffect of a neutral extract of B pilosa (NBP) on rat aorta con-tractedwithKCl (60mM) andnorepinephrine (01mM) [94]Cumulative addition of NBP relaxed the rat aorta previouslycontracted by KCl in a dose-dependent manner The EC

50

value of theNBP for vasorelaxationwas 032mgmLThedataalso showed that the NBP reduced the contraction of aortapreviously contracted by KCl irrespective of the presence ofaortic endothelium [94]

Pretreatment with glibenclamide an ATP-dependent K+channel blocker did not considerably affect the relaxant effectof the NBP on KCl-induced contraction suggesting thatthe vasodilatory effect of B pilosa was not related to theopening of this ATP-dependent K+ channel [94] On theother hand in the presence of indomethacin or pyrilaminemaleate the relaxant response induced by the plant extractwas significantly inhibited at the lower concentrations Theplant extract was able to reduce the aorta resting tone inhibitthe KCl-induced contractions by 90 at 15mgmL and theCaCl2-induced contractions by 95 at 075mgmL These

results demonstrate that B pilosa can act as a vasodilatorprobably via acting as a calcium antagonist [94]

However no specific compound for the above activityhas been identified from B pilosa to date A bioactivity-guided identification approach may be adopted to identifythe active compounds in B pilosa that possess hypotensiveand vasodilatory effects and understand their mechanism ofaction

310 Wound Healing Activity B pilosa has been traditionallyused to treat tissue injury inCameroon Brazil andVenezuela[26] Hassan and colleagues investigated the wound healingpotential of B pilosa in Wistar rats [128] Mirroring thepositive control neomycin sulfate the ethanol extract of Bpilosa had faster wound closure than control rats 3 6 and 9days after topical application Histological examination alsorevealed better collagenation angiogenesis and organizationof wound tissue seven days after application Epithelializationand total healing time in B pilosa-treated rats were compara-ble to those of neomycin sulfate Together these data suggestthat B pilosa may be a viable alternative to neomycin lotionfor the treatment of wounds

In addition to studying the wound healing effect of Bpilosa on external ulcers Tan and colleagues also examined

the effect of methanol cyclohexane and methyl chlorideextracts of B pilosa on gastric ulcers in Wistar rats fedwith 1mL HClethanol gastric necrotizing solution (150mMHCl in 60 ethanol) and macroscopically visible lesionswere scored [26] Among the three extracts methylene chlo-ride extracts exhibited the highest activity showing 464inhibition of lesion formation at a dose of 500mgkg BWand complete inhibition at 750mgkg [26] The efficacy ofthe ethylene chloride extract was followed by that of themethanol extracts which had inhibition ranging from 304to 822 at concentrations of 500mgkg and 1000mgkgBW respectively [26] The cyclohexane extracts showed thelowest activity against gastric ulcers in rats with 13340 and 797 inhibition at 500 750 and 1000mgkg BWrespectively [26] To better understand the mode of actionof the methylene chloride extract of B pilosa rats werepretreated with indomethacin a COX-2 inhibitor involved inprostaglandin synthesis Pretreatment significantly reducedthe protection against HClethanol-induced ulcers to 313inhibition at 750mgkg BW suggesting a link between theantiulcerative activity of B pilosa and prostaglandin syn-thesis Unexpectedly the methylene chloride extract of Bpilosa showed little gastric mucosal protection against gastriclesions induced by 95 ethanol (1mL) [26] Absolute alcoholis known to cause mucosalsubmucosal tissue destruction viacellular necrosis and the release of tissue-derived mediators(histamine and leukotrieneC4)Thus these data imply thatBpilosa did not prevent the generation or the necrotic action ofthese mediators on the gastric microvasculature In additionpylorus ligation can increase gastric acid secretion withoutan alteration of mucosal histamine content The methylenechloride extract of B pilosa did not possess antisecretoryactivity On the contrary it was observed that increases in thedose of the extract led to elevated gastric juice acidity [26]Results with both absolute ethanol and pylorus ligation ratmodels suggested the possibility that the ineffectiveness of Bpilosa against gastric ulcers was due to lack of antihistaminicactivity in the plant In summary overall the data suggest thatB pilosa protects against HCLethanol-mediated ulcers viainhibition of prostaglandin biosynthesis

Previous phytochemical studies showed that a groupof flavonoids acyclichalcones are present in B pilosa[129 130] and the chalcones were proposed to have anti-ulcerative activity [131] Moreover nine hydroxychalcones 10were reported to possess gastric cytoprotective effects with24-dihydroxychalcone being the most active [132] Sincemethylene chloride extracts appear to be the most active Bpilosa extracts next the specific anti-ulcerative phytochemi-cals in the methylene chloride extracts of B pilosa and theirmodes of action needs to be probed

Despite the claims listed inTable 3 relatively few scientificstudies have been conducted in vitro and in vivo to address thetraditional ethnomedical uses of B pilosa Information aboutthe use of B pilosa as a botanical therapy recorded so far isfar from complete Studies conducted thus far only serve asa starting point for further investigation of B pilosa and theultimate efficacious use of the herb in clinical applications

Evidence-Based Complementary and Alternative Medicine 47

4 Toxicology

Despite its use as an ingredient in food for human con-sumption studies on systemic toxicity (eg acute subacutechronic and subchronic toxicities) of B pilosa in humansand animals are still inadequate and insufficient So faracute andor subchronic toxicities have been evaluated inrats and mice Oral acute and 28-day toxicities of waterextract of B pilosa leaves were evaluated in Wistar rats[133] An oral dose of water extract of B pilosa leaves at10 gkg BW showed no obvious mortality or changes inthe appearance in rats [134] The same extract at 08 gkgBWday once a day showed no obvious sub-chronic toxicityin rats over 28 days as measured by survival rate bodyweight and gross examination of organs [134] These dataare consistent with our data indicating that oral deliveryof the water extract of the B pilosa whole plant at 1 gkgBWday once a day is safe in rats over 28 days (unpublisheddata) Taken together these studies suggest that ingestion ofB pilosa aqueous extract at up to at 1 gkg BWday oncea day is highly safe in rats In addition the acute toxicityof aqueous and ethanol extracts of B pilosa in mice havebeen reported [134] Five- to six-week-old mice with weightsbetween 28 and 35 g received a peritoneal injection of bothextracts at the different doses The LD

50 the dose that causes

50 lethality of the aqueous and ethanol extracts in micewas 1230 gkg BW and 615 gkg BW respectively [134] Acomplete toxicological study has not been completed forhumans Furthermore the drug interactions of B pilosa withother drugs are unknown Further safety verification andclinical trials should be performed before B pilosa can beconsidered for medicinal use

5 Conclusions

B pilosa is an erect perennial plant with green leaves whiteor yellow flowers and tiny black seeds As it is distributedworldwide and is widely used as a folk remedy B pilosacan be thought of as an extraordinary source of food andmedicine However a comprehensive up-to-date review ofresearch on B pilosa has hitherto been unavailable In thisarticle scientific studies on B pilosa have been summarizedand critically discussed from the perspectives of botanyethnomedicine phytochemistry pharmacology and toxicol-ogy B pilosa is claimed to treat more than 40 disordersand 201 compounds have been identified from this plantThe medicinal utility of B pilosa and its modes of actionin relation to its known phytochemicals were discussedherein Polyynes flavonoids phenylpropanoids fatty acidsand phenolics are the primary bioactive compounds of Bpilosa and they have been reported to be effective in thetreatment of tumors inflammationimmune modulationdiabetes viruses microbes protozoans gastrointestinal dis-eases hypertension and cardiovascular diseases Cautionshould be exercised in the therapeutic use of B pilosa forhypoglycemia hypotension bleeding and allergy

Acknowledgments

The authors thank their laboratory members for construc-tive suggestions and the authors whose publications theycited for their contributions The authors also thank MsMiranda Loney of Academia Sinica for English editing of thispaper This work was supported by Grants 99-CDA-L11 and101S0010056 from Academia Sinica Taiwan Arlene P Bar-tolome is a recipient of MECO-TECO Sandwich ScholarshipProgram between Taiwan and the Philippines

References

[1] T Shen G H Li X N Wang and H X Lou ldquoThe genusCommiphora a review of its traditional uses phytochemistryand pharmacologyrdquo Journal of Ethnopharmacology vol 142 no2 pp 319ndash330

[2] C Long P Sauleau B David et al ldquoBioactive flavonoids ofTanacetum parthenium revisitedrdquo Phytochemistry vol 64 no2 pp 567ndash569 2003

[3] P O Karis and O Ryding Asteraceae Cladistics and Classifica-tion Bremer K Eds pp 559ndash569 Timber press Portland OreUSA 1994

[4] O N Pozharitskaya A N Shikov M N Makarova et alldquoAnti-inflammatory activity of a HPLC-fingerprinted aqueousinfusion of aerial part of Bidens tripartita LrdquoPhytomedicine vol17 no 6 pp 463ndash468 2010

[5] F Q Oliveira V Andrade-Neto A U Krettli and MG L Brandao ldquoNew evidences of antimalarial activity ofBidens pilosa roots extract correlated with polyacetylene andflavonoidsrdquo Journal of Ethnopharmacology vol 93 no 1 pp 39ndash42 2004

[6] Agriculture USDA ldquoPlants databaserdquo 2013in Natural Resources Conservation Servicehttpwwwnrcsusdagovwpsportalnrcssitenationalhome

[7] Y Zhou and X Z Yan ldquoExperimental study of qi deficiencysyndrome and Codonopsis pillosulae and Astragalus injectionon the immune response in micerdquo Zhong Xi Yi Jie He Za Zhivol 9 no 5 pp 286ndash262 1989

[8] S C Chien P H Young Y J Hsu et al ldquoAnti-diabeticproperties of three common Bidens pilosa variants in TaiwanrdquoPhytochemistry vol 70 no 10 pp 1246ndash1254 2009

[9] M J Alcaraz and M J Jimenez ldquoFlavonoids as anti-inflammatory agentsrdquo Fitoterapia vol 59 no 1 pp 25ndash38 1988

[10] FAO Agriculture Food and Nutrition for AfricamdashA ResourceBook for Teachers of Agriculture Publishing ManagementGroup FAO Information Division Rome Italy 1997

[11] M B Rokaya Z Munzbergova B Timsina and K R BhattaraildquoRheum australe D Don a review of its botany ethnobotanyphytochemistry and pharmacologyrdquo Journal of Ethnopharma-cology vol 141 no 3 pp 761ndash774 2012

[12] P H Young Y J Hsu and W C Yang ldquoBidens pilosa L andits medicinal userdquo in Recent Progress in Medicinal Plants DrugPlant II A S AwaadVK Singh and JNGovil Eds StandiumPress Houston Tex USA 2010

[13] C Ge ldquoCytologic study of Bidens bipinnata LrdquoZhongguo ZhongYao Za Zhi vol 15 no 2 pp 72ndash125 1990

[14] L C Chiang J S Chang C C Chen L T Ng and C CLin ldquoAnti-herpes simplex virus activity of Bidens pilosa and

48 Evidence-Based Complementary and Alternative Medicine

Houttuynia cordatardquoAmerican Journal of ChineseMedicine vol31 no 3 pp 355ndash362 2003

[15] N P Rybalchenko V A Prykhodko S S Nagorna et alldquoIn vitro antifungal activity of phenylheptatriyne from Bidenscernua L against yeastsrdquo Fitoterapia vol 81 no 5 pp 336ndash3382010

[16] K Redl W Breu B Davis and R Bauer ldquoAnti-inflammatoryactive polyacetylenes from Bidens campylothecardquo Planta Med-ica vol 60 no 1 pp 58ndash62 1994

[17] M Ayyanar and S Ignacimuthu ldquoTraditional knowledge ofKani tribals inKouthalai of Tirunelveli hills TamilNadu IndiardquoJournal of Ethnopharmacology vol 102 no 2 pp 246ndash255 2005

[18] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant intraditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[19] AKAbbas KMMurphy andA Sher ldquoFunctional diversity ofhelper T lymphocytesrdquo Nature vol 383 no 6603 pp 787ndash7931996

[20] J H Cano and G Volpato ldquoHerbal mixtures in the traditionalmedicine of Eastern Cubardquo Journal of Ethnopharmacology vol90 no 2-3 pp 293ndash316 2004

[21] J Namukobe J M Kasenene B T Kiremire et al ldquoTraditionalplants used for medicinal purposes by local communitiesaround the Northern sector of Kibale National Park UgandardquoJournal of Ethnopharmacology vol 136 no 1 pp 236ndash245 2011

[22] C Lans ldquoComparison of plants used for skin and stomachproblems in Trinidad and Tobago with Asian ethnomedicinerdquoJournal of Ethnobiology andEthnomedicine vol 3 article 3 2007

[23] S Dharmananda ldquoA Popular Remedy Ecapes Notice ofWesternPractitionersrdquo 2013 httpwwwitmonlineorgartsbidenshtm

[24] Y J Hsu T H Lee C L T Chang Y T Huang andW C YangldquoAnti-hyperglycemic effects and mechanism of Bidens pilosawater extractrdquo Journal of Ethnopharmacology vol 122 no 2 pp379ndash383 2009

[25] E Noumi F Houngue and D Lontsi ldquoTraditional medicinesin primary health care plants used for the treatment ofhypertension in Bafia Cameroonrdquo Fitoterapia vol 70 no 2 pp134ndash139 1999

[26] P V Tan T Dimo and E Dongo ldquoEffects of methanolcyclohexane and methylene chlo ride extracts of Bidens pilosaon various gastric ulcer models in ratsrdquo Journal of Ethnophar-macology vol 73 no 3 pp 415ndash421 2000

[27] R L C Pereira T Ibrahim L Lucchetti A J R Da Silva and VL G De Moraes ldquoImmunosuppressive and anti-inflammatoryeffects of methanolic extract and the polyacetylene isolatedfrom Bidens pilosa Lrdquo Immunopharmacology vol 43 no 1 pp31ndash37 1999

[28] T Dimo J Azay P V Tan et al ldquoEffects of the aqueous andmethylene chloride extracts of Bidens pilosa leaf on fructose-hypertensive ratsrdquo Journal of Ethnopharmacology vol 76 no 3pp 215ndash221 2001

[29] CWiartMedicinal Plants of Southeast Asia Pelanduk SelabgorDarul Ehsan Malaysia 2000

[30] F L Silva D C H Fischer J F Tavares M S Silva P FDe Athayde-Filho and J M Barbosa-Filho ldquoCompilation ofsecondary metabolites from Bidens pilosa LrdquoMolecules vol 16no 2 pp 1070ndash1102 2011

[31] A H Chen S R Lin and C H Hong ldquoPhytochemical studyon Bidens pilosa L varminorrdquo Huaxue pp 38ndash42 1975

[32] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[33] M C Zulueta M Tada and C Y Ragasa ldquoA diterpene fromBidens pilosardquo Phytochemistry vol 38 no 6 pp 1449ndash14501995

[34] J Wang H Yang Z W Lin and H D Sun ldquoFlavonoids fromBidens pilosa var radiatardquo Phytochemistry vol 46 no 7 pp1275ndash1278 1997

[35] A Zhao Q Zhao L Peng et al ldquoA new chalcone glycoside fromBidens pilosardquo Acta Botanica Yunnanica vol 26 no 1 pp 121ndash126

[36] P Gao Y Wu S Cui and Y Zou ldquoSynthesis and absoluteconfiguration of the 7-phenylhepta-4 6-diyne-1 2-diol isolatedfrom Bidens pilosardquo Synthesis no 13 pp 2131ndash2135 2011

[37] S Wang B Yang D Zhu D He and L Wang ldquoActivecomponents of Bidens pilosa Lrdquo Zhongcaoyao vol 36 pp 20ndash21

[38] F Bohlmann T Burkhardt and C Zdero Naturally OccuringAcetylenes Academic Press New York NY USA 1973

[39] T M Sarg A M Ateya N M Farrag and F A AbbasldquoConstituents and biological activity of Bidens pilosa L grownin EgyptrdquoActa Pharmaceutica Hungarica vol 61 no 6 pp 317ndash323 1991

[40] H A L Valdes and H P Rego ldquoBidens pilosa Linnerdquo RevistaCubana de Plantas Medicinales vol 6 pp 28ndash33

[41] F Bohlmann H Bornowski and K M Kleine ldquoNew polyynesfrom the tribe Heliantheaerdquo Chemische Berichte vol 97 pp2135ndash2138

[42] RWang Q XWu and Y P Shi ldquoPolyacetylenes and flavonoidsfrom the aerial parts ofBidens pilosardquo PlantaMedica vol 76 no9 pp 893ndash896 2010

[43] L W Wu Y M Chiang H C Chuang et al ldquoA novelpolyacetylene significantly inhibits angiogenesis and promotesapoptosis in human endothelial cells through activation of theCDK inhibitors and caspase-7rdquo Planta Medica vol 73 no 7 pp655ndash661 2007

[44] L W Wu Y M Chiang H C Chuang et al ldquoPolyacetylenesfunction as anti-angiogenic agentsrdquo Pharmaceutical Researchvol 21 no 11 pp 2112ndash2119 2004

[45] H L Yang S C Chen N W Chang et al ldquoProtection fromoxidative damage using Bidens pilosa extracts in normal humanerythrocytesrdquo Food and Chemical Toxicology vol 44 no 9 pp1513ndash1521 2006

[46] H Q Wang S J Lu H Li and Z H Yao ldquoEDTA-enhancedphytoremediation of lead contaminated soil by Bidens maxi-mowiczianardquo Journal of Environmental Sciences vol 19 no 12pp 1496ndash1499 2007

[47] S TobinagaMK SharmaWG L Aalbersberg et al ldquoIsolationand identification of a potent antimalarial and antibacterialpolyacetylene from Bidens pilosardquo Planta Medica vol 75 no 6pp 624ndash628 2009

[48] C L T Chang H K Kuo S L Chang et al ldquoThe distincteffects of a butanol fraction of Bidens pilosa plant extract onthe development of Th1-mediated diabetes and Th2-mediatedairway inflammation in micerdquo Journal of Biomedical Sciencevol 12 no 1 pp 79ndash89 2005

[49] S L Chang C L T Chang Y M Chiang et al ldquoPolyacetyleniccompounds and butanol fraction from Bidens pilosa can mod-ulate the differentiation of helper T cells and prevent autoim-mune diabetes in non-obese diabetic micerdquo Planta Medica vol70 no 11 pp 1045ndash1051 2004

Evidence-Based Complementary and Alternative Medicine 49

[50] Y M Chiang C L T Chang S L Chang W C Yang and LF Shyur ldquoCytopiloyne a novel polyacetylenic glucoside fromBidens pilosa functions as a T helper cell modulatorrdquo Journal ofEthnopharmacology vol 110 no 3 pp 532ndash538 2007

[51] R PUbillas CDMendez S D Jolad et al ldquoAntihyperglycemicacetylenic glucosides from Bidens pilosardquo Planta Medica vol66 no 1 pp 82ndash83 2000

[52] L Alvarez S Marquina M L Villarreal D Alonso E Arandaand G Delgado ldquoBioactive polyacetylenes from Bidens pilosardquoPlanta Medica vol 62 no 4 pp 355ndash357 1996

[53] C L T Chang S L Chang Y M Lee et al ldquoCytopiloyne apolyacetylenic glucoside prevents type 1 diabetes in nonobesediabetic micerdquo Journal of Immunology vol 178 no 11 pp 6984ndash6993 2007

[54] G Kusano A Kusano and Y Seyama Novel Hypoglycemicand Anti-Inflammatory Polyacetylenic Compounds Their Com-positions Bidens Plant Extract Fractions and CompositionsContaining the Plant or Fraction JPO Tokyo Japan 2004

[55] M G L Brandao A U Krettli L S R Soares C G CNery and H C Marinuzzi ldquoAntimalarial activity of extractsand fractions from Bidens pilosa and other Bidens species(Asteraceae) correlated with the presence of acetylene andflavonoid compoundsrdquo Journal of Ethnopharmacology vol 57no 2 pp 131ndash138 1997

[56] A U Krettli V F Andrade-Neto M D G L Brandao andW M S Ferrari ldquoThe search for new anti-malarial drugs fromplants used to treat fever and malaria or plants ramdomlyselected a reviewrdquoMemorias do Instituto Oswaldo Cruz vol 96no 8 pp 1033ndash1042 2001

[57] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[58] C K Wat R K Biswas E A Graham L Bohm G H NTowers and E R Waygood ldquoUltraviolet-mediated cytotoxicactivity of phenylheptatriyne from Bidens pilosa Lrdquo Journal ofNatural Products vol 42 no 1 pp 103ndash111 1979

[59] Y Sashida K Ogawa M Kitada H Karikome Y Mimakiand H Shimomura ldquoNew aurone glucosides and new phenyl-propanoid glucosides from Bidens pilosardquo Chemical and Phar-maceutical Bulletin vol 39 no 3 pp 709ndash711 1991

[60] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens pilosa L (TFB) on animal liver injury andliver fibrosisrdquo Journal of Ethnopharmacology vol 116 no 3 pp539ndash546 2008

[61] B Hoffmann and J Holzl ldquoAcylated compounds from Bidenspilosardquo Planta Medica vol 55 pp 108ndash109

[62] B Hoffmann and J Holzl ldquoA methylated chalcone glucosidefrom Bidens pilosardquo Phytochemistry vol 27 no 11 pp 3700ndash3701 1988

[63] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[64] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[65] B Hoffmann and J Holzl ldquoWeitere acylierte chalkone ausBidens pilosardquo Planta Medica vol 54 pp 450ndash451 1988

[66] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant in11traditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[67] V V Pham V V T K P V L Hoang and V K Phan ldquoFlavonoidcompounds from the plant Bidens pilosa L (Asteraceae)rdquo TapChi Duoc Hoc vol 50 pp 48ndash53

[68] S D Sarker B Bartholomew R J Nash and N Robinson ldquo5-O-methylhoslundin an unusual flavonoid from Bidens pilosa(Asteraceae)rdquo Biochemical Systematics and Ecology vol 28 no6 pp 591ndash593 2000

[69] S L Chang Y M Chiang C L T Chang et al ldquoFlavonoidscentaurein and centaureidin from Bidens pilosa stimulate IFN-120574 expressionrdquo Journal of Ethnopharmacology vol 112 no 2 pp232ndash236 2007

[70] Y M Chiang D Y Chuang S Y Wang Y H Kuo P WTsai and L F Shyur ldquoMetabolite profiling and chemopreventivebioactivity of plant extracts from Bidens pilosardquo Journal ofEthnopharmacology vol 95 no 2-3 pp 409ndash419 2004

[71] A Kusano Y Seyama E Usami et al ldquoStudies on the antioxi-dant active constituents of the dried powder from Bidens pilosaL var radiata SchrdquoNaturalMedicines vol 57 no 3 pp 100ndash1042003

[72] I Doleckova L Rarova J Gruz et al ldquoAnti-proliferative andanti-angiogenic effects of flavone eupatorin an active con-stituent of chloroform extract ofOrthosiphon stamineus leavesrdquoFitoterapia vol 83 no 6 pp 1000ndash1007

[73] K Bairwa R Kumar R J Sharma and R K Roy ldquoAn updatedreview on Bidens pilosa Lrdquo Der Pharma Chemica vol 2 no 3pp 325ndash337

[74] Q Xia Y Liu and Y Li ldquoDetermination of hyperoside indifferent parts and different species of Herba Bidens by RP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol 50pp 48ndash53

[75] M Horiuchi and Y Seyama ldquoImprovement of the antiinflam-matory and antiallergic activity of Bidens pilosa L var radiataSCHERFF treated with enzyme (Cellulosine)rdquo Journal of HealthScience vol 54 no 3 pp 294ndash301 2008

[76] M G L Brandao C G C Nery M A S Mamao and AU Krettli ldquoTwo methoxylated flavone glycosides from Bidenspilosardquo Phytochemistry vol 48 no 2 pp 397ndash400 1998

[77] P Kumari K Misra B S Sisodia et al ldquoA promising anticancerand antimalarial component from the leaves of Bidens pilosardquoPlanta Medica vol 75 no 1 pp 59ndash61 2009

[78] G Seelinger I Merfort U Wolfle and C M Schempp ldquoAnti-carcinogenic effects of the flavonoid luteolinrdquoMolecules vol 13no 10 pp 2628ndash2651 2008

[79] G Seelinger I Merfort and C M Schempp ldquoAnti-oxidantanti-inflammatory and anti-allergic activities of luteolinrdquoPlanta Medica vol 74 no 14 pp 1667ndash1677 2008

[80] C C Yit and N P Das ldquoCytotoxic effect of butein on humancolon adenocarcinoma cell proliferationrdquo Cancer Letters vol82 no 1 pp 65ndash72 1994

[81] J A Beutler E Hamel A J Vlietinck et al ldquoStructure-activityrequirements for flavone cytotoxicity and binding to tubulinrdquoJournal of Medicinal Chemistry vol 41 no 13 pp 2333ndash23381998

[82] M R Kviecinski K B Felipe T Schoenfelder et al ldquoStudy ofthe antitumor potential of Bidens pilosa (Asteraceae) used inBrazilian folkmedicinerdquo Journal of Ethnopharmacology vol 117no 1 pp 69ndash75 2008

[83] P Sundararajan A Dey A Smith A G Doss M Rajappanand S Natarajan ldquoStudies of anticancer and antipyretic activityof Bidens pilosawhole plantrdquo African Health Sciences vol 6 no1 pp 27ndash30 2006

[84] YMChiang C P Lo Y P Chen et al ldquoEthyl caffeate suppressesNF-120581B activation and its downstream inflammatory mediatorsiNOS COX-2 and PGE2 in vitro or in mouse skinrdquo BritishJournal of Pharmacology vol 146 no 3 pp 352ndash363 2005

50 Evidence-Based Complementary and Alternative Medicine

[85] J S Kim H J Lee M H Lee J Kim C Jin and J HRyu ldquoLuteolin inhibits LPS-stimulated inducible nitric oxidesynthase expression in BV-2 microglial cellsrdquo Planta Medicavol 72 no 1 pp 65ndash68 2006

[86] P A Ruiz and D Haller ldquoFunctional diversity of flavonoidsin the inhibition of the proinflammatory NF-120581B IRF and Aktsignaling pathways in murine intestinal epithelial cellsrdquo Journalof Nutrition vol 136 no 3 pp 664ndash671 2006

[87] A Xagorari A Papapetropoulos A Mauromatis MEconomou T Fotsis and C Roussos ldquoLuteolin inhibitsan endotoxin-stimulated phosphorylation cascade andproinflammatory cytokine production in macrophagesrdquoJournal of Pharmacology and Experimental Therapeutics vol296 no 1 pp 181ndash187 2001

[88] N Yoshida T Kanekura Y Higashi and T Kanzaki ldquoBidenspilosa suppresses interleukin-1120573-induced cyclooxygenase-2expression through the inhibition of mitogen activated proteinkinases phosphorylation in normal human dermal fibroblastsrdquoJournal of Dermatology vol 33 no 10 pp 676ndash683 2006

[89] M Muchuweti C Mupure A Ndhlala T Murenje and M AN Benhura ldquoScreening of antioxidant and radical scavengingactivity of Vigna ungiculata Bidens pilosa and Cleome gynan-drardquo American Journal of Food Technology vol 2 no 3 pp 161ndash168 2007

[90] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens bipinnata L against carbon tetrachloride-induced liver fibrosis in ratsrdquo Journal of Pharmacy and Pharma-cology vol 60 no 10 pp 1393ndash1402 2008

[91] S L Chang H H Yeh Y S Lin Y M Chiang T K Wuand W C Yang ldquoThe effect of centaurein on interferon-120574 expression and Listeria infection in micerdquo Toxicology andApplied Pharmacology vol 219 no 1 pp 54ndash61 2007

[92] F Deba T D Xuan M Yasuda and S Tawata ldquoChemical com-position and antioxidant antibacterial and antifungal activitiesof the essential oils from Bidens pilosa Linn var Radiatardquo FoodControl vol 19 no 4 pp 346ndash352 2008

[93] A O T Ashafa and A J Afolayan ldquoScreening the root extractsfrom Biden pilosa L var radiata (Asteraceae) for antimicrobialpotentialsrdquo Journal of Medicinal Plant Research vol 3 no 8 pp568ndash572 2009

[94] T B Nguelefack T Dimo E P Nguelefack Mbuyo P V TanS V Rakotonirina and A Kamanyi ldquoRelaxant effects of theneutral extract of the leaves of Bidens pilosa linn on isolated ratvascular smooth musclerdquo Phytotherapy Research vol 19 no 3pp 207ndash210 2005

[95] M T Grombone-Guaratini K L Silva-Brandao V N SolferiniJ Semir and J R Trigo ldquoSesquiterpene and polyacetyleneprofile of the Bidens pilosa complex (Asteraceae Heliantheae)from Southeast of Brazilrdquo Biochemical Systematics and Ecologyvol 33 no 5 pp 479ndash486 2005

[96] L L Lin C Y Wu H C Hsiu M T Wang and H ChuangldquoStudies on diabetes mellitus I The hypoglycemic activity ofphytosterin on alloxan diabetic ratsrdquo Taiwan Yi Xue Hui Za Zhivol 66 no 2 pp 58ndash65 1967

[97] M A N Benhura and I C Chitsiku ldquoThe extractable 120573-carotene content of Guku (Bidens pilosa) leaves after cookingdrying and storagerdquo International Journal of Food Science andTechnology vol 32 no 6 pp 495ndash500 1997

[98] J S Chang L C Chiang C C Chen L T Liu K C Wangand C C Lin ldquoAtileukemic activity of Bidens pilosa l varminor (blume) sherff andHouttuynia cordata thunbrdquo AmericanJournal of Chinese Medicine vol 29 no 2 pp 303ndash312 2001

[99] F Deba T D Xuan M Yasuda and S Tawata ldquoHerbicidal andfungicidal activities and identification of potential phytotoxinsfrom Bidens pilosa L var radiata Scherff research paperrdquoWeedBiology and Management vol 7 no 2 pp 77ndash83 2007

[100] J K Kumar and A K Sinha ldquoA new disubstituted acetylacetonefrom the leaves ofBidens pilosa LinnrdquoNatural Product Researchvol 17 no 1 pp 71ndash74 2003

[101] K Ogawa and Y Sashida ldquoCaffeoyl derivatives of a sugarlactone and its hydroxy acid from the leaves of Bidens pilosardquoPhytochemistry vol 31 no 10 pp 3657ndash3658 1992

[102] Q Xia Y Liu and Y Li ldquoDetermination of gallic acid fromdifferent species and different medical parts of Herba Bidens byRP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol24 no 03 pp 308ndash310

[103] W J Lee L F Wu W K Chen C J Wang and T HTseng ldquoInhibitory effect of luteolin on hepatocyte growth fac-torscatter factor-induced HepG2 cell invasion involving bothMAPKERKs and PI3K-Akt pathwaysrdquo Chemico-BiologicalInteractions vol 160 no 2 pp 123ndash133 2006

[104] J A Beutler J H Cardellina C M Lin E Hamel G MCragg andM R Boyd ldquoCentaureidin a cytotoxic flavone fromPolymnia fruticosa inhibits tubulin polymerizationrdquo BioorganicandMedicinal Chemistry Letters vol 3 no 4 pp 581ndash584 1993

[105] A Verma A Su A M Golin B OrsquoMarrah and J K AmorosaldquoThe lateral view a screening method for knee traumardquo Aca-demic Radiology vol 8 no 5 pp 392ndash397 2001

[106] J Corren M Lemay Y Lin L Rozga and R K RandolphldquoClinical and biochemical effects of a combination botanicalproduct (ClearGuard) for allergy a pilot randomized double-blind placebo-controlled trialrdquo Nutrition Journal vol 7 no 1article 20 2008

[107] M S Gachet J S Lecaro M Kaiser et al ldquoAssessment of anti-protozoal activity of plants traditionally used in Ecuador in thetreatment of leishmaniasisrdquo Journal of Ethnopharmacology vol128 no 1 pp 184ndash197 2010

[108] W C Yang M Ghiotto B Barbarat and D Olive ldquoThe role ofTec protein-tyrosine kinase in T cell signalingrdquo The Journal ofBiological Chemistry vol 274 no 2 pp 607ndash617 1999

[109] S Tewtrakul H Miyashiro N Nakamura et al ldquoHIV-1 inte-grase inhibitory substances from Coleus parvifoliusrdquo Phytother-apy Research vol 17 no 3 pp 232ndash239 2003

[110] T Dimo T B Nguelefack P V Tan et al ldquoPossible mechanismsof action of the neutral extract from Bidens pilosa L leaves onthe cardiovascular system of anaesthetized ratsrdquo PhytotherapyResearch vol 17 no 10 pp 1135ndash1139 2003

[111] T S C Li Chinese and Related North American Herbs CRCPress New York NY USA 2002

[112] H Wu H Chen X Hua Z Shi L Zhang and J ChenldquoClinical therapeutic effect of drug-separated moxibustion onchronic diarrhea and its immunologic mechanismsrdquo Journal ofTraditional Chinese Medicine vol 17 no 4 pp 253ndash258 1997

[113] S W Wright R R Harris J S Kerr et al ldquoSynthesis chemicaland biological properties of vinylogous hydroxamic acids dualinhibitors of 5-lipoxygenase and IL-1 biosynthesisrdquo Journal ofMedicinal Chemistry vol 35 no 22 pp 4061ndash4068 1992

[114] W R Almiron and M E Brewer ldquoClassification of immaturestage habitats of Culicidae (Diptera) collected in CordobaArgentinardquo Memorias do Instituto Oswaldo Cruz vol 91 no 1pp 1ndash9 1996

[115] N L Wang J Wang X S Yao and S Kitanaka ldquoTwo newmonoterpene glycosides and a new (+)-jasmololone glucoside

Evidence-Based Complementary and Alternative Medicine 51

from Bidens parvifloraWilldrdquo Journal of Asian Natural ProductsResearch vol 9 no 5 pp 473ndash479 2007

[116] P Champagnat ldquoRole of the terminal bud in the actionexercised by the cotyledon of Bidens pilosus L var radiatus onits axillary budrdquo Comptes Rendus des Seances et Memoires de laSociete de Biologie vol 145 no 17-18 pp 1374ndash1376 1951

[117] S F Nielsen S B Christensen G Cruciani A Kharazmiand T Liljefors ldquoAntileishmaniai chalcones statistical designsynthesis and three-dimensional quantitative structure-activityrelationship analysisrdquo Journal of Medicinal Chemistry vol 41no 24 pp 4819ndash4832 1998

[118] Y C Hwang J J H Chu P L Yang W Chen and M V YatesldquoRapid identification of inhibitors that interfere with poliovirusreplication using a cell-based assayrdquo Antiviral Research vol 77no 3 pp 232ndash236 2008

[119] V F Andrade-Neto M G L Brandao F Q Oliveira et alldquoAntimalarial activity of Bidens pilosa L (Asteraceae) ethanolextracts fromwild plants collected in various localities or plantscultivated in humus soilrdquo Phytotherapy Research vol 18 no 8pp 634ndash639 2004

[120] R J Marles and N R Farnsworth ldquoAnti-diabetic plants andtheir active constituentsrdquo Phytomedicine vol 2 no 2 pp 137ndash189

[121] M Habeck ldquoDiabetes treatments get sweet help from naturerdquoNature Medicine vol 9 no 10 p 1228 2003

[122] H W Lin G Y Han and S X Liao ldquoStudies on the activeconstituents of the Chinese traditional medicine Polygonatumodoratum (Mill) DrucerdquoActa Pharmaceutica Sinica vol 29 no3 pp 215ndash222 1994

[123] C L T Chang Y C Chen H M Chen N S Yang andW C Yang ldquoNatural cures for type 1 diabetes a reviewof phytochemicals biological actions and clinical potentialrdquoCurrent Medicinal Chemistry vol 20 no 7 pp 899ndash907 2013

[124] L Dey A S Attele and C S Yuan ldquoAlternative therapies fortype 2 diabetesrdquo Alternative Medicine Review vol 7 no 1 pp45ndash58 2002

[125] C L T ChangH Y Liu T F Kuo et al ldquoAnti-diabetic effect andmode of action of cytopiloynerdquo Evidence-Based Complementaryand Alternative Medicine vol 2013 Article ID 685642 13 pages2013

[126] M R Khan M Kihara and A D Omoloso ldquoAnti-microbialactivity of Bidens pilosa Bischofia javanica Elmerillia papuanaand Sigesbekia orientalisrdquo Fitoterapia vol 72 no 6 pp 662ndash6652001

[127] T Dimo T B Nguelefack P Kamtchouing E Dongo ARakotonirina and S V Rakotonirina ldquoHypotensive effects ofmethanol extract from Bidens pilosa Linn on hypertensive ratsrdquoComptes Rendus de lrsquoAcademie des Sciences Serie III vol 322no 4 pp 323ndash329 1999

[128] A K Hassan O Deogratius J F Nyafuono O Francis and OP Engeu ldquoWound healing potential of the ethanolic extracts ofBidens pilosa and Ocimum suaverdquo African Journal of Pharmacyand Pharmacology vol 5 no 2 pp 132ndash136 2011

[129] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[130] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[131] C Kandaswami and EMiddleton ldquoFree radical scavenging andantioxidant activity of plant flavonoidsrdquo Advances in Experi-mental Medicine and Biology vol 366 pp 351ndash376 1994

[132] K Yamamoto H Kakegawa H Ueda et al ldquoGastric cytopro-tective anti-ulcerogenic actions of hydroxychalcones in ratsrdquoPlanta Medica vol 58 no 5 pp 389ndash393 1992

[133] J O C Ezeonwumelu A K Julius C N Muhoho et alldquoBiochemical and histological studies of aqueous extract ofBidens pilosa leaves from Ugandan Rift valley in ratsrdquo BritishJournal of Pharmacology and Toxicology vol 2 no 6 pp 302ndash309

[134] L Frida S Rakotonirina A Rakotonirina and J P SavineauldquoIn vivo and in vitro effects of Bidens pilosa L (Asteraceae)leaf aqueous and ethanol extracts on primed-oestrogenized ratuterine musclerdquo African Journal of Traditional Complementaryand Alternative Medicines vol 5 no 1 pp 79ndash91 2008

52 Evidence-Based Complementary and Alternative Medicine

Composition Comments

1 Please provide valid postal code to the address if

possible

2 We made the highlighted change in Table 4 for the sake

of correctness Please check

3 Please check the correctness of the highlighted part in

Table 5

4 We considered the highlighted part in Table 11 as

reference Please check similar cases throughout

5 Please check the format of Table 18

6 We reformatted Table 20 Please check

7 We made the highlighted change in Table 4 for the sake

of consistency Please check similar cases throughout the

paper

8 We made the highlighted change(s) for the sake

of consistency Please check simila highlighted cases

throughout

9 We made the highlighted change in Table 9 for the sake

of clarity Please check similar cases through out

10 We made the highlighted change(s) for the sake of

correctness Please check

11 Comment on ref [66] This reference is a repetition

of [18] Please check similar cases throughout

Author(s) Name(s)

Author 1

Author 2

Author 3

Evidence-Based Complementary and Alternative Medicine 3

Table 2 Nutritional facts about B pilosa courtesy of the United Nations Food and Agriculture Organization [7]

Plant(100 g)

Energy(kcal)

Moisture()

Protein(g)

Fat(g)

Carbohydrate(g)

Fiber(g)

Ash(g)

Calcium(mg)

Phosphorus(mg)

Iron(120583g)

Caroteneequivalent

(120583g)

Thiamine(mg)

Raw 43 851 38 05 84 39 22 340 67 mdash 1 800 mdashDried 33 886 28 06 6 13 2 111 39 23 mdash mdashldquomdashrdquo denotes not detectable

(a) (b)

(c)

Figure 1 B pilosa (a) and its flowers (b) and achenes (c)

and discuss their likelymechanisms of action (Table 3) Com-pelling evidence suggests that the various diverse bioactivitiesreported for B pilosa reflect its phytochemical complexity

B pilosa is an extraordinary source of phytochemicalsparticularly flavonoids and polyynes Plant flavonoids arecommonly reported to possess anticancer antiinflammatoryantioxidant and other bioactivities However the bioactivi-ties of only seven of the 60 flavonoids present in B pilosa havebeen studiedThe bioactivities of the remaining 53 flavonoidsare poorly understood and deserve further investigationOther classes of compounds found inB pilosa are also in needof further examination as shown in Table 11

3 Pharmacological Properties

As outlined in Table 3 B pilosa is traditionally used totreat a wide variety of ailments Different preparations ofits whole plant andor parts have been purported to treatover 40 categories of illnesses Scientific studies although notextensive have demonstrated that B pilosa extracts andorcompounds have antitumor [43 69 77ndash83] antiinflamma-tory [27 75 79 84ndash88] antidiabetic and antihyperglycemic[8 24 49 51 53] antioxidant [70 89 90] immunomodu-latory [50 69] antimalarial [47 77] antibacterial [47 9192] antifungal [92 93] antihypertensive vasodilatory [2894] and antiulcerative [26] activities In this section the

4 Evidence-Based Complementary and Alternative Medicine

Table 3 Ethnomedical information about B pilosa

Disorder Plant partb Mode of use Regioncountry References

Stomachache LE Decoction IndiaAfrica [17ndash19]

Colics WP Decoction AfricaChina [18]

Catarrh WP Juice or decoctiontaken orally Cuba [20]

Diarrhea LE andWP

Fresh leaves orDecoction

AfricaUganda [18 21]

Constipation WP Decoction Africa [18]DysenteryBacillarydysentery WP Decoction Africa

China [18]

Choleretic WP Decoction Middle America [18]

Anti-inflammatory WP Not stated ChinaCuba [20 22]

Asthma WPDecoction ormaceration takenorally

CubaChina [20 23]

Antirheumatic RT andWP Juice and decoction Hong Kong

Zulu Africa [18 22]

Acute appendicitis WP Decoction Hong Kong [23]

Enteritis WP Decoction AfricaChina [18]

Pruritus WP Decoction Hong Kong [23]

Conjunctivitis WP Decoction AfricaChina [18]

Otitis WP Decoction AfricaChina [18]

Pharyngitis WP Decoction AfricaChina [18]

Gastritis WP Juice taken orally Cuba [20]

Diabetes WP Decoction takenorally

CubaTaiwan [20 24]

Headache WP Decoction Bafia Cameroon [25]Diuretic WP Decoction Middle America [18]

Hypotensive WP Decoction takenorally Bafia Cameroon [25]

Colds LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21 23]

Yellow Fever LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21ndash23]

Influenza LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21 23]

Acute infectioushepatitis WP Decoction Hong Kong [23]

Intestinal worms WP Decoction Africa [18]

Malaria RT andWP Juice Africa

China [18 23]

Evidence-Based Complementary and Alternative Medicine 5

Table 3 Continued

Disorder Plant partb Mode of use Regioncountry References

Eye Infection LE andWP Fresh or juice Uganda

Middle America [18 21]

Antimicrobial APDecoction fordrinkingbathingexternal use

Trinidad andTobago [22]

Pulmonarytuberculosis WP

Decoction ormaceration takenorally

CubaChina [20 23]

Bacterial infectionsingastrointestinal tracts

WP Decoction Trinidad andTobago [22]

Renal infection LE Decoction takenorally Cuba [20]

Sore throat LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21 23]

Cough WP Decoction takenorally

CubaChina [20 23]

Coolness of theuterus WP Decoction taken

orally Cuba [20]

Menstrualirregularities WP Decoction taken

orally Cuba [20]

Dysmenorrhea WP Decoction Bafia Cameroon [25]Hyperemesisgravidarum(morning sickness)

WP Decoction Africa [18]

Hemorrhoids WP Decoction Hong Kong [23]

Nose bleeds LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21 23]

Stomach ulcers LE andWP

Maceration or juicetaken orally

CubaMiddle AmericaUganda

[18 20 21 23]

Cuts burns and skinproblems

LE andWP

Fresh plant ordecoction topicalapplicationbathing

Trinidad andTobagoAfricaChinaCameroonBrazilVenezuela

[18 22 26]

Wounds WP Crushed herb

ChinaAfricaCentral AmericaHawaii

[23]

Snake bites WP Pulverized herb China [23]bLE leaves ST stem FW flower WP whole plant RT roots AP aerial parts

primary pharmacological properties of B pilosa extracts andphytochemicals are presented and discussed

31 AntiCancer Activity Folkloric reports revealed the pos-sible antitumor efficacy of B pilosa and several scientific in2vitro studies have supported the claim that B pilosa extractsand isolated compounds possess anti-cancer activities againsta variety of cancer cells Several studies have used bioassay

guided isolation and fractionation methods to discover newcompounds from B pilosa For example Kviecinski andcolleagues tested hydroalcoholic crude extracts chloroformethyl acetate and methanol fractions for anti-tumor activity[82] The cytotoxicity of the extracts was assessed usingbrine shrimp hemolytic MTT and neutral red uptake(NRU) assays In vivo studies were performed using Ehrlichascites carcinoma in isogenic BALBc mice Among them

6 Evidence-Based Complementary and Alternative Medicine

Table 4 Aliphatic natural products isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

1 Heneicosane CH3(CH2)19CH3 Aerial (Tanzania) [18]2 Docosane CH3(CH2)20CH3 Aerial (Tanzania) [18]3 Tricosane CH3(CH2)21CH3 Aerial (Tanzania) [18]4 Tetracosane CH3(CH2)22CH3 Aerial (Tanzania) [18]5 Pentacosane CH3(CH2)23CH3 Aerial (Tanzania) [18]6 Hexacosane CH3(CH2)24CH3 Aerial (Tanzania) [18]7 Heptacosane CH3(CH2)25CH3 Aerial (Tanzania) [18]

8 Octacosone CH3(CH2)26CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

9 Nonacosane CH3(CH2)27CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

10 Triacontane CH3(CH2)28CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

11 Hentriacontane CH3(CH2)29CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

12 Dotriacontane CH3(CH2)30CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

13 Tritriacontane CH3(CH2)31CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

14 2-Butoxy-ethanol CH3(CH2)3OCH2CH2OH Whole (Taiwan) [32]15 Tetracosan-1-ol CH3(CH2)22CH2OH Aerial (Tanzania) [18]16 Hexacosan-1-ol CH3(CH2)24CH2OH Aerial (Tanzania) [18]17 1-Octacosanol CH3(CH2)26CH2OH Aerial (Tanzania) [18]

18 1-Hentriacontanol CH3(CH2)29CH2OHNot found(Taiwan) [31]

19 Tetradecanoic acid Myristic acid CH3(CH2)12CO2H Aerial (Tanzania) [18]20 Hexadecanoic acid Palmitic acid CH3(CH2)14CO2H Aerial (Tanzania) [18]21 Octadecanoic acid Stearic acid CH3(CH2)16CO2H Aerial (Tanzania) [18]22 Eicosanoic acid Arachidic acid CH3(CH2)18CO2H Aerial (Tanzania) [18]

23 Docosanoic acid Behenic acid CH3(CH2)20CO2HLeaves(Philippines) [33]

24 2-butenedioic acidO

OHO

HO Aerial (China) [34 35]

25 (Z)-9-Octadecenoicacid Oleic acid

O

OH Aerial (Tanzania) [18]

Evidence-Based Complementary and Alternative Medicine 7

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

26 (E)-9-Octadecenoicacid Elaidic acid

O

HOLeaves(Philippines) [33]

27 (ZZ)-912-Octadecadienoic acid

Linolic acid or Linoleicacid

O

OHAerial(Tanzania)whole (Taiwan)

[18 36]

28(ZZZ)-91215-Octadecatrienoicacid

120572-Linoleic acid

O

OH Whole (Taiwan) [32]

29(ZZ)-912-Octadecadienoic acidethyl ester

Ethyl linoleate O

O

Whole (Taiwan) [32]

30(ZZZ)-91215-Octadecatrienoicacid methyl ester

Methyl linolenate O

O

Whole (Taiwan) [32]

31(ZZZ)-91215-Octadecatrienoicacid ethyl ester

Ethyl linolenate O

O

Whole (Taiwan) [32]

32(Z)-9-Octadecenoicacid 2-butoxyethylester

2-Butoxyethyl oleate O

OO

Whole (Taiwan) [32]

33 2-Butoxyethyllinoleate O

OO Whole (Taiwan) [32]

34(ZZZ)-91215-Octadecatrienoicacid butoxyethyl ester

2-Butoxyethyl linoleate O

OO

Whole (Taiwan) [32]

3517E9E15E-Heptadecatetraene-1113-diyne

Heptadeca-2E8E10E16-tetraen-46-diyne

Not found(China) [37]

8 Evidence-Based Complementary and Alternative Medicine

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

36 111-Tridecadiene-3579-tetrayne

Roots (notstated) [38]

37 1-Tridecene-357911-pentayne Pentayneene

Leaves (notstated) andnot found(Egypt)

[38 39]

38 5-Tridecene-7911-triyne-3-ol

HO

not found(Egypt) [39]

39 21012-Tridecatriene-468-triyn-1-ol

OHPart not specified(Not stated) [40]

40 212-Tridecadiene-46810-tetrayn-1-ol

111-Tridecadiene-3579-

tetrayn-13-ol

OH Roots (Notstated) notfound(Egypt)

[38 39]

41 212-Tridecadiene-46810-tetraynal

111-Tridecadiene-3579-

tetrayn-13-al

O

Roots (Germany) [41]

42212-Tridecadiene-46810-tetrayn-1-ol1-acetate

111-Tridecadiene-3579-

tetrayn-13-acetateO

O

Roots (notstated) [38]

43(5E)-15-Tridecadiene-79-diyn-3413-triol OH

OH

HO

Aerial (China) [42]

44(6E12E)-3-oxo-tetradeca-612-dien-810-diyn-1-ol

OH

O

Aerial (China) [42]

45 (E)-5-Tridecene-7911-triyne-12-diol

12-Dihydroxy-5(E)-tridecene-7911-triyne

HO

OHWhole (Taiwan) [43]

46 (E)-6-Tetradecene-81012-triyne-13-diol

13-Dihydroxy-6(E)-tetradecene-81012-triyne

OH

OHWhole (Taiwan) [43ndash45]

47(2R3E11E)-311-Tridecadiene-579-triyne-12-diol

Safynol

OH

HO

Not found (Egyptand China) [37 39]

Evidence-Based Complementary and Alternative Medicine 9

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

4857911-Tridecatetrayne-12-diol

12-Dihydroxytrideca-57911-tetrayne

HOOH

Whole (Taiwan) [43 44 46]

49(R)-357911-Tridecapentayne-12-diol

(R)-12-Dihydroxytrideca-357911-pentayne

HO

OH

Aerial (Japan) [47]

50

(4E)-1-(Hydroxymethyl)-4-dodecene-6810-triyn-1-yl-120573-D-glucopyranoside

2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-

tridecene-7911-triyne

O

HO

H

OH

H

H

HOH

O

OH

HO

H Aerial (USA)whole (Taiwan)leaves (Taiwan)

[8 45 48ndash51]

51

(4E)-1-(2-Hydroxyethyl)-4-dodecene-6810-triyn-1-yl-120573-D-glucopyranoside

3-120573-D-Glucopyranosyloxy-1-hydroxy-6(E)-

tridecene-81012-triyne

OHO

H

OH

H

H

HOH

O

OH

OH

HAerial (USA andChina) whole(Taiwan) leaves(Taiwan)

[8 35 4245 48ndash51]

52

3-Hydroxy-6-tetradecene-81012-triynyl-120573-D-glucopyranoside

120573-D-Glucopyranosyloxy-

3-hydroxy-6E-tetradecence-81012-triyne

OH

O

OHO

H

OH

H

H

HOH

OH

HWhole (Mexico) [52]

53

1-(Hydroxymethyl)-46810-dodecatetrayne-1-yl-120573-D-glucopyranoside

Cytopiloyne2-120573-D-

Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne

O

HO

OHO

H

OH

H

H

HOH

OH

HWhole (Taiwan)leaves (Taiwan) [8 50 53]

54

2-O-D-Glucosyltrideca-11E-en-3579-tetrayn-12-diol

OH

O

O

OHH

HO

H

H

HHOH

OH

Leaves (Brazil) [27]

10 Evidence-Based Complementary and Alternative Medicine

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

55

(R)-1-(Hydroxymethyl)-246810-dodecapentayn-1-yl-a-D-glucopyranoside

2- 120573 -D-Glucopyranosyloxy-1-hydroxytrideca-357911-pentayne

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Chinaand Japan) [35 47]

56

1-[[(Carboxyacetyl)oxy]methyl]46810-dodecatetraynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Japan) [54]

57

(4E)-1-[[(Carboxyacetyl)oxy]-methyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHO

H

OH

Aerial (Japan) [54]

58

(4E)-1-[[(Carboxyacetyl)oxy]-ethyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

O

O

O

OHO

O

OHOH

H

HO

H

H

HHO

HOH

Aerial (Japan) [54]

59 (5E)-5-Heptene-13-diyn-1-yl-benzene

1-Phenylhepta-13-diyn-5en Whole (Taiwan) [32]

60 7-Phenyl-2(E)-heptene-46-diyn-1-ol

OHRoots (notstated) Aerial(China)

[38 42]

617-Phenyl-2(E)-heptene-46-diyn-1-ol-acetate

OO

Roots (notstated Brazil) [38 55 56]

62 7-Phenyl-46-heptadiyn-2-ol Pilosol A

OH

Whole (Taiwan)aerial (China) [42 57]

Evidence-Based Complementary and Alternative Medicine 11

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

63 7-Phenylhepta-46-diyn-12-diol HO

OH

Aerial (China) [42]

64 135-Heptatriyn-1-yl-benzene

1-Phenylhepta-135-triyne

Leaves (notstated) leaves oftissue culture(not stated)aerial (TanzaniaChina)Whole (Taiwan)roots (Brazil)

[18 34 3842 56ndash58]

65 7-Phenyl-246-heptatriyn-1-ol HO

Leaves (notstated) aerial(China)

[38 42]

667-Phenyl-246-heptatriyn-1-ol-acetate O

O Leaves (notstated) [38]

675-(2-Phenylethynyl)-2-thiophenemethanol

S OHAerial (China) [42]

685-(2-Phenylethynyl)-2120573-glucosylmethyl-thiophene

O

O

OH

H

OH

H

H

HOH H

O

HO

S

Aerial (China) [42]

69

3- 120573 -D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne

OH

O

OH

HOHO

H

H

HHOH

OH

Leaves(Cameroon) [28]

701-Phenyl-13-diyn-5-en-7-ol-acetate O

O

Leaves (Brazil) [27]

SN denotes serial number

the chloroform fraction was the most toxic with a halfmaximal inhibitory concentration (IC

50) of 97 plusmn 72 and 83 plusmn

52 120583gmL in NRU and MTT respectively [82] Kumari andcolleagues also reported the anti-cancer and anti-malarialactivities of B pilosa leaves [77] Based on a cytotoxicity-directed fractionation strategy they identified phenyl-135-heptatriene with IC

50values of 8plusmn001 049plusmn045 07plusmn001

and 10 plusmn 001 120583gmL against human oral liver colon andbreast cancer cell lines respectively However phenyl-135-heptatriyne showed lower activity against breast cancer celllines than the chloroform leaf extract which had an IC

50

value of 65 plusmn 001 120583gmL Moreover the positive controltaxol showed higher activity than phenyl-135-heptatriyne[77] Furthermore in vitro comet assays were performed toevaluate the toxicity of n-hexane chloroform and methanolextracts of B pilosa and its ethyl acetate acetone and waterfractions onHela andKB cellsThe ethyl acetate fraction fromthe methanol extract exhibited the highest activity with halfmaximal cytotoxic concentrations (CTC

50) of 9652120583gmL

and 5862120583gmL against Hela and KB cells respectivelyDespite the moderate toxicity these findings suggest thatthese B pilosa extractsfractions could be useful for future

12 Evidence-Based Complementary and Alternative Medicine

Table5Flavon

oids

isolated

from

Bpilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

71

2-[(34-Dihydroxyph

enyl)-

methylene]-

6-hydroxy-3(2H

)-benzofuranon

e

Sulfu

retin

O

OHO

OH

OH

Aeria

l(Ch

ina)

[35]

72

2-[(34-Dihydroxyph

enyl)-

methylene]-

67-dihydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-673101584041015840-

tetrahydroxy

maritimetin

O

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

73

2-[(34-Dihydroxyph

enyl)-

methylene]-6-

(120573-D

-glucopyrano

syloxy)-7-

hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-120573-D

-glucop

yranosyl-

673101584041015840-te

trahydroxy

maritimein

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japanand

China)aerial(Ch

ina)

[355960]

74

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

(120573-D

-glucopyrano

syloxy)-

6-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-7-O-120573-D

-glucop

yranosyl-

673101584041015840-

tetrahydroxy

O

OHO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japan)

[59]

75

6-[(6-O-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-acetyl-

120573-D

-glucopyrano

syl)

-673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OHO

H HO

H

H

HOH

HO

O

O

Leaves

(Not

stated)

aeria

l(Ch

ina)

[3559]

Evidence-Based Complementary and Alternative Medicine 13

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

76

6-[(36-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-

2-[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone(Z)-6-O

-(36-di-O

-acetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

bideno

sideA

O

O

HO

OH

OH

OHOH O

H

H

HOH

HO

O

O

O

Leaves

(China)

[60]

77

6-[(46-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(410158401015840610158401015840-

diacetyl-120573-D

-glucropyrano

syl)-

673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

HO

O

OO

Leaves

(not

stated)

aeria

l(Ch

ina)

[343561]

78

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

hydroxy-6-

[(246-tri-O

-acetyl-120573

-D-

glucop

yranosyl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(210158401015840410158401015840610158401015840-

triacetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

O

OOO

O

Leaves

(not

stated)

aeria

l(Ch

ina)

[3461]

79

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-6-

[(346-tri-O

-acetyl-120573

-D-glucopyrano

syl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(310158401015840410158401015840610158401015840-

triacetyl-120573

-D-glucrop

yranosyl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H OH

H

HOH

HO

OOO O

Aeria

l(Ch

ina)

[3435]

80

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-

6-[[6-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propenyl]-120573-

D-glucopyrano

syl]o

xy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-

p-coum

aroyl-

120573-D

-glucropyrano

syl)-

673101584041015840-te

trahydroxy

OH

O

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OLeaves

(Japan)

[59]

14 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

81

1-[2-(120573-D

-Glucopyrano

syloxy)-

4-hydroxyphenyl]-2-

hydroxy-3-(3-

hydroxyphenyl)-2-

prop

en-1-on

e

Chalcone120572

32101584041015840-

tetrahydroxy-

21015840-O

-120573-D

-glucop

yranosyl

HO

O

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[35]

821-(24-Dihydroxyph

enyl)-

3-(34-dihydroxyph

enyl)-

2-prop

en-1-on

eBu

tein

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

833-(34-D

ihydroxyph

enyl)-

1-(234-trihydroxyph

enyl)-

2-prop

en-1-on

eOkanin

O

HO

HO

OH

OH

OH

Leaves

(China)

[60]

84

3-(34-D

ihydroxyph

enyl)-

1-[3-(120573-D

-glucop

yranosyloxy)-

24-dihydroxyphenyl]-2-

prop

en-1-on

e

Okanin

31015840-O

-120573-D

-glucosid

e

O

HO

OH

OH

OH

OO

HO

H

HO

H

H

HOH

H

OH

Leaves

(Germany)

flowers(Germany)

[61ndash63]

85

3-(34-D

ihydroxyph

enyl)-

1-[4-(120573-D

-glucop

yranosyloxy)-

23-dihydroxyphenyl]-

2-prop

en-1-on

e

Okanin

41015840-O

-120573-D

-glucosid

e

OOH

OH

OH

HOO

OHO

H

HO

H

H

HOH

H

OH

Flow

ers(Germany)

leaves

(Japan)

[5963]

Evidence-Based Complementary and Alternative Medicine 15

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

86Okanin41015840-O

-120573-D

-(610158401015840-

O-acetylglucosid

e)

O

O

OH

OH

OH

OHO

H

HO

H

H

HOH

H

HO

O

O

Flow

ers(Germany)

[63]

87

1-[4-[(46-di-O

-Acetyl-

120573-D

-glucopyrano

syl)-oxy]-

23-dihydroxyphenyl]-3-

(34-dihydroxyph

enyl)-2-

prop

en-1-on

e

Okanin

41015840-O

-120573-D

-(410158401015840610158401015840-diacetyl)-

glucop

yranoside

O

O

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Aeria

l(Ch

ina)

[34]

88Okanin41015840-O

-120573-D

-(210158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[64]

89Okanin41015840-O

-120573-D

-(310158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

O

OAe

rial(Ch

ina)

[34]

16 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

90

1-[23-Dihydroxy-4-

[[6-O-[3-(4-hydroxyph

enyl)-

1-oxo-2-prop

enyl]-120573-D

-glucop

yranosyl]oxy]-ph

enyl]-

3-(34-dihydroxyph

enyl)-

2-prop

en-1on

e

Okanin

41015840-O

-120573-D

-(610158401015840-tr

ans-

p-coum

aroyl)-

glucoside

O

O

OH

OH

OH

OHOH

HO

H

H

HOH

H

HO

OO

OH

Leaves

(Germany)

[64]

91Okanin41015840-O

-120573-D

-(410158401015840-acetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Leaves

(Germany)

[65]

92

Okanin

41015840-O

-120573-D

-(210158401015840410158401015840-diacetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[65]

Evidence-Based Complementary and Alternative Medicine 17

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

93

Okanin

41015840-O

-120573-D

-(310158401015840410158401015840-diacetyl-610158401015840-

trans-p-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

OH

O O

Leaves

(Germany)

[65]

94

Okanin

41015840-O

-[120573-D

-glucopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

OOH

OH

OH

HO

OH

HO

HHO

H

HOH

HO

OH

OH

HO

HHO

H

HOH

HO

Flow

er(G

ermany)

[63]

95Okanin

3101584041015840-di-O

-120573-D

-glucosid

e

OOH

OH

OH

O OO

H HO

HHOH

HOH

HHO

OHO

H

HO

H

H

HOH

H

OH

Flow

er(G

ermany)

[63]

18 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

96

1-[3-(120573-D

-Glucopyrano

syloxy)-

24-dihydroxyphenyl]-

3-(3-hydroxy)-

4-metho

xyph

enyl)-

2-prop

en-1-on

e

Okanin

4-methylether-

31015840-O

-120573-D

-glucop

yranoside

OOH

O

OH

HO

O

H HO

HHOH

HOH

H

OHO

Leaves

(Germany)aerial

(China)

[3562]

97

Okanin4-methylether-

3101584041015840-di-O

-120573-(41015840101584061015840101584041015840101584010158406101584010158401015840-

tetraacetyl)-

glucop

yranoside

OOH

OOH

O

HH

OH

HO

HOH

H

OO

O

Ac

Ac

AcAc

OO

H

HO

H

H

HOH

H

O

Aeria

l(Ch

ina)

[42]

98Ch

alcone210158404101584061015840-tr

imetho

xy-

4-O-D

-glucopyrano

syl-

dihydro

Not

foun

dLeaves

(China)

[60]

992-(34-D

ihydroxyph

enyl)-23-

dihydro-78

-dihydroxy-

4H-1-

benzop

yran-4-one

Okanin

isoOO

OH

HO

OH

OH

Leaves

(China)

[60]

100

2-(34-D

ihydroxyph

enyl)-

23-dihydro-8-hydroxy-

7[(246-tr

i-O-acetyl-120573

-D-

glucop

yranosyl)oxy]-4H

-1-b

enzopyran-4-on

eOO

OH

OH

OH

OH

O

HHO

H

HO

HO

OAc

Ac

Ac

Aeria

l(Ch

ina)

[34]

Evidence-Based Complementary and Alternative Medicine 19

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

101

57-Dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

OO

OH

HO

OH

Aeria

l(TanzaniaC

hina)

[4266]

102

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

7-O-glucopyrano

side

OO

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Tanzania)

[66]

103

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-4H

-1-benzop

yran-4-one

Luteolin

OO

HO

OH

OH

OH

Aeria

l(TanzaniaC

hina

Vietnam)

[343542

6667]

104

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5hydroxy-4H-1-

benzop

yran-4-one

Luteolin

7-O-120573-D

-glucop

yranoside

OOOH

OH

OH

OO

HO

H

HO

H

H

HOH

OH H

Aeria

l(Tanzania)

[66]

105

57-Dim

etho

xy-6-

(5-m

etho

xy-6-m

ethyl-

4-oxo-4H

-pyran-3-yl)-

2-ph

enyl-4H-1-

benzop

yran-4-one

5-O-M

ethylhosludin

O OOO

O

O

O

Aeria

l(Ugand

a)[68]

20 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

106

3-(120573-D

-Glucopyrano

syloxy)-

57-dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Astragalin

Kaem

pferol-3-O

-120573-D

-glucopyrano

side

HO

OH

O O

OH

O

O

HO

HHO

H

H

HOH

OH

H

Aeria

l(Ch

ina)

[35]

107

Kaem

pferol3-

(23-di-E

-p-cou

maroyl-

120572-L-rhamno

pyrano

side)

Not

foun

dAe

rial(Vietnam)

[67]

108

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5-hydroxy-36-dimetho

xy-

4H-1-

benzop

yran-4-one

Axillarosid

eO O

OH

OH

OO

OH

OO

HO

H

HO

H

H

HOH

H

OH

Aeria

l(Ch

ina)

[42]

109

57-Dihydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaureidin

O

O

O

O

O

HO

OH

OH

Who

le(Taiwan)

[69]

110

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaurein

O O

OO

O OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan)w

hole

(Taiwan)

[69ndash

71]

Evidence-Based Complementary and Alternative Medicine 21

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

111

5-Hydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-

67-dimetho

xy-4H-

chromen-4-one

Eupatorin

iso

OO O

OH

O

O OH

Not

foun

d(C

hina)

[3772]

112

2-(34-D

imetho

xyph

enyl)-7-

(120573-D

-glucopyrano

syloxy)-35-

dihydroxy-

8-metho

xy-4H-1-

benzop

yran-4-one

O O

OH

O O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

113

7-(120573-D

-Glucopyrano

syloxy)-

5hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-38-

dimehtoxy-4H-1-

benzop

yran-4-one

O O

O

OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

114

Isorhamnetin

3-[O

-120572-

L-rham

nopyrano

syl-

(1-2)-120573-D

-glucop

yranoside]

Not

foun

dAe

rial(Vietnam)

[67]

115

7-[(6-deoxy-120572-L-

Manno

pyrano

syl)o

xy]-

3-120573-D

-glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-4H

-1-benzop

yran-4-one

Luteoside

O O

OH

O

OH

OH

OH

HOH

H

H HO

HO

HO

OH

HO

H

HO

OH

OH

HH

Aeria

l(Ch

ina)

[42]

22 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

116

Luteolin

3-O-120573-D

-Glucopyrano

side

O O

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

HO

Aeria

l(Tanzania)

[66]

117

57-Dihydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Quercetagetin

3631015840-tr

imethylether

OOO

O

O OH

HO

OH

Aeria

l(Ch

ina)

[42]

118

6-((2S3S4S5S)-

Tetrahydro-345-tr

ihydroxy-

6-(hydroxymethyl)-2H

-pyran-2-yloxy)-5-hydroxy-

2-(4-hydroxy-3-

metho

xyph

enyl)-

37-dimetho

xy-4H-

chromen-4-one

Quercetagetin

3731015840-

trim

ethylether-6-O

-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[42]

119

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

JaceinQ

uercetin

3631015840-

trim

ethyl

ether-7-O-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan

China)

who

le(Taiwan)

[425470]

120

2-(34-D

ihydroxyph

enyl)-

357-trihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

OO

HO

OH

OH

OH

OH

Aeria

l(Ch

ina)leaves

(China)who

le(C

hina)

[356074]

Evidence-Based Complementary and Alternative Medicine 23

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

121

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-galactopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-4-one

Quercetin

3-O-120573-D

-galactosidehyperin

hyperosid

e

O O

HO

OH

OH

OH

OOHH

HOH

H

H HO

HO

OH

Aeria

l(TanzaniaJapan)

Leaves

(China)Who

le(C

hina)

[376066

717475]

122

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-glucopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

3-O-120573-

D-glucopyrano

side

O O

HO

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

Aeria

l(TanzaniaC

hina

Japan)L

eaves(Japan)

[355966

75]

123

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-

4-oxo-4H

-1-benzop

yran-3-yl-

120573-D

-glucopyrano

siduron

icacid

Quercetin

3-O-120573-D

-glucuron

opyranoside

O O

HO

OH

OH

OH

OHOH

HOH

H

H HO

HO

OHO

Aeria

l(TanzaniaJapan)

[6671]

124

3-[[6-O-(6-

Deoxy-120572-L-m

anno

pyrano

syl)-

120573-D

-galactopyrano

syl]o

xy]-

2-(34-dihydroxyph

enyl)-57-

dihydroxy-4H

-1-benzop

yran-4-one

Quercetin

3-O-rob

inob

iosid

e

O O

HO

OH

OH

OH

O

HOHH OH

HO

H HH

OO

OHH

HOH

H

H HO

HO

Aeria

l(Japan)W

hole

(Taiwan)

[7071]

24 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

125

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-

3-metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

OH

Roots(Brazil)

[55676]

126

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-

120573-D

-glucopyrano

syl]o

xy]-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120572-

L-rham

nopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

O

HHO HHO

HO H

HH

ORo

ots(Brazil)

[5676]

127

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-120573-D

-glucop

yranosyl]oxy]-5-

hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-3-metho

xy-

4H-1-

benzop

yran-4-one

Quercetin

341015840-

dimethyl

ether-7-O-rutinoside

O OOH

OH

O

O

OH O

HOH

H

HHO

H

O

HHO HHO

HO H

HH

O

HO

Aeria

l(Ch

ina)

[3435]

128

2-(34-D

ihydroxyph

enyl)-3-

(120573-D

-glucofurano

syloxy)-57-

dihydroxy-4H

-1-benzop

yran-

4-on

e

Isoq

uercitrin

O OOH

OH

OHO

H

OHH

OH

HO

H O HOH

OH

Aeria

l(Japan

China)

[3571]

Evidence-Based Complementary and Alternative Medicine 25

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

129

Quercetin

3-O-120573-D

-galactop

yranoside

O

OH

HO

OH OH

O

O

HO

OH

OHOH

OAe

rial(no

tstated)

[18]

130

Quercetin

3-O-rutinoside

O

O

R

OO

OH

OHO

H

O

OH

OH

H3C

H3C

H3C

OH

HO

Who

le(Taiwan)

[70]

26 Evidence-Based Complementary and Alternative Medicine

Table 6 Terpenoids isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(Country) References

131371111-Tetramethylbicyclo[810]undeca-

26-dieneBicyclogermacrene Leaves

(Brazil) [95]

13241111-Trimethyl-8-

methylenebicyclo[720]undec-4-ene

E-Caryophyllene Leaves(Brazil) [95]

1331-Methyl-5-methylene-8-

(1-methylethyl)-16-cyclodecadiene

Germacrene D Leaves(Brazil) [95]

1344-(15-Dimethyl-4-hexen-1-ylidene)-1-methyl-cyclohexene

Z-120574-Bisabolene Leaves(Brazil) [95]

135Decahydro-114-trimethyl-7-methylene-1H-cycloprop[e]-

azulene120573-Gurjunene Leaves

(Brazil) [95]

136 2669-Tetramethyl-148-cycloundecatriene

120572-Humulene120572-caryophyllene

Leaves(Brazil) [95]

13712344a568a-Octahydro-1-isopropyl-7-methyl-4-methylenenaphthalene

120575-Muurolene Leaves(Brazil) [95]

138

12344a568a-Octahydro-4a8-dimethyl-2-(1-methylethylidene)-

naphthalene

Selina-37(11)-diene

Leaves(Brazil) [95]

139(2E7R11R)-371115-

Tetramethyl-2-hexadecen-1-ol

Phytol

HO

Whole(Taiwan) [57]

Evidence-Based Complementary and Alternative Medicine 27

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

140 371115-Tetramethyl-2-hexadecenoic acid Phytanic acid

O

OH

Whole(Taiwan) [57]

141371115-Tetramethyl-2-

hexadecenyl ester-heptanoicacid

Phythylheptanoate O O

Leaves(Not stated) [33]

142

(3S10R13R)-17-((2R5R)-5-Ethyl-6-methylheptan-2-yl)-

2347891011121314151617-tetradecahydro-

1013-dimethyl-1H-cyclopenta[a]phenanthren-

3-ol

Campestrol

HO

Aerial(Tanzania) [18]

143 Not found Phytosterin-B Not foundNot found(TaiwanEgypt)

[39 96]

144 Stigmast-5-en-3-ol 120573-sitosterol

HOH

H

H

Aerial(Tanzania)

whole(Taiwan)

[18 31 57]

145

1314151617-Tetradecahydro-1013-

dimethyl-1H-cyclopenta[a]phenanthren-

3-yloxy)-tetrahydro-6-(hydroxymethyl)-2H-pyran-345-triol

120573-Sitosterolglucoside

H

H

HOH

HO

HHO

H

HOHH O

OH Not found(Egypt) [39]

146 5120572-Stigmasta-7-en-3120573-ol

HO

Whole(Taiwan) [57]

147 5120572-Stigmasta-722t-dien-3120573-ol

HO

Whole(Taiwan) [57]

148 Stigmasta-522-dien-3-ol Stigmasterol

HOH

H

H

Not found(Taiwan)aerial

(Tanzania)leaves (notstated)whole

(Taiwan)

[18 31 3357]

28 Evidence-Based Complementary and Alternative Medicine

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

149 Lup-20(29)-en-3-ol Lupeol

H

HH

HOH

Not found(Egypt) [39]

150 Lup-20(29)-en-3-ol acetate Lupeol acetate

H

HH

O

OH

Not found(Egypt) [39]

151 Olean-12-en-3-ol 120573-amyrin

H

HOH

Not found(Egypt) [39]

152 5913-Trimethyl-242526-trinoroleanan-3-ol Friedelan-3120573-ol

HO

Aerial(Tanzania) [18]

153 5913-Trimethyl-242526-trinoroleanan-3-one

Friedelinfriedelan-3-one

O

Aerial(Tanzania) [18]

1542610151923-

Hexamethyl-2610141822-tetracosahexaene

Squalene

Aerial(Tanzania)Leaves (Notstated)Whole(Taiwan)

[18 33 57]

155 120573120573-Carotene 120573-Carotene Leaves (Notstated) [97]

Evidence-Based Complementary and Alternative Medicine 29

studies [83] Hot water extracts of B pilosa varminor Sheriffwere also assessed for its antileukemic effects on leukemiccell lines L1210 U937 K562 Raji and P3HR1 using XTT-based colorimetric assays The extract inhibited the five celllines with IC

50values ranging from 145120583gmL to 586120583gmL

L1210 K562 Raji and P3HR1 were more sensitive to B pilosaextract with IC

50values below 200120583gmL [98]

Consistent with the antitumor activities of B pilosaextracts and fractions some of its phytochemicals alsoshowed anticancer activity as outlined in Table 11 Amongthem luteolin (103) a well-studied flavonoid with multiplebioactivities was more effective against tumor cell prolifera-tion than its derivatives with IC

50values ranging from 3 120583M

to 50120583M in cells and 5 to 10mgkg in animals Luteolin wasalso found to fight cancer as a food additive at concentrationsof 50 to 200 ppm [78] and prevent skin cancer [78] and cancer3invasion [103] Lee and colleagues reported that luteolinprevents cancer by inhibiting cell adhesion and invasion[103] Significant inhibition concentration was reported to be5 120583M and complete inhibition concentration was reported tobe 40120583M Moreover luteolin was reported to inhibit hepa-tocyte growth factor (HGF)-induced cell scattering as wellas cytoskeleton changes such as filopodia and lamellipodiawhichwas determined using phase-contrast and fluorescencemicroscopy Furthermore luteolin also inhibited the HGF-induced phosphorylation of c-Met ERK 12 and Akt aswell as the MAPKERK and P13K-Akt pathways [78 103]Other mechanisms underlying the anticancer activities ofluteolin are the inhibition of topoisomerase I and II whichinhibits cell replication and DNA repair thus promotingapoptosis regulation of PI-3-KinaseAktMAPKERKJNKactivation of apoptosis in the mitochondrial pathway byactivating caspase 9 and caspase 3 which were found inmalignant cells but not in normal human peripheral bloodmononuclear cells death receptor-induced apoptosis and acell-cycle arrest mechanism inhibition of fatty acid synthasewhich is upregulated in many cancer cells and sensitizationto chemotherapywhereby luteolin increases the susceptibilityof cancer cells to chemotherapy [78] Butein (82) is anotherflavonoid that showed a cytotoxic effect on human colonadenocarcinoma cell proliferation with a reported IC

50value

of 175 120583M Butein at 2 120583M affected the incorporation of[14C]-labeled leucine thymidine and uridine which cancause the inhibition of DNA RNA and protein synthesis ofhuman colon cancer cells Moreover butein also exhibitednoncompetitive inhibition of 1-chloro-24-dinitrobenzene(CDNB) in glutathione S-transferase (GST) activity Tumorresistance was correlated with high levels of GST thus buteininhibited proliferation of cancer cells [80] Another flavonoidpresent in B pilosa centaureidin (109) also showed anti-cancer activity in B lymphoma cells Centaureidin isolatedfrom Polymnia fruticosa inhibited tubulin polymerization invitro and induced mitotic figure formation in CA46 Burkittlymphoma cells Using turbidimetric assay the IC

50value of

centaureidin in inhibition of mitosis was 3120583M [104] Cyto-toxicity of centaureidin was further analyzed using AmericanNational Cancer Institute (NCI) 60 human tumor cell linesThe cytotoxicity potency of centaureidin expressed as GI

50

(50 growth inhibition in the NCI tumor line panel) was024120583M [81] These data mark centaureidin as a promisingantimitotic agent for tumor therapy

In addition to anti-tumor flavones polyynes found in B 4pilosa have also been shown to possess anti-tumor proper-ties Based on a bioactivity-directed isolation approach Wuand colleagues identified two polyyne aglycones from theethyl acetate fraction of B pilosa [44] 12-Dihydroxytrideca-57911-tetrayne (48) and 13-Dihydroxy-6(E)- tetradecene-81012-triyne (46) exhibited significant anticell prolifera-tion activity in primary human umbilical vein endotheliumcells (HUVEC) with IC

50values of 125 120583M and 173 120583M

respectively They also decreased angiogenesis and pro-moted apoptosis in human endothelial cells Their anti-angiogenic and cytotoxic effects correlated with activationof the CDK inhibitors and caspase-7 [44] In addition 12-Dihyroxy-5(E)-tridecene-7911-triyne (45) showed antian-giogenic effects in HUVECs with an IC

50value of 124 120583M as

evidenced by a decrease in the tube formation and migrationof HUVECs [43] The IC

50value of compound 45 in the

inhibition of basic fibroblast growth factor-induced HUVECgrowth was 282120583M However it had higher IC

50values

than those for lung carcinoma cells and keratinocytes Thiscompound could also inhibit cell proliferation of HUVECslung carcinoma A549 cells and HACAT keratinocytes Themechanism by which compound 45 inhibits HUVEC growthand angiogenesis is complicated and includes decreasing theexpression of cell cycle regulators (CDK4 cyclins D1 and Aretinoblastoma (Rb) and vascular endothelial growth factorreceptor 1) caspase-mediated activation of CDK inhibitorsp21 (Cip1) and p27 (Kip) upregulation of Fas ligand expres-sion downregulation of Bcl-2 expression and activation ofcaspase-7 and poly (ADP-ribose) polymerase [43]

32 Anti-Inflammatory Activity B pilosa is commonly usedto treat inflammatory disorders The anti-inflammatory phy-tochemicals present in B pilosa are listed in Table 11 5Cyclooxygenase-2 (COX-2) is a physiologically importantenzyme that converts arachidonic acid to prostaglandin(PGE2) Its expression is induced by a wide variety of external

stimuli indicating its involvement in inflammatory diseasesand it is used as an inflammatory marker [88] Yoshida andcolleagues studied the effects of the aqueous extracts of Bpilosa aerial parts in the production of COX-2 and PGE

2as

well as on the activation of mitogen activated protein kinases(MAPKs) in normal human dermal fibroblasts (HDFs) inresponse to inflammatory cytokine IL-1120573 This work showedthat IL-1120573 activated MAPKs such as ERK12 p38 and JNKto different extents and induced COX-2 expression TheCOX-2 expression in HDFs was regulated mainly by p38following IL-1120573 stimulation Consistently the p38 inhibitorSB203580 blocked this expression Using this cell platformB pilosa extracts were tested for inhibition of inflammationThe extract dose-dependently suppressed the activation ofp38 and JNK and moderately suppressed ERK12 as wellas suppressing COX-2 expression and PGE

2production

30 Evidence-Based Complementary and Alternative Medicine

Table7Ph

enylprop

anoids

isolatedfro

mB

pilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

156

3-(4-H

ydroxyph

enyl)-2-prop

enoica

cid

p-Cou

maricacid

OHO

OH

Who

le(Ja

pan)

[99]

157

2-Metho

xy-4-(2-prop

en-1-yl)-ph

enol

Eugeno

lOOH

Leaves

androots(Japan)

[99]

158

3-(4-H

ydroxy-3-m

etho

xyph

enyl)-

2-prop

enoica

cid

Ferulic

acid

O

OH

O

HO

Who

le(Ja

pan)

[99]

159

3-(34-D

ihydroxyph

enyl)-2-prop

enoic

acid

Caffeicacid

O

HO

OH

OH

Who

leandaeria

l(Japan)

[7599]

160

3-Prop

yl-3[(245-trim

etho

xyph

enyl)-

metho

xy]-24-pentanedione

3-Prop

yl-3-(245-

trim

etho

xy)benzyloxy-

pentan-24-dione

O

O

O

O

O

O

Leaves

(India)

[100]

161

3-(34-D

ihydroxyph

enyl)-2-

prop

enoica

cidethyleste

rCa

ffeateethyl

O

O

OH

OH

Not

foun

d(Taiwan)

who

le(Taiwan)

[434584]

Evidence-Based Complementary and Alternative Medicine 31Ta

ble7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

162

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

d-Erythron

icacid

2-O-caffeoyl-2-C

-methyl

O

HO

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

163

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

2-O-caffeoyl-2-C

-methyl

O

O

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

164

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-24-

dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

3-O-caffeoyl-2-C

-methyl

O

O

HO

HO

O

O

OH

OH

Leaves

(Japan)

[101]

165

4-(A

cetyloxy)-3-[[3-(34-

dihydroxyphenyl)-

1-oxo-2-propen-1-y

l-]oxy]-2-hydroxy-

2-methyl-b

utanoica

cid

O

HO

OH

O OHO

HO

OO

Not

foun

d(Ja

pan)

[73]

166

3-(34-D

ihydroxyph

enyl)-tetrahydro-4-

hydroxy-4-

methyl-5

-oxo-3-fu

ranyl

ester-2-prop

enoica

cid

3-O-C

affeoyl-2-C

-methyl-

D-erythrono

-14-la

cton

eHO

HO

O

OO

OOH

Leaves

(Japan)

[101]

167

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]-oxy]-145-

trihydroxy-cyclohexanecarboxylicacid

Chlorogenica

cid

O

HO

HO

OH

O

OH

O

OH

HO

Aeria

l(Japan)w

hole

(Taiwan)

[707175]

32 Evidence-Based Complementary and Alternative Medicine

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

168

4-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]oxy]-

135-tr

ihydroxy-cyclohexanecarboxylic

acid

4-O-C

affeoylqu

inicacid

OHO

HO

OH

OH

O

O

OH

OH

Aeria

l(Japan)

[71]

169

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-15-

dihydroxy-cyclo

hexanecarboxylicacid

34-Di-O

-caffeoylqu

inic

acid

HO

HO

O

O

O

O

OH

OH

HO

OH O

H

O

Aeria

l(Japan)W

hole

(Taiwan)

[45497071]

170

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-14-dihydroxy-

cyclo

hexanecarboxylicacid

35-Di-O

-caffeoylqu

inic

acid

HO

HO

O O

HO

O

HO

OH OO

OHOH

Aeria

l(Japan)w

hole

(Taiwan)

[45497071]

Evidence-Based Complementary and Alternative Medicine 33

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

171

34-bis[[(2E

)-3-(34-D

ihydroxyph

enyl)-1-

oxo-2-prop

en-1-

yl]oxo]-15

-dihydroxy-

cyclo

hexanecarboxylicacid

45-Di-O

-caffeoylqu

inic

acid

OHO

OH

HO

O

O

OH

OH

O

OH

HO

OWho

le(Taiwan)

[454970]

172

3-[4-[[-6-O-[3-(4-H

ydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]-phenyl]-2-prop

enoica

cid

120573-D

-p-C

oumaricacid4-O

-(6-O

-p-caoum

aroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HOH

HO

O

O

Leaves

(Japan)

[59]

173

3-[4-[[2-O

-Acetyl-6

-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]ph

enyl]2-propeno

icacid

120573-D

-p-C

oumaricacid4-O

-(2-O

-acetyl-6

-O-p-

caou

maroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HO

HO

O

O

Leaves

(Japan)aerial

(China)

[3459]

174

67-Dihydroxy-2-chrom

enon

eEsculetin

cicho

rigenin

OHO HO

O

Not

foun

d(Egypt)

[39]

34 Evidence-Based Complementary and Alternative Medicine

Table 8 Aromatic compounds isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

175 12-Benzenediol PyrocatechinHO

HOWhole(Japan) [99]

176 4-Ethyl-12-benzenediol Pyrocatechol

OH

OH

Whole(Japan) [99]

177 Dimethoxyphenol26-dimethoxyphenol

O

HO

O

Roots (Japan) [99]

178 4-Ethenyl-2-methoxy-phenol p-Vinylguaiacol

O

OHWhole(Japan) [99]

179 2-Hydroxy-6-methylbenzaldehyde

6-Methyl-salicylaldehyde O

OH

Whole(Japan) [99]

180 Benzene-ethanol 2-Phenyl-ethanolHO

Whole(Japan) [57]

181 4-Hydroxy-3-methoxy-benzaldehyde Vanillin O

O

HOAerial(Japan) [99]

182 3-Hydroxy-4-methoxy-benzaldehyde Vanillin iso

OO

OH

Leaves(Japan) [99]

183 4-Hydroxybenzoic acid p-Hydroxybenzoicacid

O

HOOH Whole

(Japan) [99]

184 2-Hydroxybenzoic acid Salicylic acidO

HO

HO

Stem androots (Japan) [99]

185 34-Dihydroxybenzoicacid Protocatechuic acid

O

HOOH

OH

Whole(Japan) [99]

1864-Hydroxy-

3methoxybenzoicacid

Vanillic acid

OOH

O

OH

Aerial(Uganda)

Roots (Japan)[68 99]

Evidence-Based Complementary and Alternative Medicine 35

Table 8 Continued

S N IUPAC names Common names Structure Plant part(country) References

187 345-Trihydroxybenzoicacid Gallic acid

O

OHHO

HO

HO

Whole(China) [102]

[88] This work supports the use of B pilosa as an anti-inflammatory agent however no compounds responsible forthe anti-inflammatory activity of B pilosa were identified

A further study also reported the anti-inflammatoryactivity as well as the antiallergic activity of B pilosa [75] Inthis study dried powder of the aerial part of B pilosa whichhad been pretreated with the enzyme cellulosine was usedfor further tests The results showed that oral administrationof the cellulosine-treated B pilosa lowered the level of serumIgE in mice 10 days after immunization with DNP (24-dintrophenyl)-Ascaris as an antigen This treatment alsoreduced dye exudation in skin induced by passive cutaneousanaphylaxis and production of inflammatory mediators his-tamine and substance P in rats [75] Phytochemical analysisshowed that cellulosine treatment increased the percentage ofcaffeic acid and flavonoids This study suggests that B pilosaand its phenolics have anti-inflammatory functions

Phenolics and polyynes are major anti-inflammatoryphytochemicals present in B pilosa (Table 11) Unsurpris-6ingly phenolics such as luteolin (103) and ethyl caffeate(161) that are major constituents of B pilosa have alsobeen reported to possess anti-inflammatory activity Lute-olin was reported to exhibit anti-inflammatory activity inmacrophages Xagorari and colleagues showed that luteolininhibited the release of inflammatory cytokines TNF-120572 andinterleukin-6 in RAW 2647 cells following LPS stimulation[87] It inhibited TNF-120572 production with an IC

50value

of 1120583M The underlying anti-inflammatory mechanism ofluteolin was reported to be the inactivation of Akt andNF-120581B activation [87] In addition luteolin was reportedto confer anti-inflammatory activity through inhibition ofLPS-stimulated iNOS expression in BV-2 microglial cellsIt inhibited LPS-activated microglia in a dose-dependentmanner with an IC

50value of 69 120583MMoreover immunoblot

and RT-PCR data proved that luteolin suppressed I120581B-120572degradation and iNOS expression in LPS-activated microglia[85] Kim and colleagues stated that luteolin may havebeneficial effects on inflammatory neural diseases throughinhibition of iNOS expression [85] A related study revealedthat luteolin decreased the transcriptional activity of NF-120581BRelA via partial inhibition of TNF-mediated NF-120581B DNAbinding activity Luteolin also inhibited Akt phosphorylationand induced degradation of a transcription factor interferonregulatory factor (IRF) [86]

Chiang and colleagues showed that ethyl caffeate(161) significantly inhibited NO production in mousemacrophages RAW 2647 cells [84] Based on MTT assays

they concluded that this inhibition was not due to thecytotoxicity of ethyl caffeate The IC

50value of ethyl caffeate

in the inhibition of NO production was 55 120583gmL slightlylower than curcumin (positive control) which has anIC50

value of 65120583gmL They demonstrated that ethylcaffeate exerted anti-inflammatory activity via the reducedtranscription and translation of iNOS (inducible nitric oxidesynthase) in RAW 2467 cells In addition this compoundalso suppressed COX-2 expression in RAW 2467 cellsand MCF-7 cells The in vivo anti-inflammatory effectof ethyl caffeate was verified by testing in TPA-treatedmouse skin Like celecoxib the positive control ethylcaffeate significantly abolished COX-2 expression in adose-dependent manner Ethyl caffeate at 1mg200120583Lsite(24mM) inhibited COX-2 expression at a level comparableto celecoxib at 1mg200120583Lsite (13mM) Remarkably ethylcaffeate at 48mM (2mg200120583Lsite) was more effectivethan celecoxib at 131mM (10mg200120583Lsite) Moreoverthis compound inhibited the activation of nuclear factor-120581B(NF-120581B) by LPS via the prevention of NF-120581B binding toDNA [84] In addition 3 ethyl caffeate analogs (ethyl 34-dihydroxyhydrocinnamate ethyl cinnamate and catechol)also showed different degrees of NF-120581B binding to DNA aslisted in Table 12 Ethyl cinnamate lacks a catechol moietywhich results in ineffective inhibition of NF-120581B binding toDNA [84]

Pereira and colleagues assessed the anti- 7inflammatory and immunomodulatory activities ofB pilosa methanol extract as well as one polyyne2-O-120573-glucosyltrideca-11(E)-en-3 579-tetrayn-12-diol(54) in T lymphocytes and a zymosan-induced arthritismouse model [27] They first examined the in vitro effect of 8the B pilosa extract and compound 54 on cell proliferation ofhuman T cells stimulated with 5120583gmL phytohemagglutinin(PHA) or 100 nM 12-O-tetradecanoyl phorbol-13-acetate(TPA) plus 15 120583M ionomycin and on cell proliferation ofmouse T cells stimulated with 5 120583gmL concanavalin A(Con A) The data demonstrated that both methanol extractand compound 54 suppressed T-cell proliferation in adose-dependent manner The estimated IC

50values of the B

pilosa extract against human T cells stimulated with 5120583gmLPHA and 100 nM TPA plus 15120583M ionomycin were 125 and25 120583gmL respectively In comparison with the methanolextract compound 54 showed 10-fold more inhibition ofhuman T-cell proliferation with an estimated IC

50value of

15 120583gmL Accordingly the B pilosa extract and compound54 dose-dependently suppressed mouse T-cell proliferation

36 Evidence-Based Complementary and Alternative Medicine

Table 9 Porphyrins isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

188

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(15S16S)-10-ethenyl-5-ethyl-1161820-tetrahydro-6111522-tetramethyl-1820-dioxo-15H-912-imino-212-

metheno-471714-dinitrilopyrano[43-

b]azacyclononadecine-16-propanoic

acid

AristophyllC

NH N

HNN

O

O OOOPhytyl

O

Leaves(Taiwan) [100]

189

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -20b-(methoxycarbonyl)-9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-dinitrilo-12-

dioxeto-[310158404101584034]-cyclo-pent[1 2]azacyclononadecine-

19-propanoic acid

Bidenphytin A

NH N

HNN

OPhytylO O O

O O

Leaves(Taiwan) [100]

190

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -2a-

hydroxy-20b-(methoxycarbonyl)-

9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-

dinitrilo-12-dioxeto[310158404101584034]-cyclo-pent[12-b]-

azacyclononadecine-19-propanoic acid

Bidenphytin B

NH N

HNN

OPhytylO O O

O O

OH

Leaves(Taiwan) [100]

191

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO O

HOOO

Leaves(Taiwan) [100]

192

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

Evidence-Based Complementary and Alternative Medicine 37

Table 9 Continued

S N IUPAC names Common names Structure Plant part(country) References

193

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

Leaves(Taiwan) [100]

194

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

195

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3S4S21R)-9-ethenyl-14-ethyl-21-(methoxycarbonyl)-

481318-tetramethyl-20-oxo-3-phorbinepropanoic acid

Pheophytin A

NH N

HNN

PhytylO

O

OO O

Leaves(Taiwan) [100]

with estimated IC50values of 30 and 25 120583gmL respectively

Taken together the data indicate that the B pilosa extractand compound 54 act on human and mouse T cells Totest the in vivo effect of the B pilosa extract and compound54 a zymosan-induced arthritis mouse model was usedThis model was established from B10ASgSnJ mice with aninjection of zymosan (015mg) The zymosan-injected mice9received an intraperitoneal injection of the B pilosa extract(1 5 or 10mg) at one dose a day for 5 days Popliteal lymphnode (PLN) weight was monitored to check the developmentof arthritis The results revealed that 10mg of the methanolextract of B pilosa extract could significantly diminishinflammation as evidenced by PLN weight [27] This worksuggests that B pilosa (and compound 54) can suppressimmune response and inflammation

33 Antidiabetic Activity Anti-diabetic agents are primarilydeveloped from plants and other natural resources [8 24 4951 53] B pilosa is one of 1200 plant species that have beeninvestigated for antidiabetic activity [120 121]B pilosa is usedas an anti-diabetic herb in America Africa and Asia [51 120

122] Many studies have indicated that B pilosa could treattype 1 diabetes (T1D) and type 2 diabetes (T2D) in animals

Etiologically speaking T1D is caused by autoimmune-mediated destruction of pancreatic 120573 cells leading to insulindeficiency hyperglycemia and complications Currentlythere is no cure for T1D Polarization ofTh cell differentiationcontrols the development of T1D Suppression of Th1 celldifferentiation and promotion of Th2 cell differentiationameliorate T1D [123] One study showed that the butanolfraction of B pilosa inhibited T-cell proliferation decreasedTh1 cells and cytokines and increased Th2 cells andcytokines leading to prevention of T1D in nonobesediabetic (NOD) mice [49] Based on a bioactivity-directedisolation strategy 3 polyynes 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne (53) also known ascytopiloyne 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D- Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne (50) were identifiedfrom B pilosa [49 53] The IC

50value of the butanol fraction

was 200120583gmL This inhibition was reported to be partiallyattributed to cytotoxicity because the butanol fraction at

38 Evidence-Based Complementary and Alternative Medicine

Table 10 Other compounds isolated from B pilosa [30]

SN IUPAC names Common names Structure Plant part(country) References

19637-Dihydro-137-

trimethyl-1H-purine-26-dione

CaffeineN

NN

NO

O

Aerial(Uganda) [68]

197

1-((2R4S5R)-Tetrahydro-4-hydroxy-5-(hydroxymethyl)furan-2-yl)-5-

methylpyrimidine-24(1H3H)-dione

ThymidineHO

HO

ON

O

NHO Not found

(China) [37]

198 1-(2-Thienyl)-ethanone 2-Acetyl-thiophene

S

O

Roots(Germany) [41]

199

(2R3S4S5S)-2-(Heptan-2-yloxy)-tetrahydro-6-

((tetrahydro-34-dihydroxy-5-

(hydroxymethyl)furan-2-yloxy)methyl)-

2H-pyran-345-triol

Heptanyl2-O-120573-xylofuranosyl-

(1rarr 6)-120573-glucopyranoside

O

OOH

OH

HO

OHO

H

HO

H

H

HOHH

O

Whole(Taiwan) [70]

200

2-[(3R7R11R)-3-Hydroxy-371115-

tetramethylhexadecyl]-356-trimethyl-

25-cyclohexadiene-14-dione

120572-Tocopherylquinone

OO

HO

Whole(Taiwan) [32]

2017-O-(410158401015840610158401015840-Diacetyl)-

120573-D-glucopyranoside

Not found Leaves(China) [60]

180 120583gmL could cause 50 death of Th1 cells Moreoverthis study suggested that the butanol fraction may preventdiabetes in NOD mice in vivo via downregulation of Th1cells or upregulation Th2 cells that have effects which areantagonistic of those of Th1 cells [49] This was proven byintraperitoneal injection of the butanol fraction at a doseof 3mgkg BW 3 times a week to NOD mice from 4 to 27weeks This dosage resulted in lower incidence of diabetes(33) At a dose of 10mgkg the butanol fraction of Bpilosa totally eliminated (0) the initiation of the diseaseTo further support this result assessment of IgG2a and IgEproduction was performed in the serum of NOD mice Asin vivo results obtained from intracellular cytokine stainingexperiments were not very conclusive levels of IgG2a and IgE

were measured since Th1 cytokine IFN120574 and Th2 cytokineIL-4 favor the production of IgG2a and IgE respectively Asexpected high levels of IgE and some decline in the levels ofIgG2a were observed in the serum Profiling of the butanolextract revealed five compounds 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne(50) 45-Di-O-caffeoyl- quinic acid 35-Di-O-caffeoylquinicacid and 34-Di-O-caffeoylquinic acid Only the first twocompounds showed similar effects on the prevention ofdiabetes in NOD mice as the B pilosa butanol fractionMoreover compound 50 showed greater activity thancompound 69 in terms of enhancement (by 34 comparedto 8) of differentiation of Th0 to Th2 at 15 120583gmL (both

Evidence-Based Complementary and Alternative Medicine 39

Table 11 Chemical constituents of B pilosa and their biological activities

SN Name Classification Molecularformula Biological activities

109 Centaureidin [91] Flavonoid C18H16O8Anti-listerial [69 91]

Cytotoxic [10]

110 Centaurein [91] Flavonoid C24H26O13

Anti-listerial [69 91]Cytotoxic [10]Anti-viral [105]

103 Luteolin [106] Flavonoid C15H10O6

Anti-viral [107 108]Cytotoxic [77]

Anti-inflammatory [109]Anti-allergic [109]

82 Butein [110] Flavonoid C15H12O5Anti-leishmanial [111]

Cytotoxic [78]48 12-Dihydroxytrideca-57911-tetrayne [112] Polyyne C13H12O2 Anti-angiogeneic [112]46 13-Dihyroxy-6(E)-tetradecene-81012-triyne [112] Polyyne C14H16O2 Anti-angiogeneic [112]

45 12-Dihyroxy-5(E)-tridecene-7911-triyne [113] Polyyne C13H14O2Anti-angiogeneic [113]Anti-proliferative [113]

64 1-Phenylhepta-135-triyne [114] Polyyne C13H8

Anti-microbial [115]Anti-malarial [3]Cytotoxic [3]Antifungal [15]

27 Linoleic acid [79] Fatty acid C18H32O2Anti-viral (100)Cytotoxic [17]

161 Ethyl caffeate [84] Phenylpropanoid C11H12O4 Anti-inflammatory [84]

54 2-O-120573-Glucosyltrideca-11(E)-en-3579-tetrayn-12-diol [26] Polyyne C19H20O7Immunosuppressive andAnti-inflammatory [26]

53 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne [50] Polyyne C19H22O7Anti-diabetic [116]

Anti-inflammatory [50]

69 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne [28] Polyyne C20H26O7Anti-diabetic [28]

Anti-inflammatory [94]

50 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne [28] Polyyne C19H24O7

Anti-diabetic [28]Anti-inflammatory [94]

Anti-malarial andantibacterial [47]

129 Quercetin 3-O-120573-D-galactopyranoside [18] Flavonoid C21H20O12 Anti-inflammatory [117]

170 35-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

171 45-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

169 34-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

126 Quercetin 331015840-dimethyl ether7-O-120572-L-rhamnopyranosyl-(1rarr 6)-120573-D-glucopyranoside [118] Flavonoid C21H20O11 Anti-malarial [119]

125 Quercetin 331015840-dimethyl ether-7-O-120573-D-glucopyranoside [118] Flavonoid C21H20O12 Anti-malarial [119]70 1-Phenyl-13-diyn-5-en-7-ol-acetate [27] Polyyne C15H12O2 Anti-malarial [27]199 Heptanyl 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside [70] Miscellaneous C18H44O10 Antioxidant [70]124 3-O-Rabinobioside [70] Saccharide C27H38O15 Antioxidant [70]130 Quercetin 3-O-rutinoside [70] Flavonoid C27H38O15 Antioxidant [70]167 Chlorogenic acid [70] Phenolic C16H26O9 Antioxidant [70]119 Jacein [70] Flavonoid C24H26O13 Antioxidant [70]

49 (R)-12-dihydroxytrideca-357911-pentayne [47] Polyyne C13H8O2Anti-malarial andAntibacterial [47]

40 Evidence-Based Complementary and Alternative Medicine

Table 12 Structure and activity relationship studies of ethyl caffeateusing in vitro NF-120581BDNA binding assays [84]

Compound Concentration(120583M)

NF-120581BDNA binding

Ethyl caffeate 50 100inhibition

Ethyl 34-dihydroxyhydrocinnamate 100 100inhibition

Catechol 400 100inhibition

Ethyl cinnamate 400 Noinhibition

Table 13 Apoptosis in cocultures of T cells and pancreatic 120573 cells[53]

Cellmedium apoptosis andnecrosis

CD4+ T cellscontrol medium lt4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 2

CD4+ T cellswith cytopiloyne-treated 120573 cells 18CD4+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 7

CD8+ T cellscontrol medium 4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 4

CD8+ T cellswith cytopiloyne-treated 120573 cells 4CD8+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 4

Table 14 Radical scavenging activities of B pilosa extracts [70]

Extractscontrol DPPH assayIC50 (120583gmL)

NBThypoxanthinesuperoxide assayIC50 (120583gmL)

Quercetin 198 15Ascorbic acid 634 Not determined120572-tocopherol 897 Not determinedEthyl acetateextract 1383 597

Butanol extract 1669 114Water extract gt100 gt100

compounds) and inhibition (by 40 compared to 10) ofdifferentiation toTh1 at the same concentration [49]

Among the three polyynes found in B pilosa cytopiloyne(53) had the most potent anti-T1D activity [53] To test the invivo effect of cytopiloyne NODmice received intraperitonealor intramuscular injection of cytopiloyne at 25120583gkg BW3 times per week Twelve-week-old NOD mice started todevelop T1D and 70 of NOD mice aged 23 weeks andover developed T1D Remarkably 12- to 30-week-old NODmice treated with cytopiloyne showed normal levels of bloodglucose (lt200mgdL) and insulin (1-2 ngmL) Consistent

Table 15 Radical scavenging activity of secondarymetabolites fromB pilosa [70]

Metabolite (Table 11)control DPPH assay IC50 (120583gmL)199 Not determined124 53130 68167 105169 33171 38119 Not determined110 Not determinedQuercetin 256Caffeic acid 890

Table 16 Antioxidant activity of the essential oils and water extractsfrom B pilosa [92]

Extract IC50 (120583gmL)Leaf essential oils 47Flower essential oils 50Leaf extract 61Flower extract 172

with T1D incidence cytopiloyne delayed and reduced theinvasion of CD4+ T cells into the pancreatic islets [53]

In vitro study showed that cytopiloyne (53) inhibited thedifferentiation of naıveTh (Th0) cells (ie CD4+ T cells) intoTh1 cells and promoted differentiation of Th0 cells into Th2cells [50] The in vitro data are consistent with the in vivoresults indicating that cytopiloyne reducedTh1differentiationand increased Th2 differentiation as shown by intracellularcytokine staining and FACS analysis [53] In line with theskewing of Th differentiation the level of serum IFN-120574 andIgG2c decreased while that of serum IL-4 and serum IgEincreased compared to the negative controls (PBS-treatedmice) Cytopiloyne also enhanced the expression of GATA-3 a master gene for Th2 cell differentiation but not theexpression of T-bet a master gene forTh1 cell differentiationfurther supporting its role in skewingTh differentiation [53]

Also importantly cytopiloyne partially depleted CD4+rather than CD8+ T cells in NOD mice [53] As shown inTable 13 coculture assays showed that the depletion of CD4+T cells was mediated through the induction of Fas ligandexpression on pancreatic islet cells by cytopiloyne leading toapoptosis of infiltrating CD4+ T cells in the pancreas via theFas and Fas ligand pathways However cytopiloyne did notinduce the expression of TNF-120572 in pancreatic islet cells andthus had no effect on CD8+ T cells [53]

In addition Chang and colleagues showed that cytopi-loyne dose-dependently inhibited T-cell proliferation stimu-lated by IL-2 plus Con A or anti-CD3 antibody using [3H]thymidine incorporation assay [53]

Overall the mechanism of action of cytopiloyne andprobably its derivatives in T1D includes inhibition of T-cellproliferation skewing of Th cell differentiation and partialdepletion of Th cells Due to the anti-diabetic mechanisms

Evidence-Based Complementary and Alternative Medicine 41

Table 17 Antibacterial activity of essential oils and flower extracts from B pilosa [92]

Strain Mean zone of inhibition (mm)Leaf essential oil Flower essential oil Leaf extract Flower extract

Micrococcus flavus 127 plusmn 03 87 plusmn 03 102 plusmn 02 108 plusmn 03Bacillus subtilis 173 plusmn 19 117 plusmn 02 109 plusmn 02 103 plusmn 02Bacillus cereus 190 plusmn 14 112 plusmn 03 118 plusmn 04 185 plusmn 10Bacillus pumilus 123 plusmn 07 108 plusmn 02 105 plusmn 04 77 plusmn 02Escherichia coli 137 plusmn 04 203 plusmn 07 102 plusmn 11 140 plusmn 13Pseudomonas ovalis 125 plusmn 08 137 plusmn 15 102 plusmn 06 125 plusmn 06

Table 18 Antibacterial activity of root extracts from B pilosa [93]

Strain MIC50 (mgmL)Methanol extract Acetone extract

Bacillus cereus 10 mdashEscherichia coli 5 5Klebsilla pneumonia 5 10Micrococcus kristinae 10 mdashPseudomonas aeruginosa 10 10Staphylococcus aureus 5 10Sraphylococcus epidermidis 5 5Serratia marcescens 10 mdashShigelea flexneri 10 mdashStreptococcus faecalis 10 mdash

Table 19 Antibacterial activity of B pilosa of compound 29 [47]

Strain MIC50 (120583gmL)Escherichia coli NIHJ 1Escherichia coli ATCC25922 1Klebsiella pneumoniae ATCC700603 128Serratia marcescens ATCC13880 16Pseudomonas aeruginosa ATCC27853 8Staphylococcus aureus FDA209P 05Staphylococcus aureus ATCC29213 025Staphylococcus aureus N315 (MRSA) 05Enterococcus faecalis ATCC29212 2Enterococcus faecalis NCTC12201 (VRE) 1Bacillus subtilis ATCC6633 05Candida albicans ATCC10231 025

of action it was hypothesized that cytopiloyne protectsNOD mice from diabetes by a generalized suppression ofadaptive immunity To evaluate this hypothesis ovalbumin(Ova) was used as a T-cell dependent antigen to primeNOD mice which had already received cytopiloyne orPBS vehicle Ova priming boosted similar anti-Ova titersin cytopiloyne-treated mice and PBS-treated mice but adifference in immunoglobulin isotype was observed in thetwo groups Thus it was concluded that cytopiloyne is animmunomodulatory compound rather than an immunosup-pressive compound [50 53]

T2D is a chronic metabolic disease with serious com-plications resulting from defects in either insulin secretion

insulin action or both [124] A study by Ubillas et alshowed that the aqueous ethanol extract of the aerial partof B pilosa at 1 gkg body weight (BW) lowered bloodglucose in dbdb mice a T2D mouse model [51] Basedon a bioactivity-guided identification compounds 69 and50 were identified Further the mixture of the compounds(69 50) in a 2 3 ratio significantly decreased blood glucoseconcentration and reduced food intake on the second dayof treatment when administered at doses of 250mgkg twicea day to C5BLKs-dbdb mice When tested at 500mgkga more substantial drop in blood glucose level as wellas the stronger anorexic effect (food intake reduced from58 gmouseday to 25 gmouseday) was observed [51] Inthis study it was suggested that the blood glucose lower-ing effect of B pilosa was caused in part by the hungersuppressing effect of its polyynes [51] However the hungersuppressing effect of the ethanol extract of B pilosa wasnot found in the studies described below In another study[24] water extracts of B pilosa (BPWE) were used indiabetic dbdb mice aged 6ndash8 weeks with postprandialblood glucose levels of 350 to 400mgdL Like oral anti-diabetic glimepiride which stimulates insulin release onesingle dose of BPWE reduced blood glucose levels from374 to 144mgdL The antihyperglycemic effect of BPWEwas inversely correlated to an increase in serum insulinlevels suggesting that BPWE acts to lower blood glucose viaincreased insulin production However BPWE had differentinsulin secretion kinetics to glimepiride [24] One flaw incurrent anti-diabetics is their decreasing efficacy over timeThe authors investigated the long term anti-diabetic effectof BPWE in dbdb mice BPWE reduced blood glucoseincreased blood insulin improved glucose tolerance andreduced the percentage of glycosylated hemoglobin (HbA1c)Both long-term and one-time experiments strongly supportthe anti-diabetic action of BPWE [24] In sharp contrast toglimepiride BPWE protected against islet atrophy in mousepancreas The investigators further evaluated anti-diabeticproperties of 3 B pilosa varieties B pilosa L var radiate(BPR) Bpilosa L var pilosa (BPP) and B pilosa L varminor(BPM) in dbdb mice [8] One single oral dose (10 50 and250mgkg body weight) of BPR BPP or BPM crude extractsdecreased postprandial blood glucose levels in dbdb micefor up to four hours and the reduction of glucose levels inthe blood appeared to be dose-dependent Comparing thethree variants BPR extract resulted in a higher reduction inblood glucose levels when administered at the same dose as

42 Evidence-Based Complementary and Alternative Medicine

Table 20 Antifungal activity of B pilosa [92]

Partextract Concentration (ppm) Strain InhibitionCortiicum rolfsii Fusarium solani Fusarium oxysporum

Leaves

Essentialoils

100 857 plusmn 09 682 plusmn 0 745 plusmn 17250 960 plusmn 08 779 plusmn 18 879 plusmn 04

AqueousExtracts

100 446 plusmn 17 605 plusmn 21 716 plusmn 07250 942 plusmn 03 689 plusmn 07 824 plusmn 19

Flowers

Essentialoils

100 604 plusmn 09 892 plusmn 04 869 plusmn 05250 894 plusmn 12 980 plusmn 03 949 plusmn 06

AqueousExtracts

100 331 plusmn 11 714 plusmn 07 573 plusmn 22250 661 plusmn 14 912 plusmn 0 900 plusmn 07

Table 21 Antifungal activity of B pilosa root extracts [93]

Strain LC50 (mgmL)Acetoneextracts

Methanolextracts

Waterextracts

Aspergillus niger 014 006 007Aspergillus flavus 1091 658 0Penicillium notatum 005 005 005

the other two varieties In terms of serum insulin levels a doseof 50mgkg of each extract was used and the BPR extracttogether with the three polyynes significantly increased theserum insulin level in dbdb mice Long-term experiments(28-day treatment) were then conducted using diabetic micewith postprandial glucose levels from 370 to 420mgdL andglimepiride was used as positive controlThe range of dosagesapplied was from 10mgkg BW to 250mgkg BW Resultsshowed that the positive control as well as the crude extractsof the three varieties lowered the blood glucose levels indbdb mice However only BPR extract containing a higherpercentage of cytopiloyne (53) reduced blood glucose levelsand augmented blood insulin levelsmore than BPP andBPMThe percentage of glycosylated hemoglobin A1c (HbA1c) wasalso measured and found to be 79 plusmn 05 in mice aged 10ndash12 weeks and 66 plusmn 02 61 plusmn 03 and 62 plusmn 03 inthe blood of age-matchedmice following treatment with BPRcrude extract (50mgkg) glimepiride (1mgkg) and com-pound 53 (05mgkg) respectively [8] Among the polyynesfound in B pilosa cytopiloyne was the most effective againstT2D Hence cytopiloyne was used for further study on anti-diabetic action and mechanism [125] The data confirmedthat cytopiloyne reduced postprandial blood glucose lev-els increased blood insulin improved glucose tolerancesuppressed HbA1c level and protected pancreatic islets indbdb mice Nevertheless cytopiloyne failed to decreaseblood glucose in streptoztocin (STZ)-treated mice whose bcells were already destroyed Additionally cytopiloyne dose-dependently increased insulin secretion and expression in bcells as well as calcium influx diacylglycerol and activationof protein kinase C120572 Collectively the mechanistic studiessuggest that cytopiloyne treats T2D via regulation of insulin

production involving the calciumDAGPKC120572 cascade in bcells

The studies detailed above point to the conclusion thatcytopiloyne and related polyynes (compounds 69 and 49)are anti-diabetics in animal models The data uncover a newbiological action of polyynes It should be noted that likeall anti-diabetic drugs cytopiloyne failed to prevent or curediabetes completely but reduced diabetic complications [125]Intriguingly 34 polyynes have been found in B pilosa so farIt remains to be seen whether all the polyynes present in thisplant have anti-diabetic activities

34 Antioxidant Activity Free radicals can damage cellularcomponents via a series of chemical reactions [89] leadingto development and progression of cardiovascular diseasecancer neurodegenerative diseases and ageing [70] Freeradicals nitric oxide (NO) and superoxide anions can beproduced in macrophages to kill microbes However anexcessive generation of the free radicals under pathologicalconditions is associated with a wide range of illnessesPlants are known to be rich in antioxidant phytochemicalsChiang and colleagues evaluated the free radical scaveng-ing activity of crude extract fractions and compounds ofB pilosa using 11-diphenyl-2-picrylhydrazyl (DPPH) andhypoxanthinexanthine oxidase assays [70] Using DPPHand hypoxanthinexanthine oxidase assays they found thatthe B pilosa crude extract and the ethyl acetate butanoland water fractions had free radical scavenging activityNine compounds Heptyl- 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside (199) 3-O-Rabinobioside (124) Quercetin3-O-rutinoside (130) Chlorogenic acid (167) 34-Di-O-caffeoylquinic acid (169) 35-Di-O-caffeoylquinic acid (170)45-Di-O-caffeoylquinic acid (171) Jacein (119) and Cen-taurein (110) had DPPH radical scavenging activity [70]The IC

50values of the B pilosa crude extractfractions and

compounds are summarized in Tables 14 and 15 respectivelyMeasurement of free radical scavenging activities is one

way of assessing the antioxidant activities of B pilosa andits fractions and compounds It is interesting that the ethylacetate and butanol fractions are more active than the waterfraction and B pilosa crude extract [70] Of the secondarymetabolites only phenolic compounds 124 130 167 169 and171 showed significant DPPH-radical scavenging activities

Evidence-Based Complementary and Alternative Medicine 43

Further analysis of the structure-activity relationship of thecompounds suggested that substitution of the C3 hydroxylgroup with glycosides increased the activity approximately2-fold (for example in compounds 124 and 130) relative toquercetin (-OH in its C3) [70] Further modification of thestructures of the active compounds needs to be performed totest the effects of various substituents on activity The reasonwhy most of the antioxidant compounds contain phenolmoieties in their structure could be that the reduction-oxidation (redox) properties of phenols allow them to actas reducing agents singlet oxygen quenchers and hydrogendonors

A complementary study by Muchuweti and colleaguesdetermined phenolic content antioxidant activity and thephenolic profile of B pilosa methanol extract [89] Theyestimated that the phenolic content of the methanol extractof B pilosawas 11028plusmn22mgg [89] Vanillin hydroxyben-zaldehyde caffeic acid coumaric acid and ferulic acid werefound in this extractThe B pilosa extract also showed DPPHradical scavenging activity Furthermore the antioxidantactivity of the flavonoids found in B pilosa was correlatedwith its hepatoprotective effects through their inhibitionof NF-120581B activation which may lessen the oxidative stresscaused by the production of free radicals during liver injury[60]This activity might also be due to the anti-inflammatoryeffects of the aqueous extracts of B pilosa aerial parts on theinhibition of COX-2 and PGE

2production [88]

Essential oils from B pilosa flowers and leaves are alsoreported to possess antioxidant activity With the aim ofreplacing chemically synthesized additives Deba and col-leagues [92] worked on the antioxidant antibacterial andantifungal activities of essential oils and water extracts of Bpilosarsquos leaves and flowers Table 16 summarizes the resultsobtained from DPPH free radical scavenging assay

It can be inferred fromTable 16 that essential oils from theleaves possessed the highest activity It is reported elsewherethat monoterpenes present in essential oils such that of Bpilosa have protective effects and antioxidant properties [92]Beta-carotene bleaching method was also performed Leavesessential oils and aqueous extracts of the leaves and flowersshowed higher activity than the flower essential oils This isdue to the volatility of the flower essential oils The activityexhibited by the aqueous extracts was accounted to thepresence of phenolic compounds that are reported to donatea hydrogen atom to free radicals such that the propagation ofthe chain reaction during lipid oxidation is terminated [92]Overall essential oils and phenolics present in B pilosa canbe thought of as major antioxidant compounds

35 Immunomodulatory Activity B pilosa is thought to bean immunomodulatory plant and is reported to be effectivein the treatment of immune disorders such as allergy [75]arthritis [75] and T1D [48 50 53]

As pointed out in the discussion of its anti-diabeticactivities (Section 33) a combination of phytochemicalsmethod and T-cell activation assays was used to studyimmunomodulatory properties of B pilosa IFN-120574 is a keycytokine released by T and NK cells that mediates immune

cells and sustains immunity against pathogens Defects inIFN-120574 expression regulation and activation result in vulner-ability to diseases caused by bacteria and viruses [69] Anelegant study performed byChang and colleagues using IFN-120574 promoter-driven luciferase reporter construct in JurkatT cells showed that hot water crude extracts of B pilosaincreased IFN-120574 promoter activity two-fold [69] Out of thesubfractions of this extract the butanol fraction but not thewater or ethyl acetate fractions increased IFN-120574 promoteractivity six-fold Centaurein (110) and centaureidin (109)were identified from the butanol fraction and were stated tocause a four-fold increase in IFN-120574 promoter activity withEC50

values of 75 120583gmL and 09120583gmL respectively Themechanism of action of centaurein was determined usingtranscription factors such as AP-1 NFAT and NF120581B whichare reported to bind to IFN-120574 promoter and regulate IFN-120574transcription Unlike with the activity of the positive controlPHA centaurein caused a four-fold increase in NFAT a3-fold increase in NF120581B and had little if any effect onAP-1 enhancer activities (23-fold three-fold and ten-foldincreases respectively were seenwith PHA) [69]The authorsconcluded that centaureinmodulates IFN-120574 expression by theNFAT and NF120581B pathways The article only determined themechanism of action of centaurein Its aglycone centaureidinmay act through the samemechanism though this conclusionneeds to be further verified

B pilosa extract and its compounds are reported to inhibitdifferentiation of naıve CD4+ helper T (Th0) cells into Th1cells [49] Using theThcell differentiation assay as a screeningplatform 3 polyynes 2-120573-D-glucopyranosyloxy-1 -hydroxytrideca-57911-tetrayne (53) 3-120573-D-glucopyranosyl-1-hydroxy-6(E)- tetradecene-81012-triyne (69) and 2-120573-D-glucopyranosyloxy-1-hydroxy-5(E)- tridecene-7911-triyne(49) were discovered from B pilosa [49 53] The data showsthat cytopiloyne and other two polyynes suppressed thedifferentiation of type 1 helper T (Th1) cells and productionof Th1 cytokines and promoted that of type 2 helper T (Th2)cells and production of Th2 cytokines thus explaining theimmunomodulatory and anti-inflammatory effects of Bpilosa and its polyynes

Chang and colleagues were the first to report the effect ofthe butanol extract of B pilosa on the autoimmune diabetesand airway inflammation inmice [48] Imbalance in the levelsofTh1 andTh2 and of various cytokines leads to autoimmunediseases T1D and other autoimmune diseases (rheumatoidarthritis Crohnrsquos disease among others) are exacerbated byan increase Th1 levels (specifically CD4+ Th1 cells) whileTh2 cells antagonize this effect [19] Moreover Th2 cellsmediate asthma in ovalbumin-induced hypersensitivity inBALBc mice [48] In their work [48] Chang and colleaguesshowed that 10mgkg butanol extracts with 15 (ww)compound 69 and 11 (ww) compound 49 (Table 11)ameliorated the development of Th1-mediated diabetes inNOD mice through inhibition of 120573 cell death and leukocyteinfiltration At the same dosage the butanol extracts alsoexacerbated ovalbumin-induced pulmonary inflammationin BALBc mice with an increase in the infiltration ofeosinophils and mast cells into the airway of the mice [48]Despite the different outcomes both mouse models proved

44 Evidence-Based Complementary and Alternative Medicine

the concept that control over the Th1Th2 shift is associatedwith autoimmune diseases and that the B pilosa butanolextract can shift the differentiation of Th0 cells to Th2 cells[48 49]

An extended study presented by Chiang and colleaguesshowed that compound 53 modulates T-cell functions [50]Using CD4+ T cells from BALBc mice they demonstratedthat cytopiloyne decreased levels of IFN-120574 producing cells(Th1) by 122 (from 72 to 598) Since Th1 and Th2 celldifferentiation is antagonistic it was expected that compound53 increased the percentage of mouse IL-4 producing cells(Th2) by 72 (from 237 to 309) Subsequent assess-ment of effect of compound 53 on the modulation of thetranscription of IL-4 and IFN-120574 showed that cytopiloyne asexpected decreased the splenocyte levels of IFN-120574 mRNAand increasing that of IL-4 in a dose-dependent manner Inthe range of 01 to 3 120583gmL of compound 53 the effects werenot attributed to its cytotoxicity Consequently using 3120583gmLcytopiloyne for 72 and 96 hours the protein concentration ofIFN-120574 decreased to 186 and 444 respectively Under thesame conditions cytopiloyne increased IL-4 concentrationsto 1985 and 2470 (for 72 and 96 hours resp) Thismodulation of T-cell differentiation exhibited by compound53 was used to explain its anti-diabetic activity [50]

The anti-diabetic role of cytopiloyne was extensivelydiscussed above (Section 33 anti-diabetic activity) Themolecular basis of the regulation of cytokine expression bycytopiloyne has been described Cytopiloyne directly elevatedthe expression level of IL-4 via GATA-3 upregulation inT cells [53] However reduction of IFN-120574 expression in Tcells seemed to come from the indirect opposing effect IL-4cytokine because the expression level of T-bet was unaltered[53] In this way cytopiloyne skewedTh1 polarization into theTh2 state conferring protection against T1D in NOD miceAside frompolarization ofThcell differentiation cytopiloynealso activated the expression of Fas ligand in pancreaticb cells this increase leading to the partial depletion of Tcells and reduction of immune response in local areas suchas the pancreas Of note cytopiloyne also inhibited T-cellproliferation and activation By targeting T cells from threeimmunomodulatory actions cytopiloyne protects againstT1D and probably other Th1-mediated autoimmune diseases[19]

The phytochemical constituents of B pilosa exert theirfunctions on different immune cells to modulate immuneresponse It is possible that some of the compounds mayhave agonistic or antagonistic effects on immune responseImmune function of B pilosa may depend on its compo-sition and amount of compounds which could explain theapparently conflicting report of B pilosa butanol extractaggravating allergy in mice [48] while cellulosine-treatedextract ameliorated allergy [48 75] IFN-120574 promoter reporterassays and T-cell differentiation assays were used to isolate2 flavonoids [69] and 3 polyynes [49 50] as immunomod-ulatory compounds from the butanol fraction of B pilosaInterestingly the flavonoids promote IFN-120574 expression inNK and T cells In marked contrast the polyynes promoteIL-4 expression and indirectly inhibit IFN-120574 expression indifferentiating T cells Sensitivity appears to be the key to

identifying structure- and bioactivity-related phytochemicalsfrom medicinal plants

36 Antimalarial Activity The use of chemical drugs againstpathogens has resulted in drug-resistant mutants Examplesof drug resistance can be found in the species of thePlasmodium that cause malaria It is important to search fornew compounds to combat Plasmodium parasites [47] Astudy of the anti-malarial activity of the leaf extracts of Bpilosa using a combination of phytochemistry and bioassaysshowed that compound 49 (Table 11) showed activity againsta malaria parasite (P falciparum NF54 strain) with an IC

50

value of 60 120583gmL [77] In addition compound 49 isolatedfrom the aerial parts of B pilosa inhibited growth of the Pfalciparum FCR-3 strain with an IC

50value of 035 120583gmL

This compound was tested for its in vivo effect in miceinfected with P berghei NK-65 strain Results showed thatcompound 49 decreased the average parasitemia in the redblood cells by 207 (from 328 of that of the control to121) after an intravenous injection of 08mgkg BWday forfour days [47] Further studies addressing the anti-malarialmechanism underlying both polyynes and clinical studies areneeded

37 Antibacterial Activity Emergence of multiple antibiotic-resistantmicrobes is becoming a global threat to public healthand a challenge to disease treatment For instance penicillinis commonly used to combat a food-borne intracellularbacteriumListeria however penicillin-resistant bacteria havebeen discovered recently [69] Chang and coworkers iso-lated centaurein (110) and centaureidin (109) from B pilosaextract [69 70] Centaurein enhances expression of IFN-120574a key cytokine for macrophage activation and consequentlyenhances bactericidal activity in macrophages [69 70] Inagreement with observed in vitro effects centaurein wasreported to prevent and treat Listeria infection in C57BL6Jmice [91] Mechanistic studies confirmed that centaureinexerted antilisterial action via IFN-120574 expression in wild-typemice but not IFN-120574 knockout mice [91] Further in vitrostudies on centaurein showed that this compound increasedIFN-120574 expression by 13 (from 17 to 20) 20 (from 21to 41) and 11 (from 6 to 17) in CD4+ T cells CD8+Tcells and NK cells respectively That is to say there wasan increase in IFN-120574 producing immune cells As expectedcentaurein also enhanced the expression level of T-bet a keynuclear factor for IFN-120574 expression Consistently centaureinaugmented the serum IFN-120574 levels inC57BL6Jmice and thisaugmentation peaked 24 hours after compound injectionThe quantity of mouse serum IFN-120574 was sufficient to activatemacrophages in vitro and eradicated GFP-producing Listeriainside macrophages However the entry of Listeria intomacrophages was not affected by centaurein-treated mousesera Centaurein treatment at 20120583g per mouse rescued 30of the mice infected with a lethal dose of Listeria (2 times106 CFU) It is noteworthy that in the presence of ampicillin(5 120583gmouse) centaurein (20 120583gmouse) rescued 70 of themice suggesting an additive effect between ampicillin andcentaurein [91] Despite lower abundance centaureidin was

Evidence-Based Complementary and Alternative Medicine 45

30 times more active than centaurein in terms of IFN-120574production [91]

Aside from the indirect antibacterial action mentionedabove extract andor compounds of B pilosa also showeddirect bacteriostatic andor bactericidal action One studyreported that essential oils and leafflower extracts of Bpilosa could suppress the growth of gram positive and gramnegative bacteria as evidenced by zone of inhibition assaysIn this study antibacterial activity of the essential oils fromthe leaves and flowers of B pilosa was determined in anattempt to identify natural products as food preservatives forprevention of microbial multiplication and food oxidationThe essential oils and extracts of B pilosa leaves and flow-ers showed moderate but different extents of antibacterialactivity (Table 17) In general essential oils had higherantibacterial activity than crude extracts One explanation forthis could be that monoterpenes in the essential oils destroycellular integrity and subsequently inhibit the respirationand ion transport processes The presence of antibacterial120573-caryophyllene could be another explanation as reportedelsewhere [92] Another study reported that the methanoland acetone extract of B pilosa roots displayed antibacterialactivities against the bacteria listed in Table 18 [93] andmethanol extracts from the roots seemed to be the mosteffective

Another study indicated that the polyyne (R)-12-dihydroxytrideca-357911-pentayne (49) from this plantalso suppressed bacterial growth as shown by the minimuminhibitory concentration required to inhibit 50 bacterialgrowth (MIC

50) in Table 19 This compound was highly

effective against several Gram positive and Gram negativebacteria including the drug-resistant bacteria Staphylococcusaureus N315 (MRSA) and Enterococcus faecalis NCTC12201(VRE) [47] Strikingly compound 49 had a similar MIC

50

value to antibiotics (ampicillin tetracycline norfloxacin andamphotericin B) in most of the bacteria tested

Antibacterial activity of B pilosa extracts and compo-nents expressed as MIC

50and the mean zone of inhibition

is tabulated in Tables 17 18 and 19The zone of inhibition forAmpicilline (positive control) ranges from 153 plusmn 03mm to443 plusmn 02mm [92]

38 Antifungal Activity B pilosa has traditionally been usedto treat microbial infection Recently different parts of Bpilosa have been tested for antifungal activities Deba andcolleagues first evaluated the antifungal effect of the hotwater extracts of the B pilosa roots stems and leaves againstCorticium rolfsii Fusarium solani and Fusarium oxysporumThey discovered that C rolfsii was most suppressed bytreatment with B pilosa as its growth was reduced at almostall the tested doses followed by F oxysporum and F solani[99] However the fungicidal activities of the stems androots were greater than the leaves [99] Moreover the samegroup assessed the antifungal activity of the essential oilsand aqueous extracts from B pilosa flowers and leaves [92]They showed that the extracts and oils had antifungal activityagainst C rolfsii F solani and F oxysporum Essential oils

appeared to have better fungicidal activity thanwater extractsas summarized in Table 20

Another study by Ashafa and colleagues showed thatacetone methanol and water extracts of the B pilosa rootsshowed antifungal activities against Aspergillus niger Aflavus and Penicillium notatum using the agar dilutionmethod The results are tabulated in Table 21 [93] Negativecontrols showed 0 growth inhibitionThe methanol extractof the B pilosa roots at 10mgmL was also effective againstCandida albicans [93]Of note B pilosa obtained fromPapuaNew Guinea had no activity against A niger and C albicans[126] but the SouthAfrican (EasternCape) ecotype exhibitedmoderate activity against C albicans [93] This discrepancymay depend on extraction solvents extraction procedureassay techniques different plant parts and abundance ofactive compounds

B pilosa produces a variety of secondary metabolitessuch as flavonoids phenylacetylenes alkaloids sterols ter-penoids and tannis [92 93] However none of them havebeen confirmed as active compounds against fungi Furtherinvestigation of active compounds from B pilosa is necessaryto further understand the antifungal efficacy of this plant

39 Hypotensive and Vasodilatory Activities In early studiesDimo and colleagues used three rat models normotensiveWistar rats (NTR) salt-loading hypertensive rats (SLHR)and spontaneous hypertensive rats (SHR) to investigate thehypotensive effect of the methanol crude extract of B pilosaleaves [28 127] The extract lowered systolic blood pressurein hypertensive rats (SLHR and SHR) to a greater degreethan NTR [28 127] In addition a decrease in urinarysodium ions and an increase in urinary potassium ions wereobserved after treatment with the methanol extract of Bpilosa leaves although neither differences were statisticallysignificant [28 127] Taking the data together the studyproposed that B pilosa leaf extract reduced blood pressurevia vasodilation [28 127] The same group continued totest the antihypertensive effect of aqueous and methylenechloride extracts of B pilosa leaves in a hypertensive ratmodel [28 127] To establish a fructose-induced hypertensionmodel male Wistar rats were given 10 fructose solutionto drink ad libitum for three weeks In addition to freeaccess to 10 fructose the rats were treated with the aqueous(150mgkg) or methyl chloride (350mgkg) extracts of Bpilosa for additional three weeks [28 127] Both extracts ofB pilosa leaves had a hypotensive effect on rats Howeverneither extracts reversed the elevation of serum insulin infructose-fed rats Therefore B pilosa lowered blood pressureirrespective of insulin [28 127] To better understand thehypotensive mechanism the authors investigated the effectof a neutral extract of B pilosa (NBP) a mixture of methanolandmethylene chloride (1 1) extract after neutralization withNaOH and HCl on the heart and the blood pressure ofNTR and SHR [110] This study showed that an intravenousinjection of the NBP resulted in a biphasic reduction insystolic blood pressure In addition one intravenous dose ofthe extract at 10 20 and 30mgkg BW decreased systolicblood pressure in normal rats by 183 425 and 30

46 Evidence-Based Complementary and Alternative Medicine

respectively and the same doses reduced the blood pressurein hypertensive rats by 258 389 and 286 respectivelyOnly the highest dose (30mgkg) affected the force of thecontraction of the heart Atropine and propranolol were usedto interfere with the hypotensive action of the NBP Atropinereduced the initial phase of the hypotensive response in NBPand completely abolished the second phase of hypotensiveresponse in NBP In contrast propranolol increased thefirst hypotensive response but partially abolished the secondhypotensive response provoked by the NBP [110]This mech-anistic study suggested that B pilosa invokes the biphasichypotensive responses via targeting cardiac pump efficiencyduring the first phase and vasodilation at the second phase[110]

A further study was performed to investigate the relaxingeffect of a neutral extract of B pilosa (NBP) on rat aorta con-tractedwithKCl (60mM) andnorepinephrine (01mM) [94]Cumulative addition of NBP relaxed the rat aorta previouslycontracted by KCl in a dose-dependent manner The EC

50

value of theNBP for vasorelaxationwas 032mgmLThedataalso showed that the NBP reduced the contraction of aortapreviously contracted by KCl irrespective of the presence ofaortic endothelium [94]

Pretreatment with glibenclamide an ATP-dependent K+channel blocker did not considerably affect the relaxant effectof the NBP on KCl-induced contraction suggesting thatthe vasodilatory effect of B pilosa was not related to theopening of this ATP-dependent K+ channel [94] On theother hand in the presence of indomethacin or pyrilaminemaleate the relaxant response induced by the plant extractwas significantly inhibited at the lower concentrations Theplant extract was able to reduce the aorta resting tone inhibitthe KCl-induced contractions by 90 at 15mgmL and theCaCl2-induced contractions by 95 at 075mgmL These

results demonstrate that B pilosa can act as a vasodilatorprobably via acting as a calcium antagonist [94]

However no specific compound for the above activityhas been identified from B pilosa to date A bioactivity-guided identification approach may be adopted to identifythe active compounds in B pilosa that possess hypotensiveand vasodilatory effects and understand their mechanism ofaction

310 Wound Healing Activity B pilosa has been traditionallyused to treat tissue injury inCameroon Brazil andVenezuela[26] Hassan and colleagues investigated the wound healingpotential of B pilosa in Wistar rats [128] Mirroring thepositive control neomycin sulfate the ethanol extract of Bpilosa had faster wound closure than control rats 3 6 and 9days after topical application Histological examination alsorevealed better collagenation angiogenesis and organizationof wound tissue seven days after application Epithelializationand total healing time in B pilosa-treated rats were compara-ble to those of neomycin sulfate Together these data suggestthat B pilosa may be a viable alternative to neomycin lotionfor the treatment of wounds

In addition to studying the wound healing effect of Bpilosa on external ulcers Tan and colleagues also examined

the effect of methanol cyclohexane and methyl chlorideextracts of B pilosa on gastric ulcers in Wistar rats fedwith 1mL HClethanol gastric necrotizing solution (150mMHCl in 60 ethanol) and macroscopically visible lesionswere scored [26] Among the three extracts methylene chlo-ride extracts exhibited the highest activity showing 464inhibition of lesion formation at a dose of 500mgkg BWand complete inhibition at 750mgkg [26] The efficacy ofthe ethylene chloride extract was followed by that of themethanol extracts which had inhibition ranging from 304to 822 at concentrations of 500mgkg and 1000mgkgBW respectively [26] The cyclohexane extracts showed thelowest activity against gastric ulcers in rats with 13340 and 797 inhibition at 500 750 and 1000mgkg BWrespectively [26] To better understand the mode of actionof the methylene chloride extract of B pilosa rats werepretreated with indomethacin a COX-2 inhibitor involved inprostaglandin synthesis Pretreatment significantly reducedthe protection against HClethanol-induced ulcers to 313inhibition at 750mgkg BW suggesting a link between theantiulcerative activity of B pilosa and prostaglandin syn-thesis Unexpectedly the methylene chloride extract of Bpilosa showed little gastric mucosal protection against gastriclesions induced by 95 ethanol (1mL) [26] Absolute alcoholis known to cause mucosalsubmucosal tissue destruction viacellular necrosis and the release of tissue-derived mediators(histamine and leukotrieneC4)Thus these data imply thatBpilosa did not prevent the generation or the necrotic action ofthese mediators on the gastric microvasculature In additionpylorus ligation can increase gastric acid secretion withoutan alteration of mucosal histamine content The methylenechloride extract of B pilosa did not possess antisecretoryactivity On the contrary it was observed that increases in thedose of the extract led to elevated gastric juice acidity [26]Results with both absolute ethanol and pylorus ligation ratmodels suggested the possibility that the ineffectiveness of Bpilosa against gastric ulcers was due to lack of antihistaminicactivity in the plant In summary overall the data suggest thatB pilosa protects against HCLethanol-mediated ulcers viainhibition of prostaglandin biosynthesis

Previous phytochemical studies showed that a groupof flavonoids acyclichalcones are present in B pilosa[129 130] and the chalcones were proposed to have anti-ulcerative activity [131] Moreover nine hydroxychalcones 10were reported to possess gastric cytoprotective effects with24-dihydroxychalcone being the most active [132] Sincemethylene chloride extracts appear to be the most active Bpilosa extracts next the specific anti-ulcerative phytochemi-cals in the methylene chloride extracts of B pilosa and theirmodes of action needs to be probed

Despite the claims listed inTable 3 relatively few scientificstudies have been conducted in vitro and in vivo to address thetraditional ethnomedical uses of B pilosa Information aboutthe use of B pilosa as a botanical therapy recorded so far isfar from complete Studies conducted thus far only serve asa starting point for further investigation of B pilosa and theultimate efficacious use of the herb in clinical applications

Evidence-Based Complementary and Alternative Medicine 47

4 Toxicology

Despite its use as an ingredient in food for human con-sumption studies on systemic toxicity (eg acute subacutechronic and subchronic toxicities) of B pilosa in humansand animals are still inadequate and insufficient So faracute andor subchronic toxicities have been evaluated inrats and mice Oral acute and 28-day toxicities of waterextract of B pilosa leaves were evaluated in Wistar rats[133] An oral dose of water extract of B pilosa leaves at10 gkg BW showed no obvious mortality or changes inthe appearance in rats [134] The same extract at 08 gkgBWday once a day showed no obvious sub-chronic toxicityin rats over 28 days as measured by survival rate bodyweight and gross examination of organs [134] These dataare consistent with our data indicating that oral deliveryof the water extract of the B pilosa whole plant at 1 gkgBWday once a day is safe in rats over 28 days (unpublisheddata) Taken together these studies suggest that ingestion ofB pilosa aqueous extract at up to at 1 gkg BWday oncea day is highly safe in rats In addition the acute toxicityof aqueous and ethanol extracts of B pilosa in mice havebeen reported [134] Five- to six-week-old mice with weightsbetween 28 and 35 g received a peritoneal injection of bothextracts at the different doses The LD

50 the dose that causes

50 lethality of the aqueous and ethanol extracts in micewas 1230 gkg BW and 615 gkg BW respectively [134] Acomplete toxicological study has not been completed forhumans Furthermore the drug interactions of B pilosa withother drugs are unknown Further safety verification andclinical trials should be performed before B pilosa can beconsidered for medicinal use

5 Conclusions

B pilosa is an erect perennial plant with green leaves whiteor yellow flowers and tiny black seeds As it is distributedworldwide and is widely used as a folk remedy B pilosacan be thought of as an extraordinary source of food andmedicine However a comprehensive up-to-date review ofresearch on B pilosa has hitherto been unavailable In thisarticle scientific studies on B pilosa have been summarizedand critically discussed from the perspectives of botanyethnomedicine phytochemistry pharmacology and toxicol-ogy B pilosa is claimed to treat more than 40 disordersand 201 compounds have been identified from this plantThe medicinal utility of B pilosa and its modes of actionin relation to its known phytochemicals were discussedherein Polyynes flavonoids phenylpropanoids fatty acidsand phenolics are the primary bioactive compounds of Bpilosa and they have been reported to be effective in thetreatment of tumors inflammationimmune modulationdiabetes viruses microbes protozoans gastrointestinal dis-eases hypertension and cardiovascular diseases Cautionshould be exercised in the therapeutic use of B pilosa forhypoglycemia hypotension bleeding and allergy

Acknowledgments

The authors thank their laboratory members for construc-tive suggestions and the authors whose publications theycited for their contributions The authors also thank MsMiranda Loney of Academia Sinica for English editing of thispaper This work was supported by Grants 99-CDA-L11 and101S0010056 from Academia Sinica Taiwan Arlene P Bar-tolome is a recipient of MECO-TECO Sandwich ScholarshipProgram between Taiwan and the Philippines

References

[1] T Shen G H Li X N Wang and H X Lou ldquoThe genusCommiphora a review of its traditional uses phytochemistryand pharmacologyrdquo Journal of Ethnopharmacology vol 142 no2 pp 319ndash330

[2] C Long P Sauleau B David et al ldquoBioactive flavonoids ofTanacetum parthenium revisitedrdquo Phytochemistry vol 64 no2 pp 567ndash569 2003

[3] P O Karis and O Ryding Asteraceae Cladistics and Classifica-tion Bremer K Eds pp 559ndash569 Timber press Portland OreUSA 1994

[4] O N Pozharitskaya A N Shikov M N Makarova et alldquoAnti-inflammatory activity of a HPLC-fingerprinted aqueousinfusion of aerial part of Bidens tripartita LrdquoPhytomedicine vol17 no 6 pp 463ndash468 2010

[5] F Q Oliveira V Andrade-Neto A U Krettli and MG L Brandao ldquoNew evidences of antimalarial activity ofBidens pilosa roots extract correlated with polyacetylene andflavonoidsrdquo Journal of Ethnopharmacology vol 93 no 1 pp 39ndash42 2004

[6] Agriculture USDA ldquoPlants databaserdquo 2013in Natural Resources Conservation Servicehttpwwwnrcsusdagovwpsportalnrcssitenationalhome

[7] Y Zhou and X Z Yan ldquoExperimental study of qi deficiencysyndrome and Codonopsis pillosulae and Astragalus injectionon the immune response in micerdquo Zhong Xi Yi Jie He Za Zhivol 9 no 5 pp 286ndash262 1989

[8] S C Chien P H Young Y J Hsu et al ldquoAnti-diabeticproperties of three common Bidens pilosa variants in TaiwanrdquoPhytochemistry vol 70 no 10 pp 1246ndash1254 2009

[9] M J Alcaraz and M J Jimenez ldquoFlavonoids as anti-inflammatory agentsrdquo Fitoterapia vol 59 no 1 pp 25ndash38 1988

[10] FAO Agriculture Food and Nutrition for AfricamdashA ResourceBook for Teachers of Agriculture Publishing ManagementGroup FAO Information Division Rome Italy 1997

[11] M B Rokaya Z Munzbergova B Timsina and K R BhattaraildquoRheum australe D Don a review of its botany ethnobotanyphytochemistry and pharmacologyrdquo Journal of Ethnopharma-cology vol 141 no 3 pp 761ndash774 2012

[12] P H Young Y J Hsu and W C Yang ldquoBidens pilosa L andits medicinal userdquo in Recent Progress in Medicinal Plants DrugPlant II A S AwaadVK Singh and JNGovil Eds StandiumPress Houston Tex USA 2010

[13] C Ge ldquoCytologic study of Bidens bipinnata LrdquoZhongguo ZhongYao Za Zhi vol 15 no 2 pp 72ndash125 1990

[14] L C Chiang J S Chang C C Chen L T Ng and C CLin ldquoAnti-herpes simplex virus activity of Bidens pilosa and

48 Evidence-Based Complementary and Alternative Medicine

Houttuynia cordatardquoAmerican Journal of ChineseMedicine vol31 no 3 pp 355ndash362 2003

[15] N P Rybalchenko V A Prykhodko S S Nagorna et alldquoIn vitro antifungal activity of phenylheptatriyne from Bidenscernua L against yeastsrdquo Fitoterapia vol 81 no 5 pp 336ndash3382010

[16] K Redl W Breu B Davis and R Bauer ldquoAnti-inflammatoryactive polyacetylenes from Bidens campylothecardquo Planta Med-ica vol 60 no 1 pp 58ndash62 1994

[17] M Ayyanar and S Ignacimuthu ldquoTraditional knowledge ofKani tribals inKouthalai of Tirunelveli hills TamilNadu IndiardquoJournal of Ethnopharmacology vol 102 no 2 pp 246ndash255 2005

[18] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant intraditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[19] AKAbbas KMMurphy andA Sher ldquoFunctional diversity ofhelper T lymphocytesrdquo Nature vol 383 no 6603 pp 787ndash7931996

[20] J H Cano and G Volpato ldquoHerbal mixtures in the traditionalmedicine of Eastern Cubardquo Journal of Ethnopharmacology vol90 no 2-3 pp 293ndash316 2004

[21] J Namukobe J M Kasenene B T Kiremire et al ldquoTraditionalplants used for medicinal purposes by local communitiesaround the Northern sector of Kibale National Park UgandardquoJournal of Ethnopharmacology vol 136 no 1 pp 236ndash245 2011

[22] C Lans ldquoComparison of plants used for skin and stomachproblems in Trinidad and Tobago with Asian ethnomedicinerdquoJournal of Ethnobiology andEthnomedicine vol 3 article 3 2007

[23] S Dharmananda ldquoA Popular Remedy Ecapes Notice ofWesternPractitionersrdquo 2013 httpwwwitmonlineorgartsbidenshtm

[24] Y J Hsu T H Lee C L T Chang Y T Huang andW C YangldquoAnti-hyperglycemic effects and mechanism of Bidens pilosawater extractrdquo Journal of Ethnopharmacology vol 122 no 2 pp379ndash383 2009

[25] E Noumi F Houngue and D Lontsi ldquoTraditional medicinesin primary health care plants used for the treatment ofhypertension in Bafia Cameroonrdquo Fitoterapia vol 70 no 2 pp134ndash139 1999

[26] P V Tan T Dimo and E Dongo ldquoEffects of methanolcyclohexane and methylene chlo ride extracts of Bidens pilosaon various gastric ulcer models in ratsrdquo Journal of Ethnophar-macology vol 73 no 3 pp 415ndash421 2000

[27] R L C Pereira T Ibrahim L Lucchetti A J R Da Silva and VL G De Moraes ldquoImmunosuppressive and anti-inflammatoryeffects of methanolic extract and the polyacetylene isolatedfrom Bidens pilosa Lrdquo Immunopharmacology vol 43 no 1 pp31ndash37 1999

[28] T Dimo J Azay P V Tan et al ldquoEffects of the aqueous andmethylene chloride extracts of Bidens pilosa leaf on fructose-hypertensive ratsrdquo Journal of Ethnopharmacology vol 76 no 3pp 215ndash221 2001

[29] CWiartMedicinal Plants of Southeast Asia Pelanduk SelabgorDarul Ehsan Malaysia 2000

[30] F L Silva D C H Fischer J F Tavares M S Silva P FDe Athayde-Filho and J M Barbosa-Filho ldquoCompilation ofsecondary metabolites from Bidens pilosa LrdquoMolecules vol 16no 2 pp 1070ndash1102 2011

[31] A H Chen S R Lin and C H Hong ldquoPhytochemical studyon Bidens pilosa L varminorrdquo Huaxue pp 38ndash42 1975

[32] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[33] M C Zulueta M Tada and C Y Ragasa ldquoA diterpene fromBidens pilosardquo Phytochemistry vol 38 no 6 pp 1449ndash14501995

[34] J Wang H Yang Z W Lin and H D Sun ldquoFlavonoids fromBidens pilosa var radiatardquo Phytochemistry vol 46 no 7 pp1275ndash1278 1997

[35] A Zhao Q Zhao L Peng et al ldquoA new chalcone glycoside fromBidens pilosardquo Acta Botanica Yunnanica vol 26 no 1 pp 121ndash126

[36] P Gao Y Wu S Cui and Y Zou ldquoSynthesis and absoluteconfiguration of the 7-phenylhepta-4 6-diyne-1 2-diol isolatedfrom Bidens pilosardquo Synthesis no 13 pp 2131ndash2135 2011

[37] S Wang B Yang D Zhu D He and L Wang ldquoActivecomponents of Bidens pilosa Lrdquo Zhongcaoyao vol 36 pp 20ndash21

[38] F Bohlmann T Burkhardt and C Zdero Naturally OccuringAcetylenes Academic Press New York NY USA 1973

[39] T M Sarg A M Ateya N M Farrag and F A AbbasldquoConstituents and biological activity of Bidens pilosa L grownin EgyptrdquoActa Pharmaceutica Hungarica vol 61 no 6 pp 317ndash323 1991

[40] H A L Valdes and H P Rego ldquoBidens pilosa Linnerdquo RevistaCubana de Plantas Medicinales vol 6 pp 28ndash33

[41] F Bohlmann H Bornowski and K M Kleine ldquoNew polyynesfrom the tribe Heliantheaerdquo Chemische Berichte vol 97 pp2135ndash2138

[42] RWang Q XWu and Y P Shi ldquoPolyacetylenes and flavonoidsfrom the aerial parts ofBidens pilosardquo PlantaMedica vol 76 no9 pp 893ndash896 2010

[43] L W Wu Y M Chiang H C Chuang et al ldquoA novelpolyacetylene significantly inhibits angiogenesis and promotesapoptosis in human endothelial cells through activation of theCDK inhibitors and caspase-7rdquo Planta Medica vol 73 no 7 pp655ndash661 2007

[44] L W Wu Y M Chiang H C Chuang et al ldquoPolyacetylenesfunction as anti-angiogenic agentsrdquo Pharmaceutical Researchvol 21 no 11 pp 2112ndash2119 2004

[45] H L Yang S C Chen N W Chang et al ldquoProtection fromoxidative damage using Bidens pilosa extracts in normal humanerythrocytesrdquo Food and Chemical Toxicology vol 44 no 9 pp1513ndash1521 2006

[46] H Q Wang S J Lu H Li and Z H Yao ldquoEDTA-enhancedphytoremediation of lead contaminated soil by Bidens maxi-mowiczianardquo Journal of Environmental Sciences vol 19 no 12pp 1496ndash1499 2007

[47] S TobinagaMK SharmaWG L Aalbersberg et al ldquoIsolationand identification of a potent antimalarial and antibacterialpolyacetylene from Bidens pilosardquo Planta Medica vol 75 no 6pp 624ndash628 2009

[48] C L T Chang H K Kuo S L Chang et al ldquoThe distincteffects of a butanol fraction of Bidens pilosa plant extract onthe development of Th1-mediated diabetes and Th2-mediatedairway inflammation in micerdquo Journal of Biomedical Sciencevol 12 no 1 pp 79ndash89 2005

[49] S L Chang C L T Chang Y M Chiang et al ldquoPolyacetyleniccompounds and butanol fraction from Bidens pilosa can mod-ulate the differentiation of helper T cells and prevent autoim-mune diabetes in non-obese diabetic micerdquo Planta Medica vol70 no 11 pp 1045ndash1051 2004

Evidence-Based Complementary and Alternative Medicine 49

[50] Y M Chiang C L T Chang S L Chang W C Yang and LF Shyur ldquoCytopiloyne a novel polyacetylenic glucoside fromBidens pilosa functions as a T helper cell modulatorrdquo Journal ofEthnopharmacology vol 110 no 3 pp 532ndash538 2007

[51] R PUbillas CDMendez S D Jolad et al ldquoAntihyperglycemicacetylenic glucosides from Bidens pilosardquo Planta Medica vol66 no 1 pp 82ndash83 2000

[52] L Alvarez S Marquina M L Villarreal D Alonso E Arandaand G Delgado ldquoBioactive polyacetylenes from Bidens pilosardquoPlanta Medica vol 62 no 4 pp 355ndash357 1996

[53] C L T Chang S L Chang Y M Lee et al ldquoCytopiloyne apolyacetylenic glucoside prevents type 1 diabetes in nonobesediabetic micerdquo Journal of Immunology vol 178 no 11 pp 6984ndash6993 2007

[54] G Kusano A Kusano and Y Seyama Novel Hypoglycemicand Anti-Inflammatory Polyacetylenic Compounds Their Com-positions Bidens Plant Extract Fractions and CompositionsContaining the Plant or Fraction JPO Tokyo Japan 2004

[55] M G L Brandao A U Krettli L S R Soares C G CNery and H C Marinuzzi ldquoAntimalarial activity of extractsand fractions from Bidens pilosa and other Bidens species(Asteraceae) correlated with the presence of acetylene andflavonoid compoundsrdquo Journal of Ethnopharmacology vol 57no 2 pp 131ndash138 1997

[56] A U Krettli V F Andrade-Neto M D G L Brandao andW M S Ferrari ldquoThe search for new anti-malarial drugs fromplants used to treat fever and malaria or plants ramdomlyselected a reviewrdquoMemorias do Instituto Oswaldo Cruz vol 96no 8 pp 1033ndash1042 2001

[57] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[58] C K Wat R K Biswas E A Graham L Bohm G H NTowers and E R Waygood ldquoUltraviolet-mediated cytotoxicactivity of phenylheptatriyne from Bidens pilosa Lrdquo Journal ofNatural Products vol 42 no 1 pp 103ndash111 1979

[59] Y Sashida K Ogawa M Kitada H Karikome Y Mimakiand H Shimomura ldquoNew aurone glucosides and new phenyl-propanoid glucosides from Bidens pilosardquo Chemical and Phar-maceutical Bulletin vol 39 no 3 pp 709ndash711 1991

[60] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens pilosa L (TFB) on animal liver injury andliver fibrosisrdquo Journal of Ethnopharmacology vol 116 no 3 pp539ndash546 2008

[61] B Hoffmann and J Holzl ldquoAcylated compounds from Bidenspilosardquo Planta Medica vol 55 pp 108ndash109

[62] B Hoffmann and J Holzl ldquoA methylated chalcone glucosidefrom Bidens pilosardquo Phytochemistry vol 27 no 11 pp 3700ndash3701 1988

[63] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[64] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[65] B Hoffmann and J Holzl ldquoWeitere acylierte chalkone ausBidens pilosardquo Planta Medica vol 54 pp 450ndash451 1988

[66] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant in11traditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[67] V V Pham V V T K P V L Hoang and V K Phan ldquoFlavonoidcompounds from the plant Bidens pilosa L (Asteraceae)rdquo TapChi Duoc Hoc vol 50 pp 48ndash53

[68] S D Sarker B Bartholomew R J Nash and N Robinson ldquo5-O-methylhoslundin an unusual flavonoid from Bidens pilosa(Asteraceae)rdquo Biochemical Systematics and Ecology vol 28 no6 pp 591ndash593 2000

[69] S L Chang Y M Chiang C L T Chang et al ldquoFlavonoidscentaurein and centaureidin from Bidens pilosa stimulate IFN-120574 expressionrdquo Journal of Ethnopharmacology vol 112 no 2 pp232ndash236 2007

[70] Y M Chiang D Y Chuang S Y Wang Y H Kuo P WTsai and L F Shyur ldquoMetabolite profiling and chemopreventivebioactivity of plant extracts from Bidens pilosardquo Journal ofEthnopharmacology vol 95 no 2-3 pp 409ndash419 2004

[71] A Kusano Y Seyama E Usami et al ldquoStudies on the antioxi-dant active constituents of the dried powder from Bidens pilosaL var radiata SchrdquoNaturalMedicines vol 57 no 3 pp 100ndash1042003

[72] I Doleckova L Rarova J Gruz et al ldquoAnti-proliferative andanti-angiogenic effects of flavone eupatorin an active con-stituent of chloroform extract ofOrthosiphon stamineus leavesrdquoFitoterapia vol 83 no 6 pp 1000ndash1007

[73] K Bairwa R Kumar R J Sharma and R K Roy ldquoAn updatedreview on Bidens pilosa Lrdquo Der Pharma Chemica vol 2 no 3pp 325ndash337

[74] Q Xia Y Liu and Y Li ldquoDetermination of hyperoside indifferent parts and different species of Herba Bidens by RP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol 50pp 48ndash53

[75] M Horiuchi and Y Seyama ldquoImprovement of the antiinflam-matory and antiallergic activity of Bidens pilosa L var radiataSCHERFF treated with enzyme (Cellulosine)rdquo Journal of HealthScience vol 54 no 3 pp 294ndash301 2008

[76] M G L Brandao C G C Nery M A S Mamao and AU Krettli ldquoTwo methoxylated flavone glycosides from Bidenspilosardquo Phytochemistry vol 48 no 2 pp 397ndash400 1998

[77] P Kumari K Misra B S Sisodia et al ldquoA promising anticancerand antimalarial component from the leaves of Bidens pilosardquoPlanta Medica vol 75 no 1 pp 59ndash61 2009

[78] G Seelinger I Merfort U Wolfle and C M Schempp ldquoAnti-carcinogenic effects of the flavonoid luteolinrdquoMolecules vol 13no 10 pp 2628ndash2651 2008

[79] G Seelinger I Merfort and C M Schempp ldquoAnti-oxidantanti-inflammatory and anti-allergic activities of luteolinrdquoPlanta Medica vol 74 no 14 pp 1667ndash1677 2008

[80] C C Yit and N P Das ldquoCytotoxic effect of butein on humancolon adenocarcinoma cell proliferationrdquo Cancer Letters vol82 no 1 pp 65ndash72 1994

[81] J A Beutler E Hamel A J Vlietinck et al ldquoStructure-activityrequirements for flavone cytotoxicity and binding to tubulinrdquoJournal of Medicinal Chemistry vol 41 no 13 pp 2333ndash23381998

[82] M R Kviecinski K B Felipe T Schoenfelder et al ldquoStudy ofthe antitumor potential of Bidens pilosa (Asteraceae) used inBrazilian folkmedicinerdquo Journal of Ethnopharmacology vol 117no 1 pp 69ndash75 2008

[83] P Sundararajan A Dey A Smith A G Doss M Rajappanand S Natarajan ldquoStudies of anticancer and antipyretic activityof Bidens pilosawhole plantrdquo African Health Sciences vol 6 no1 pp 27ndash30 2006

[84] YMChiang C P Lo Y P Chen et al ldquoEthyl caffeate suppressesNF-120581B activation and its downstream inflammatory mediatorsiNOS COX-2 and PGE2 in vitro or in mouse skinrdquo BritishJournal of Pharmacology vol 146 no 3 pp 352ndash363 2005

50 Evidence-Based Complementary and Alternative Medicine

[85] J S Kim H J Lee M H Lee J Kim C Jin and J HRyu ldquoLuteolin inhibits LPS-stimulated inducible nitric oxidesynthase expression in BV-2 microglial cellsrdquo Planta Medicavol 72 no 1 pp 65ndash68 2006

[86] P A Ruiz and D Haller ldquoFunctional diversity of flavonoidsin the inhibition of the proinflammatory NF-120581B IRF and Aktsignaling pathways in murine intestinal epithelial cellsrdquo Journalof Nutrition vol 136 no 3 pp 664ndash671 2006

[87] A Xagorari A Papapetropoulos A Mauromatis MEconomou T Fotsis and C Roussos ldquoLuteolin inhibitsan endotoxin-stimulated phosphorylation cascade andproinflammatory cytokine production in macrophagesrdquoJournal of Pharmacology and Experimental Therapeutics vol296 no 1 pp 181ndash187 2001

[88] N Yoshida T Kanekura Y Higashi and T Kanzaki ldquoBidenspilosa suppresses interleukin-1120573-induced cyclooxygenase-2expression through the inhibition of mitogen activated proteinkinases phosphorylation in normal human dermal fibroblastsrdquoJournal of Dermatology vol 33 no 10 pp 676ndash683 2006

[89] M Muchuweti C Mupure A Ndhlala T Murenje and M AN Benhura ldquoScreening of antioxidant and radical scavengingactivity of Vigna ungiculata Bidens pilosa and Cleome gynan-drardquo American Journal of Food Technology vol 2 no 3 pp 161ndash168 2007

[90] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens bipinnata L against carbon tetrachloride-induced liver fibrosis in ratsrdquo Journal of Pharmacy and Pharma-cology vol 60 no 10 pp 1393ndash1402 2008

[91] S L Chang H H Yeh Y S Lin Y M Chiang T K Wuand W C Yang ldquoThe effect of centaurein on interferon-120574 expression and Listeria infection in micerdquo Toxicology andApplied Pharmacology vol 219 no 1 pp 54ndash61 2007

[92] F Deba T D Xuan M Yasuda and S Tawata ldquoChemical com-position and antioxidant antibacterial and antifungal activitiesof the essential oils from Bidens pilosa Linn var Radiatardquo FoodControl vol 19 no 4 pp 346ndash352 2008

[93] A O T Ashafa and A J Afolayan ldquoScreening the root extractsfrom Biden pilosa L var radiata (Asteraceae) for antimicrobialpotentialsrdquo Journal of Medicinal Plant Research vol 3 no 8 pp568ndash572 2009

[94] T B Nguelefack T Dimo E P Nguelefack Mbuyo P V TanS V Rakotonirina and A Kamanyi ldquoRelaxant effects of theneutral extract of the leaves of Bidens pilosa linn on isolated ratvascular smooth musclerdquo Phytotherapy Research vol 19 no 3pp 207ndash210 2005

[95] M T Grombone-Guaratini K L Silva-Brandao V N SolferiniJ Semir and J R Trigo ldquoSesquiterpene and polyacetyleneprofile of the Bidens pilosa complex (Asteraceae Heliantheae)from Southeast of Brazilrdquo Biochemical Systematics and Ecologyvol 33 no 5 pp 479ndash486 2005

[96] L L Lin C Y Wu H C Hsiu M T Wang and H ChuangldquoStudies on diabetes mellitus I The hypoglycemic activity ofphytosterin on alloxan diabetic ratsrdquo Taiwan Yi Xue Hui Za Zhivol 66 no 2 pp 58ndash65 1967

[97] M A N Benhura and I C Chitsiku ldquoThe extractable 120573-carotene content of Guku (Bidens pilosa) leaves after cookingdrying and storagerdquo International Journal of Food Science andTechnology vol 32 no 6 pp 495ndash500 1997

[98] J S Chang L C Chiang C C Chen L T Liu K C Wangand C C Lin ldquoAtileukemic activity of Bidens pilosa l varminor (blume) sherff andHouttuynia cordata thunbrdquo AmericanJournal of Chinese Medicine vol 29 no 2 pp 303ndash312 2001

[99] F Deba T D Xuan M Yasuda and S Tawata ldquoHerbicidal andfungicidal activities and identification of potential phytotoxinsfrom Bidens pilosa L var radiata Scherff research paperrdquoWeedBiology and Management vol 7 no 2 pp 77ndash83 2007

[100] J K Kumar and A K Sinha ldquoA new disubstituted acetylacetonefrom the leaves ofBidens pilosa LinnrdquoNatural Product Researchvol 17 no 1 pp 71ndash74 2003

[101] K Ogawa and Y Sashida ldquoCaffeoyl derivatives of a sugarlactone and its hydroxy acid from the leaves of Bidens pilosardquoPhytochemistry vol 31 no 10 pp 3657ndash3658 1992

[102] Q Xia Y Liu and Y Li ldquoDetermination of gallic acid fromdifferent species and different medical parts of Herba Bidens byRP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol24 no 03 pp 308ndash310

[103] W J Lee L F Wu W K Chen C J Wang and T HTseng ldquoInhibitory effect of luteolin on hepatocyte growth fac-torscatter factor-induced HepG2 cell invasion involving bothMAPKERKs and PI3K-Akt pathwaysrdquo Chemico-BiologicalInteractions vol 160 no 2 pp 123ndash133 2006

[104] J A Beutler J H Cardellina C M Lin E Hamel G MCragg andM R Boyd ldquoCentaureidin a cytotoxic flavone fromPolymnia fruticosa inhibits tubulin polymerizationrdquo BioorganicandMedicinal Chemistry Letters vol 3 no 4 pp 581ndash584 1993

[105] A Verma A Su A M Golin B OrsquoMarrah and J K AmorosaldquoThe lateral view a screening method for knee traumardquo Aca-demic Radiology vol 8 no 5 pp 392ndash397 2001

[106] J Corren M Lemay Y Lin L Rozga and R K RandolphldquoClinical and biochemical effects of a combination botanicalproduct (ClearGuard) for allergy a pilot randomized double-blind placebo-controlled trialrdquo Nutrition Journal vol 7 no 1article 20 2008

[107] M S Gachet J S Lecaro M Kaiser et al ldquoAssessment of anti-protozoal activity of plants traditionally used in Ecuador in thetreatment of leishmaniasisrdquo Journal of Ethnopharmacology vol128 no 1 pp 184ndash197 2010

[108] W C Yang M Ghiotto B Barbarat and D Olive ldquoThe role ofTec protein-tyrosine kinase in T cell signalingrdquo The Journal ofBiological Chemistry vol 274 no 2 pp 607ndash617 1999

[109] S Tewtrakul H Miyashiro N Nakamura et al ldquoHIV-1 inte-grase inhibitory substances from Coleus parvifoliusrdquo Phytother-apy Research vol 17 no 3 pp 232ndash239 2003

[110] T Dimo T B Nguelefack P V Tan et al ldquoPossible mechanismsof action of the neutral extract from Bidens pilosa L leaves onthe cardiovascular system of anaesthetized ratsrdquo PhytotherapyResearch vol 17 no 10 pp 1135ndash1139 2003

[111] T S C Li Chinese and Related North American Herbs CRCPress New York NY USA 2002

[112] H Wu H Chen X Hua Z Shi L Zhang and J ChenldquoClinical therapeutic effect of drug-separated moxibustion onchronic diarrhea and its immunologic mechanismsrdquo Journal ofTraditional Chinese Medicine vol 17 no 4 pp 253ndash258 1997

[113] S W Wright R R Harris J S Kerr et al ldquoSynthesis chemicaland biological properties of vinylogous hydroxamic acids dualinhibitors of 5-lipoxygenase and IL-1 biosynthesisrdquo Journal ofMedicinal Chemistry vol 35 no 22 pp 4061ndash4068 1992

[114] W R Almiron and M E Brewer ldquoClassification of immaturestage habitats of Culicidae (Diptera) collected in CordobaArgentinardquo Memorias do Instituto Oswaldo Cruz vol 91 no 1pp 1ndash9 1996

[115] N L Wang J Wang X S Yao and S Kitanaka ldquoTwo newmonoterpene glycosides and a new (+)-jasmololone glucoside

Evidence-Based Complementary and Alternative Medicine 51

from Bidens parvifloraWilldrdquo Journal of Asian Natural ProductsResearch vol 9 no 5 pp 473ndash479 2007

[116] P Champagnat ldquoRole of the terminal bud in the actionexercised by the cotyledon of Bidens pilosus L var radiatus onits axillary budrdquo Comptes Rendus des Seances et Memoires de laSociete de Biologie vol 145 no 17-18 pp 1374ndash1376 1951

[117] S F Nielsen S B Christensen G Cruciani A Kharazmiand T Liljefors ldquoAntileishmaniai chalcones statistical designsynthesis and three-dimensional quantitative structure-activityrelationship analysisrdquo Journal of Medicinal Chemistry vol 41no 24 pp 4819ndash4832 1998

[118] Y C Hwang J J H Chu P L Yang W Chen and M V YatesldquoRapid identification of inhibitors that interfere with poliovirusreplication using a cell-based assayrdquo Antiviral Research vol 77no 3 pp 232ndash236 2008

[119] V F Andrade-Neto M G L Brandao F Q Oliveira et alldquoAntimalarial activity of Bidens pilosa L (Asteraceae) ethanolextracts fromwild plants collected in various localities or plantscultivated in humus soilrdquo Phytotherapy Research vol 18 no 8pp 634ndash639 2004

[120] R J Marles and N R Farnsworth ldquoAnti-diabetic plants andtheir active constituentsrdquo Phytomedicine vol 2 no 2 pp 137ndash189

[121] M Habeck ldquoDiabetes treatments get sweet help from naturerdquoNature Medicine vol 9 no 10 p 1228 2003

[122] H W Lin G Y Han and S X Liao ldquoStudies on the activeconstituents of the Chinese traditional medicine Polygonatumodoratum (Mill) DrucerdquoActa Pharmaceutica Sinica vol 29 no3 pp 215ndash222 1994

[123] C L T Chang Y C Chen H M Chen N S Yang andW C Yang ldquoNatural cures for type 1 diabetes a reviewof phytochemicals biological actions and clinical potentialrdquoCurrent Medicinal Chemistry vol 20 no 7 pp 899ndash907 2013

[124] L Dey A S Attele and C S Yuan ldquoAlternative therapies fortype 2 diabetesrdquo Alternative Medicine Review vol 7 no 1 pp45ndash58 2002

[125] C L T ChangH Y Liu T F Kuo et al ldquoAnti-diabetic effect andmode of action of cytopiloynerdquo Evidence-Based Complementaryand Alternative Medicine vol 2013 Article ID 685642 13 pages2013

[126] M R Khan M Kihara and A D Omoloso ldquoAnti-microbialactivity of Bidens pilosa Bischofia javanica Elmerillia papuanaand Sigesbekia orientalisrdquo Fitoterapia vol 72 no 6 pp 662ndash6652001

[127] T Dimo T B Nguelefack P Kamtchouing E Dongo ARakotonirina and S V Rakotonirina ldquoHypotensive effects ofmethanol extract from Bidens pilosa Linn on hypertensive ratsrdquoComptes Rendus de lrsquoAcademie des Sciences Serie III vol 322no 4 pp 323ndash329 1999

[128] A K Hassan O Deogratius J F Nyafuono O Francis and OP Engeu ldquoWound healing potential of the ethanolic extracts ofBidens pilosa and Ocimum suaverdquo African Journal of Pharmacyand Pharmacology vol 5 no 2 pp 132ndash136 2011

[129] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[130] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[131] C Kandaswami and EMiddleton ldquoFree radical scavenging andantioxidant activity of plant flavonoidsrdquo Advances in Experi-mental Medicine and Biology vol 366 pp 351ndash376 1994

[132] K Yamamoto H Kakegawa H Ueda et al ldquoGastric cytopro-tective anti-ulcerogenic actions of hydroxychalcones in ratsrdquoPlanta Medica vol 58 no 5 pp 389ndash393 1992

[133] J O C Ezeonwumelu A K Julius C N Muhoho et alldquoBiochemical and histological studies of aqueous extract ofBidens pilosa leaves from Ugandan Rift valley in ratsrdquo BritishJournal of Pharmacology and Toxicology vol 2 no 6 pp 302ndash309

[134] L Frida S Rakotonirina A Rakotonirina and J P SavineauldquoIn vivo and in vitro effects of Bidens pilosa L (Asteraceae)leaf aqueous and ethanol extracts on primed-oestrogenized ratuterine musclerdquo African Journal of Traditional Complementaryand Alternative Medicines vol 5 no 1 pp 79ndash91 2008

52 Evidence-Based Complementary and Alternative Medicine

Composition Comments

1 Please provide valid postal code to the address if

possible

2 We made the highlighted change in Table 4 for the sake

of correctness Please check

3 Please check the correctness of the highlighted part in

Table 5

4 We considered the highlighted part in Table 11 as

reference Please check similar cases throughout

5 Please check the format of Table 18

6 We reformatted Table 20 Please check

7 We made the highlighted change in Table 4 for the sake

of consistency Please check similar cases throughout the

paper

8 We made the highlighted change(s) for the sake

of consistency Please check simila highlighted cases

throughout

9 We made the highlighted change in Table 9 for the sake

of clarity Please check similar cases through out

10 We made the highlighted change(s) for the sake of

correctness Please check

11 Comment on ref [66] This reference is a repetition

of [18] Please check similar cases throughout

Author(s) Name(s)

Author 1

Author 2

Author 3

4 Evidence-Based Complementary and Alternative Medicine

Table 3 Ethnomedical information about B pilosa

Disorder Plant partb Mode of use Regioncountry References

Stomachache LE Decoction IndiaAfrica [17ndash19]

Colics WP Decoction AfricaChina [18]

Catarrh WP Juice or decoctiontaken orally Cuba [20]

Diarrhea LE andWP

Fresh leaves orDecoction

AfricaUganda [18 21]

Constipation WP Decoction Africa [18]DysenteryBacillarydysentery WP Decoction Africa

China [18]

Choleretic WP Decoction Middle America [18]

Anti-inflammatory WP Not stated ChinaCuba [20 22]

Asthma WPDecoction ormaceration takenorally

CubaChina [20 23]

Antirheumatic RT andWP Juice and decoction Hong Kong

Zulu Africa [18 22]

Acute appendicitis WP Decoction Hong Kong [23]

Enteritis WP Decoction AfricaChina [18]

Pruritus WP Decoction Hong Kong [23]

Conjunctivitis WP Decoction AfricaChina [18]

Otitis WP Decoction AfricaChina [18]

Pharyngitis WP Decoction AfricaChina [18]

Gastritis WP Juice taken orally Cuba [20]

Diabetes WP Decoction takenorally

CubaTaiwan [20 24]

Headache WP Decoction Bafia Cameroon [25]Diuretic WP Decoction Middle America [18]

Hypotensive WP Decoction takenorally Bafia Cameroon [25]

Colds LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21 23]

Yellow Fever LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21ndash23]

Influenza LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21 23]

Acute infectioushepatitis WP Decoction Hong Kong [23]

Intestinal worms WP Decoction Africa [18]

Malaria RT andWP Juice Africa

China [18 23]

Evidence-Based Complementary and Alternative Medicine 5

Table 3 Continued

Disorder Plant partb Mode of use Regioncountry References

Eye Infection LE andWP Fresh or juice Uganda

Middle America [18 21]

Antimicrobial APDecoction fordrinkingbathingexternal use

Trinidad andTobago [22]

Pulmonarytuberculosis WP

Decoction ormaceration takenorally

CubaChina [20 23]

Bacterial infectionsingastrointestinal tracts

WP Decoction Trinidad andTobago [22]

Renal infection LE Decoction takenorally Cuba [20]

Sore throat LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21 23]

Cough WP Decoction takenorally

CubaChina [20 23]

Coolness of theuterus WP Decoction taken

orally Cuba [20]

Menstrualirregularities WP Decoction taken

orally Cuba [20]

Dysmenorrhea WP Decoction Bafia Cameroon [25]Hyperemesisgravidarum(morning sickness)

WP Decoction Africa [18]

Hemorrhoids WP Decoction Hong Kong [23]

Nose bleeds LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21 23]

Stomach ulcers LE andWP

Maceration or juicetaken orally

CubaMiddle AmericaUganda

[18 20 21 23]

Cuts burns and skinproblems

LE andWP

Fresh plant ordecoction topicalapplicationbathing

Trinidad andTobagoAfricaChinaCameroonBrazilVenezuela

[18 22 26]

Wounds WP Crushed herb

ChinaAfricaCentral AmericaHawaii

[23]

Snake bites WP Pulverized herb China [23]bLE leaves ST stem FW flower WP whole plant RT roots AP aerial parts

primary pharmacological properties of B pilosa extracts andphytochemicals are presented and discussed

31 AntiCancer Activity Folkloric reports revealed the pos-sible antitumor efficacy of B pilosa and several scientific in2vitro studies have supported the claim that B pilosa extractsand isolated compounds possess anti-cancer activities againsta variety of cancer cells Several studies have used bioassay

guided isolation and fractionation methods to discover newcompounds from B pilosa For example Kviecinski andcolleagues tested hydroalcoholic crude extracts chloroformethyl acetate and methanol fractions for anti-tumor activity[82] The cytotoxicity of the extracts was assessed usingbrine shrimp hemolytic MTT and neutral red uptake(NRU) assays In vivo studies were performed using Ehrlichascites carcinoma in isogenic BALBc mice Among them

6 Evidence-Based Complementary and Alternative Medicine

Table 4 Aliphatic natural products isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

1 Heneicosane CH3(CH2)19CH3 Aerial (Tanzania) [18]2 Docosane CH3(CH2)20CH3 Aerial (Tanzania) [18]3 Tricosane CH3(CH2)21CH3 Aerial (Tanzania) [18]4 Tetracosane CH3(CH2)22CH3 Aerial (Tanzania) [18]5 Pentacosane CH3(CH2)23CH3 Aerial (Tanzania) [18]6 Hexacosane CH3(CH2)24CH3 Aerial (Tanzania) [18]7 Heptacosane CH3(CH2)25CH3 Aerial (Tanzania) [18]

8 Octacosone CH3(CH2)26CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

9 Nonacosane CH3(CH2)27CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

10 Triacontane CH3(CH2)28CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

11 Hentriacontane CH3(CH2)29CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

12 Dotriacontane CH3(CH2)30CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

13 Tritriacontane CH3(CH2)31CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

14 2-Butoxy-ethanol CH3(CH2)3OCH2CH2OH Whole (Taiwan) [32]15 Tetracosan-1-ol CH3(CH2)22CH2OH Aerial (Tanzania) [18]16 Hexacosan-1-ol CH3(CH2)24CH2OH Aerial (Tanzania) [18]17 1-Octacosanol CH3(CH2)26CH2OH Aerial (Tanzania) [18]

18 1-Hentriacontanol CH3(CH2)29CH2OHNot found(Taiwan) [31]

19 Tetradecanoic acid Myristic acid CH3(CH2)12CO2H Aerial (Tanzania) [18]20 Hexadecanoic acid Palmitic acid CH3(CH2)14CO2H Aerial (Tanzania) [18]21 Octadecanoic acid Stearic acid CH3(CH2)16CO2H Aerial (Tanzania) [18]22 Eicosanoic acid Arachidic acid CH3(CH2)18CO2H Aerial (Tanzania) [18]

23 Docosanoic acid Behenic acid CH3(CH2)20CO2HLeaves(Philippines) [33]

24 2-butenedioic acidO

OHO

HO Aerial (China) [34 35]

25 (Z)-9-Octadecenoicacid Oleic acid

O

OH Aerial (Tanzania) [18]

Evidence-Based Complementary and Alternative Medicine 7

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

26 (E)-9-Octadecenoicacid Elaidic acid

O

HOLeaves(Philippines) [33]

27 (ZZ)-912-Octadecadienoic acid

Linolic acid or Linoleicacid

O

OHAerial(Tanzania)whole (Taiwan)

[18 36]

28(ZZZ)-91215-Octadecatrienoicacid

120572-Linoleic acid

O

OH Whole (Taiwan) [32]

29(ZZ)-912-Octadecadienoic acidethyl ester

Ethyl linoleate O

O

Whole (Taiwan) [32]

30(ZZZ)-91215-Octadecatrienoicacid methyl ester

Methyl linolenate O

O

Whole (Taiwan) [32]

31(ZZZ)-91215-Octadecatrienoicacid ethyl ester

Ethyl linolenate O

O

Whole (Taiwan) [32]

32(Z)-9-Octadecenoicacid 2-butoxyethylester

2-Butoxyethyl oleate O

OO

Whole (Taiwan) [32]

33 2-Butoxyethyllinoleate O

OO Whole (Taiwan) [32]

34(ZZZ)-91215-Octadecatrienoicacid butoxyethyl ester

2-Butoxyethyl linoleate O

OO

Whole (Taiwan) [32]

3517E9E15E-Heptadecatetraene-1113-diyne

Heptadeca-2E8E10E16-tetraen-46-diyne

Not found(China) [37]

8 Evidence-Based Complementary and Alternative Medicine

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

36 111-Tridecadiene-3579-tetrayne

Roots (notstated) [38]

37 1-Tridecene-357911-pentayne Pentayneene

Leaves (notstated) andnot found(Egypt)

[38 39]

38 5-Tridecene-7911-triyne-3-ol

HO

not found(Egypt) [39]

39 21012-Tridecatriene-468-triyn-1-ol

OHPart not specified(Not stated) [40]

40 212-Tridecadiene-46810-tetrayn-1-ol

111-Tridecadiene-3579-

tetrayn-13-ol

OH Roots (Notstated) notfound(Egypt)

[38 39]

41 212-Tridecadiene-46810-tetraynal

111-Tridecadiene-3579-

tetrayn-13-al

O

Roots (Germany) [41]

42212-Tridecadiene-46810-tetrayn-1-ol1-acetate

111-Tridecadiene-3579-

tetrayn-13-acetateO

O

Roots (notstated) [38]

43(5E)-15-Tridecadiene-79-diyn-3413-triol OH

OH

HO

Aerial (China) [42]

44(6E12E)-3-oxo-tetradeca-612-dien-810-diyn-1-ol

OH

O

Aerial (China) [42]

45 (E)-5-Tridecene-7911-triyne-12-diol

12-Dihydroxy-5(E)-tridecene-7911-triyne

HO

OHWhole (Taiwan) [43]

46 (E)-6-Tetradecene-81012-triyne-13-diol

13-Dihydroxy-6(E)-tetradecene-81012-triyne

OH

OHWhole (Taiwan) [43ndash45]

47(2R3E11E)-311-Tridecadiene-579-triyne-12-diol

Safynol

OH

HO

Not found (Egyptand China) [37 39]

Evidence-Based Complementary and Alternative Medicine 9

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

4857911-Tridecatetrayne-12-diol

12-Dihydroxytrideca-57911-tetrayne

HOOH

Whole (Taiwan) [43 44 46]

49(R)-357911-Tridecapentayne-12-diol

(R)-12-Dihydroxytrideca-357911-pentayne

HO

OH

Aerial (Japan) [47]

50

(4E)-1-(Hydroxymethyl)-4-dodecene-6810-triyn-1-yl-120573-D-glucopyranoside

2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-

tridecene-7911-triyne

O

HO

H

OH

H

H

HOH

O

OH

HO

H Aerial (USA)whole (Taiwan)leaves (Taiwan)

[8 45 48ndash51]

51

(4E)-1-(2-Hydroxyethyl)-4-dodecene-6810-triyn-1-yl-120573-D-glucopyranoside

3-120573-D-Glucopyranosyloxy-1-hydroxy-6(E)-

tridecene-81012-triyne

OHO

H

OH

H

H

HOH

O

OH

OH

HAerial (USA andChina) whole(Taiwan) leaves(Taiwan)

[8 35 4245 48ndash51]

52

3-Hydroxy-6-tetradecene-81012-triynyl-120573-D-glucopyranoside

120573-D-Glucopyranosyloxy-

3-hydroxy-6E-tetradecence-81012-triyne

OH

O

OHO

H

OH

H

H

HOH

OH

HWhole (Mexico) [52]

53

1-(Hydroxymethyl)-46810-dodecatetrayne-1-yl-120573-D-glucopyranoside

Cytopiloyne2-120573-D-

Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne

O

HO

OHO

H

OH

H

H

HOH

OH

HWhole (Taiwan)leaves (Taiwan) [8 50 53]

54

2-O-D-Glucosyltrideca-11E-en-3579-tetrayn-12-diol

OH

O

O

OHH

HO

H

H

HHOH

OH

Leaves (Brazil) [27]

10 Evidence-Based Complementary and Alternative Medicine

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

55

(R)-1-(Hydroxymethyl)-246810-dodecapentayn-1-yl-a-D-glucopyranoside

2- 120573 -D-Glucopyranosyloxy-1-hydroxytrideca-357911-pentayne

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Chinaand Japan) [35 47]

56

1-[[(Carboxyacetyl)oxy]methyl]46810-dodecatetraynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Japan) [54]

57

(4E)-1-[[(Carboxyacetyl)oxy]-methyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHO

H

OH

Aerial (Japan) [54]

58

(4E)-1-[[(Carboxyacetyl)oxy]-ethyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

O

O

O

OHO

O

OHOH

H

HO

H

H

HHO

HOH

Aerial (Japan) [54]

59 (5E)-5-Heptene-13-diyn-1-yl-benzene

1-Phenylhepta-13-diyn-5en Whole (Taiwan) [32]

60 7-Phenyl-2(E)-heptene-46-diyn-1-ol

OHRoots (notstated) Aerial(China)

[38 42]

617-Phenyl-2(E)-heptene-46-diyn-1-ol-acetate

OO

Roots (notstated Brazil) [38 55 56]

62 7-Phenyl-46-heptadiyn-2-ol Pilosol A

OH

Whole (Taiwan)aerial (China) [42 57]

Evidence-Based Complementary and Alternative Medicine 11

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

63 7-Phenylhepta-46-diyn-12-diol HO

OH

Aerial (China) [42]

64 135-Heptatriyn-1-yl-benzene

1-Phenylhepta-135-triyne

Leaves (notstated) leaves oftissue culture(not stated)aerial (TanzaniaChina)Whole (Taiwan)roots (Brazil)

[18 34 3842 56ndash58]

65 7-Phenyl-246-heptatriyn-1-ol HO

Leaves (notstated) aerial(China)

[38 42]

667-Phenyl-246-heptatriyn-1-ol-acetate O

O Leaves (notstated) [38]

675-(2-Phenylethynyl)-2-thiophenemethanol

S OHAerial (China) [42]

685-(2-Phenylethynyl)-2120573-glucosylmethyl-thiophene

O

O

OH

H

OH

H

H

HOH H

O

HO

S

Aerial (China) [42]

69

3- 120573 -D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne

OH

O

OH

HOHO

H

H

HHOH

OH

Leaves(Cameroon) [28]

701-Phenyl-13-diyn-5-en-7-ol-acetate O

O

Leaves (Brazil) [27]

SN denotes serial number

the chloroform fraction was the most toxic with a halfmaximal inhibitory concentration (IC

50) of 97 plusmn 72 and 83 plusmn

52 120583gmL in NRU and MTT respectively [82] Kumari andcolleagues also reported the anti-cancer and anti-malarialactivities of B pilosa leaves [77] Based on a cytotoxicity-directed fractionation strategy they identified phenyl-135-heptatriene with IC

50values of 8plusmn001 049plusmn045 07plusmn001

and 10 plusmn 001 120583gmL against human oral liver colon andbreast cancer cell lines respectively However phenyl-135-heptatriyne showed lower activity against breast cancer celllines than the chloroform leaf extract which had an IC

50

value of 65 plusmn 001 120583gmL Moreover the positive controltaxol showed higher activity than phenyl-135-heptatriyne[77] Furthermore in vitro comet assays were performed toevaluate the toxicity of n-hexane chloroform and methanolextracts of B pilosa and its ethyl acetate acetone and waterfractions onHela andKB cellsThe ethyl acetate fraction fromthe methanol extract exhibited the highest activity with halfmaximal cytotoxic concentrations (CTC

50) of 9652120583gmL

and 5862120583gmL against Hela and KB cells respectivelyDespite the moderate toxicity these findings suggest thatthese B pilosa extractsfractions could be useful for future

12 Evidence-Based Complementary and Alternative Medicine

Table5Flavon

oids

isolated

from

Bpilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

71

2-[(34-Dihydroxyph

enyl)-

methylene]-

6-hydroxy-3(2H

)-benzofuranon

e

Sulfu

retin

O

OHO

OH

OH

Aeria

l(Ch

ina)

[35]

72

2-[(34-Dihydroxyph

enyl)-

methylene]-

67-dihydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-673101584041015840-

tetrahydroxy

maritimetin

O

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

73

2-[(34-Dihydroxyph

enyl)-

methylene]-6-

(120573-D

-glucopyrano

syloxy)-7-

hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-120573-D

-glucop

yranosyl-

673101584041015840-te

trahydroxy

maritimein

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japanand

China)aerial(Ch

ina)

[355960]

74

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

(120573-D

-glucopyrano

syloxy)-

6-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-7-O-120573-D

-glucop

yranosyl-

673101584041015840-

tetrahydroxy

O

OHO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japan)

[59]

75

6-[(6-O-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-acetyl-

120573-D

-glucopyrano

syl)

-673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OHO

H HO

H

H

HOH

HO

O

O

Leaves

(Not

stated)

aeria

l(Ch

ina)

[3559]

Evidence-Based Complementary and Alternative Medicine 13

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

76

6-[(36-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-

2-[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone(Z)-6-O

-(36-di-O

-acetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

bideno

sideA

O

O

HO

OH

OH

OHOH O

H

H

HOH

HO

O

O

O

Leaves

(China)

[60]

77

6-[(46-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(410158401015840610158401015840-

diacetyl-120573-D

-glucropyrano

syl)-

673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

HO

O

OO

Leaves

(not

stated)

aeria

l(Ch

ina)

[343561]

78

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

hydroxy-6-

[(246-tri-O

-acetyl-120573

-D-

glucop

yranosyl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(210158401015840410158401015840610158401015840-

triacetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

O

OOO

O

Leaves

(not

stated)

aeria

l(Ch

ina)

[3461]

79

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-6-

[(346-tri-O

-acetyl-120573

-D-glucopyrano

syl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(310158401015840410158401015840610158401015840-

triacetyl-120573

-D-glucrop

yranosyl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H OH

H

HOH

HO

OOO O

Aeria

l(Ch

ina)

[3435]

80

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-

6-[[6-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propenyl]-120573-

D-glucopyrano

syl]o

xy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-

p-coum

aroyl-

120573-D

-glucropyrano

syl)-

673101584041015840-te

trahydroxy

OH

O

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OLeaves

(Japan)

[59]

14 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

81

1-[2-(120573-D

-Glucopyrano

syloxy)-

4-hydroxyphenyl]-2-

hydroxy-3-(3-

hydroxyphenyl)-2-

prop

en-1-on

e

Chalcone120572

32101584041015840-

tetrahydroxy-

21015840-O

-120573-D

-glucop

yranosyl

HO

O

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[35]

821-(24-Dihydroxyph

enyl)-

3-(34-dihydroxyph

enyl)-

2-prop

en-1-on

eBu

tein

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

833-(34-D

ihydroxyph

enyl)-

1-(234-trihydroxyph

enyl)-

2-prop

en-1-on

eOkanin

O

HO

HO

OH

OH

OH

Leaves

(China)

[60]

84

3-(34-D

ihydroxyph

enyl)-

1-[3-(120573-D

-glucop

yranosyloxy)-

24-dihydroxyphenyl]-2-

prop

en-1-on

e

Okanin

31015840-O

-120573-D

-glucosid

e

O

HO

OH

OH

OH

OO

HO

H

HO

H

H

HOH

H

OH

Leaves

(Germany)

flowers(Germany)

[61ndash63]

85

3-(34-D

ihydroxyph

enyl)-

1-[4-(120573-D

-glucop

yranosyloxy)-

23-dihydroxyphenyl]-

2-prop

en-1-on

e

Okanin

41015840-O

-120573-D

-glucosid

e

OOH

OH

OH

HOO

OHO

H

HO

H

H

HOH

H

OH

Flow

ers(Germany)

leaves

(Japan)

[5963]

Evidence-Based Complementary and Alternative Medicine 15

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

86Okanin41015840-O

-120573-D

-(610158401015840-

O-acetylglucosid

e)

O

O

OH

OH

OH

OHO

H

HO

H

H

HOH

H

HO

O

O

Flow

ers(Germany)

[63]

87

1-[4-[(46-di-O

-Acetyl-

120573-D

-glucopyrano

syl)-oxy]-

23-dihydroxyphenyl]-3-

(34-dihydroxyph

enyl)-2-

prop

en-1-on

e

Okanin

41015840-O

-120573-D

-(410158401015840610158401015840-diacetyl)-

glucop

yranoside

O

O

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Aeria

l(Ch

ina)

[34]

88Okanin41015840-O

-120573-D

-(210158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[64]

89Okanin41015840-O

-120573-D

-(310158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

O

OAe

rial(Ch

ina)

[34]

16 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

90

1-[23-Dihydroxy-4-

[[6-O-[3-(4-hydroxyph

enyl)-

1-oxo-2-prop

enyl]-120573-D

-glucop

yranosyl]oxy]-ph

enyl]-

3-(34-dihydroxyph

enyl)-

2-prop

en-1on

e

Okanin

41015840-O

-120573-D

-(610158401015840-tr

ans-

p-coum

aroyl)-

glucoside

O

O

OH

OH

OH

OHOH

HO

H

H

HOH

H

HO

OO

OH

Leaves

(Germany)

[64]

91Okanin41015840-O

-120573-D

-(410158401015840-acetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Leaves

(Germany)

[65]

92

Okanin

41015840-O

-120573-D

-(210158401015840410158401015840-diacetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[65]

Evidence-Based Complementary and Alternative Medicine 17

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

93

Okanin

41015840-O

-120573-D

-(310158401015840410158401015840-diacetyl-610158401015840-

trans-p-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

OH

O O

Leaves

(Germany)

[65]

94

Okanin

41015840-O

-[120573-D

-glucopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

OOH

OH

OH

HO

OH

HO

HHO

H

HOH

HO

OH

OH

HO

HHO

H

HOH

HO

Flow

er(G

ermany)

[63]

95Okanin

3101584041015840-di-O

-120573-D

-glucosid

e

OOH

OH

OH

O OO

H HO

HHOH

HOH

HHO

OHO

H

HO

H

H

HOH

H

OH

Flow

er(G

ermany)

[63]

18 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

96

1-[3-(120573-D

-Glucopyrano

syloxy)-

24-dihydroxyphenyl]-

3-(3-hydroxy)-

4-metho

xyph

enyl)-

2-prop

en-1-on

e

Okanin

4-methylether-

31015840-O

-120573-D

-glucop

yranoside

OOH

O

OH

HO

O

H HO

HHOH

HOH

H

OHO

Leaves

(Germany)aerial

(China)

[3562]

97

Okanin4-methylether-

3101584041015840-di-O

-120573-(41015840101584061015840101584041015840101584010158406101584010158401015840-

tetraacetyl)-

glucop

yranoside

OOH

OOH

O

HH

OH

HO

HOH

H

OO

O

Ac

Ac

AcAc

OO

H

HO

H

H

HOH

H

O

Aeria

l(Ch

ina)

[42]

98Ch

alcone210158404101584061015840-tr

imetho

xy-

4-O-D

-glucopyrano

syl-

dihydro

Not

foun

dLeaves

(China)

[60]

992-(34-D

ihydroxyph

enyl)-23-

dihydro-78

-dihydroxy-

4H-1-

benzop

yran-4-one

Okanin

isoOO

OH

HO

OH

OH

Leaves

(China)

[60]

100

2-(34-D

ihydroxyph

enyl)-

23-dihydro-8-hydroxy-

7[(246-tr

i-O-acetyl-120573

-D-

glucop

yranosyl)oxy]-4H

-1-b

enzopyran-4-on

eOO

OH

OH

OH

OH

O

HHO

H

HO

HO

OAc

Ac

Ac

Aeria

l(Ch

ina)

[34]

Evidence-Based Complementary and Alternative Medicine 19

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

101

57-Dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

OO

OH

HO

OH

Aeria

l(TanzaniaC

hina)

[4266]

102

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

7-O-glucopyrano

side

OO

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Tanzania)

[66]

103

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-4H

-1-benzop

yran-4-one

Luteolin

OO

HO

OH

OH

OH

Aeria

l(TanzaniaC

hina

Vietnam)

[343542

6667]

104

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5hydroxy-4H-1-

benzop

yran-4-one

Luteolin

7-O-120573-D

-glucop

yranoside

OOOH

OH

OH

OO

HO

H

HO

H

H

HOH

OH H

Aeria

l(Tanzania)

[66]

105

57-Dim

etho

xy-6-

(5-m

etho

xy-6-m

ethyl-

4-oxo-4H

-pyran-3-yl)-

2-ph

enyl-4H-1-

benzop

yran-4-one

5-O-M

ethylhosludin

O OOO

O

O

O

Aeria

l(Ugand

a)[68]

20 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

106

3-(120573-D

-Glucopyrano

syloxy)-

57-dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Astragalin

Kaem

pferol-3-O

-120573-D

-glucopyrano

side

HO

OH

O O

OH

O

O

HO

HHO

H

H

HOH

OH

H

Aeria

l(Ch

ina)

[35]

107

Kaem

pferol3-

(23-di-E

-p-cou

maroyl-

120572-L-rhamno

pyrano

side)

Not

foun

dAe

rial(Vietnam)

[67]

108

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5-hydroxy-36-dimetho

xy-

4H-1-

benzop

yran-4-one

Axillarosid

eO O

OH

OH

OO

OH

OO

HO

H

HO

H

H

HOH

H

OH

Aeria

l(Ch

ina)

[42]

109

57-Dihydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaureidin

O

O

O

O

O

HO

OH

OH

Who

le(Taiwan)

[69]

110

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaurein

O O

OO

O OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan)w

hole

(Taiwan)

[69ndash

71]

Evidence-Based Complementary and Alternative Medicine 21

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

111

5-Hydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-

67-dimetho

xy-4H-

chromen-4-one

Eupatorin

iso

OO O

OH

O

O OH

Not

foun

d(C

hina)

[3772]

112

2-(34-D

imetho

xyph

enyl)-7-

(120573-D

-glucopyrano

syloxy)-35-

dihydroxy-

8-metho

xy-4H-1-

benzop

yran-4-one

O O

OH

O O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

113

7-(120573-D

-Glucopyrano

syloxy)-

5hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-38-

dimehtoxy-4H-1-

benzop

yran-4-one

O O

O

OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

114

Isorhamnetin

3-[O

-120572-

L-rham

nopyrano

syl-

(1-2)-120573-D

-glucop

yranoside]

Not

foun

dAe

rial(Vietnam)

[67]

115

7-[(6-deoxy-120572-L-

Manno

pyrano

syl)o

xy]-

3-120573-D

-glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-4H

-1-benzop

yran-4-one

Luteoside

O O

OH

O

OH

OH

OH

HOH

H

H HO

HO

HO

OH

HO

H

HO

OH

OH

HH

Aeria

l(Ch

ina)

[42]

22 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

116

Luteolin

3-O-120573-D

-Glucopyrano

side

O O

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

HO

Aeria

l(Tanzania)

[66]

117

57-Dihydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Quercetagetin

3631015840-tr

imethylether

OOO

O

O OH

HO

OH

Aeria

l(Ch

ina)

[42]

118

6-((2S3S4S5S)-

Tetrahydro-345-tr

ihydroxy-

6-(hydroxymethyl)-2H

-pyran-2-yloxy)-5-hydroxy-

2-(4-hydroxy-3-

metho

xyph

enyl)-

37-dimetho

xy-4H-

chromen-4-one

Quercetagetin

3731015840-

trim

ethylether-6-O

-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[42]

119

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

JaceinQ

uercetin

3631015840-

trim

ethyl

ether-7-O-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan

China)

who

le(Taiwan)

[425470]

120

2-(34-D

ihydroxyph

enyl)-

357-trihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

OO

HO

OH

OH

OH

OH

Aeria

l(Ch

ina)leaves

(China)who

le(C

hina)

[356074]

Evidence-Based Complementary and Alternative Medicine 23

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

121

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-galactopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-4-one

Quercetin

3-O-120573-D

-galactosidehyperin

hyperosid

e

O O

HO

OH

OH

OH

OOHH

HOH

H

H HO

HO

OH

Aeria

l(TanzaniaJapan)

Leaves

(China)Who

le(C

hina)

[376066

717475]

122

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-glucopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

3-O-120573-

D-glucopyrano

side

O O

HO

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

Aeria

l(TanzaniaC

hina

Japan)L

eaves(Japan)

[355966

75]

123

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-

4-oxo-4H

-1-benzop

yran-3-yl-

120573-D

-glucopyrano

siduron

icacid

Quercetin

3-O-120573-D

-glucuron

opyranoside

O O

HO

OH

OH

OH

OHOH

HOH

H

H HO

HO

OHO

Aeria

l(TanzaniaJapan)

[6671]

124

3-[[6-O-(6-

Deoxy-120572-L-m

anno

pyrano

syl)-

120573-D

-galactopyrano

syl]o

xy]-

2-(34-dihydroxyph

enyl)-57-

dihydroxy-4H

-1-benzop

yran-4-one

Quercetin

3-O-rob

inob

iosid

e

O O

HO

OH

OH

OH

O

HOHH OH

HO

H HH

OO

OHH

HOH

H

H HO

HO

Aeria

l(Japan)W

hole

(Taiwan)

[7071]

24 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

125

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-

3-metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

OH

Roots(Brazil)

[55676]

126

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-

120573-D

-glucopyrano

syl]o

xy]-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120572-

L-rham

nopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

O

HHO HHO

HO H

HH

ORo

ots(Brazil)

[5676]

127

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-120573-D

-glucop

yranosyl]oxy]-5-

hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-3-metho

xy-

4H-1-

benzop

yran-4-one

Quercetin

341015840-

dimethyl

ether-7-O-rutinoside

O OOH

OH

O

O

OH O

HOH

H

HHO

H

O

HHO HHO

HO H

HH

O

HO

Aeria

l(Ch

ina)

[3435]

128

2-(34-D

ihydroxyph

enyl)-3-

(120573-D

-glucofurano

syloxy)-57-

dihydroxy-4H

-1-benzop

yran-

4-on

e

Isoq

uercitrin

O OOH

OH

OHO

H

OHH

OH

HO

H O HOH

OH

Aeria

l(Japan

China)

[3571]

Evidence-Based Complementary and Alternative Medicine 25

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

129

Quercetin

3-O-120573-D

-galactop

yranoside

O

OH

HO

OH OH

O

O

HO

OH

OHOH

OAe

rial(no

tstated)

[18]

130

Quercetin

3-O-rutinoside

O

O

R

OO

OH

OHO

H

O

OH

OH

H3C

H3C

H3C

OH

HO

Who

le(Taiwan)

[70]

26 Evidence-Based Complementary and Alternative Medicine

Table 6 Terpenoids isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(Country) References

131371111-Tetramethylbicyclo[810]undeca-

26-dieneBicyclogermacrene Leaves

(Brazil) [95]

13241111-Trimethyl-8-

methylenebicyclo[720]undec-4-ene

E-Caryophyllene Leaves(Brazil) [95]

1331-Methyl-5-methylene-8-

(1-methylethyl)-16-cyclodecadiene

Germacrene D Leaves(Brazil) [95]

1344-(15-Dimethyl-4-hexen-1-ylidene)-1-methyl-cyclohexene

Z-120574-Bisabolene Leaves(Brazil) [95]

135Decahydro-114-trimethyl-7-methylene-1H-cycloprop[e]-

azulene120573-Gurjunene Leaves

(Brazil) [95]

136 2669-Tetramethyl-148-cycloundecatriene

120572-Humulene120572-caryophyllene

Leaves(Brazil) [95]

13712344a568a-Octahydro-1-isopropyl-7-methyl-4-methylenenaphthalene

120575-Muurolene Leaves(Brazil) [95]

138

12344a568a-Octahydro-4a8-dimethyl-2-(1-methylethylidene)-

naphthalene

Selina-37(11)-diene

Leaves(Brazil) [95]

139(2E7R11R)-371115-

Tetramethyl-2-hexadecen-1-ol

Phytol

HO

Whole(Taiwan) [57]

Evidence-Based Complementary and Alternative Medicine 27

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

140 371115-Tetramethyl-2-hexadecenoic acid Phytanic acid

O

OH

Whole(Taiwan) [57]

141371115-Tetramethyl-2-

hexadecenyl ester-heptanoicacid

Phythylheptanoate O O

Leaves(Not stated) [33]

142

(3S10R13R)-17-((2R5R)-5-Ethyl-6-methylheptan-2-yl)-

2347891011121314151617-tetradecahydro-

1013-dimethyl-1H-cyclopenta[a]phenanthren-

3-ol

Campestrol

HO

Aerial(Tanzania) [18]

143 Not found Phytosterin-B Not foundNot found(TaiwanEgypt)

[39 96]

144 Stigmast-5-en-3-ol 120573-sitosterol

HOH

H

H

Aerial(Tanzania)

whole(Taiwan)

[18 31 57]

145

1314151617-Tetradecahydro-1013-

dimethyl-1H-cyclopenta[a]phenanthren-

3-yloxy)-tetrahydro-6-(hydroxymethyl)-2H-pyran-345-triol

120573-Sitosterolglucoside

H

H

HOH

HO

HHO

H

HOHH O

OH Not found(Egypt) [39]

146 5120572-Stigmasta-7-en-3120573-ol

HO

Whole(Taiwan) [57]

147 5120572-Stigmasta-722t-dien-3120573-ol

HO

Whole(Taiwan) [57]

148 Stigmasta-522-dien-3-ol Stigmasterol

HOH

H

H

Not found(Taiwan)aerial

(Tanzania)leaves (notstated)whole

(Taiwan)

[18 31 3357]

28 Evidence-Based Complementary and Alternative Medicine

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

149 Lup-20(29)-en-3-ol Lupeol

H

HH

HOH

Not found(Egypt) [39]

150 Lup-20(29)-en-3-ol acetate Lupeol acetate

H

HH

O

OH

Not found(Egypt) [39]

151 Olean-12-en-3-ol 120573-amyrin

H

HOH

Not found(Egypt) [39]

152 5913-Trimethyl-242526-trinoroleanan-3-ol Friedelan-3120573-ol

HO

Aerial(Tanzania) [18]

153 5913-Trimethyl-242526-trinoroleanan-3-one

Friedelinfriedelan-3-one

O

Aerial(Tanzania) [18]

1542610151923-

Hexamethyl-2610141822-tetracosahexaene

Squalene

Aerial(Tanzania)Leaves (Notstated)Whole(Taiwan)

[18 33 57]

155 120573120573-Carotene 120573-Carotene Leaves (Notstated) [97]

Evidence-Based Complementary and Alternative Medicine 29

studies [83] Hot water extracts of B pilosa varminor Sheriffwere also assessed for its antileukemic effects on leukemiccell lines L1210 U937 K562 Raji and P3HR1 using XTT-based colorimetric assays The extract inhibited the five celllines with IC

50values ranging from 145120583gmL to 586120583gmL

L1210 K562 Raji and P3HR1 were more sensitive to B pilosaextract with IC

50values below 200120583gmL [98]

Consistent with the antitumor activities of B pilosaextracts and fractions some of its phytochemicals alsoshowed anticancer activity as outlined in Table 11 Amongthem luteolin (103) a well-studied flavonoid with multiplebioactivities was more effective against tumor cell prolifera-tion than its derivatives with IC

50values ranging from 3 120583M

to 50120583M in cells and 5 to 10mgkg in animals Luteolin wasalso found to fight cancer as a food additive at concentrationsof 50 to 200 ppm [78] and prevent skin cancer [78] and cancer3invasion [103] Lee and colleagues reported that luteolinprevents cancer by inhibiting cell adhesion and invasion[103] Significant inhibition concentration was reported to be5 120583M and complete inhibition concentration was reported tobe 40120583M Moreover luteolin was reported to inhibit hepa-tocyte growth factor (HGF)-induced cell scattering as wellas cytoskeleton changes such as filopodia and lamellipodiawhichwas determined using phase-contrast and fluorescencemicroscopy Furthermore luteolin also inhibited the HGF-induced phosphorylation of c-Met ERK 12 and Akt aswell as the MAPKERK and P13K-Akt pathways [78 103]Other mechanisms underlying the anticancer activities ofluteolin are the inhibition of topoisomerase I and II whichinhibits cell replication and DNA repair thus promotingapoptosis regulation of PI-3-KinaseAktMAPKERKJNKactivation of apoptosis in the mitochondrial pathway byactivating caspase 9 and caspase 3 which were found inmalignant cells but not in normal human peripheral bloodmononuclear cells death receptor-induced apoptosis and acell-cycle arrest mechanism inhibition of fatty acid synthasewhich is upregulated in many cancer cells and sensitizationto chemotherapywhereby luteolin increases the susceptibilityof cancer cells to chemotherapy [78] Butein (82) is anotherflavonoid that showed a cytotoxic effect on human colonadenocarcinoma cell proliferation with a reported IC

50value

of 175 120583M Butein at 2 120583M affected the incorporation of[14C]-labeled leucine thymidine and uridine which cancause the inhibition of DNA RNA and protein synthesis ofhuman colon cancer cells Moreover butein also exhibitednoncompetitive inhibition of 1-chloro-24-dinitrobenzene(CDNB) in glutathione S-transferase (GST) activity Tumorresistance was correlated with high levels of GST thus buteininhibited proliferation of cancer cells [80] Another flavonoidpresent in B pilosa centaureidin (109) also showed anti-cancer activity in B lymphoma cells Centaureidin isolatedfrom Polymnia fruticosa inhibited tubulin polymerization invitro and induced mitotic figure formation in CA46 Burkittlymphoma cells Using turbidimetric assay the IC

50value of

centaureidin in inhibition of mitosis was 3120583M [104] Cyto-toxicity of centaureidin was further analyzed using AmericanNational Cancer Institute (NCI) 60 human tumor cell linesThe cytotoxicity potency of centaureidin expressed as GI

50

(50 growth inhibition in the NCI tumor line panel) was024120583M [81] These data mark centaureidin as a promisingantimitotic agent for tumor therapy

In addition to anti-tumor flavones polyynes found in B 4pilosa have also been shown to possess anti-tumor proper-ties Based on a bioactivity-directed isolation approach Wuand colleagues identified two polyyne aglycones from theethyl acetate fraction of B pilosa [44] 12-Dihydroxytrideca-57911-tetrayne (48) and 13-Dihydroxy-6(E)- tetradecene-81012-triyne (46) exhibited significant anticell prolifera-tion activity in primary human umbilical vein endotheliumcells (HUVEC) with IC

50values of 125 120583M and 173 120583M

respectively They also decreased angiogenesis and pro-moted apoptosis in human endothelial cells Their anti-angiogenic and cytotoxic effects correlated with activationof the CDK inhibitors and caspase-7 [44] In addition 12-Dihyroxy-5(E)-tridecene-7911-triyne (45) showed antian-giogenic effects in HUVECs with an IC

50value of 124 120583M as

evidenced by a decrease in the tube formation and migrationof HUVECs [43] The IC

50value of compound 45 in the

inhibition of basic fibroblast growth factor-induced HUVECgrowth was 282120583M However it had higher IC

50values

than those for lung carcinoma cells and keratinocytes Thiscompound could also inhibit cell proliferation of HUVECslung carcinoma A549 cells and HACAT keratinocytes Themechanism by which compound 45 inhibits HUVEC growthand angiogenesis is complicated and includes decreasing theexpression of cell cycle regulators (CDK4 cyclins D1 and Aretinoblastoma (Rb) and vascular endothelial growth factorreceptor 1) caspase-mediated activation of CDK inhibitorsp21 (Cip1) and p27 (Kip) upregulation of Fas ligand expres-sion downregulation of Bcl-2 expression and activation ofcaspase-7 and poly (ADP-ribose) polymerase [43]

32 Anti-Inflammatory Activity B pilosa is commonly usedto treat inflammatory disorders The anti-inflammatory phy-tochemicals present in B pilosa are listed in Table 11 5Cyclooxygenase-2 (COX-2) is a physiologically importantenzyme that converts arachidonic acid to prostaglandin(PGE2) Its expression is induced by a wide variety of external

stimuli indicating its involvement in inflammatory diseasesand it is used as an inflammatory marker [88] Yoshida andcolleagues studied the effects of the aqueous extracts of Bpilosa aerial parts in the production of COX-2 and PGE

2as

well as on the activation of mitogen activated protein kinases(MAPKs) in normal human dermal fibroblasts (HDFs) inresponse to inflammatory cytokine IL-1120573 This work showedthat IL-1120573 activated MAPKs such as ERK12 p38 and JNKto different extents and induced COX-2 expression TheCOX-2 expression in HDFs was regulated mainly by p38following IL-1120573 stimulation Consistently the p38 inhibitorSB203580 blocked this expression Using this cell platformB pilosa extracts were tested for inhibition of inflammationThe extract dose-dependently suppressed the activation ofp38 and JNK and moderately suppressed ERK12 as wellas suppressing COX-2 expression and PGE

2production

30 Evidence-Based Complementary and Alternative Medicine

Table7Ph

enylprop

anoids

isolatedfro

mB

pilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

156

3-(4-H

ydroxyph

enyl)-2-prop

enoica

cid

p-Cou

maricacid

OHO

OH

Who

le(Ja

pan)

[99]

157

2-Metho

xy-4-(2-prop

en-1-yl)-ph

enol

Eugeno

lOOH

Leaves

androots(Japan)

[99]

158

3-(4-H

ydroxy-3-m

etho

xyph

enyl)-

2-prop

enoica

cid

Ferulic

acid

O

OH

O

HO

Who

le(Ja

pan)

[99]

159

3-(34-D

ihydroxyph

enyl)-2-prop

enoic

acid

Caffeicacid

O

HO

OH

OH

Who

leandaeria

l(Japan)

[7599]

160

3-Prop

yl-3[(245-trim

etho

xyph

enyl)-

metho

xy]-24-pentanedione

3-Prop

yl-3-(245-

trim

etho

xy)benzyloxy-

pentan-24-dione

O

O

O

O

O

O

Leaves

(India)

[100]

161

3-(34-D

ihydroxyph

enyl)-2-

prop

enoica

cidethyleste

rCa

ffeateethyl

O

O

OH

OH

Not

foun

d(Taiwan)

who

le(Taiwan)

[434584]

Evidence-Based Complementary and Alternative Medicine 31Ta

ble7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

162

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

d-Erythron

icacid

2-O-caffeoyl-2-C

-methyl

O

HO

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

163

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

2-O-caffeoyl-2-C

-methyl

O

O

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

164

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-24-

dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

3-O-caffeoyl-2-C

-methyl

O

O

HO

HO

O

O

OH

OH

Leaves

(Japan)

[101]

165

4-(A

cetyloxy)-3-[[3-(34-

dihydroxyphenyl)-

1-oxo-2-propen-1-y

l-]oxy]-2-hydroxy-

2-methyl-b

utanoica

cid

O

HO

OH

O OHO

HO

OO

Not

foun

d(Ja

pan)

[73]

166

3-(34-D

ihydroxyph

enyl)-tetrahydro-4-

hydroxy-4-

methyl-5

-oxo-3-fu

ranyl

ester-2-prop

enoica

cid

3-O-C

affeoyl-2-C

-methyl-

D-erythrono

-14-la

cton

eHO

HO

O

OO

OOH

Leaves

(Japan)

[101]

167

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]-oxy]-145-

trihydroxy-cyclohexanecarboxylicacid

Chlorogenica

cid

O

HO

HO

OH

O

OH

O

OH

HO

Aeria

l(Japan)w

hole

(Taiwan)

[707175]

32 Evidence-Based Complementary and Alternative Medicine

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

168

4-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]oxy]-

135-tr

ihydroxy-cyclohexanecarboxylic

acid

4-O-C

affeoylqu

inicacid

OHO

HO

OH

OH

O

O

OH

OH

Aeria

l(Japan)

[71]

169

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-15-

dihydroxy-cyclo

hexanecarboxylicacid

34-Di-O

-caffeoylqu

inic

acid

HO

HO

O

O

O

O

OH

OH

HO

OH O

H

O

Aeria

l(Japan)W

hole

(Taiwan)

[45497071]

170

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-14-dihydroxy-

cyclo

hexanecarboxylicacid

35-Di-O

-caffeoylqu

inic

acid

HO

HO

O O

HO

O

HO

OH OO

OHOH

Aeria

l(Japan)w

hole

(Taiwan)

[45497071]

Evidence-Based Complementary and Alternative Medicine 33

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

171

34-bis[[(2E

)-3-(34-D

ihydroxyph

enyl)-1-

oxo-2-prop

en-1-

yl]oxo]-15

-dihydroxy-

cyclo

hexanecarboxylicacid

45-Di-O

-caffeoylqu

inic

acid

OHO

OH

HO

O

O

OH

OH

O

OH

HO

OWho

le(Taiwan)

[454970]

172

3-[4-[[-6-O-[3-(4-H

ydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]-phenyl]-2-prop

enoica

cid

120573-D

-p-C

oumaricacid4-O

-(6-O

-p-caoum

aroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HOH

HO

O

O

Leaves

(Japan)

[59]

173

3-[4-[[2-O

-Acetyl-6

-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]ph

enyl]2-propeno

icacid

120573-D

-p-C

oumaricacid4-O

-(2-O

-acetyl-6

-O-p-

caou

maroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HO

HO

O

O

Leaves

(Japan)aerial

(China)

[3459]

174

67-Dihydroxy-2-chrom

enon

eEsculetin

cicho

rigenin

OHO HO

O

Not

foun

d(Egypt)

[39]

34 Evidence-Based Complementary and Alternative Medicine

Table 8 Aromatic compounds isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

175 12-Benzenediol PyrocatechinHO

HOWhole(Japan) [99]

176 4-Ethyl-12-benzenediol Pyrocatechol

OH

OH

Whole(Japan) [99]

177 Dimethoxyphenol26-dimethoxyphenol

O

HO

O

Roots (Japan) [99]

178 4-Ethenyl-2-methoxy-phenol p-Vinylguaiacol

O

OHWhole(Japan) [99]

179 2-Hydroxy-6-methylbenzaldehyde

6-Methyl-salicylaldehyde O

OH

Whole(Japan) [99]

180 Benzene-ethanol 2-Phenyl-ethanolHO

Whole(Japan) [57]

181 4-Hydroxy-3-methoxy-benzaldehyde Vanillin O

O

HOAerial(Japan) [99]

182 3-Hydroxy-4-methoxy-benzaldehyde Vanillin iso

OO

OH

Leaves(Japan) [99]

183 4-Hydroxybenzoic acid p-Hydroxybenzoicacid

O

HOOH Whole

(Japan) [99]

184 2-Hydroxybenzoic acid Salicylic acidO

HO

HO

Stem androots (Japan) [99]

185 34-Dihydroxybenzoicacid Protocatechuic acid

O

HOOH

OH

Whole(Japan) [99]

1864-Hydroxy-

3methoxybenzoicacid

Vanillic acid

OOH

O

OH

Aerial(Uganda)

Roots (Japan)[68 99]

Evidence-Based Complementary and Alternative Medicine 35

Table 8 Continued

S N IUPAC names Common names Structure Plant part(country) References

187 345-Trihydroxybenzoicacid Gallic acid

O

OHHO

HO

HO

Whole(China) [102]

[88] This work supports the use of B pilosa as an anti-inflammatory agent however no compounds responsible forthe anti-inflammatory activity of B pilosa were identified

A further study also reported the anti-inflammatoryactivity as well as the antiallergic activity of B pilosa [75] Inthis study dried powder of the aerial part of B pilosa whichhad been pretreated with the enzyme cellulosine was usedfor further tests The results showed that oral administrationof the cellulosine-treated B pilosa lowered the level of serumIgE in mice 10 days after immunization with DNP (24-dintrophenyl)-Ascaris as an antigen This treatment alsoreduced dye exudation in skin induced by passive cutaneousanaphylaxis and production of inflammatory mediators his-tamine and substance P in rats [75] Phytochemical analysisshowed that cellulosine treatment increased the percentage ofcaffeic acid and flavonoids This study suggests that B pilosaand its phenolics have anti-inflammatory functions

Phenolics and polyynes are major anti-inflammatoryphytochemicals present in B pilosa (Table 11) Unsurpris-6ingly phenolics such as luteolin (103) and ethyl caffeate(161) that are major constituents of B pilosa have alsobeen reported to possess anti-inflammatory activity Lute-olin was reported to exhibit anti-inflammatory activity inmacrophages Xagorari and colleagues showed that luteolininhibited the release of inflammatory cytokines TNF-120572 andinterleukin-6 in RAW 2647 cells following LPS stimulation[87] It inhibited TNF-120572 production with an IC

50value

of 1120583M The underlying anti-inflammatory mechanism ofluteolin was reported to be the inactivation of Akt andNF-120581B activation [87] In addition luteolin was reportedto confer anti-inflammatory activity through inhibition ofLPS-stimulated iNOS expression in BV-2 microglial cellsIt inhibited LPS-activated microglia in a dose-dependentmanner with an IC

50value of 69 120583MMoreover immunoblot

and RT-PCR data proved that luteolin suppressed I120581B-120572degradation and iNOS expression in LPS-activated microglia[85] Kim and colleagues stated that luteolin may havebeneficial effects on inflammatory neural diseases throughinhibition of iNOS expression [85] A related study revealedthat luteolin decreased the transcriptional activity of NF-120581BRelA via partial inhibition of TNF-mediated NF-120581B DNAbinding activity Luteolin also inhibited Akt phosphorylationand induced degradation of a transcription factor interferonregulatory factor (IRF) [86]

Chiang and colleagues showed that ethyl caffeate(161) significantly inhibited NO production in mousemacrophages RAW 2647 cells [84] Based on MTT assays

they concluded that this inhibition was not due to thecytotoxicity of ethyl caffeate The IC

50value of ethyl caffeate

in the inhibition of NO production was 55 120583gmL slightlylower than curcumin (positive control) which has anIC50

value of 65120583gmL They demonstrated that ethylcaffeate exerted anti-inflammatory activity via the reducedtranscription and translation of iNOS (inducible nitric oxidesynthase) in RAW 2467 cells In addition this compoundalso suppressed COX-2 expression in RAW 2467 cellsand MCF-7 cells The in vivo anti-inflammatory effectof ethyl caffeate was verified by testing in TPA-treatedmouse skin Like celecoxib the positive control ethylcaffeate significantly abolished COX-2 expression in adose-dependent manner Ethyl caffeate at 1mg200120583Lsite(24mM) inhibited COX-2 expression at a level comparableto celecoxib at 1mg200120583Lsite (13mM) Remarkably ethylcaffeate at 48mM (2mg200120583Lsite) was more effectivethan celecoxib at 131mM (10mg200120583Lsite) Moreoverthis compound inhibited the activation of nuclear factor-120581B(NF-120581B) by LPS via the prevention of NF-120581B binding toDNA [84] In addition 3 ethyl caffeate analogs (ethyl 34-dihydroxyhydrocinnamate ethyl cinnamate and catechol)also showed different degrees of NF-120581B binding to DNA aslisted in Table 12 Ethyl cinnamate lacks a catechol moietywhich results in ineffective inhibition of NF-120581B binding toDNA [84]

Pereira and colleagues assessed the anti- 7inflammatory and immunomodulatory activities ofB pilosa methanol extract as well as one polyyne2-O-120573-glucosyltrideca-11(E)-en-3 579-tetrayn-12-diol(54) in T lymphocytes and a zymosan-induced arthritismouse model [27] They first examined the in vitro effect of 8the B pilosa extract and compound 54 on cell proliferation ofhuman T cells stimulated with 5120583gmL phytohemagglutinin(PHA) or 100 nM 12-O-tetradecanoyl phorbol-13-acetate(TPA) plus 15 120583M ionomycin and on cell proliferation ofmouse T cells stimulated with 5 120583gmL concanavalin A(Con A) The data demonstrated that both methanol extractand compound 54 suppressed T-cell proliferation in adose-dependent manner The estimated IC

50values of the B

pilosa extract against human T cells stimulated with 5120583gmLPHA and 100 nM TPA plus 15120583M ionomycin were 125 and25 120583gmL respectively In comparison with the methanolextract compound 54 showed 10-fold more inhibition ofhuman T-cell proliferation with an estimated IC

50value of

15 120583gmL Accordingly the B pilosa extract and compound54 dose-dependently suppressed mouse T-cell proliferation

36 Evidence-Based Complementary and Alternative Medicine

Table 9 Porphyrins isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

188

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(15S16S)-10-ethenyl-5-ethyl-1161820-tetrahydro-6111522-tetramethyl-1820-dioxo-15H-912-imino-212-

metheno-471714-dinitrilopyrano[43-

b]azacyclononadecine-16-propanoic

acid

AristophyllC

NH N

HNN

O

O OOOPhytyl

O

Leaves(Taiwan) [100]

189

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -20b-(methoxycarbonyl)-9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-dinitrilo-12-

dioxeto-[310158404101584034]-cyclo-pent[1 2]azacyclononadecine-

19-propanoic acid

Bidenphytin A

NH N

HNN

OPhytylO O O

O O

Leaves(Taiwan) [100]

190

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -2a-

hydroxy-20b-(methoxycarbonyl)-

9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-

dinitrilo-12-dioxeto[310158404101584034]-cyclo-pent[12-b]-

azacyclononadecine-19-propanoic acid

Bidenphytin B

NH N

HNN

OPhytylO O O

O O

OH

Leaves(Taiwan) [100]

191

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO O

HOOO

Leaves(Taiwan) [100]

192

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

Evidence-Based Complementary and Alternative Medicine 37

Table 9 Continued

S N IUPAC names Common names Structure Plant part(country) References

193

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

Leaves(Taiwan) [100]

194

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

195

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3S4S21R)-9-ethenyl-14-ethyl-21-(methoxycarbonyl)-

481318-tetramethyl-20-oxo-3-phorbinepropanoic acid

Pheophytin A

NH N

HNN

PhytylO

O

OO O

Leaves(Taiwan) [100]

with estimated IC50values of 30 and 25 120583gmL respectively

Taken together the data indicate that the B pilosa extractand compound 54 act on human and mouse T cells Totest the in vivo effect of the B pilosa extract and compound54 a zymosan-induced arthritis mouse model was usedThis model was established from B10ASgSnJ mice with aninjection of zymosan (015mg) The zymosan-injected mice9received an intraperitoneal injection of the B pilosa extract(1 5 or 10mg) at one dose a day for 5 days Popliteal lymphnode (PLN) weight was monitored to check the developmentof arthritis The results revealed that 10mg of the methanolextract of B pilosa extract could significantly diminishinflammation as evidenced by PLN weight [27] This worksuggests that B pilosa (and compound 54) can suppressimmune response and inflammation

33 Antidiabetic Activity Anti-diabetic agents are primarilydeveloped from plants and other natural resources [8 24 4951 53] B pilosa is one of 1200 plant species that have beeninvestigated for antidiabetic activity [120 121]B pilosa is usedas an anti-diabetic herb in America Africa and Asia [51 120

122] Many studies have indicated that B pilosa could treattype 1 diabetes (T1D) and type 2 diabetes (T2D) in animals

Etiologically speaking T1D is caused by autoimmune-mediated destruction of pancreatic 120573 cells leading to insulindeficiency hyperglycemia and complications Currentlythere is no cure for T1D Polarization ofTh cell differentiationcontrols the development of T1D Suppression of Th1 celldifferentiation and promotion of Th2 cell differentiationameliorate T1D [123] One study showed that the butanolfraction of B pilosa inhibited T-cell proliferation decreasedTh1 cells and cytokines and increased Th2 cells andcytokines leading to prevention of T1D in nonobesediabetic (NOD) mice [49] Based on a bioactivity-directedisolation strategy 3 polyynes 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne (53) also known ascytopiloyne 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D- Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne (50) were identifiedfrom B pilosa [49 53] The IC

50value of the butanol fraction

was 200120583gmL This inhibition was reported to be partiallyattributed to cytotoxicity because the butanol fraction at

38 Evidence-Based Complementary and Alternative Medicine

Table 10 Other compounds isolated from B pilosa [30]

SN IUPAC names Common names Structure Plant part(country) References

19637-Dihydro-137-

trimethyl-1H-purine-26-dione

CaffeineN

NN

NO

O

Aerial(Uganda) [68]

197

1-((2R4S5R)-Tetrahydro-4-hydroxy-5-(hydroxymethyl)furan-2-yl)-5-

methylpyrimidine-24(1H3H)-dione

ThymidineHO

HO

ON

O

NHO Not found

(China) [37]

198 1-(2-Thienyl)-ethanone 2-Acetyl-thiophene

S

O

Roots(Germany) [41]

199

(2R3S4S5S)-2-(Heptan-2-yloxy)-tetrahydro-6-

((tetrahydro-34-dihydroxy-5-

(hydroxymethyl)furan-2-yloxy)methyl)-

2H-pyran-345-triol

Heptanyl2-O-120573-xylofuranosyl-

(1rarr 6)-120573-glucopyranoside

O

OOH

OH

HO

OHO

H

HO

H

H

HOHH

O

Whole(Taiwan) [70]

200

2-[(3R7R11R)-3-Hydroxy-371115-

tetramethylhexadecyl]-356-trimethyl-

25-cyclohexadiene-14-dione

120572-Tocopherylquinone

OO

HO

Whole(Taiwan) [32]

2017-O-(410158401015840610158401015840-Diacetyl)-

120573-D-glucopyranoside

Not found Leaves(China) [60]

180 120583gmL could cause 50 death of Th1 cells Moreoverthis study suggested that the butanol fraction may preventdiabetes in NOD mice in vivo via downregulation of Th1cells or upregulation Th2 cells that have effects which areantagonistic of those of Th1 cells [49] This was proven byintraperitoneal injection of the butanol fraction at a doseof 3mgkg BW 3 times a week to NOD mice from 4 to 27weeks This dosage resulted in lower incidence of diabetes(33) At a dose of 10mgkg the butanol fraction of Bpilosa totally eliminated (0) the initiation of the diseaseTo further support this result assessment of IgG2a and IgEproduction was performed in the serum of NOD mice Asin vivo results obtained from intracellular cytokine stainingexperiments were not very conclusive levels of IgG2a and IgE

were measured since Th1 cytokine IFN120574 and Th2 cytokineIL-4 favor the production of IgG2a and IgE respectively Asexpected high levels of IgE and some decline in the levels ofIgG2a were observed in the serum Profiling of the butanolextract revealed five compounds 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne(50) 45-Di-O-caffeoyl- quinic acid 35-Di-O-caffeoylquinicacid and 34-Di-O-caffeoylquinic acid Only the first twocompounds showed similar effects on the prevention ofdiabetes in NOD mice as the B pilosa butanol fractionMoreover compound 50 showed greater activity thancompound 69 in terms of enhancement (by 34 comparedto 8) of differentiation of Th0 to Th2 at 15 120583gmL (both

Evidence-Based Complementary and Alternative Medicine 39

Table 11 Chemical constituents of B pilosa and their biological activities

SN Name Classification Molecularformula Biological activities

109 Centaureidin [91] Flavonoid C18H16O8Anti-listerial [69 91]

Cytotoxic [10]

110 Centaurein [91] Flavonoid C24H26O13

Anti-listerial [69 91]Cytotoxic [10]Anti-viral [105]

103 Luteolin [106] Flavonoid C15H10O6

Anti-viral [107 108]Cytotoxic [77]

Anti-inflammatory [109]Anti-allergic [109]

82 Butein [110] Flavonoid C15H12O5Anti-leishmanial [111]

Cytotoxic [78]48 12-Dihydroxytrideca-57911-tetrayne [112] Polyyne C13H12O2 Anti-angiogeneic [112]46 13-Dihyroxy-6(E)-tetradecene-81012-triyne [112] Polyyne C14H16O2 Anti-angiogeneic [112]

45 12-Dihyroxy-5(E)-tridecene-7911-triyne [113] Polyyne C13H14O2Anti-angiogeneic [113]Anti-proliferative [113]

64 1-Phenylhepta-135-triyne [114] Polyyne C13H8

Anti-microbial [115]Anti-malarial [3]Cytotoxic [3]Antifungal [15]

27 Linoleic acid [79] Fatty acid C18H32O2Anti-viral (100)Cytotoxic [17]

161 Ethyl caffeate [84] Phenylpropanoid C11H12O4 Anti-inflammatory [84]

54 2-O-120573-Glucosyltrideca-11(E)-en-3579-tetrayn-12-diol [26] Polyyne C19H20O7Immunosuppressive andAnti-inflammatory [26]

53 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne [50] Polyyne C19H22O7Anti-diabetic [116]

Anti-inflammatory [50]

69 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne [28] Polyyne C20H26O7Anti-diabetic [28]

Anti-inflammatory [94]

50 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne [28] Polyyne C19H24O7

Anti-diabetic [28]Anti-inflammatory [94]

Anti-malarial andantibacterial [47]

129 Quercetin 3-O-120573-D-galactopyranoside [18] Flavonoid C21H20O12 Anti-inflammatory [117]

170 35-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

171 45-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

169 34-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

126 Quercetin 331015840-dimethyl ether7-O-120572-L-rhamnopyranosyl-(1rarr 6)-120573-D-glucopyranoside [118] Flavonoid C21H20O11 Anti-malarial [119]

125 Quercetin 331015840-dimethyl ether-7-O-120573-D-glucopyranoside [118] Flavonoid C21H20O12 Anti-malarial [119]70 1-Phenyl-13-diyn-5-en-7-ol-acetate [27] Polyyne C15H12O2 Anti-malarial [27]199 Heptanyl 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside [70] Miscellaneous C18H44O10 Antioxidant [70]124 3-O-Rabinobioside [70] Saccharide C27H38O15 Antioxidant [70]130 Quercetin 3-O-rutinoside [70] Flavonoid C27H38O15 Antioxidant [70]167 Chlorogenic acid [70] Phenolic C16H26O9 Antioxidant [70]119 Jacein [70] Flavonoid C24H26O13 Antioxidant [70]

49 (R)-12-dihydroxytrideca-357911-pentayne [47] Polyyne C13H8O2Anti-malarial andAntibacterial [47]

40 Evidence-Based Complementary and Alternative Medicine

Table 12 Structure and activity relationship studies of ethyl caffeateusing in vitro NF-120581BDNA binding assays [84]

Compound Concentration(120583M)

NF-120581BDNA binding

Ethyl caffeate 50 100inhibition

Ethyl 34-dihydroxyhydrocinnamate 100 100inhibition

Catechol 400 100inhibition

Ethyl cinnamate 400 Noinhibition

Table 13 Apoptosis in cocultures of T cells and pancreatic 120573 cells[53]

Cellmedium apoptosis andnecrosis

CD4+ T cellscontrol medium lt4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 2

CD4+ T cellswith cytopiloyne-treated 120573 cells 18CD4+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 7

CD8+ T cellscontrol medium 4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 4

CD8+ T cellswith cytopiloyne-treated 120573 cells 4CD8+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 4

Table 14 Radical scavenging activities of B pilosa extracts [70]

Extractscontrol DPPH assayIC50 (120583gmL)

NBThypoxanthinesuperoxide assayIC50 (120583gmL)

Quercetin 198 15Ascorbic acid 634 Not determined120572-tocopherol 897 Not determinedEthyl acetateextract 1383 597

Butanol extract 1669 114Water extract gt100 gt100

compounds) and inhibition (by 40 compared to 10) ofdifferentiation toTh1 at the same concentration [49]

Among the three polyynes found in B pilosa cytopiloyne(53) had the most potent anti-T1D activity [53] To test the invivo effect of cytopiloyne NODmice received intraperitonealor intramuscular injection of cytopiloyne at 25120583gkg BW3 times per week Twelve-week-old NOD mice started todevelop T1D and 70 of NOD mice aged 23 weeks andover developed T1D Remarkably 12- to 30-week-old NODmice treated with cytopiloyne showed normal levels of bloodglucose (lt200mgdL) and insulin (1-2 ngmL) Consistent

Table 15 Radical scavenging activity of secondarymetabolites fromB pilosa [70]

Metabolite (Table 11)control DPPH assay IC50 (120583gmL)199 Not determined124 53130 68167 105169 33171 38119 Not determined110 Not determinedQuercetin 256Caffeic acid 890

Table 16 Antioxidant activity of the essential oils and water extractsfrom B pilosa [92]

Extract IC50 (120583gmL)Leaf essential oils 47Flower essential oils 50Leaf extract 61Flower extract 172

with T1D incidence cytopiloyne delayed and reduced theinvasion of CD4+ T cells into the pancreatic islets [53]

In vitro study showed that cytopiloyne (53) inhibited thedifferentiation of naıveTh (Th0) cells (ie CD4+ T cells) intoTh1 cells and promoted differentiation of Th0 cells into Th2cells [50] The in vitro data are consistent with the in vivoresults indicating that cytopiloyne reducedTh1differentiationand increased Th2 differentiation as shown by intracellularcytokine staining and FACS analysis [53] In line with theskewing of Th differentiation the level of serum IFN-120574 andIgG2c decreased while that of serum IL-4 and serum IgEincreased compared to the negative controls (PBS-treatedmice) Cytopiloyne also enhanced the expression of GATA-3 a master gene for Th2 cell differentiation but not theexpression of T-bet a master gene forTh1 cell differentiationfurther supporting its role in skewingTh differentiation [53]

Also importantly cytopiloyne partially depleted CD4+rather than CD8+ T cells in NOD mice [53] As shown inTable 13 coculture assays showed that the depletion of CD4+T cells was mediated through the induction of Fas ligandexpression on pancreatic islet cells by cytopiloyne leading toapoptosis of infiltrating CD4+ T cells in the pancreas via theFas and Fas ligand pathways However cytopiloyne did notinduce the expression of TNF-120572 in pancreatic islet cells andthus had no effect on CD8+ T cells [53]

In addition Chang and colleagues showed that cytopi-loyne dose-dependently inhibited T-cell proliferation stimu-lated by IL-2 plus Con A or anti-CD3 antibody using [3H]thymidine incorporation assay [53]

Overall the mechanism of action of cytopiloyne andprobably its derivatives in T1D includes inhibition of T-cellproliferation skewing of Th cell differentiation and partialdepletion of Th cells Due to the anti-diabetic mechanisms

Evidence-Based Complementary and Alternative Medicine 41

Table 17 Antibacterial activity of essential oils and flower extracts from B pilosa [92]

Strain Mean zone of inhibition (mm)Leaf essential oil Flower essential oil Leaf extract Flower extract

Micrococcus flavus 127 plusmn 03 87 plusmn 03 102 plusmn 02 108 plusmn 03Bacillus subtilis 173 plusmn 19 117 plusmn 02 109 plusmn 02 103 plusmn 02Bacillus cereus 190 plusmn 14 112 plusmn 03 118 plusmn 04 185 plusmn 10Bacillus pumilus 123 plusmn 07 108 plusmn 02 105 plusmn 04 77 plusmn 02Escherichia coli 137 plusmn 04 203 plusmn 07 102 plusmn 11 140 plusmn 13Pseudomonas ovalis 125 plusmn 08 137 plusmn 15 102 plusmn 06 125 plusmn 06

Table 18 Antibacterial activity of root extracts from B pilosa [93]

Strain MIC50 (mgmL)Methanol extract Acetone extract

Bacillus cereus 10 mdashEscherichia coli 5 5Klebsilla pneumonia 5 10Micrococcus kristinae 10 mdashPseudomonas aeruginosa 10 10Staphylococcus aureus 5 10Sraphylococcus epidermidis 5 5Serratia marcescens 10 mdashShigelea flexneri 10 mdashStreptococcus faecalis 10 mdash

Table 19 Antibacterial activity of B pilosa of compound 29 [47]

Strain MIC50 (120583gmL)Escherichia coli NIHJ 1Escherichia coli ATCC25922 1Klebsiella pneumoniae ATCC700603 128Serratia marcescens ATCC13880 16Pseudomonas aeruginosa ATCC27853 8Staphylococcus aureus FDA209P 05Staphylococcus aureus ATCC29213 025Staphylococcus aureus N315 (MRSA) 05Enterococcus faecalis ATCC29212 2Enterococcus faecalis NCTC12201 (VRE) 1Bacillus subtilis ATCC6633 05Candida albicans ATCC10231 025

of action it was hypothesized that cytopiloyne protectsNOD mice from diabetes by a generalized suppression ofadaptive immunity To evaluate this hypothesis ovalbumin(Ova) was used as a T-cell dependent antigen to primeNOD mice which had already received cytopiloyne orPBS vehicle Ova priming boosted similar anti-Ova titersin cytopiloyne-treated mice and PBS-treated mice but adifference in immunoglobulin isotype was observed in thetwo groups Thus it was concluded that cytopiloyne is animmunomodulatory compound rather than an immunosup-pressive compound [50 53]

T2D is a chronic metabolic disease with serious com-plications resulting from defects in either insulin secretion

insulin action or both [124] A study by Ubillas et alshowed that the aqueous ethanol extract of the aerial partof B pilosa at 1 gkg body weight (BW) lowered bloodglucose in dbdb mice a T2D mouse model [51] Basedon a bioactivity-guided identification compounds 69 and50 were identified Further the mixture of the compounds(69 50) in a 2 3 ratio significantly decreased blood glucoseconcentration and reduced food intake on the second dayof treatment when administered at doses of 250mgkg twicea day to C5BLKs-dbdb mice When tested at 500mgkga more substantial drop in blood glucose level as wellas the stronger anorexic effect (food intake reduced from58 gmouseday to 25 gmouseday) was observed [51] Inthis study it was suggested that the blood glucose lower-ing effect of B pilosa was caused in part by the hungersuppressing effect of its polyynes [51] However the hungersuppressing effect of the ethanol extract of B pilosa wasnot found in the studies described below In another study[24] water extracts of B pilosa (BPWE) were used indiabetic dbdb mice aged 6ndash8 weeks with postprandialblood glucose levels of 350 to 400mgdL Like oral anti-diabetic glimepiride which stimulates insulin release onesingle dose of BPWE reduced blood glucose levels from374 to 144mgdL The antihyperglycemic effect of BPWEwas inversely correlated to an increase in serum insulinlevels suggesting that BPWE acts to lower blood glucose viaincreased insulin production However BPWE had differentinsulin secretion kinetics to glimepiride [24] One flaw incurrent anti-diabetics is their decreasing efficacy over timeThe authors investigated the long term anti-diabetic effectof BPWE in dbdb mice BPWE reduced blood glucoseincreased blood insulin improved glucose tolerance andreduced the percentage of glycosylated hemoglobin (HbA1c)Both long-term and one-time experiments strongly supportthe anti-diabetic action of BPWE [24] In sharp contrast toglimepiride BPWE protected against islet atrophy in mousepancreas The investigators further evaluated anti-diabeticproperties of 3 B pilosa varieties B pilosa L var radiate(BPR) Bpilosa L var pilosa (BPP) and B pilosa L varminor(BPM) in dbdb mice [8] One single oral dose (10 50 and250mgkg body weight) of BPR BPP or BPM crude extractsdecreased postprandial blood glucose levels in dbdb micefor up to four hours and the reduction of glucose levels inthe blood appeared to be dose-dependent Comparing thethree variants BPR extract resulted in a higher reduction inblood glucose levels when administered at the same dose as

42 Evidence-Based Complementary and Alternative Medicine

Table 20 Antifungal activity of B pilosa [92]

Partextract Concentration (ppm) Strain InhibitionCortiicum rolfsii Fusarium solani Fusarium oxysporum

Leaves

Essentialoils

100 857 plusmn 09 682 plusmn 0 745 plusmn 17250 960 plusmn 08 779 plusmn 18 879 plusmn 04

AqueousExtracts

100 446 plusmn 17 605 plusmn 21 716 plusmn 07250 942 plusmn 03 689 plusmn 07 824 plusmn 19

Flowers

Essentialoils

100 604 plusmn 09 892 plusmn 04 869 plusmn 05250 894 plusmn 12 980 plusmn 03 949 plusmn 06

AqueousExtracts

100 331 plusmn 11 714 plusmn 07 573 plusmn 22250 661 plusmn 14 912 plusmn 0 900 plusmn 07

Table 21 Antifungal activity of B pilosa root extracts [93]

Strain LC50 (mgmL)Acetoneextracts

Methanolextracts

Waterextracts

Aspergillus niger 014 006 007Aspergillus flavus 1091 658 0Penicillium notatum 005 005 005

the other two varieties In terms of serum insulin levels a doseof 50mgkg of each extract was used and the BPR extracttogether with the three polyynes significantly increased theserum insulin level in dbdb mice Long-term experiments(28-day treatment) were then conducted using diabetic micewith postprandial glucose levels from 370 to 420mgdL andglimepiride was used as positive controlThe range of dosagesapplied was from 10mgkg BW to 250mgkg BW Resultsshowed that the positive control as well as the crude extractsof the three varieties lowered the blood glucose levels indbdb mice However only BPR extract containing a higherpercentage of cytopiloyne (53) reduced blood glucose levelsand augmented blood insulin levelsmore than BPP andBPMThe percentage of glycosylated hemoglobin A1c (HbA1c) wasalso measured and found to be 79 plusmn 05 in mice aged 10ndash12 weeks and 66 plusmn 02 61 plusmn 03 and 62 plusmn 03 inthe blood of age-matchedmice following treatment with BPRcrude extract (50mgkg) glimepiride (1mgkg) and com-pound 53 (05mgkg) respectively [8] Among the polyynesfound in B pilosa cytopiloyne was the most effective againstT2D Hence cytopiloyne was used for further study on anti-diabetic action and mechanism [125] The data confirmedthat cytopiloyne reduced postprandial blood glucose lev-els increased blood insulin improved glucose tolerancesuppressed HbA1c level and protected pancreatic islets indbdb mice Nevertheless cytopiloyne failed to decreaseblood glucose in streptoztocin (STZ)-treated mice whose bcells were already destroyed Additionally cytopiloyne dose-dependently increased insulin secretion and expression in bcells as well as calcium influx diacylglycerol and activationof protein kinase C120572 Collectively the mechanistic studiessuggest that cytopiloyne treats T2D via regulation of insulin

production involving the calciumDAGPKC120572 cascade in bcells

The studies detailed above point to the conclusion thatcytopiloyne and related polyynes (compounds 69 and 49)are anti-diabetics in animal models The data uncover a newbiological action of polyynes It should be noted that likeall anti-diabetic drugs cytopiloyne failed to prevent or curediabetes completely but reduced diabetic complications [125]Intriguingly 34 polyynes have been found in B pilosa so farIt remains to be seen whether all the polyynes present in thisplant have anti-diabetic activities

34 Antioxidant Activity Free radicals can damage cellularcomponents via a series of chemical reactions [89] leadingto development and progression of cardiovascular diseasecancer neurodegenerative diseases and ageing [70] Freeradicals nitric oxide (NO) and superoxide anions can beproduced in macrophages to kill microbes However anexcessive generation of the free radicals under pathologicalconditions is associated with a wide range of illnessesPlants are known to be rich in antioxidant phytochemicalsChiang and colleagues evaluated the free radical scaveng-ing activity of crude extract fractions and compounds ofB pilosa using 11-diphenyl-2-picrylhydrazyl (DPPH) andhypoxanthinexanthine oxidase assays [70] Using DPPHand hypoxanthinexanthine oxidase assays they found thatthe B pilosa crude extract and the ethyl acetate butanoland water fractions had free radical scavenging activityNine compounds Heptyl- 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside (199) 3-O-Rabinobioside (124) Quercetin3-O-rutinoside (130) Chlorogenic acid (167) 34-Di-O-caffeoylquinic acid (169) 35-Di-O-caffeoylquinic acid (170)45-Di-O-caffeoylquinic acid (171) Jacein (119) and Cen-taurein (110) had DPPH radical scavenging activity [70]The IC

50values of the B pilosa crude extractfractions and

compounds are summarized in Tables 14 and 15 respectivelyMeasurement of free radical scavenging activities is one

way of assessing the antioxidant activities of B pilosa andits fractions and compounds It is interesting that the ethylacetate and butanol fractions are more active than the waterfraction and B pilosa crude extract [70] Of the secondarymetabolites only phenolic compounds 124 130 167 169 and171 showed significant DPPH-radical scavenging activities

Evidence-Based Complementary and Alternative Medicine 43

Further analysis of the structure-activity relationship of thecompounds suggested that substitution of the C3 hydroxylgroup with glycosides increased the activity approximately2-fold (for example in compounds 124 and 130) relative toquercetin (-OH in its C3) [70] Further modification of thestructures of the active compounds needs to be performed totest the effects of various substituents on activity The reasonwhy most of the antioxidant compounds contain phenolmoieties in their structure could be that the reduction-oxidation (redox) properties of phenols allow them to actas reducing agents singlet oxygen quenchers and hydrogendonors

A complementary study by Muchuweti and colleaguesdetermined phenolic content antioxidant activity and thephenolic profile of B pilosa methanol extract [89] Theyestimated that the phenolic content of the methanol extractof B pilosawas 11028plusmn22mgg [89] Vanillin hydroxyben-zaldehyde caffeic acid coumaric acid and ferulic acid werefound in this extractThe B pilosa extract also showed DPPHradical scavenging activity Furthermore the antioxidantactivity of the flavonoids found in B pilosa was correlatedwith its hepatoprotective effects through their inhibitionof NF-120581B activation which may lessen the oxidative stresscaused by the production of free radicals during liver injury[60]This activity might also be due to the anti-inflammatoryeffects of the aqueous extracts of B pilosa aerial parts on theinhibition of COX-2 and PGE

2production [88]

Essential oils from B pilosa flowers and leaves are alsoreported to possess antioxidant activity With the aim ofreplacing chemically synthesized additives Deba and col-leagues [92] worked on the antioxidant antibacterial andantifungal activities of essential oils and water extracts of Bpilosarsquos leaves and flowers Table 16 summarizes the resultsobtained from DPPH free radical scavenging assay

It can be inferred fromTable 16 that essential oils from theleaves possessed the highest activity It is reported elsewherethat monoterpenes present in essential oils such that of Bpilosa have protective effects and antioxidant properties [92]Beta-carotene bleaching method was also performed Leavesessential oils and aqueous extracts of the leaves and flowersshowed higher activity than the flower essential oils This isdue to the volatility of the flower essential oils The activityexhibited by the aqueous extracts was accounted to thepresence of phenolic compounds that are reported to donatea hydrogen atom to free radicals such that the propagation ofthe chain reaction during lipid oxidation is terminated [92]Overall essential oils and phenolics present in B pilosa canbe thought of as major antioxidant compounds

35 Immunomodulatory Activity B pilosa is thought to bean immunomodulatory plant and is reported to be effectivein the treatment of immune disorders such as allergy [75]arthritis [75] and T1D [48 50 53]

As pointed out in the discussion of its anti-diabeticactivities (Section 33) a combination of phytochemicalsmethod and T-cell activation assays was used to studyimmunomodulatory properties of B pilosa IFN-120574 is a keycytokine released by T and NK cells that mediates immune

cells and sustains immunity against pathogens Defects inIFN-120574 expression regulation and activation result in vulner-ability to diseases caused by bacteria and viruses [69] Anelegant study performed byChang and colleagues using IFN-120574 promoter-driven luciferase reporter construct in JurkatT cells showed that hot water crude extracts of B pilosaincreased IFN-120574 promoter activity two-fold [69] Out of thesubfractions of this extract the butanol fraction but not thewater or ethyl acetate fractions increased IFN-120574 promoteractivity six-fold Centaurein (110) and centaureidin (109)were identified from the butanol fraction and were stated tocause a four-fold increase in IFN-120574 promoter activity withEC50

values of 75 120583gmL and 09120583gmL respectively Themechanism of action of centaurein was determined usingtranscription factors such as AP-1 NFAT and NF120581B whichare reported to bind to IFN-120574 promoter and regulate IFN-120574transcription Unlike with the activity of the positive controlPHA centaurein caused a four-fold increase in NFAT a3-fold increase in NF120581B and had little if any effect onAP-1 enhancer activities (23-fold three-fold and ten-foldincreases respectively were seenwith PHA) [69]The authorsconcluded that centaureinmodulates IFN-120574 expression by theNFAT and NF120581B pathways The article only determined themechanism of action of centaurein Its aglycone centaureidinmay act through the samemechanism though this conclusionneeds to be further verified

B pilosa extract and its compounds are reported to inhibitdifferentiation of naıve CD4+ helper T (Th0) cells into Th1cells [49] Using theThcell differentiation assay as a screeningplatform 3 polyynes 2-120573-D-glucopyranosyloxy-1 -hydroxytrideca-57911-tetrayne (53) 3-120573-D-glucopyranosyl-1-hydroxy-6(E)- tetradecene-81012-triyne (69) and 2-120573-D-glucopyranosyloxy-1-hydroxy-5(E)- tridecene-7911-triyne(49) were discovered from B pilosa [49 53] The data showsthat cytopiloyne and other two polyynes suppressed thedifferentiation of type 1 helper T (Th1) cells and productionof Th1 cytokines and promoted that of type 2 helper T (Th2)cells and production of Th2 cytokines thus explaining theimmunomodulatory and anti-inflammatory effects of Bpilosa and its polyynes

Chang and colleagues were the first to report the effect ofthe butanol extract of B pilosa on the autoimmune diabetesand airway inflammation inmice [48] Imbalance in the levelsofTh1 andTh2 and of various cytokines leads to autoimmunediseases T1D and other autoimmune diseases (rheumatoidarthritis Crohnrsquos disease among others) are exacerbated byan increase Th1 levels (specifically CD4+ Th1 cells) whileTh2 cells antagonize this effect [19] Moreover Th2 cellsmediate asthma in ovalbumin-induced hypersensitivity inBALBc mice [48] In their work [48] Chang and colleaguesshowed that 10mgkg butanol extracts with 15 (ww)compound 69 and 11 (ww) compound 49 (Table 11)ameliorated the development of Th1-mediated diabetes inNOD mice through inhibition of 120573 cell death and leukocyteinfiltration At the same dosage the butanol extracts alsoexacerbated ovalbumin-induced pulmonary inflammationin BALBc mice with an increase in the infiltration ofeosinophils and mast cells into the airway of the mice [48]Despite the different outcomes both mouse models proved

44 Evidence-Based Complementary and Alternative Medicine

the concept that control over the Th1Th2 shift is associatedwith autoimmune diseases and that the B pilosa butanolextract can shift the differentiation of Th0 cells to Th2 cells[48 49]

An extended study presented by Chiang and colleaguesshowed that compound 53 modulates T-cell functions [50]Using CD4+ T cells from BALBc mice they demonstratedthat cytopiloyne decreased levels of IFN-120574 producing cells(Th1) by 122 (from 72 to 598) Since Th1 and Th2 celldifferentiation is antagonistic it was expected that compound53 increased the percentage of mouse IL-4 producing cells(Th2) by 72 (from 237 to 309) Subsequent assess-ment of effect of compound 53 on the modulation of thetranscription of IL-4 and IFN-120574 showed that cytopiloyne asexpected decreased the splenocyte levels of IFN-120574 mRNAand increasing that of IL-4 in a dose-dependent manner Inthe range of 01 to 3 120583gmL of compound 53 the effects werenot attributed to its cytotoxicity Consequently using 3120583gmLcytopiloyne for 72 and 96 hours the protein concentration ofIFN-120574 decreased to 186 and 444 respectively Under thesame conditions cytopiloyne increased IL-4 concentrationsto 1985 and 2470 (for 72 and 96 hours resp) Thismodulation of T-cell differentiation exhibited by compound53 was used to explain its anti-diabetic activity [50]

The anti-diabetic role of cytopiloyne was extensivelydiscussed above (Section 33 anti-diabetic activity) Themolecular basis of the regulation of cytokine expression bycytopiloyne has been described Cytopiloyne directly elevatedthe expression level of IL-4 via GATA-3 upregulation inT cells [53] However reduction of IFN-120574 expression in Tcells seemed to come from the indirect opposing effect IL-4cytokine because the expression level of T-bet was unaltered[53] In this way cytopiloyne skewedTh1 polarization into theTh2 state conferring protection against T1D in NOD miceAside frompolarization ofThcell differentiation cytopiloynealso activated the expression of Fas ligand in pancreaticb cells this increase leading to the partial depletion of Tcells and reduction of immune response in local areas suchas the pancreas Of note cytopiloyne also inhibited T-cellproliferation and activation By targeting T cells from threeimmunomodulatory actions cytopiloyne protects againstT1D and probably other Th1-mediated autoimmune diseases[19]

The phytochemical constituents of B pilosa exert theirfunctions on different immune cells to modulate immuneresponse It is possible that some of the compounds mayhave agonistic or antagonistic effects on immune responseImmune function of B pilosa may depend on its compo-sition and amount of compounds which could explain theapparently conflicting report of B pilosa butanol extractaggravating allergy in mice [48] while cellulosine-treatedextract ameliorated allergy [48 75] IFN-120574 promoter reporterassays and T-cell differentiation assays were used to isolate2 flavonoids [69] and 3 polyynes [49 50] as immunomod-ulatory compounds from the butanol fraction of B pilosaInterestingly the flavonoids promote IFN-120574 expression inNK and T cells In marked contrast the polyynes promoteIL-4 expression and indirectly inhibit IFN-120574 expression indifferentiating T cells Sensitivity appears to be the key to

identifying structure- and bioactivity-related phytochemicalsfrom medicinal plants

36 Antimalarial Activity The use of chemical drugs againstpathogens has resulted in drug-resistant mutants Examplesof drug resistance can be found in the species of thePlasmodium that cause malaria It is important to search fornew compounds to combat Plasmodium parasites [47] Astudy of the anti-malarial activity of the leaf extracts of Bpilosa using a combination of phytochemistry and bioassaysshowed that compound 49 (Table 11) showed activity againsta malaria parasite (P falciparum NF54 strain) with an IC

50

value of 60 120583gmL [77] In addition compound 49 isolatedfrom the aerial parts of B pilosa inhibited growth of the Pfalciparum FCR-3 strain with an IC

50value of 035 120583gmL

This compound was tested for its in vivo effect in miceinfected with P berghei NK-65 strain Results showed thatcompound 49 decreased the average parasitemia in the redblood cells by 207 (from 328 of that of the control to121) after an intravenous injection of 08mgkg BWday forfour days [47] Further studies addressing the anti-malarialmechanism underlying both polyynes and clinical studies areneeded

37 Antibacterial Activity Emergence of multiple antibiotic-resistantmicrobes is becoming a global threat to public healthand a challenge to disease treatment For instance penicillinis commonly used to combat a food-borne intracellularbacteriumListeria however penicillin-resistant bacteria havebeen discovered recently [69] Chang and coworkers iso-lated centaurein (110) and centaureidin (109) from B pilosaextract [69 70] Centaurein enhances expression of IFN-120574a key cytokine for macrophage activation and consequentlyenhances bactericidal activity in macrophages [69 70] Inagreement with observed in vitro effects centaurein wasreported to prevent and treat Listeria infection in C57BL6Jmice [91] Mechanistic studies confirmed that centaureinexerted antilisterial action via IFN-120574 expression in wild-typemice but not IFN-120574 knockout mice [91] Further in vitrostudies on centaurein showed that this compound increasedIFN-120574 expression by 13 (from 17 to 20) 20 (from 21to 41) and 11 (from 6 to 17) in CD4+ T cells CD8+Tcells and NK cells respectively That is to say there wasan increase in IFN-120574 producing immune cells As expectedcentaurein also enhanced the expression level of T-bet a keynuclear factor for IFN-120574 expression Consistently centaureinaugmented the serum IFN-120574 levels inC57BL6Jmice and thisaugmentation peaked 24 hours after compound injectionThe quantity of mouse serum IFN-120574 was sufficient to activatemacrophages in vitro and eradicated GFP-producing Listeriainside macrophages However the entry of Listeria intomacrophages was not affected by centaurein-treated mousesera Centaurein treatment at 20120583g per mouse rescued 30of the mice infected with a lethal dose of Listeria (2 times106 CFU) It is noteworthy that in the presence of ampicillin(5 120583gmouse) centaurein (20 120583gmouse) rescued 70 of themice suggesting an additive effect between ampicillin andcentaurein [91] Despite lower abundance centaureidin was

Evidence-Based Complementary and Alternative Medicine 45

30 times more active than centaurein in terms of IFN-120574production [91]

Aside from the indirect antibacterial action mentionedabove extract andor compounds of B pilosa also showeddirect bacteriostatic andor bactericidal action One studyreported that essential oils and leafflower extracts of Bpilosa could suppress the growth of gram positive and gramnegative bacteria as evidenced by zone of inhibition assaysIn this study antibacterial activity of the essential oils fromthe leaves and flowers of B pilosa was determined in anattempt to identify natural products as food preservatives forprevention of microbial multiplication and food oxidationThe essential oils and extracts of B pilosa leaves and flow-ers showed moderate but different extents of antibacterialactivity (Table 17) In general essential oils had higherantibacterial activity than crude extracts One explanation forthis could be that monoterpenes in the essential oils destroycellular integrity and subsequently inhibit the respirationand ion transport processes The presence of antibacterial120573-caryophyllene could be another explanation as reportedelsewhere [92] Another study reported that the methanoland acetone extract of B pilosa roots displayed antibacterialactivities against the bacteria listed in Table 18 [93] andmethanol extracts from the roots seemed to be the mosteffective

Another study indicated that the polyyne (R)-12-dihydroxytrideca-357911-pentayne (49) from this plantalso suppressed bacterial growth as shown by the minimuminhibitory concentration required to inhibit 50 bacterialgrowth (MIC

50) in Table 19 This compound was highly

effective against several Gram positive and Gram negativebacteria including the drug-resistant bacteria Staphylococcusaureus N315 (MRSA) and Enterococcus faecalis NCTC12201(VRE) [47] Strikingly compound 49 had a similar MIC

50

value to antibiotics (ampicillin tetracycline norfloxacin andamphotericin B) in most of the bacteria tested

Antibacterial activity of B pilosa extracts and compo-nents expressed as MIC

50and the mean zone of inhibition

is tabulated in Tables 17 18 and 19The zone of inhibition forAmpicilline (positive control) ranges from 153 plusmn 03mm to443 plusmn 02mm [92]

38 Antifungal Activity B pilosa has traditionally been usedto treat microbial infection Recently different parts of Bpilosa have been tested for antifungal activities Deba andcolleagues first evaluated the antifungal effect of the hotwater extracts of the B pilosa roots stems and leaves againstCorticium rolfsii Fusarium solani and Fusarium oxysporumThey discovered that C rolfsii was most suppressed bytreatment with B pilosa as its growth was reduced at almostall the tested doses followed by F oxysporum and F solani[99] However the fungicidal activities of the stems androots were greater than the leaves [99] Moreover the samegroup assessed the antifungal activity of the essential oilsand aqueous extracts from B pilosa flowers and leaves [92]They showed that the extracts and oils had antifungal activityagainst C rolfsii F solani and F oxysporum Essential oils

appeared to have better fungicidal activity thanwater extractsas summarized in Table 20

Another study by Ashafa and colleagues showed thatacetone methanol and water extracts of the B pilosa rootsshowed antifungal activities against Aspergillus niger Aflavus and Penicillium notatum using the agar dilutionmethod The results are tabulated in Table 21 [93] Negativecontrols showed 0 growth inhibitionThe methanol extractof the B pilosa roots at 10mgmL was also effective againstCandida albicans [93]Of note B pilosa obtained fromPapuaNew Guinea had no activity against A niger and C albicans[126] but the SouthAfrican (EasternCape) ecotype exhibitedmoderate activity against C albicans [93] This discrepancymay depend on extraction solvents extraction procedureassay techniques different plant parts and abundance ofactive compounds

B pilosa produces a variety of secondary metabolitessuch as flavonoids phenylacetylenes alkaloids sterols ter-penoids and tannis [92 93] However none of them havebeen confirmed as active compounds against fungi Furtherinvestigation of active compounds from B pilosa is necessaryto further understand the antifungal efficacy of this plant

39 Hypotensive and Vasodilatory Activities In early studiesDimo and colleagues used three rat models normotensiveWistar rats (NTR) salt-loading hypertensive rats (SLHR)and spontaneous hypertensive rats (SHR) to investigate thehypotensive effect of the methanol crude extract of B pilosaleaves [28 127] The extract lowered systolic blood pressurein hypertensive rats (SLHR and SHR) to a greater degreethan NTR [28 127] In addition a decrease in urinarysodium ions and an increase in urinary potassium ions wereobserved after treatment with the methanol extract of Bpilosa leaves although neither differences were statisticallysignificant [28 127] Taking the data together the studyproposed that B pilosa leaf extract reduced blood pressurevia vasodilation [28 127] The same group continued totest the antihypertensive effect of aqueous and methylenechloride extracts of B pilosa leaves in a hypertensive ratmodel [28 127] To establish a fructose-induced hypertensionmodel male Wistar rats were given 10 fructose solutionto drink ad libitum for three weeks In addition to freeaccess to 10 fructose the rats were treated with the aqueous(150mgkg) or methyl chloride (350mgkg) extracts of Bpilosa for additional three weeks [28 127] Both extracts ofB pilosa leaves had a hypotensive effect on rats Howeverneither extracts reversed the elevation of serum insulin infructose-fed rats Therefore B pilosa lowered blood pressureirrespective of insulin [28 127] To better understand thehypotensive mechanism the authors investigated the effectof a neutral extract of B pilosa (NBP) a mixture of methanolandmethylene chloride (1 1) extract after neutralization withNaOH and HCl on the heart and the blood pressure ofNTR and SHR [110] This study showed that an intravenousinjection of the NBP resulted in a biphasic reduction insystolic blood pressure In addition one intravenous dose ofthe extract at 10 20 and 30mgkg BW decreased systolicblood pressure in normal rats by 183 425 and 30

46 Evidence-Based Complementary and Alternative Medicine

respectively and the same doses reduced the blood pressurein hypertensive rats by 258 389 and 286 respectivelyOnly the highest dose (30mgkg) affected the force of thecontraction of the heart Atropine and propranolol were usedto interfere with the hypotensive action of the NBP Atropinereduced the initial phase of the hypotensive response in NBPand completely abolished the second phase of hypotensiveresponse in NBP In contrast propranolol increased thefirst hypotensive response but partially abolished the secondhypotensive response provoked by the NBP [110]This mech-anistic study suggested that B pilosa invokes the biphasichypotensive responses via targeting cardiac pump efficiencyduring the first phase and vasodilation at the second phase[110]

A further study was performed to investigate the relaxingeffect of a neutral extract of B pilosa (NBP) on rat aorta con-tractedwithKCl (60mM) andnorepinephrine (01mM) [94]Cumulative addition of NBP relaxed the rat aorta previouslycontracted by KCl in a dose-dependent manner The EC

50

value of theNBP for vasorelaxationwas 032mgmLThedataalso showed that the NBP reduced the contraction of aortapreviously contracted by KCl irrespective of the presence ofaortic endothelium [94]

Pretreatment with glibenclamide an ATP-dependent K+channel blocker did not considerably affect the relaxant effectof the NBP on KCl-induced contraction suggesting thatthe vasodilatory effect of B pilosa was not related to theopening of this ATP-dependent K+ channel [94] On theother hand in the presence of indomethacin or pyrilaminemaleate the relaxant response induced by the plant extractwas significantly inhibited at the lower concentrations Theplant extract was able to reduce the aorta resting tone inhibitthe KCl-induced contractions by 90 at 15mgmL and theCaCl2-induced contractions by 95 at 075mgmL These

results demonstrate that B pilosa can act as a vasodilatorprobably via acting as a calcium antagonist [94]

However no specific compound for the above activityhas been identified from B pilosa to date A bioactivity-guided identification approach may be adopted to identifythe active compounds in B pilosa that possess hypotensiveand vasodilatory effects and understand their mechanism ofaction

310 Wound Healing Activity B pilosa has been traditionallyused to treat tissue injury inCameroon Brazil andVenezuela[26] Hassan and colleagues investigated the wound healingpotential of B pilosa in Wistar rats [128] Mirroring thepositive control neomycin sulfate the ethanol extract of Bpilosa had faster wound closure than control rats 3 6 and 9days after topical application Histological examination alsorevealed better collagenation angiogenesis and organizationof wound tissue seven days after application Epithelializationand total healing time in B pilosa-treated rats were compara-ble to those of neomycin sulfate Together these data suggestthat B pilosa may be a viable alternative to neomycin lotionfor the treatment of wounds

In addition to studying the wound healing effect of Bpilosa on external ulcers Tan and colleagues also examined

the effect of methanol cyclohexane and methyl chlorideextracts of B pilosa on gastric ulcers in Wistar rats fedwith 1mL HClethanol gastric necrotizing solution (150mMHCl in 60 ethanol) and macroscopically visible lesionswere scored [26] Among the three extracts methylene chlo-ride extracts exhibited the highest activity showing 464inhibition of lesion formation at a dose of 500mgkg BWand complete inhibition at 750mgkg [26] The efficacy ofthe ethylene chloride extract was followed by that of themethanol extracts which had inhibition ranging from 304to 822 at concentrations of 500mgkg and 1000mgkgBW respectively [26] The cyclohexane extracts showed thelowest activity against gastric ulcers in rats with 13340 and 797 inhibition at 500 750 and 1000mgkg BWrespectively [26] To better understand the mode of actionof the methylene chloride extract of B pilosa rats werepretreated with indomethacin a COX-2 inhibitor involved inprostaglandin synthesis Pretreatment significantly reducedthe protection against HClethanol-induced ulcers to 313inhibition at 750mgkg BW suggesting a link between theantiulcerative activity of B pilosa and prostaglandin syn-thesis Unexpectedly the methylene chloride extract of Bpilosa showed little gastric mucosal protection against gastriclesions induced by 95 ethanol (1mL) [26] Absolute alcoholis known to cause mucosalsubmucosal tissue destruction viacellular necrosis and the release of tissue-derived mediators(histamine and leukotrieneC4)Thus these data imply thatBpilosa did not prevent the generation or the necrotic action ofthese mediators on the gastric microvasculature In additionpylorus ligation can increase gastric acid secretion withoutan alteration of mucosal histamine content The methylenechloride extract of B pilosa did not possess antisecretoryactivity On the contrary it was observed that increases in thedose of the extract led to elevated gastric juice acidity [26]Results with both absolute ethanol and pylorus ligation ratmodels suggested the possibility that the ineffectiveness of Bpilosa against gastric ulcers was due to lack of antihistaminicactivity in the plant In summary overall the data suggest thatB pilosa protects against HCLethanol-mediated ulcers viainhibition of prostaglandin biosynthesis

Previous phytochemical studies showed that a groupof flavonoids acyclichalcones are present in B pilosa[129 130] and the chalcones were proposed to have anti-ulcerative activity [131] Moreover nine hydroxychalcones 10were reported to possess gastric cytoprotective effects with24-dihydroxychalcone being the most active [132] Sincemethylene chloride extracts appear to be the most active Bpilosa extracts next the specific anti-ulcerative phytochemi-cals in the methylene chloride extracts of B pilosa and theirmodes of action needs to be probed

Despite the claims listed inTable 3 relatively few scientificstudies have been conducted in vitro and in vivo to address thetraditional ethnomedical uses of B pilosa Information aboutthe use of B pilosa as a botanical therapy recorded so far isfar from complete Studies conducted thus far only serve asa starting point for further investigation of B pilosa and theultimate efficacious use of the herb in clinical applications

Evidence-Based Complementary and Alternative Medicine 47

4 Toxicology

Despite its use as an ingredient in food for human con-sumption studies on systemic toxicity (eg acute subacutechronic and subchronic toxicities) of B pilosa in humansand animals are still inadequate and insufficient So faracute andor subchronic toxicities have been evaluated inrats and mice Oral acute and 28-day toxicities of waterextract of B pilosa leaves were evaluated in Wistar rats[133] An oral dose of water extract of B pilosa leaves at10 gkg BW showed no obvious mortality or changes inthe appearance in rats [134] The same extract at 08 gkgBWday once a day showed no obvious sub-chronic toxicityin rats over 28 days as measured by survival rate bodyweight and gross examination of organs [134] These dataare consistent with our data indicating that oral deliveryof the water extract of the B pilosa whole plant at 1 gkgBWday once a day is safe in rats over 28 days (unpublisheddata) Taken together these studies suggest that ingestion ofB pilosa aqueous extract at up to at 1 gkg BWday oncea day is highly safe in rats In addition the acute toxicityof aqueous and ethanol extracts of B pilosa in mice havebeen reported [134] Five- to six-week-old mice with weightsbetween 28 and 35 g received a peritoneal injection of bothextracts at the different doses The LD

50 the dose that causes

50 lethality of the aqueous and ethanol extracts in micewas 1230 gkg BW and 615 gkg BW respectively [134] Acomplete toxicological study has not been completed forhumans Furthermore the drug interactions of B pilosa withother drugs are unknown Further safety verification andclinical trials should be performed before B pilosa can beconsidered for medicinal use

5 Conclusions

B pilosa is an erect perennial plant with green leaves whiteor yellow flowers and tiny black seeds As it is distributedworldwide and is widely used as a folk remedy B pilosacan be thought of as an extraordinary source of food andmedicine However a comprehensive up-to-date review ofresearch on B pilosa has hitherto been unavailable In thisarticle scientific studies on B pilosa have been summarizedand critically discussed from the perspectives of botanyethnomedicine phytochemistry pharmacology and toxicol-ogy B pilosa is claimed to treat more than 40 disordersand 201 compounds have been identified from this plantThe medicinal utility of B pilosa and its modes of actionin relation to its known phytochemicals were discussedherein Polyynes flavonoids phenylpropanoids fatty acidsand phenolics are the primary bioactive compounds of Bpilosa and they have been reported to be effective in thetreatment of tumors inflammationimmune modulationdiabetes viruses microbes protozoans gastrointestinal dis-eases hypertension and cardiovascular diseases Cautionshould be exercised in the therapeutic use of B pilosa forhypoglycemia hypotension bleeding and allergy

Acknowledgments

The authors thank their laboratory members for construc-tive suggestions and the authors whose publications theycited for their contributions The authors also thank MsMiranda Loney of Academia Sinica for English editing of thispaper This work was supported by Grants 99-CDA-L11 and101S0010056 from Academia Sinica Taiwan Arlene P Bar-tolome is a recipient of MECO-TECO Sandwich ScholarshipProgram between Taiwan and the Philippines

References

[1] T Shen G H Li X N Wang and H X Lou ldquoThe genusCommiphora a review of its traditional uses phytochemistryand pharmacologyrdquo Journal of Ethnopharmacology vol 142 no2 pp 319ndash330

[2] C Long P Sauleau B David et al ldquoBioactive flavonoids ofTanacetum parthenium revisitedrdquo Phytochemistry vol 64 no2 pp 567ndash569 2003

[3] P O Karis and O Ryding Asteraceae Cladistics and Classifica-tion Bremer K Eds pp 559ndash569 Timber press Portland OreUSA 1994

[4] O N Pozharitskaya A N Shikov M N Makarova et alldquoAnti-inflammatory activity of a HPLC-fingerprinted aqueousinfusion of aerial part of Bidens tripartita LrdquoPhytomedicine vol17 no 6 pp 463ndash468 2010

[5] F Q Oliveira V Andrade-Neto A U Krettli and MG L Brandao ldquoNew evidences of antimalarial activity ofBidens pilosa roots extract correlated with polyacetylene andflavonoidsrdquo Journal of Ethnopharmacology vol 93 no 1 pp 39ndash42 2004

[6] Agriculture USDA ldquoPlants databaserdquo 2013in Natural Resources Conservation Servicehttpwwwnrcsusdagovwpsportalnrcssitenationalhome

[7] Y Zhou and X Z Yan ldquoExperimental study of qi deficiencysyndrome and Codonopsis pillosulae and Astragalus injectionon the immune response in micerdquo Zhong Xi Yi Jie He Za Zhivol 9 no 5 pp 286ndash262 1989

[8] S C Chien P H Young Y J Hsu et al ldquoAnti-diabeticproperties of three common Bidens pilosa variants in TaiwanrdquoPhytochemistry vol 70 no 10 pp 1246ndash1254 2009

[9] M J Alcaraz and M J Jimenez ldquoFlavonoids as anti-inflammatory agentsrdquo Fitoterapia vol 59 no 1 pp 25ndash38 1988

[10] FAO Agriculture Food and Nutrition for AfricamdashA ResourceBook for Teachers of Agriculture Publishing ManagementGroup FAO Information Division Rome Italy 1997

[11] M B Rokaya Z Munzbergova B Timsina and K R BhattaraildquoRheum australe D Don a review of its botany ethnobotanyphytochemistry and pharmacologyrdquo Journal of Ethnopharma-cology vol 141 no 3 pp 761ndash774 2012

[12] P H Young Y J Hsu and W C Yang ldquoBidens pilosa L andits medicinal userdquo in Recent Progress in Medicinal Plants DrugPlant II A S AwaadVK Singh and JNGovil Eds StandiumPress Houston Tex USA 2010

[13] C Ge ldquoCytologic study of Bidens bipinnata LrdquoZhongguo ZhongYao Za Zhi vol 15 no 2 pp 72ndash125 1990

[14] L C Chiang J S Chang C C Chen L T Ng and C CLin ldquoAnti-herpes simplex virus activity of Bidens pilosa and

48 Evidence-Based Complementary and Alternative Medicine

Houttuynia cordatardquoAmerican Journal of ChineseMedicine vol31 no 3 pp 355ndash362 2003

[15] N P Rybalchenko V A Prykhodko S S Nagorna et alldquoIn vitro antifungal activity of phenylheptatriyne from Bidenscernua L against yeastsrdquo Fitoterapia vol 81 no 5 pp 336ndash3382010

[16] K Redl W Breu B Davis and R Bauer ldquoAnti-inflammatoryactive polyacetylenes from Bidens campylothecardquo Planta Med-ica vol 60 no 1 pp 58ndash62 1994

[17] M Ayyanar and S Ignacimuthu ldquoTraditional knowledge ofKani tribals inKouthalai of Tirunelveli hills TamilNadu IndiardquoJournal of Ethnopharmacology vol 102 no 2 pp 246ndash255 2005

[18] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant intraditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[19] AKAbbas KMMurphy andA Sher ldquoFunctional diversity ofhelper T lymphocytesrdquo Nature vol 383 no 6603 pp 787ndash7931996

[20] J H Cano and G Volpato ldquoHerbal mixtures in the traditionalmedicine of Eastern Cubardquo Journal of Ethnopharmacology vol90 no 2-3 pp 293ndash316 2004

[21] J Namukobe J M Kasenene B T Kiremire et al ldquoTraditionalplants used for medicinal purposes by local communitiesaround the Northern sector of Kibale National Park UgandardquoJournal of Ethnopharmacology vol 136 no 1 pp 236ndash245 2011

[22] C Lans ldquoComparison of plants used for skin and stomachproblems in Trinidad and Tobago with Asian ethnomedicinerdquoJournal of Ethnobiology andEthnomedicine vol 3 article 3 2007

[23] S Dharmananda ldquoA Popular Remedy Ecapes Notice ofWesternPractitionersrdquo 2013 httpwwwitmonlineorgartsbidenshtm

[24] Y J Hsu T H Lee C L T Chang Y T Huang andW C YangldquoAnti-hyperglycemic effects and mechanism of Bidens pilosawater extractrdquo Journal of Ethnopharmacology vol 122 no 2 pp379ndash383 2009

[25] E Noumi F Houngue and D Lontsi ldquoTraditional medicinesin primary health care plants used for the treatment ofhypertension in Bafia Cameroonrdquo Fitoterapia vol 70 no 2 pp134ndash139 1999

[26] P V Tan T Dimo and E Dongo ldquoEffects of methanolcyclohexane and methylene chlo ride extracts of Bidens pilosaon various gastric ulcer models in ratsrdquo Journal of Ethnophar-macology vol 73 no 3 pp 415ndash421 2000

[27] R L C Pereira T Ibrahim L Lucchetti A J R Da Silva and VL G De Moraes ldquoImmunosuppressive and anti-inflammatoryeffects of methanolic extract and the polyacetylene isolatedfrom Bidens pilosa Lrdquo Immunopharmacology vol 43 no 1 pp31ndash37 1999

[28] T Dimo J Azay P V Tan et al ldquoEffects of the aqueous andmethylene chloride extracts of Bidens pilosa leaf on fructose-hypertensive ratsrdquo Journal of Ethnopharmacology vol 76 no 3pp 215ndash221 2001

[29] CWiartMedicinal Plants of Southeast Asia Pelanduk SelabgorDarul Ehsan Malaysia 2000

[30] F L Silva D C H Fischer J F Tavares M S Silva P FDe Athayde-Filho and J M Barbosa-Filho ldquoCompilation ofsecondary metabolites from Bidens pilosa LrdquoMolecules vol 16no 2 pp 1070ndash1102 2011

[31] A H Chen S R Lin and C H Hong ldquoPhytochemical studyon Bidens pilosa L varminorrdquo Huaxue pp 38ndash42 1975

[32] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[33] M C Zulueta M Tada and C Y Ragasa ldquoA diterpene fromBidens pilosardquo Phytochemistry vol 38 no 6 pp 1449ndash14501995

[34] J Wang H Yang Z W Lin and H D Sun ldquoFlavonoids fromBidens pilosa var radiatardquo Phytochemistry vol 46 no 7 pp1275ndash1278 1997

[35] A Zhao Q Zhao L Peng et al ldquoA new chalcone glycoside fromBidens pilosardquo Acta Botanica Yunnanica vol 26 no 1 pp 121ndash126

[36] P Gao Y Wu S Cui and Y Zou ldquoSynthesis and absoluteconfiguration of the 7-phenylhepta-4 6-diyne-1 2-diol isolatedfrom Bidens pilosardquo Synthesis no 13 pp 2131ndash2135 2011

[37] S Wang B Yang D Zhu D He and L Wang ldquoActivecomponents of Bidens pilosa Lrdquo Zhongcaoyao vol 36 pp 20ndash21

[38] F Bohlmann T Burkhardt and C Zdero Naturally OccuringAcetylenes Academic Press New York NY USA 1973

[39] T M Sarg A M Ateya N M Farrag and F A AbbasldquoConstituents and biological activity of Bidens pilosa L grownin EgyptrdquoActa Pharmaceutica Hungarica vol 61 no 6 pp 317ndash323 1991

[40] H A L Valdes and H P Rego ldquoBidens pilosa Linnerdquo RevistaCubana de Plantas Medicinales vol 6 pp 28ndash33

[41] F Bohlmann H Bornowski and K M Kleine ldquoNew polyynesfrom the tribe Heliantheaerdquo Chemische Berichte vol 97 pp2135ndash2138

[42] RWang Q XWu and Y P Shi ldquoPolyacetylenes and flavonoidsfrom the aerial parts ofBidens pilosardquo PlantaMedica vol 76 no9 pp 893ndash896 2010

[43] L W Wu Y M Chiang H C Chuang et al ldquoA novelpolyacetylene significantly inhibits angiogenesis and promotesapoptosis in human endothelial cells through activation of theCDK inhibitors and caspase-7rdquo Planta Medica vol 73 no 7 pp655ndash661 2007

[44] L W Wu Y M Chiang H C Chuang et al ldquoPolyacetylenesfunction as anti-angiogenic agentsrdquo Pharmaceutical Researchvol 21 no 11 pp 2112ndash2119 2004

[45] H L Yang S C Chen N W Chang et al ldquoProtection fromoxidative damage using Bidens pilosa extracts in normal humanerythrocytesrdquo Food and Chemical Toxicology vol 44 no 9 pp1513ndash1521 2006

[46] H Q Wang S J Lu H Li and Z H Yao ldquoEDTA-enhancedphytoremediation of lead contaminated soil by Bidens maxi-mowiczianardquo Journal of Environmental Sciences vol 19 no 12pp 1496ndash1499 2007

[47] S TobinagaMK SharmaWG L Aalbersberg et al ldquoIsolationand identification of a potent antimalarial and antibacterialpolyacetylene from Bidens pilosardquo Planta Medica vol 75 no 6pp 624ndash628 2009

[48] C L T Chang H K Kuo S L Chang et al ldquoThe distincteffects of a butanol fraction of Bidens pilosa plant extract onthe development of Th1-mediated diabetes and Th2-mediatedairway inflammation in micerdquo Journal of Biomedical Sciencevol 12 no 1 pp 79ndash89 2005

[49] S L Chang C L T Chang Y M Chiang et al ldquoPolyacetyleniccompounds and butanol fraction from Bidens pilosa can mod-ulate the differentiation of helper T cells and prevent autoim-mune diabetes in non-obese diabetic micerdquo Planta Medica vol70 no 11 pp 1045ndash1051 2004

Evidence-Based Complementary and Alternative Medicine 49

[50] Y M Chiang C L T Chang S L Chang W C Yang and LF Shyur ldquoCytopiloyne a novel polyacetylenic glucoside fromBidens pilosa functions as a T helper cell modulatorrdquo Journal ofEthnopharmacology vol 110 no 3 pp 532ndash538 2007

[51] R PUbillas CDMendez S D Jolad et al ldquoAntihyperglycemicacetylenic glucosides from Bidens pilosardquo Planta Medica vol66 no 1 pp 82ndash83 2000

[52] L Alvarez S Marquina M L Villarreal D Alonso E Arandaand G Delgado ldquoBioactive polyacetylenes from Bidens pilosardquoPlanta Medica vol 62 no 4 pp 355ndash357 1996

[53] C L T Chang S L Chang Y M Lee et al ldquoCytopiloyne apolyacetylenic glucoside prevents type 1 diabetes in nonobesediabetic micerdquo Journal of Immunology vol 178 no 11 pp 6984ndash6993 2007

[54] G Kusano A Kusano and Y Seyama Novel Hypoglycemicand Anti-Inflammatory Polyacetylenic Compounds Their Com-positions Bidens Plant Extract Fractions and CompositionsContaining the Plant or Fraction JPO Tokyo Japan 2004

[55] M G L Brandao A U Krettli L S R Soares C G CNery and H C Marinuzzi ldquoAntimalarial activity of extractsand fractions from Bidens pilosa and other Bidens species(Asteraceae) correlated with the presence of acetylene andflavonoid compoundsrdquo Journal of Ethnopharmacology vol 57no 2 pp 131ndash138 1997

[56] A U Krettli V F Andrade-Neto M D G L Brandao andW M S Ferrari ldquoThe search for new anti-malarial drugs fromplants used to treat fever and malaria or plants ramdomlyselected a reviewrdquoMemorias do Instituto Oswaldo Cruz vol 96no 8 pp 1033ndash1042 2001

[57] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[58] C K Wat R K Biswas E A Graham L Bohm G H NTowers and E R Waygood ldquoUltraviolet-mediated cytotoxicactivity of phenylheptatriyne from Bidens pilosa Lrdquo Journal ofNatural Products vol 42 no 1 pp 103ndash111 1979

[59] Y Sashida K Ogawa M Kitada H Karikome Y Mimakiand H Shimomura ldquoNew aurone glucosides and new phenyl-propanoid glucosides from Bidens pilosardquo Chemical and Phar-maceutical Bulletin vol 39 no 3 pp 709ndash711 1991

[60] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens pilosa L (TFB) on animal liver injury andliver fibrosisrdquo Journal of Ethnopharmacology vol 116 no 3 pp539ndash546 2008

[61] B Hoffmann and J Holzl ldquoAcylated compounds from Bidenspilosardquo Planta Medica vol 55 pp 108ndash109

[62] B Hoffmann and J Holzl ldquoA methylated chalcone glucosidefrom Bidens pilosardquo Phytochemistry vol 27 no 11 pp 3700ndash3701 1988

[63] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[64] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[65] B Hoffmann and J Holzl ldquoWeitere acylierte chalkone ausBidens pilosardquo Planta Medica vol 54 pp 450ndash451 1988

[66] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant in11traditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[67] V V Pham V V T K P V L Hoang and V K Phan ldquoFlavonoidcompounds from the plant Bidens pilosa L (Asteraceae)rdquo TapChi Duoc Hoc vol 50 pp 48ndash53

[68] S D Sarker B Bartholomew R J Nash and N Robinson ldquo5-O-methylhoslundin an unusual flavonoid from Bidens pilosa(Asteraceae)rdquo Biochemical Systematics and Ecology vol 28 no6 pp 591ndash593 2000

[69] S L Chang Y M Chiang C L T Chang et al ldquoFlavonoidscentaurein and centaureidin from Bidens pilosa stimulate IFN-120574 expressionrdquo Journal of Ethnopharmacology vol 112 no 2 pp232ndash236 2007

[70] Y M Chiang D Y Chuang S Y Wang Y H Kuo P WTsai and L F Shyur ldquoMetabolite profiling and chemopreventivebioactivity of plant extracts from Bidens pilosardquo Journal ofEthnopharmacology vol 95 no 2-3 pp 409ndash419 2004

[71] A Kusano Y Seyama E Usami et al ldquoStudies on the antioxi-dant active constituents of the dried powder from Bidens pilosaL var radiata SchrdquoNaturalMedicines vol 57 no 3 pp 100ndash1042003

[72] I Doleckova L Rarova J Gruz et al ldquoAnti-proliferative andanti-angiogenic effects of flavone eupatorin an active con-stituent of chloroform extract ofOrthosiphon stamineus leavesrdquoFitoterapia vol 83 no 6 pp 1000ndash1007

[73] K Bairwa R Kumar R J Sharma and R K Roy ldquoAn updatedreview on Bidens pilosa Lrdquo Der Pharma Chemica vol 2 no 3pp 325ndash337

[74] Q Xia Y Liu and Y Li ldquoDetermination of hyperoside indifferent parts and different species of Herba Bidens by RP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol 50pp 48ndash53

[75] M Horiuchi and Y Seyama ldquoImprovement of the antiinflam-matory and antiallergic activity of Bidens pilosa L var radiataSCHERFF treated with enzyme (Cellulosine)rdquo Journal of HealthScience vol 54 no 3 pp 294ndash301 2008

[76] M G L Brandao C G C Nery M A S Mamao and AU Krettli ldquoTwo methoxylated flavone glycosides from Bidenspilosardquo Phytochemistry vol 48 no 2 pp 397ndash400 1998

[77] P Kumari K Misra B S Sisodia et al ldquoA promising anticancerand antimalarial component from the leaves of Bidens pilosardquoPlanta Medica vol 75 no 1 pp 59ndash61 2009

[78] G Seelinger I Merfort U Wolfle and C M Schempp ldquoAnti-carcinogenic effects of the flavonoid luteolinrdquoMolecules vol 13no 10 pp 2628ndash2651 2008

[79] G Seelinger I Merfort and C M Schempp ldquoAnti-oxidantanti-inflammatory and anti-allergic activities of luteolinrdquoPlanta Medica vol 74 no 14 pp 1667ndash1677 2008

[80] C C Yit and N P Das ldquoCytotoxic effect of butein on humancolon adenocarcinoma cell proliferationrdquo Cancer Letters vol82 no 1 pp 65ndash72 1994

[81] J A Beutler E Hamel A J Vlietinck et al ldquoStructure-activityrequirements for flavone cytotoxicity and binding to tubulinrdquoJournal of Medicinal Chemistry vol 41 no 13 pp 2333ndash23381998

[82] M R Kviecinski K B Felipe T Schoenfelder et al ldquoStudy ofthe antitumor potential of Bidens pilosa (Asteraceae) used inBrazilian folkmedicinerdquo Journal of Ethnopharmacology vol 117no 1 pp 69ndash75 2008

[83] P Sundararajan A Dey A Smith A G Doss M Rajappanand S Natarajan ldquoStudies of anticancer and antipyretic activityof Bidens pilosawhole plantrdquo African Health Sciences vol 6 no1 pp 27ndash30 2006

[84] YMChiang C P Lo Y P Chen et al ldquoEthyl caffeate suppressesNF-120581B activation and its downstream inflammatory mediatorsiNOS COX-2 and PGE2 in vitro or in mouse skinrdquo BritishJournal of Pharmacology vol 146 no 3 pp 352ndash363 2005

50 Evidence-Based Complementary and Alternative Medicine

[85] J S Kim H J Lee M H Lee J Kim C Jin and J HRyu ldquoLuteolin inhibits LPS-stimulated inducible nitric oxidesynthase expression in BV-2 microglial cellsrdquo Planta Medicavol 72 no 1 pp 65ndash68 2006

[86] P A Ruiz and D Haller ldquoFunctional diversity of flavonoidsin the inhibition of the proinflammatory NF-120581B IRF and Aktsignaling pathways in murine intestinal epithelial cellsrdquo Journalof Nutrition vol 136 no 3 pp 664ndash671 2006

[87] A Xagorari A Papapetropoulos A Mauromatis MEconomou T Fotsis and C Roussos ldquoLuteolin inhibitsan endotoxin-stimulated phosphorylation cascade andproinflammatory cytokine production in macrophagesrdquoJournal of Pharmacology and Experimental Therapeutics vol296 no 1 pp 181ndash187 2001

[88] N Yoshida T Kanekura Y Higashi and T Kanzaki ldquoBidenspilosa suppresses interleukin-1120573-induced cyclooxygenase-2expression through the inhibition of mitogen activated proteinkinases phosphorylation in normal human dermal fibroblastsrdquoJournal of Dermatology vol 33 no 10 pp 676ndash683 2006

[89] M Muchuweti C Mupure A Ndhlala T Murenje and M AN Benhura ldquoScreening of antioxidant and radical scavengingactivity of Vigna ungiculata Bidens pilosa and Cleome gynan-drardquo American Journal of Food Technology vol 2 no 3 pp 161ndash168 2007

[90] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens bipinnata L against carbon tetrachloride-induced liver fibrosis in ratsrdquo Journal of Pharmacy and Pharma-cology vol 60 no 10 pp 1393ndash1402 2008

[91] S L Chang H H Yeh Y S Lin Y M Chiang T K Wuand W C Yang ldquoThe effect of centaurein on interferon-120574 expression and Listeria infection in micerdquo Toxicology andApplied Pharmacology vol 219 no 1 pp 54ndash61 2007

[92] F Deba T D Xuan M Yasuda and S Tawata ldquoChemical com-position and antioxidant antibacterial and antifungal activitiesof the essential oils from Bidens pilosa Linn var Radiatardquo FoodControl vol 19 no 4 pp 346ndash352 2008

[93] A O T Ashafa and A J Afolayan ldquoScreening the root extractsfrom Biden pilosa L var radiata (Asteraceae) for antimicrobialpotentialsrdquo Journal of Medicinal Plant Research vol 3 no 8 pp568ndash572 2009

[94] T B Nguelefack T Dimo E P Nguelefack Mbuyo P V TanS V Rakotonirina and A Kamanyi ldquoRelaxant effects of theneutral extract of the leaves of Bidens pilosa linn on isolated ratvascular smooth musclerdquo Phytotherapy Research vol 19 no 3pp 207ndash210 2005

[95] M T Grombone-Guaratini K L Silva-Brandao V N SolferiniJ Semir and J R Trigo ldquoSesquiterpene and polyacetyleneprofile of the Bidens pilosa complex (Asteraceae Heliantheae)from Southeast of Brazilrdquo Biochemical Systematics and Ecologyvol 33 no 5 pp 479ndash486 2005

[96] L L Lin C Y Wu H C Hsiu M T Wang and H ChuangldquoStudies on diabetes mellitus I The hypoglycemic activity ofphytosterin on alloxan diabetic ratsrdquo Taiwan Yi Xue Hui Za Zhivol 66 no 2 pp 58ndash65 1967

[97] M A N Benhura and I C Chitsiku ldquoThe extractable 120573-carotene content of Guku (Bidens pilosa) leaves after cookingdrying and storagerdquo International Journal of Food Science andTechnology vol 32 no 6 pp 495ndash500 1997

[98] J S Chang L C Chiang C C Chen L T Liu K C Wangand C C Lin ldquoAtileukemic activity of Bidens pilosa l varminor (blume) sherff andHouttuynia cordata thunbrdquo AmericanJournal of Chinese Medicine vol 29 no 2 pp 303ndash312 2001

[99] F Deba T D Xuan M Yasuda and S Tawata ldquoHerbicidal andfungicidal activities and identification of potential phytotoxinsfrom Bidens pilosa L var radiata Scherff research paperrdquoWeedBiology and Management vol 7 no 2 pp 77ndash83 2007

[100] J K Kumar and A K Sinha ldquoA new disubstituted acetylacetonefrom the leaves ofBidens pilosa LinnrdquoNatural Product Researchvol 17 no 1 pp 71ndash74 2003

[101] K Ogawa and Y Sashida ldquoCaffeoyl derivatives of a sugarlactone and its hydroxy acid from the leaves of Bidens pilosardquoPhytochemistry vol 31 no 10 pp 3657ndash3658 1992

[102] Q Xia Y Liu and Y Li ldquoDetermination of gallic acid fromdifferent species and different medical parts of Herba Bidens byRP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol24 no 03 pp 308ndash310

[103] W J Lee L F Wu W K Chen C J Wang and T HTseng ldquoInhibitory effect of luteolin on hepatocyte growth fac-torscatter factor-induced HepG2 cell invasion involving bothMAPKERKs and PI3K-Akt pathwaysrdquo Chemico-BiologicalInteractions vol 160 no 2 pp 123ndash133 2006

[104] J A Beutler J H Cardellina C M Lin E Hamel G MCragg andM R Boyd ldquoCentaureidin a cytotoxic flavone fromPolymnia fruticosa inhibits tubulin polymerizationrdquo BioorganicandMedicinal Chemistry Letters vol 3 no 4 pp 581ndash584 1993

[105] A Verma A Su A M Golin B OrsquoMarrah and J K AmorosaldquoThe lateral view a screening method for knee traumardquo Aca-demic Radiology vol 8 no 5 pp 392ndash397 2001

[106] J Corren M Lemay Y Lin L Rozga and R K RandolphldquoClinical and biochemical effects of a combination botanicalproduct (ClearGuard) for allergy a pilot randomized double-blind placebo-controlled trialrdquo Nutrition Journal vol 7 no 1article 20 2008

[107] M S Gachet J S Lecaro M Kaiser et al ldquoAssessment of anti-protozoal activity of plants traditionally used in Ecuador in thetreatment of leishmaniasisrdquo Journal of Ethnopharmacology vol128 no 1 pp 184ndash197 2010

[108] W C Yang M Ghiotto B Barbarat and D Olive ldquoThe role ofTec protein-tyrosine kinase in T cell signalingrdquo The Journal ofBiological Chemistry vol 274 no 2 pp 607ndash617 1999

[109] S Tewtrakul H Miyashiro N Nakamura et al ldquoHIV-1 inte-grase inhibitory substances from Coleus parvifoliusrdquo Phytother-apy Research vol 17 no 3 pp 232ndash239 2003

[110] T Dimo T B Nguelefack P V Tan et al ldquoPossible mechanismsof action of the neutral extract from Bidens pilosa L leaves onthe cardiovascular system of anaesthetized ratsrdquo PhytotherapyResearch vol 17 no 10 pp 1135ndash1139 2003

[111] T S C Li Chinese and Related North American Herbs CRCPress New York NY USA 2002

[112] H Wu H Chen X Hua Z Shi L Zhang and J ChenldquoClinical therapeutic effect of drug-separated moxibustion onchronic diarrhea and its immunologic mechanismsrdquo Journal ofTraditional Chinese Medicine vol 17 no 4 pp 253ndash258 1997

[113] S W Wright R R Harris J S Kerr et al ldquoSynthesis chemicaland biological properties of vinylogous hydroxamic acids dualinhibitors of 5-lipoxygenase and IL-1 biosynthesisrdquo Journal ofMedicinal Chemistry vol 35 no 22 pp 4061ndash4068 1992

[114] W R Almiron and M E Brewer ldquoClassification of immaturestage habitats of Culicidae (Diptera) collected in CordobaArgentinardquo Memorias do Instituto Oswaldo Cruz vol 91 no 1pp 1ndash9 1996

[115] N L Wang J Wang X S Yao and S Kitanaka ldquoTwo newmonoterpene glycosides and a new (+)-jasmololone glucoside

Evidence-Based Complementary and Alternative Medicine 51

from Bidens parvifloraWilldrdquo Journal of Asian Natural ProductsResearch vol 9 no 5 pp 473ndash479 2007

[116] P Champagnat ldquoRole of the terminal bud in the actionexercised by the cotyledon of Bidens pilosus L var radiatus onits axillary budrdquo Comptes Rendus des Seances et Memoires de laSociete de Biologie vol 145 no 17-18 pp 1374ndash1376 1951

[117] S F Nielsen S B Christensen G Cruciani A Kharazmiand T Liljefors ldquoAntileishmaniai chalcones statistical designsynthesis and three-dimensional quantitative structure-activityrelationship analysisrdquo Journal of Medicinal Chemistry vol 41no 24 pp 4819ndash4832 1998

[118] Y C Hwang J J H Chu P L Yang W Chen and M V YatesldquoRapid identification of inhibitors that interfere with poliovirusreplication using a cell-based assayrdquo Antiviral Research vol 77no 3 pp 232ndash236 2008

[119] V F Andrade-Neto M G L Brandao F Q Oliveira et alldquoAntimalarial activity of Bidens pilosa L (Asteraceae) ethanolextracts fromwild plants collected in various localities or plantscultivated in humus soilrdquo Phytotherapy Research vol 18 no 8pp 634ndash639 2004

[120] R J Marles and N R Farnsworth ldquoAnti-diabetic plants andtheir active constituentsrdquo Phytomedicine vol 2 no 2 pp 137ndash189

[121] M Habeck ldquoDiabetes treatments get sweet help from naturerdquoNature Medicine vol 9 no 10 p 1228 2003

[122] H W Lin G Y Han and S X Liao ldquoStudies on the activeconstituents of the Chinese traditional medicine Polygonatumodoratum (Mill) DrucerdquoActa Pharmaceutica Sinica vol 29 no3 pp 215ndash222 1994

[123] C L T Chang Y C Chen H M Chen N S Yang andW C Yang ldquoNatural cures for type 1 diabetes a reviewof phytochemicals biological actions and clinical potentialrdquoCurrent Medicinal Chemistry vol 20 no 7 pp 899ndash907 2013

[124] L Dey A S Attele and C S Yuan ldquoAlternative therapies fortype 2 diabetesrdquo Alternative Medicine Review vol 7 no 1 pp45ndash58 2002

[125] C L T ChangH Y Liu T F Kuo et al ldquoAnti-diabetic effect andmode of action of cytopiloynerdquo Evidence-Based Complementaryand Alternative Medicine vol 2013 Article ID 685642 13 pages2013

[126] M R Khan M Kihara and A D Omoloso ldquoAnti-microbialactivity of Bidens pilosa Bischofia javanica Elmerillia papuanaand Sigesbekia orientalisrdquo Fitoterapia vol 72 no 6 pp 662ndash6652001

[127] T Dimo T B Nguelefack P Kamtchouing E Dongo ARakotonirina and S V Rakotonirina ldquoHypotensive effects ofmethanol extract from Bidens pilosa Linn on hypertensive ratsrdquoComptes Rendus de lrsquoAcademie des Sciences Serie III vol 322no 4 pp 323ndash329 1999

[128] A K Hassan O Deogratius J F Nyafuono O Francis and OP Engeu ldquoWound healing potential of the ethanolic extracts ofBidens pilosa and Ocimum suaverdquo African Journal of Pharmacyand Pharmacology vol 5 no 2 pp 132ndash136 2011

[129] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[130] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[131] C Kandaswami and EMiddleton ldquoFree radical scavenging andantioxidant activity of plant flavonoidsrdquo Advances in Experi-mental Medicine and Biology vol 366 pp 351ndash376 1994

[132] K Yamamoto H Kakegawa H Ueda et al ldquoGastric cytopro-tective anti-ulcerogenic actions of hydroxychalcones in ratsrdquoPlanta Medica vol 58 no 5 pp 389ndash393 1992

[133] J O C Ezeonwumelu A K Julius C N Muhoho et alldquoBiochemical and histological studies of aqueous extract ofBidens pilosa leaves from Ugandan Rift valley in ratsrdquo BritishJournal of Pharmacology and Toxicology vol 2 no 6 pp 302ndash309

[134] L Frida S Rakotonirina A Rakotonirina and J P SavineauldquoIn vivo and in vitro effects of Bidens pilosa L (Asteraceae)leaf aqueous and ethanol extracts on primed-oestrogenized ratuterine musclerdquo African Journal of Traditional Complementaryand Alternative Medicines vol 5 no 1 pp 79ndash91 2008

52 Evidence-Based Complementary and Alternative Medicine

Composition Comments

1 Please provide valid postal code to the address if

possible

2 We made the highlighted change in Table 4 for the sake

of correctness Please check

3 Please check the correctness of the highlighted part in

Table 5

4 We considered the highlighted part in Table 11 as

reference Please check similar cases throughout

5 Please check the format of Table 18

6 We reformatted Table 20 Please check

7 We made the highlighted change in Table 4 for the sake

of consistency Please check similar cases throughout the

paper

8 We made the highlighted change(s) for the sake

of consistency Please check simila highlighted cases

throughout

9 We made the highlighted change in Table 9 for the sake

of clarity Please check similar cases through out

10 We made the highlighted change(s) for the sake of

correctness Please check

11 Comment on ref [66] This reference is a repetition

of [18] Please check similar cases throughout

Author(s) Name(s)

Author 1

Author 2

Author 3

Evidence-Based Complementary and Alternative Medicine 5

Table 3 Continued

Disorder Plant partb Mode of use Regioncountry References

Eye Infection LE andWP Fresh or juice Uganda

Middle America [18 21]

Antimicrobial APDecoction fordrinkingbathingexternal use

Trinidad andTobago [22]

Pulmonarytuberculosis WP

Decoction ormaceration takenorally

CubaChina [20 23]

Bacterial infectionsingastrointestinal tracts

WP Decoction Trinidad andTobago [22]

Renal infection LE Decoction takenorally Cuba [20]

Sore throat LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21 23]

Cough WP Decoction takenorally

CubaChina [20 23]

Coolness of theuterus WP Decoction taken

orally Cuba [20]

Menstrualirregularities WP Decoction taken

orally Cuba [20]

Dysmenorrhea WP Decoction Bafia Cameroon [25]Hyperemesisgravidarum(morning sickness)

WP Decoction Africa [18]

Hemorrhoids WP Decoction Hong Kong [23]

Nose bleeds LE andWP Fresh or decoction

ChinaMiddle AmericaUganda

[18 21 23]

Stomach ulcers LE andWP

Maceration or juicetaken orally

CubaMiddle AmericaUganda

[18 20 21 23]

Cuts burns and skinproblems

LE andWP

Fresh plant ordecoction topicalapplicationbathing

Trinidad andTobagoAfricaChinaCameroonBrazilVenezuela

[18 22 26]

Wounds WP Crushed herb

ChinaAfricaCentral AmericaHawaii

[23]

Snake bites WP Pulverized herb China [23]bLE leaves ST stem FW flower WP whole plant RT roots AP aerial parts

primary pharmacological properties of B pilosa extracts andphytochemicals are presented and discussed

31 AntiCancer Activity Folkloric reports revealed the pos-sible antitumor efficacy of B pilosa and several scientific in2vitro studies have supported the claim that B pilosa extractsand isolated compounds possess anti-cancer activities againsta variety of cancer cells Several studies have used bioassay

guided isolation and fractionation methods to discover newcompounds from B pilosa For example Kviecinski andcolleagues tested hydroalcoholic crude extracts chloroformethyl acetate and methanol fractions for anti-tumor activity[82] The cytotoxicity of the extracts was assessed usingbrine shrimp hemolytic MTT and neutral red uptake(NRU) assays In vivo studies were performed using Ehrlichascites carcinoma in isogenic BALBc mice Among them

6 Evidence-Based Complementary and Alternative Medicine

Table 4 Aliphatic natural products isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

1 Heneicosane CH3(CH2)19CH3 Aerial (Tanzania) [18]2 Docosane CH3(CH2)20CH3 Aerial (Tanzania) [18]3 Tricosane CH3(CH2)21CH3 Aerial (Tanzania) [18]4 Tetracosane CH3(CH2)22CH3 Aerial (Tanzania) [18]5 Pentacosane CH3(CH2)23CH3 Aerial (Tanzania) [18]6 Hexacosane CH3(CH2)24CH3 Aerial (Tanzania) [18]7 Heptacosane CH3(CH2)25CH3 Aerial (Tanzania) [18]

8 Octacosone CH3(CH2)26CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

9 Nonacosane CH3(CH2)27CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

10 Triacontane CH3(CH2)28CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

11 Hentriacontane CH3(CH2)29CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

12 Dotriacontane CH3(CH2)30CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

13 Tritriacontane CH3(CH2)31CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

14 2-Butoxy-ethanol CH3(CH2)3OCH2CH2OH Whole (Taiwan) [32]15 Tetracosan-1-ol CH3(CH2)22CH2OH Aerial (Tanzania) [18]16 Hexacosan-1-ol CH3(CH2)24CH2OH Aerial (Tanzania) [18]17 1-Octacosanol CH3(CH2)26CH2OH Aerial (Tanzania) [18]

18 1-Hentriacontanol CH3(CH2)29CH2OHNot found(Taiwan) [31]

19 Tetradecanoic acid Myristic acid CH3(CH2)12CO2H Aerial (Tanzania) [18]20 Hexadecanoic acid Palmitic acid CH3(CH2)14CO2H Aerial (Tanzania) [18]21 Octadecanoic acid Stearic acid CH3(CH2)16CO2H Aerial (Tanzania) [18]22 Eicosanoic acid Arachidic acid CH3(CH2)18CO2H Aerial (Tanzania) [18]

23 Docosanoic acid Behenic acid CH3(CH2)20CO2HLeaves(Philippines) [33]

24 2-butenedioic acidO

OHO

HO Aerial (China) [34 35]

25 (Z)-9-Octadecenoicacid Oleic acid

O

OH Aerial (Tanzania) [18]

Evidence-Based Complementary and Alternative Medicine 7

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

26 (E)-9-Octadecenoicacid Elaidic acid

O

HOLeaves(Philippines) [33]

27 (ZZ)-912-Octadecadienoic acid

Linolic acid or Linoleicacid

O

OHAerial(Tanzania)whole (Taiwan)

[18 36]

28(ZZZ)-91215-Octadecatrienoicacid

120572-Linoleic acid

O

OH Whole (Taiwan) [32]

29(ZZ)-912-Octadecadienoic acidethyl ester

Ethyl linoleate O

O

Whole (Taiwan) [32]

30(ZZZ)-91215-Octadecatrienoicacid methyl ester

Methyl linolenate O

O

Whole (Taiwan) [32]

31(ZZZ)-91215-Octadecatrienoicacid ethyl ester

Ethyl linolenate O

O

Whole (Taiwan) [32]

32(Z)-9-Octadecenoicacid 2-butoxyethylester

2-Butoxyethyl oleate O

OO

Whole (Taiwan) [32]

33 2-Butoxyethyllinoleate O

OO Whole (Taiwan) [32]

34(ZZZ)-91215-Octadecatrienoicacid butoxyethyl ester

2-Butoxyethyl linoleate O

OO

Whole (Taiwan) [32]

3517E9E15E-Heptadecatetraene-1113-diyne

Heptadeca-2E8E10E16-tetraen-46-diyne

Not found(China) [37]

8 Evidence-Based Complementary and Alternative Medicine

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

36 111-Tridecadiene-3579-tetrayne

Roots (notstated) [38]

37 1-Tridecene-357911-pentayne Pentayneene

Leaves (notstated) andnot found(Egypt)

[38 39]

38 5-Tridecene-7911-triyne-3-ol

HO

not found(Egypt) [39]

39 21012-Tridecatriene-468-triyn-1-ol

OHPart not specified(Not stated) [40]

40 212-Tridecadiene-46810-tetrayn-1-ol

111-Tridecadiene-3579-

tetrayn-13-ol

OH Roots (Notstated) notfound(Egypt)

[38 39]

41 212-Tridecadiene-46810-tetraynal

111-Tridecadiene-3579-

tetrayn-13-al

O

Roots (Germany) [41]

42212-Tridecadiene-46810-tetrayn-1-ol1-acetate

111-Tridecadiene-3579-

tetrayn-13-acetateO

O

Roots (notstated) [38]

43(5E)-15-Tridecadiene-79-diyn-3413-triol OH

OH

HO

Aerial (China) [42]

44(6E12E)-3-oxo-tetradeca-612-dien-810-diyn-1-ol

OH

O

Aerial (China) [42]

45 (E)-5-Tridecene-7911-triyne-12-diol

12-Dihydroxy-5(E)-tridecene-7911-triyne

HO

OHWhole (Taiwan) [43]

46 (E)-6-Tetradecene-81012-triyne-13-diol

13-Dihydroxy-6(E)-tetradecene-81012-triyne

OH

OHWhole (Taiwan) [43ndash45]

47(2R3E11E)-311-Tridecadiene-579-triyne-12-diol

Safynol

OH

HO

Not found (Egyptand China) [37 39]

Evidence-Based Complementary and Alternative Medicine 9

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

4857911-Tridecatetrayne-12-diol

12-Dihydroxytrideca-57911-tetrayne

HOOH

Whole (Taiwan) [43 44 46]

49(R)-357911-Tridecapentayne-12-diol

(R)-12-Dihydroxytrideca-357911-pentayne

HO

OH

Aerial (Japan) [47]

50

(4E)-1-(Hydroxymethyl)-4-dodecene-6810-triyn-1-yl-120573-D-glucopyranoside

2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-

tridecene-7911-triyne

O

HO

H

OH

H

H

HOH

O

OH

HO

H Aerial (USA)whole (Taiwan)leaves (Taiwan)

[8 45 48ndash51]

51

(4E)-1-(2-Hydroxyethyl)-4-dodecene-6810-triyn-1-yl-120573-D-glucopyranoside

3-120573-D-Glucopyranosyloxy-1-hydroxy-6(E)-

tridecene-81012-triyne

OHO

H

OH

H

H

HOH

O

OH

OH

HAerial (USA andChina) whole(Taiwan) leaves(Taiwan)

[8 35 4245 48ndash51]

52

3-Hydroxy-6-tetradecene-81012-triynyl-120573-D-glucopyranoside

120573-D-Glucopyranosyloxy-

3-hydroxy-6E-tetradecence-81012-triyne

OH

O

OHO

H

OH

H

H

HOH

OH

HWhole (Mexico) [52]

53

1-(Hydroxymethyl)-46810-dodecatetrayne-1-yl-120573-D-glucopyranoside

Cytopiloyne2-120573-D-

Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne

O

HO

OHO

H

OH

H

H

HOH

OH

HWhole (Taiwan)leaves (Taiwan) [8 50 53]

54

2-O-D-Glucosyltrideca-11E-en-3579-tetrayn-12-diol

OH

O

O

OHH

HO

H

H

HHOH

OH

Leaves (Brazil) [27]

10 Evidence-Based Complementary and Alternative Medicine

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

55

(R)-1-(Hydroxymethyl)-246810-dodecapentayn-1-yl-a-D-glucopyranoside

2- 120573 -D-Glucopyranosyloxy-1-hydroxytrideca-357911-pentayne

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Chinaand Japan) [35 47]

56

1-[[(Carboxyacetyl)oxy]methyl]46810-dodecatetraynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Japan) [54]

57

(4E)-1-[[(Carboxyacetyl)oxy]-methyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHO

H

OH

Aerial (Japan) [54]

58

(4E)-1-[[(Carboxyacetyl)oxy]-ethyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

O

O

O

OHO

O

OHOH

H

HO

H

H

HHO

HOH

Aerial (Japan) [54]

59 (5E)-5-Heptene-13-diyn-1-yl-benzene

1-Phenylhepta-13-diyn-5en Whole (Taiwan) [32]

60 7-Phenyl-2(E)-heptene-46-diyn-1-ol

OHRoots (notstated) Aerial(China)

[38 42]

617-Phenyl-2(E)-heptene-46-diyn-1-ol-acetate

OO

Roots (notstated Brazil) [38 55 56]

62 7-Phenyl-46-heptadiyn-2-ol Pilosol A

OH

Whole (Taiwan)aerial (China) [42 57]

Evidence-Based Complementary and Alternative Medicine 11

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

63 7-Phenylhepta-46-diyn-12-diol HO

OH

Aerial (China) [42]

64 135-Heptatriyn-1-yl-benzene

1-Phenylhepta-135-triyne

Leaves (notstated) leaves oftissue culture(not stated)aerial (TanzaniaChina)Whole (Taiwan)roots (Brazil)

[18 34 3842 56ndash58]

65 7-Phenyl-246-heptatriyn-1-ol HO

Leaves (notstated) aerial(China)

[38 42]

667-Phenyl-246-heptatriyn-1-ol-acetate O

O Leaves (notstated) [38]

675-(2-Phenylethynyl)-2-thiophenemethanol

S OHAerial (China) [42]

685-(2-Phenylethynyl)-2120573-glucosylmethyl-thiophene

O

O

OH

H

OH

H

H

HOH H

O

HO

S

Aerial (China) [42]

69

3- 120573 -D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne

OH

O

OH

HOHO

H

H

HHOH

OH

Leaves(Cameroon) [28]

701-Phenyl-13-diyn-5-en-7-ol-acetate O

O

Leaves (Brazil) [27]

SN denotes serial number

the chloroform fraction was the most toxic with a halfmaximal inhibitory concentration (IC

50) of 97 plusmn 72 and 83 plusmn

52 120583gmL in NRU and MTT respectively [82] Kumari andcolleagues also reported the anti-cancer and anti-malarialactivities of B pilosa leaves [77] Based on a cytotoxicity-directed fractionation strategy they identified phenyl-135-heptatriene with IC

50values of 8plusmn001 049plusmn045 07plusmn001

and 10 plusmn 001 120583gmL against human oral liver colon andbreast cancer cell lines respectively However phenyl-135-heptatriyne showed lower activity against breast cancer celllines than the chloroform leaf extract which had an IC

50

value of 65 plusmn 001 120583gmL Moreover the positive controltaxol showed higher activity than phenyl-135-heptatriyne[77] Furthermore in vitro comet assays were performed toevaluate the toxicity of n-hexane chloroform and methanolextracts of B pilosa and its ethyl acetate acetone and waterfractions onHela andKB cellsThe ethyl acetate fraction fromthe methanol extract exhibited the highest activity with halfmaximal cytotoxic concentrations (CTC

50) of 9652120583gmL

and 5862120583gmL against Hela and KB cells respectivelyDespite the moderate toxicity these findings suggest thatthese B pilosa extractsfractions could be useful for future

12 Evidence-Based Complementary and Alternative Medicine

Table5Flavon

oids

isolated

from

Bpilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

71

2-[(34-Dihydroxyph

enyl)-

methylene]-

6-hydroxy-3(2H

)-benzofuranon

e

Sulfu

retin

O

OHO

OH

OH

Aeria

l(Ch

ina)

[35]

72

2-[(34-Dihydroxyph

enyl)-

methylene]-

67-dihydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-673101584041015840-

tetrahydroxy

maritimetin

O

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

73

2-[(34-Dihydroxyph

enyl)-

methylene]-6-

(120573-D

-glucopyrano

syloxy)-7-

hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-120573-D

-glucop

yranosyl-

673101584041015840-te

trahydroxy

maritimein

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japanand

China)aerial(Ch

ina)

[355960]

74

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

(120573-D

-glucopyrano

syloxy)-

6-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-7-O-120573-D

-glucop

yranosyl-

673101584041015840-

tetrahydroxy

O

OHO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japan)

[59]

75

6-[(6-O-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-acetyl-

120573-D

-glucopyrano

syl)

-673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OHO

H HO

H

H

HOH

HO

O

O

Leaves

(Not

stated)

aeria

l(Ch

ina)

[3559]

Evidence-Based Complementary and Alternative Medicine 13

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

76

6-[(36-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-

2-[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone(Z)-6-O

-(36-di-O

-acetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

bideno

sideA

O

O

HO

OH

OH

OHOH O

H

H

HOH

HO

O

O

O

Leaves

(China)

[60]

77

6-[(46-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(410158401015840610158401015840-

diacetyl-120573-D

-glucropyrano

syl)-

673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

HO

O

OO

Leaves

(not

stated)

aeria

l(Ch

ina)

[343561]

78

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

hydroxy-6-

[(246-tri-O

-acetyl-120573

-D-

glucop

yranosyl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(210158401015840410158401015840610158401015840-

triacetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

O

OOO

O

Leaves

(not

stated)

aeria

l(Ch

ina)

[3461]

79

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-6-

[(346-tri-O

-acetyl-120573

-D-glucopyrano

syl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(310158401015840410158401015840610158401015840-

triacetyl-120573

-D-glucrop

yranosyl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H OH

H

HOH

HO

OOO O

Aeria

l(Ch

ina)

[3435]

80

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-

6-[[6-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propenyl]-120573-

D-glucopyrano

syl]o

xy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-

p-coum

aroyl-

120573-D

-glucropyrano

syl)-

673101584041015840-te

trahydroxy

OH

O

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OLeaves

(Japan)

[59]

14 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

81

1-[2-(120573-D

-Glucopyrano

syloxy)-

4-hydroxyphenyl]-2-

hydroxy-3-(3-

hydroxyphenyl)-2-

prop

en-1-on

e

Chalcone120572

32101584041015840-

tetrahydroxy-

21015840-O

-120573-D

-glucop

yranosyl

HO

O

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[35]

821-(24-Dihydroxyph

enyl)-

3-(34-dihydroxyph

enyl)-

2-prop

en-1-on

eBu

tein

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

833-(34-D

ihydroxyph

enyl)-

1-(234-trihydroxyph

enyl)-

2-prop

en-1-on

eOkanin

O

HO

HO

OH

OH

OH

Leaves

(China)

[60]

84

3-(34-D

ihydroxyph

enyl)-

1-[3-(120573-D

-glucop

yranosyloxy)-

24-dihydroxyphenyl]-2-

prop

en-1-on

e

Okanin

31015840-O

-120573-D

-glucosid

e

O

HO

OH

OH

OH

OO

HO

H

HO

H

H

HOH

H

OH

Leaves

(Germany)

flowers(Germany)

[61ndash63]

85

3-(34-D

ihydroxyph

enyl)-

1-[4-(120573-D

-glucop

yranosyloxy)-

23-dihydroxyphenyl]-

2-prop

en-1-on

e

Okanin

41015840-O

-120573-D

-glucosid

e

OOH

OH

OH

HOO

OHO

H

HO

H

H

HOH

H

OH

Flow

ers(Germany)

leaves

(Japan)

[5963]

Evidence-Based Complementary and Alternative Medicine 15

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

86Okanin41015840-O

-120573-D

-(610158401015840-

O-acetylglucosid

e)

O

O

OH

OH

OH

OHO

H

HO

H

H

HOH

H

HO

O

O

Flow

ers(Germany)

[63]

87

1-[4-[(46-di-O

-Acetyl-

120573-D

-glucopyrano

syl)-oxy]-

23-dihydroxyphenyl]-3-

(34-dihydroxyph

enyl)-2-

prop

en-1-on

e

Okanin

41015840-O

-120573-D

-(410158401015840610158401015840-diacetyl)-

glucop

yranoside

O

O

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Aeria

l(Ch

ina)

[34]

88Okanin41015840-O

-120573-D

-(210158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[64]

89Okanin41015840-O

-120573-D

-(310158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

O

OAe

rial(Ch

ina)

[34]

16 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

90

1-[23-Dihydroxy-4-

[[6-O-[3-(4-hydroxyph

enyl)-

1-oxo-2-prop

enyl]-120573-D

-glucop

yranosyl]oxy]-ph

enyl]-

3-(34-dihydroxyph

enyl)-

2-prop

en-1on

e

Okanin

41015840-O

-120573-D

-(610158401015840-tr

ans-

p-coum

aroyl)-

glucoside

O

O

OH

OH

OH

OHOH

HO

H

H

HOH

H

HO

OO

OH

Leaves

(Germany)

[64]

91Okanin41015840-O

-120573-D

-(410158401015840-acetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Leaves

(Germany)

[65]

92

Okanin

41015840-O

-120573-D

-(210158401015840410158401015840-diacetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[65]

Evidence-Based Complementary and Alternative Medicine 17

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

93

Okanin

41015840-O

-120573-D

-(310158401015840410158401015840-diacetyl-610158401015840-

trans-p-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

OH

O O

Leaves

(Germany)

[65]

94

Okanin

41015840-O

-[120573-D

-glucopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

OOH

OH

OH

HO

OH

HO

HHO

H

HOH

HO

OH

OH

HO

HHO

H

HOH

HO

Flow

er(G

ermany)

[63]

95Okanin

3101584041015840-di-O

-120573-D

-glucosid

e

OOH

OH

OH

O OO

H HO

HHOH

HOH

HHO

OHO

H

HO

H

H

HOH

H

OH

Flow

er(G

ermany)

[63]

18 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

96

1-[3-(120573-D

-Glucopyrano

syloxy)-

24-dihydroxyphenyl]-

3-(3-hydroxy)-

4-metho

xyph

enyl)-

2-prop

en-1-on

e

Okanin

4-methylether-

31015840-O

-120573-D

-glucop

yranoside

OOH

O

OH

HO

O

H HO

HHOH

HOH

H

OHO

Leaves

(Germany)aerial

(China)

[3562]

97

Okanin4-methylether-

3101584041015840-di-O

-120573-(41015840101584061015840101584041015840101584010158406101584010158401015840-

tetraacetyl)-

glucop

yranoside

OOH

OOH

O

HH

OH

HO

HOH

H

OO

O

Ac

Ac

AcAc

OO

H

HO

H

H

HOH

H

O

Aeria

l(Ch

ina)

[42]

98Ch

alcone210158404101584061015840-tr

imetho

xy-

4-O-D

-glucopyrano

syl-

dihydro

Not

foun

dLeaves

(China)

[60]

992-(34-D

ihydroxyph

enyl)-23-

dihydro-78

-dihydroxy-

4H-1-

benzop

yran-4-one

Okanin

isoOO

OH

HO

OH

OH

Leaves

(China)

[60]

100

2-(34-D

ihydroxyph

enyl)-

23-dihydro-8-hydroxy-

7[(246-tr

i-O-acetyl-120573

-D-

glucop

yranosyl)oxy]-4H

-1-b

enzopyran-4-on

eOO

OH

OH

OH

OH

O

HHO

H

HO

HO

OAc

Ac

Ac

Aeria

l(Ch

ina)

[34]

Evidence-Based Complementary and Alternative Medicine 19

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

101

57-Dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

OO

OH

HO

OH

Aeria

l(TanzaniaC

hina)

[4266]

102

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

7-O-glucopyrano

side

OO

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Tanzania)

[66]

103

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-4H

-1-benzop

yran-4-one

Luteolin

OO

HO

OH

OH

OH

Aeria

l(TanzaniaC

hina

Vietnam)

[343542

6667]

104

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5hydroxy-4H-1-

benzop

yran-4-one

Luteolin

7-O-120573-D

-glucop

yranoside

OOOH

OH

OH

OO

HO

H

HO

H

H

HOH

OH H

Aeria

l(Tanzania)

[66]

105

57-Dim

etho

xy-6-

(5-m

etho

xy-6-m

ethyl-

4-oxo-4H

-pyran-3-yl)-

2-ph

enyl-4H-1-

benzop

yran-4-one

5-O-M

ethylhosludin

O OOO

O

O

O

Aeria

l(Ugand

a)[68]

20 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

106

3-(120573-D

-Glucopyrano

syloxy)-

57-dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Astragalin

Kaem

pferol-3-O

-120573-D

-glucopyrano

side

HO

OH

O O

OH

O

O

HO

HHO

H

H

HOH

OH

H

Aeria

l(Ch

ina)

[35]

107

Kaem

pferol3-

(23-di-E

-p-cou

maroyl-

120572-L-rhamno

pyrano

side)

Not

foun

dAe

rial(Vietnam)

[67]

108

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5-hydroxy-36-dimetho

xy-

4H-1-

benzop

yran-4-one

Axillarosid

eO O

OH

OH

OO

OH

OO

HO

H

HO

H

H

HOH

H

OH

Aeria

l(Ch

ina)

[42]

109

57-Dihydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaureidin

O

O

O

O

O

HO

OH

OH

Who

le(Taiwan)

[69]

110

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaurein

O O

OO

O OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan)w

hole

(Taiwan)

[69ndash

71]

Evidence-Based Complementary and Alternative Medicine 21

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

111

5-Hydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-

67-dimetho

xy-4H-

chromen-4-one

Eupatorin

iso

OO O

OH

O

O OH

Not

foun

d(C

hina)

[3772]

112

2-(34-D

imetho

xyph

enyl)-7-

(120573-D

-glucopyrano

syloxy)-35-

dihydroxy-

8-metho

xy-4H-1-

benzop

yran-4-one

O O

OH

O O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

113

7-(120573-D

-Glucopyrano

syloxy)-

5hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-38-

dimehtoxy-4H-1-

benzop

yran-4-one

O O

O

OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

114

Isorhamnetin

3-[O

-120572-

L-rham

nopyrano

syl-

(1-2)-120573-D

-glucop

yranoside]

Not

foun

dAe

rial(Vietnam)

[67]

115

7-[(6-deoxy-120572-L-

Manno

pyrano

syl)o

xy]-

3-120573-D

-glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-4H

-1-benzop

yran-4-one

Luteoside

O O

OH

O

OH

OH

OH

HOH

H

H HO

HO

HO

OH

HO

H

HO

OH

OH

HH

Aeria

l(Ch

ina)

[42]

22 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

116

Luteolin

3-O-120573-D

-Glucopyrano

side

O O

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

HO

Aeria

l(Tanzania)

[66]

117

57-Dihydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Quercetagetin

3631015840-tr

imethylether

OOO

O

O OH

HO

OH

Aeria

l(Ch

ina)

[42]

118

6-((2S3S4S5S)-

Tetrahydro-345-tr

ihydroxy-

6-(hydroxymethyl)-2H

-pyran-2-yloxy)-5-hydroxy-

2-(4-hydroxy-3-

metho

xyph

enyl)-

37-dimetho

xy-4H-

chromen-4-one

Quercetagetin

3731015840-

trim

ethylether-6-O

-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[42]

119

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

JaceinQ

uercetin

3631015840-

trim

ethyl

ether-7-O-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan

China)

who

le(Taiwan)

[425470]

120

2-(34-D

ihydroxyph

enyl)-

357-trihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

OO

HO

OH

OH

OH

OH

Aeria

l(Ch

ina)leaves

(China)who

le(C

hina)

[356074]

Evidence-Based Complementary and Alternative Medicine 23

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

121

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-galactopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-4-one

Quercetin

3-O-120573-D

-galactosidehyperin

hyperosid

e

O O

HO

OH

OH

OH

OOHH

HOH

H

H HO

HO

OH

Aeria

l(TanzaniaJapan)

Leaves

(China)Who

le(C

hina)

[376066

717475]

122

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-glucopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

3-O-120573-

D-glucopyrano

side

O O

HO

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

Aeria

l(TanzaniaC

hina

Japan)L

eaves(Japan)

[355966

75]

123

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-

4-oxo-4H

-1-benzop

yran-3-yl-

120573-D

-glucopyrano

siduron

icacid

Quercetin

3-O-120573-D

-glucuron

opyranoside

O O

HO

OH

OH

OH

OHOH

HOH

H

H HO

HO

OHO

Aeria

l(TanzaniaJapan)

[6671]

124

3-[[6-O-(6-

Deoxy-120572-L-m

anno

pyrano

syl)-

120573-D

-galactopyrano

syl]o

xy]-

2-(34-dihydroxyph

enyl)-57-

dihydroxy-4H

-1-benzop

yran-4-one

Quercetin

3-O-rob

inob

iosid

e

O O

HO

OH

OH

OH

O

HOHH OH

HO

H HH

OO

OHH

HOH

H

H HO

HO

Aeria

l(Japan)W

hole

(Taiwan)

[7071]

24 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

125

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-

3-metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

OH

Roots(Brazil)

[55676]

126

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-

120573-D

-glucopyrano

syl]o

xy]-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120572-

L-rham

nopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

O

HHO HHO

HO H

HH

ORo

ots(Brazil)

[5676]

127

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-120573-D

-glucop

yranosyl]oxy]-5-

hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-3-metho

xy-

4H-1-

benzop

yran-4-one

Quercetin

341015840-

dimethyl

ether-7-O-rutinoside

O OOH

OH

O

O

OH O

HOH

H

HHO

H

O

HHO HHO

HO H

HH

O

HO

Aeria

l(Ch

ina)

[3435]

128

2-(34-D

ihydroxyph

enyl)-3-

(120573-D

-glucofurano

syloxy)-57-

dihydroxy-4H

-1-benzop

yran-

4-on

e

Isoq

uercitrin

O OOH

OH

OHO

H

OHH

OH

HO

H O HOH

OH

Aeria

l(Japan

China)

[3571]

Evidence-Based Complementary and Alternative Medicine 25

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

129

Quercetin

3-O-120573-D

-galactop

yranoside

O

OH

HO

OH OH

O

O

HO

OH

OHOH

OAe

rial(no

tstated)

[18]

130

Quercetin

3-O-rutinoside

O

O

R

OO

OH

OHO

H

O

OH

OH

H3C

H3C

H3C

OH

HO

Who

le(Taiwan)

[70]

26 Evidence-Based Complementary and Alternative Medicine

Table 6 Terpenoids isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(Country) References

131371111-Tetramethylbicyclo[810]undeca-

26-dieneBicyclogermacrene Leaves

(Brazil) [95]

13241111-Trimethyl-8-

methylenebicyclo[720]undec-4-ene

E-Caryophyllene Leaves(Brazil) [95]

1331-Methyl-5-methylene-8-

(1-methylethyl)-16-cyclodecadiene

Germacrene D Leaves(Brazil) [95]

1344-(15-Dimethyl-4-hexen-1-ylidene)-1-methyl-cyclohexene

Z-120574-Bisabolene Leaves(Brazil) [95]

135Decahydro-114-trimethyl-7-methylene-1H-cycloprop[e]-

azulene120573-Gurjunene Leaves

(Brazil) [95]

136 2669-Tetramethyl-148-cycloundecatriene

120572-Humulene120572-caryophyllene

Leaves(Brazil) [95]

13712344a568a-Octahydro-1-isopropyl-7-methyl-4-methylenenaphthalene

120575-Muurolene Leaves(Brazil) [95]

138

12344a568a-Octahydro-4a8-dimethyl-2-(1-methylethylidene)-

naphthalene

Selina-37(11)-diene

Leaves(Brazil) [95]

139(2E7R11R)-371115-

Tetramethyl-2-hexadecen-1-ol

Phytol

HO

Whole(Taiwan) [57]

Evidence-Based Complementary and Alternative Medicine 27

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

140 371115-Tetramethyl-2-hexadecenoic acid Phytanic acid

O

OH

Whole(Taiwan) [57]

141371115-Tetramethyl-2-

hexadecenyl ester-heptanoicacid

Phythylheptanoate O O

Leaves(Not stated) [33]

142

(3S10R13R)-17-((2R5R)-5-Ethyl-6-methylheptan-2-yl)-

2347891011121314151617-tetradecahydro-

1013-dimethyl-1H-cyclopenta[a]phenanthren-

3-ol

Campestrol

HO

Aerial(Tanzania) [18]

143 Not found Phytosterin-B Not foundNot found(TaiwanEgypt)

[39 96]

144 Stigmast-5-en-3-ol 120573-sitosterol

HOH

H

H

Aerial(Tanzania)

whole(Taiwan)

[18 31 57]

145

1314151617-Tetradecahydro-1013-

dimethyl-1H-cyclopenta[a]phenanthren-

3-yloxy)-tetrahydro-6-(hydroxymethyl)-2H-pyran-345-triol

120573-Sitosterolglucoside

H

H

HOH

HO

HHO

H

HOHH O

OH Not found(Egypt) [39]

146 5120572-Stigmasta-7-en-3120573-ol

HO

Whole(Taiwan) [57]

147 5120572-Stigmasta-722t-dien-3120573-ol

HO

Whole(Taiwan) [57]

148 Stigmasta-522-dien-3-ol Stigmasterol

HOH

H

H

Not found(Taiwan)aerial

(Tanzania)leaves (notstated)whole

(Taiwan)

[18 31 3357]

28 Evidence-Based Complementary and Alternative Medicine

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

149 Lup-20(29)-en-3-ol Lupeol

H

HH

HOH

Not found(Egypt) [39]

150 Lup-20(29)-en-3-ol acetate Lupeol acetate

H

HH

O

OH

Not found(Egypt) [39]

151 Olean-12-en-3-ol 120573-amyrin

H

HOH

Not found(Egypt) [39]

152 5913-Trimethyl-242526-trinoroleanan-3-ol Friedelan-3120573-ol

HO

Aerial(Tanzania) [18]

153 5913-Trimethyl-242526-trinoroleanan-3-one

Friedelinfriedelan-3-one

O

Aerial(Tanzania) [18]

1542610151923-

Hexamethyl-2610141822-tetracosahexaene

Squalene

Aerial(Tanzania)Leaves (Notstated)Whole(Taiwan)

[18 33 57]

155 120573120573-Carotene 120573-Carotene Leaves (Notstated) [97]

Evidence-Based Complementary and Alternative Medicine 29

studies [83] Hot water extracts of B pilosa varminor Sheriffwere also assessed for its antileukemic effects on leukemiccell lines L1210 U937 K562 Raji and P3HR1 using XTT-based colorimetric assays The extract inhibited the five celllines with IC

50values ranging from 145120583gmL to 586120583gmL

L1210 K562 Raji and P3HR1 were more sensitive to B pilosaextract with IC

50values below 200120583gmL [98]

Consistent with the antitumor activities of B pilosaextracts and fractions some of its phytochemicals alsoshowed anticancer activity as outlined in Table 11 Amongthem luteolin (103) a well-studied flavonoid with multiplebioactivities was more effective against tumor cell prolifera-tion than its derivatives with IC

50values ranging from 3 120583M

to 50120583M in cells and 5 to 10mgkg in animals Luteolin wasalso found to fight cancer as a food additive at concentrationsof 50 to 200 ppm [78] and prevent skin cancer [78] and cancer3invasion [103] Lee and colleagues reported that luteolinprevents cancer by inhibiting cell adhesion and invasion[103] Significant inhibition concentration was reported to be5 120583M and complete inhibition concentration was reported tobe 40120583M Moreover luteolin was reported to inhibit hepa-tocyte growth factor (HGF)-induced cell scattering as wellas cytoskeleton changes such as filopodia and lamellipodiawhichwas determined using phase-contrast and fluorescencemicroscopy Furthermore luteolin also inhibited the HGF-induced phosphorylation of c-Met ERK 12 and Akt aswell as the MAPKERK and P13K-Akt pathways [78 103]Other mechanisms underlying the anticancer activities ofluteolin are the inhibition of topoisomerase I and II whichinhibits cell replication and DNA repair thus promotingapoptosis regulation of PI-3-KinaseAktMAPKERKJNKactivation of apoptosis in the mitochondrial pathway byactivating caspase 9 and caspase 3 which were found inmalignant cells but not in normal human peripheral bloodmononuclear cells death receptor-induced apoptosis and acell-cycle arrest mechanism inhibition of fatty acid synthasewhich is upregulated in many cancer cells and sensitizationto chemotherapywhereby luteolin increases the susceptibilityof cancer cells to chemotherapy [78] Butein (82) is anotherflavonoid that showed a cytotoxic effect on human colonadenocarcinoma cell proliferation with a reported IC

50value

of 175 120583M Butein at 2 120583M affected the incorporation of[14C]-labeled leucine thymidine and uridine which cancause the inhibition of DNA RNA and protein synthesis ofhuman colon cancer cells Moreover butein also exhibitednoncompetitive inhibition of 1-chloro-24-dinitrobenzene(CDNB) in glutathione S-transferase (GST) activity Tumorresistance was correlated with high levels of GST thus buteininhibited proliferation of cancer cells [80] Another flavonoidpresent in B pilosa centaureidin (109) also showed anti-cancer activity in B lymphoma cells Centaureidin isolatedfrom Polymnia fruticosa inhibited tubulin polymerization invitro and induced mitotic figure formation in CA46 Burkittlymphoma cells Using turbidimetric assay the IC

50value of

centaureidin in inhibition of mitosis was 3120583M [104] Cyto-toxicity of centaureidin was further analyzed using AmericanNational Cancer Institute (NCI) 60 human tumor cell linesThe cytotoxicity potency of centaureidin expressed as GI

50

(50 growth inhibition in the NCI tumor line panel) was024120583M [81] These data mark centaureidin as a promisingantimitotic agent for tumor therapy

In addition to anti-tumor flavones polyynes found in B 4pilosa have also been shown to possess anti-tumor proper-ties Based on a bioactivity-directed isolation approach Wuand colleagues identified two polyyne aglycones from theethyl acetate fraction of B pilosa [44] 12-Dihydroxytrideca-57911-tetrayne (48) and 13-Dihydroxy-6(E)- tetradecene-81012-triyne (46) exhibited significant anticell prolifera-tion activity in primary human umbilical vein endotheliumcells (HUVEC) with IC

50values of 125 120583M and 173 120583M

respectively They also decreased angiogenesis and pro-moted apoptosis in human endothelial cells Their anti-angiogenic and cytotoxic effects correlated with activationof the CDK inhibitors and caspase-7 [44] In addition 12-Dihyroxy-5(E)-tridecene-7911-triyne (45) showed antian-giogenic effects in HUVECs with an IC

50value of 124 120583M as

evidenced by a decrease in the tube formation and migrationof HUVECs [43] The IC

50value of compound 45 in the

inhibition of basic fibroblast growth factor-induced HUVECgrowth was 282120583M However it had higher IC

50values

than those for lung carcinoma cells and keratinocytes Thiscompound could also inhibit cell proliferation of HUVECslung carcinoma A549 cells and HACAT keratinocytes Themechanism by which compound 45 inhibits HUVEC growthand angiogenesis is complicated and includes decreasing theexpression of cell cycle regulators (CDK4 cyclins D1 and Aretinoblastoma (Rb) and vascular endothelial growth factorreceptor 1) caspase-mediated activation of CDK inhibitorsp21 (Cip1) and p27 (Kip) upregulation of Fas ligand expres-sion downregulation of Bcl-2 expression and activation ofcaspase-7 and poly (ADP-ribose) polymerase [43]

32 Anti-Inflammatory Activity B pilosa is commonly usedto treat inflammatory disorders The anti-inflammatory phy-tochemicals present in B pilosa are listed in Table 11 5Cyclooxygenase-2 (COX-2) is a physiologically importantenzyme that converts arachidonic acid to prostaglandin(PGE2) Its expression is induced by a wide variety of external

stimuli indicating its involvement in inflammatory diseasesand it is used as an inflammatory marker [88] Yoshida andcolleagues studied the effects of the aqueous extracts of Bpilosa aerial parts in the production of COX-2 and PGE

2as

well as on the activation of mitogen activated protein kinases(MAPKs) in normal human dermal fibroblasts (HDFs) inresponse to inflammatory cytokine IL-1120573 This work showedthat IL-1120573 activated MAPKs such as ERK12 p38 and JNKto different extents and induced COX-2 expression TheCOX-2 expression in HDFs was regulated mainly by p38following IL-1120573 stimulation Consistently the p38 inhibitorSB203580 blocked this expression Using this cell platformB pilosa extracts were tested for inhibition of inflammationThe extract dose-dependently suppressed the activation ofp38 and JNK and moderately suppressed ERK12 as wellas suppressing COX-2 expression and PGE

2production

30 Evidence-Based Complementary and Alternative Medicine

Table7Ph

enylprop

anoids

isolatedfro

mB

pilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

156

3-(4-H

ydroxyph

enyl)-2-prop

enoica

cid

p-Cou

maricacid

OHO

OH

Who

le(Ja

pan)

[99]

157

2-Metho

xy-4-(2-prop

en-1-yl)-ph

enol

Eugeno

lOOH

Leaves

androots(Japan)

[99]

158

3-(4-H

ydroxy-3-m

etho

xyph

enyl)-

2-prop

enoica

cid

Ferulic

acid

O

OH

O

HO

Who

le(Ja

pan)

[99]

159

3-(34-D

ihydroxyph

enyl)-2-prop

enoic

acid

Caffeicacid

O

HO

OH

OH

Who

leandaeria

l(Japan)

[7599]

160

3-Prop

yl-3[(245-trim

etho

xyph

enyl)-

metho

xy]-24-pentanedione

3-Prop

yl-3-(245-

trim

etho

xy)benzyloxy-

pentan-24-dione

O

O

O

O

O

O

Leaves

(India)

[100]

161

3-(34-D

ihydroxyph

enyl)-2-

prop

enoica

cidethyleste

rCa

ffeateethyl

O

O

OH

OH

Not

foun

d(Taiwan)

who

le(Taiwan)

[434584]

Evidence-Based Complementary and Alternative Medicine 31Ta

ble7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

162

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

d-Erythron

icacid

2-O-caffeoyl-2-C

-methyl

O

HO

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

163

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

2-O-caffeoyl-2-C

-methyl

O

O

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

164

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-24-

dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

3-O-caffeoyl-2-C

-methyl

O

O

HO

HO

O

O

OH

OH

Leaves

(Japan)

[101]

165

4-(A

cetyloxy)-3-[[3-(34-

dihydroxyphenyl)-

1-oxo-2-propen-1-y

l-]oxy]-2-hydroxy-

2-methyl-b

utanoica

cid

O

HO

OH

O OHO

HO

OO

Not

foun

d(Ja

pan)

[73]

166

3-(34-D

ihydroxyph

enyl)-tetrahydro-4-

hydroxy-4-

methyl-5

-oxo-3-fu

ranyl

ester-2-prop

enoica

cid

3-O-C

affeoyl-2-C

-methyl-

D-erythrono

-14-la

cton

eHO

HO

O

OO

OOH

Leaves

(Japan)

[101]

167

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]-oxy]-145-

trihydroxy-cyclohexanecarboxylicacid

Chlorogenica

cid

O

HO

HO

OH

O

OH

O

OH

HO

Aeria

l(Japan)w

hole

(Taiwan)

[707175]

32 Evidence-Based Complementary and Alternative Medicine

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

168

4-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]oxy]-

135-tr

ihydroxy-cyclohexanecarboxylic

acid

4-O-C

affeoylqu

inicacid

OHO

HO

OH

OH

O

O

OH

OH

Aeria

l(Japan)

[71]

169

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-15-

dihydroxy-cyclo

hexanecarboxylicacid

34-Di-O

-caffeoylqu

inic

acid

HO

HO

O

O

O

O

OH

OH

HO

OH O

H

O

Aeria

l(Japan)W

hole

(Taiwan)

[45497071]

170

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-14-dihydroxy-

cyclo

hexanecarboxylicacid

35-Di-O

-caffeoylqu

inic

acid

HO

HO

O O

HO

O

HO

OH OO

OHOH

Aeria

l(Japan)w

hole

(Taiwan)

[45497071]

Evidence-Based Complementary and Alternative Medicine 33

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

171

34-bis[[(2E

)-3-(34-D

ihydroxyph

enyl)-1-

oxo-2-prop

en-1-

yl]oxo]-15

-dihydroxy-

cyclo

hexanecarboxylicacid

45-Di-O

-caffeoylqu

inic

acid

OHO

OH

HO

O

O

OH

OH

O

OH

HO

OWho

le(Taiwan)

[454970]

172

3-[4-[[-6-O-[3-(4-H

ydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]-phenyl]-2-prop

enoica

cid

120573-D

-p-C

oumaricacid4-O

-(6-O

-p-caoum

aroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HOH

HO

O

O

Leaves

(Japan)

[59]

173

3-[4-[[2-O

-Acetyl-6

-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]ph

enyl]2-propeno

icacid

120573-D

-p-C

oumaricacid4-O

-(2-O

-acetyl-6

-O-p-

caou

maroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HO

HO

O

O

Leaves

(Japan)aerial

(China)

[3459]

174

67-Dihydroxy-2-chrom

enon

eEsculetin

cicho

rigenin

OHO HO

O

Not

foun

d(Egypt)

[39]

34 Evidence-Based Complementary and Alternative Medicine

Table 8 Aromatic compounds isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

175 12-Benzenediol PyrocatechinHO

HOWhole(Japan) [99]

176 4-Ethyl-12-benzenediol Pyrocatechol

OH

OH

Whole(Japan) [99]

177 Dimethoxyphenol26-dimethoxyphenol

O

HO

O

Roots (Japan) [99]

178 4-Ethenyl-2-methoxy-phenol p-Vinylguaiacol

O

OHWhole(Japan) [99]

179 2-Hydroxy-6-methylbenzaldehyde

6-Methyl-salicylaldehyde O

OH

Whole(Japan) [99]

180 Benzene-ethanol 2-Phenyl-ethanolHO

Whole(Japan) [57]

181 4-Hydroxy-3-methoxy-benzaldehyde Vanillin O

O

HOAerial(Japan) [99]

182 3-Hydroxy-4-methoxy-benzaldehyde Vanillin iso

OO

OH

Leaves(Japan) [99]

183 4-Hydroxybenzoic acid p-Hydroxybenzoicacid

O

HOOH Whole

(Japan) [99]

184 2-Hydroxybenzoic acid Salicylic acidO

HO

HO

Stem androots (Japan) [99]

185 34-Dihydroxybenzoicacid Protocatechuic acid

O

HOOH

OH

Whole(Japan) [99]

1864-Hydroxy-

3methoxybenzoicacid

Vanillic acid

OOH

O

OH

Aerial(Uganda)

Roots (Japan)[68 99]

Evidence-Based Complementary and Alternative Medicine 35

Table 8 Continued

S N IUPAC names Common names Structure Plant part(country) References

187 345-Trihydroxybenzoicacid Gallic acid

O

OHHO

HO

HO

Whole(China) [102]

[88] This work supports the use of B pilosa as an anti-inflammatory agent however no compounds responsible forthe anti-inflammatory activity of B pilosa were identified

A further study also reported the anti-inflammatoryactivity as well as the antiallergic activity of B pilosa [75] Inthis study dried powder of the aerial part of B pilosa whichhad been pretreated with the enzyme cellulosine was usedfor further tests The results showed that oral administrationof the cellulosine-treated B pilosa lowered the level of serumIgE in mice 10 days after immunization with DNP (24-dintrophenyl)-Ascaris as an antigen This treatment alsoreduced dye exudation in skin induced by passive cutaneousanaphylaxis and production of inflammatory mediators his-tamine and substance P in rats [75] Phytochemical analysisshowed that cellulosine treatment increased the percentage ofcaffeic acid and flavonoids This study suggests that B pilosaand its phenolics have anti-inflammatory functions

Phenolics and polyynes are major anti-inflammatoryphytochemicals present in B pilosa (Table 11) Unsurpris-6ingly phenolics such as luteolin (103) and ethyl caffeate(161) that are major constituents of B pilosa have alsobeen reported to possess anti-inflammatory activity Lute-olin was reported to exhibit anti-inflammatory activity inmacrophages Xagorari and colleagues showed that luteolininhibited the release of inflammatory cytokines TNF-120572 andinterleukin-6 in RAW 2647 cells following LPS stimulation[87] It inhibited TNF-120572 production with an IC

50value

of 1120583M The underlying anti-inflammatory mechanism ofluteolin was reported to be the inactivation of Akt andNF-120581B activation [87] In addition luteolin was reportedto confer anti-inflammatory activity through inhibition ofLPS-stimulated iNOS expression in BV-2 microglial cellsIt inhibited LPS-activated microglia in a dose-dependentmanner with an IC

50value of 69 120583MMoreover immunoblot

and RT-PCR data proved that luteolin suppressed I120581B-120572degradation and iNOS expression in LPS-activated microglia[85] Kim and colleagues stated that luteolin may havebeneficial effects on inflammatory neural diseases throughinhibition of iNOS expression [85] A related study revealedthat luteolin decreased the transcriptional activity of NF-120581BRelA via partial inhibition of TNF-mediated NF-120581B DNAbinding activity Luteolin also inhibited Akt phosphorylationand induced degradation of a transcription factor interferonregulatory factor (IRF) [86]

Chiang and colleagues showed that ethyl caffeate(161) significantly inhibited NO production in mousemacrophages RAW 2647 cells [84] Based on MTT assays

they concluded that this inhibition was not due to thecytotoxicity of ethyl caffeate The IC

50value of ethyl caffeate

in the inhibition of NO production was 55 120583gmL slightlylower than curcumin (positive control) which has anIC50

value of 65120583gmL They demonstrated that ethylcaffeate exerted anti-inflammatory activity via the reducedtranscription and translation of iNOS (inducible nitric oxidesynthase) in RAW 2467 cells In addition this compoundalso suppressed COX-2 expression in RAW 2467 cellsand MCF-7 cells The in vivo anti-inflammatory effectof ethyl caffeate was verified by testing in TPA-treatedmouse skin Like celecoxib the positive control ethylcaffeate significantly abolished COX-2 expression in adose-dependent manner Ethyl caffeate at 1mg200120583Lsite(24mM) inhibited COX-2 expression at a level comparableto celecoxib at 1mg200120583Lsite (13mM) Remarkably ethylcaffeate at 48mM (2mg200120583Lsite) was more effectivethan celecoxib at 131mM (10mg200120583Lsite) Moreoverthis compound inhibited the activation of nuclear factor-120581B(NF-120581B) by LPS via the prevention of NF-120581B binding toDNA [84] In addition 3 ethyl caffeate analogs (ethyl 34-dihydroxyhydrocinnamate ethyl cinnamate and catechol)also showed different degrees of NF-120581B binding to DNA aslisted in Table 12 Ethyl cinnamate lacks a catechol moietywhich results in ineffective inhibition of NF-120581B binding toDNA [84]

Pereira and colleagues assessed the anti- 7inflammatory and immunomodulatory activities ofB pilosa methanol extract as well as one polyyne2-O-120573-glucosyltrideca-11(E)-en-3 579-tetrayn-12-diol(54) in T lymphocytes and a zymosan-induced arthritismouse model [27] They first examined the in vitro effect of 8the B pilosa extract and compound 54 on cell proliferation ofhuman T cells stimulated with 5120583gmL phytohemagglutinin(PHA) or 100 nM 12-O-tetradecanoyl phorbol-13-acetate(TPA) plus 15 120583M ionomycin and on cell proliferation ofmouse T cells stimulated with 5 120583gmL concanavalin A(Con A) The data demonstrated that both methanol extractand compound 54 suppressed T-cell proliferation in adose-dependent manner The estimated IC

50values of the B

pilosa extract against human T cells stimulated with 5120583gmLPHA and 100 nM TPA plus 15120583M ionomycin were 125 and25 120583gmL respectively In comparison with the methanolextract compound 54 showed 10-fold more inhibition ofhuman T-cell proliferation with an estimated IC

50value of

15 120583gmL Accordingly the B pilosa extract and compound54 dose-dependently suppressed mouse T-cell proliferation

36 Evidence-Based Complementary and Alternative Medicine

Table 9 Porphyrins isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

188

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(15S16S)-10-ethenyl-5-ethyl-1161820-tetrahydro-6111522-tetramethyl-1820-dioxo-15H-912-imino-212-

metheno-471714-dinitrilopyrano[43-

b]azacyclononadecine-16-propanoic

acid

AristophyllC

NH N

HNN

O

O OOOPhytyl

O

Leaves(Taiwan) [100]

189

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -20b-(methoxycarbonyl)-9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-dinitrilo-12-

dioxeto-[310158404101584034]-cyclo-pent[1 2]azacyclononadecine-

19-propanoic acid

Bidenphytin A

NH N

HNN

OPhytylO O O

O O

Leaves(Taiwan) [100]

190

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -2a-

hydroxy-20b-(methoxycarbonyl)-

9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-

dinitrilo-12-dioxeto[310158404101584034]-cyclo-pent[12-b]-

azacyclononadecine-19-propanoic acid

Bidenphytin B

NH N

HNN

OPhytylO O O

O O

OH

Leaves(Taiwan) [100]

191

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO O

HOOO

Leaves(Taiwan) [100]

192

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

Evidence-Based Complementary and Alternative Medicine 37

Table 9 Continued

S N IUPAC names Common names Structure Plant part(country) References

193

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

Leaves(Taiwan) [100]

194

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

195

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3S4S21R)-9-ethenyl-14-ethyl-21-(methoxycarbonyl)-

481318-tetramethyl-20-oxo-3-phorbinepropanoic acid

Pheophytin A

NH N

HNN

PhytylO

O

OO O

Leaves(Taiwan) [100]

with estimated IC50values of 30 and 25 120583gmL respectively

Taken together the data indicate that the B pilosa extractand compound 54 act on human and mouse T cells Totest the in vivo effect of the B pilosa extract and compound54 a zymosan-induced arthritis mouse model was usedThis model was established from B10ASgSnJ mice with aninjection of zymosan (015mg) The zymosan-injected mice9received an intraperitoneal injection of the B pilosa extract(1 5 or 10mg) at one dose a day for 5 days Popliteal lymphnode (PLN) weight was monitored to check the developmentof arthritis The results revealed that 10mg of the methanolextract of B pilosa extract could significantly diminishinflammation as evidenced by PLN weight [27] This worksuggests that B pilosa (and compound 54) can suppressimmune response and inflammation

33 Antidiabetic Activity Anti-diabetic agents are primarilydeveloped from plants and other natural resources [8 24 4951 53] B pilosa is one of 1200 plant species that have beeninvestigated for antidiabetic activity [120 121]B pilosa is usedas an anti-diabetic herb in America Africa and Asia [51 120

122] Many studies have indicated that B pilosa could treattype 1 diabetes (T1D) and type 2 diabetes (T2D) in animals

Etiologically speaking T1D is caused by autoimmune-mediated destruction of pancreatic 120573 cells leading to insulindeficiency hyperglycemia and complications Currentlythere is no cure for T1D Polarization ofTh cell differentiationcontrols the development of T1D Suppression of Th1 celldifferentiation and promotion of Th2 cell differentiationameliorate T1D [123] One study showed that the butanolfraction of B pilosa inhibited T-cell proliferation decreasedTh1 cells and cytokines and increased Th2 cells andcytokines leading to prevention of T1D in nonobesediabetic (NOD) mice [49] Based on a bioactivity-directedisolation strategy 3 polyynes 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne (53) also known ascytopiloyne 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D- Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne (50) were identifiedfrom B pilosa [49 53] The IC

50value of the butanol fraction

was 200120583gmL This inhibition was reported to be partiallyattributed to cytotoxicity because the butanol fraction at

38 Evidence-Based Complementary and Alternative Medicine

Table 10 Other compounds isolated from B pilosa [30]

SN IUPAC names Common names Structure Plant part(country) References

19637-Dihydro-137-

trimethyl-1H-purine-26-dione

CaffeineN

NN

NO

O

Aerial(Uganda) [68]

197

1-((2R4S5R)-Tetrahydro-4-hydroxy-5-(hydroxymethyl)furan-2-yl)-5-

methylpyrimidine-24(1H3H)-dione

ThymidineHO

HO

ON

O

NHO Not found

(China) [37]

198 1-(2-Thienyl)-ethanone 2-Acetyl-thiophene

S

O

Roots(Germany) [41]

199

(2R3S4S5S)-2-(Heptan-2-yloxy)-tetrahydro-6-

((tetrahydro-34-dihydroxy-5-

(hydroxymethyl)furan-2-yloxy)methyl)-

2H-pyran-345-triol

Heptanyl2-O-120573-xylofuranosyl-

(1rarr 6)-120573-glucopyranoside

O

OOH

OH

HO

OHO

H

HO

H

H

HOHH

O

Whole(Taiwan) [70]

200

2-[(3R7R11R)-3-Hydroxy-371115-

tetramethylhexadecyl]-356-trimethyl-

25-cyclohexadiene-14-dione

120572-Tocopherylquinone

OO

HO

Whole(Taiwan) [32]

2017-O-(410158401015840610158401015840-Diacetyl)-

120573-D-glucopyranoside

Not found Leaves(China) [60]

180 120583gmL could cause 50 death of Th1 cells Moreoverthis study suggested that the butanol fraction may preventdiabetes in NOD mice in vivo via downregulation of Th1cells or upregulation Th2 cells that have effects which areantagonistic of those of Th1 cells [49] This was proven byintraperitoneal injection of the butanol fraction at a doseof 3mgkg BW 3 times a week to NOD mice from 4 to 27weeks This dosage resulted in lower incidence of diabetes(33) At a dose of 10mgkg the butanol fraction of Bpilosa totally eliminated (0) the initiation of the diseaseTo further support this result assessment of IgG2a and IgEproduction was performed in the serum of NOD mice Asin vivo results obtained from intracellular cytokine stainingexperiments were not very conclusive levels of IgG2a and IgE

were measured since Th1 cytokine IFN120574 and Th2 cytokineIL-4 favor the production of IgG2a and IgE respectively Asexpected high levels of IgE and some decline in the levels ofIgG2a were observed in the serum Profiling of the butanolextract revealed five compounds 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne(50) 45-Di-O-caffeoyl- quinic acid 35-Di-O-caffeoylquinicacid and 34-Di-O-caffeoylquinic acid Only the first twocompounds showed similar effects on the prevention ofdiabetes in NOD mice as the B pilosa butanol fractionMoreover compound 50 showed greater activity thancompound 69 in terms of enhancement (by 34 comparedto 8) of differentiation of Th0 to Th2 at 15 120583gmL (both

Evidence-Based Complementary and Alternative Medicine 39

Table 11 Chemical constituents of B pilosa and their biological activities

SN Name Classification Molecularformula Biological activities

109 Centaureidin [91] Flavonoid C18H16O8Anti-listerial [69 91]

Cytotoxic [10]

110 Centaurein [91] Flavonoid C24H26O13

Anti-listerial [69 91]Cytotoxic [10]Anti-viral [105]

103 Luteolin [106] Flavonoid C15H10O6

Anti-viral [107 108]Cytotoxic [77]

Anti-inflammatory [109]Anti-allergic [109]

82 Butein [110] Flavonoid C15H12O5Anti-leishmanial [111]

Cytotoxic [78]48 12-Dihydroxytrideca-57911-tetrayne [112] Polyyne C13H12O2 Anti-angiogeneic [112]46 13-Dihyroxy-6(E)-tetradecene-81012-triyne [112] Polyyne C14H16O2 Anti-angiogeneic [112]

45 12-Dihyroxy-5(E)-tridecene-7911-triyne [113] Polyyne C13H14O2Anti-angiogeneic [113]Anti-proliferative [113]

64 1-Phenylhepta-135-triyne [114] Polyyne C13H8

Anti-microbial [115]Anti-malarial [3]Cytotoxic [3]Antifungal [15]

27 Linoleic acid [79] Fatty acid C18H32O2Anti-viral (100)Cytotoxic [17]

161 Ethyl caffeate [84] Phenylpropanoid C11H12O4 Anti-inflammatory [84]

54 2-O-120573-Glucosyltrideca-11(E)-en-3579-tetrayn-12-diol [26] Polyyne C19H20O7Immunosuppressive andAnti-inflammatory [26]

53 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne [50] Polyyne C19H22O7Anti-diabetic [116]

Anti-inflammatory [50]

69 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne [28] Polyyne C20H26O7Anti-diabetic [28]

Anti-inflammatory [94]

50 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne [28] Polyyne C19H24O7

Anti-diabetic [28]Anti-inflammatory [94]

Anti-malarial andantibacterial [47]

129 Quercetin 3-O-120573-D-galactopyranoside [18] Flavonoid C21H20O12 Anti-inflammatory [117]

170 35-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

171 45-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

169 34-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

126 Quercetin 331015840-dimethyl ether7-O-120572-L-rhamnopyranosyl-(1rarr 6)-120573-D-glucopyranoside [118] Flavonoid C21H20O11 Anti-malarial [119]

125 Quercetin 331015840-dimethyl ether-7-O-120573-D-glucopyranoside [118] Flavonoid C21H20O12 Anti-malarial [119]70 1-Phenyl-13-diyn-5-en-7-ol-acetate [27] Polyyne C15H12O2 Anti-malarial [27]199 Heptanyl 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside [70] Miscellaneous C18H44O10 Antioxidant [70]124 3-O-Rabinobioside [70] Saccharide C27H38O15 Antioxidant [70]130 Quercetin 3-O-rutinoside [70] Flavonoid C27H38O15 Antioxidant [70]167 Chlorogenic acid [70] Phenolic C16H26O9 Antioxidant [70]119 Jacein [70] Flavonoid C24H26O13 Antioxidant [70]

49 (R)-12-dihydroxytrideca-357911-pentayne [47] Polyyne C13H8O2Anti-malarial andAntibacterial [47]

40 Evidence-Based Complementary and Alternative Medicine

Table 12 Structure and activity relationship studies of ethyl caffeateusing in vitro NF-120581BDNA binding assays [84]

Compound Concentration(120583M)

NF-120581BDNA binding

Ethyl caffeate 50 100inhibition

Ethyl 34-dihydroxyhydrocinnamate 100 100inhibition

Catechol 400 100inhibition

Ethyl cinnamate 400 Noinhibition

Table 13 Apoptosis in cocultures of T cells and pancreatic 120573 cells[53]

Cellmedium apoptosis andnecrosis

CD4+ T cellscontrol medium lt4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 2

CD4+ T cellswith cytopiloyne-treated 120573 cells 18CD4+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 7

CD8+ T cellscontrol medium 4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 4

CD8+ T cellswith cytopiloyne-treated 120573 cells 4CD8+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 4

Table 14 Radical scavenging activities of B pilosa extracts [70]

Extractscontrol DPPH assayIC50 (120583gmL)

NBThypoxanthinesuperoxide assayIC50 (120583gmL)

Quercetin 198 15Ascorbic acid 634 Not determined120572-tocopherol 897 Not determinedEthyl acetateextract 1383 597

Butanol extract 1669 114Water extract gt100 gt100

compounds) and inhibition (by 40 compared to 10) ofdifferentiation toTh1 at the same concentration [49]

Among the three polyynes found in B pilosa cytopiloyne(53) had the most potent anti-T1D activity [53] To test the invivo effect of cytopiloyne NODmice received intraperitonealor intramuscular injection of cytopiloyne at 25120583gkg BW3 times per week Twelve-week-old NOD mice started todevelop T1D and 70 of NOD mice aged 23 weeks andover developed T1D Remarkably 12- to 30-week-old NODmice treated with cytopiloyne showed normal levels of bloodglucose (lt200mgdL) and insulin (1-2 ngmL) Consistent

Table 15 Radical scavenging activity of secondarymetabolites fromB pilosa [70]

Metabolite (Table 11)control DPPH assay IC50 (120583gmL)199 Not determined124 53130 68167 105169 33171 38119 Not determined110 Not determinedQuercetin 256Caffeic acid 890

Table 16 Antioxidant activity of the essential oils and water extractsfrom B pilosa [92]

Extract IC50 (120583gmL)Leaf essential oils 47Flower essential oils 50Leaf extract 61Flower extract 172

with T1D incidence cytopiloyne delayed and reduced theinvasion of CD4+ T cells into the pancreatic islets [53]

In vitro study showed that cytopiloyne (53) inhibited thedifferentiation of naıveTh (Th0) cells (ie CD4+ T cells) intoTh1 cells and promoted differentiation of Th0 cells into Th2cells [50] The in vitro data are consistent with the in vivoresults indicating that cytopiloyne reducedTh1differentiationand increased Th2 differentiation as shown by intracellularcytokine staining and FACS analysis [53] In line with theskewing of Th differentiation the level of serum IFN-120574 andIgG2c decreased while that of serum IL-4 and serum IgEincreased compared to the negative controls (PBS-treatedmice) Cytopiloyne also enhanced the expression of GATA-3 a master gene for Th2 cell differentiation but not theexpression of T-bet a master gene forTh1 cell differentiationfurther supporting its role in skewingTh differentiation [53]

Also importantly cytopiloyne partially depleted CD4+rather than CD8+ T cells in NOD mice [53] As shown inTable 13 coculture assays showed that the depletion of CD4+T cells was mediated through the induction of Fas ligandexpression on pancreatic islet cells by cytopiloyne leading toapoptosis of infiltrating CD4+ T cells in the pancreas via theFas and Fas ligand pathways However cytopiloyne did notinduce the expression of TNF-120572 in pancreatic islet cells andthus had no effect on CD8+ T cells [53]

In addition Chang and colleagues showed that cytopi-loyne dose-dependently inhibited T-cell proliferation stimu-lated by IL-2 plus Con A or anti-CD3 antibody using [3H]thymidine incorporation assay [53]

Overall the mechanism of action of cytopiloyne andprobably its derivatives in T1D includes inhibition of T-cellproliferation skewing of Th cell differentiation and partialdepletion of Th cells Due to the anti-diabetic mechanisms

Evidence-Based Complementary and Alternative Medicine 41

Table 17 Antibacterial activity of essential oils and flower extracts from B pilosa [92]

Strain Mean zone of inhibition (mm)Leaf essential oil Flower essential oil Leaf extract Flower extract

Micrococcus flavus 127 plusmn 03 87 plusmn 03 102 plusmn 02 108 plusmn 03Bacillus subtilis 173 plusmn 19 117 plusmn 02 109 plusmn 02 103 plusmn 02Bacillus cereus 190 plusmn 14 112 plusmn 03 118 plusmn 04 185 plusmn 10Bacillus pumilus 123 plusmn 07 108 plusmn 02 105 plusmn 04 77 plusmn 02Escherichia coli 137 plusmn 04 203 plusmn 07 102 plusmn 11 140 plusmn 13Pseudomonas ovalis 125 plusmn 08 137 plusmn 15 102 plusmn 06 125 plusmn 06

Table 18 Antibacterial activity of root extracts from B pilosa [93]

Strain MIC50 (mgmL)Methanol extract Acetone extract

Bacillus cereus 10 mdashEscherichia coli 5 5Klebsilla pneumonia 5 10Micrococcus kristinae 10 mdashPseudomonas aeruginosa 10 10Staphylococcus aureus 5 10Sraphylococcus epidermidis 5 5Serratia marcescens 10 mdashShigelea flexneri 10 mdashStreptococcus faecalis 10 mdash

Table 19 Antibacterial activity of B pilosa of compound 29 [47]

Strain MIC50 (120583gmL)Escherichia coli NIHJ 1Escherichia coli ATCC25922 1Klebsiella pneumoniae ATCC700603 128Serratia marcescens ATCC13880 16Pseudomonas aeruginosa ATCC27853 8Staphylococcus aureus FDA209P 05Staphylococcus aureus ATCC29213 025Staphylococcus aureus N315 (MRSA) 05Enterococcus faecalis ATCC29212 2Enterococcus faecalis NCTC12201 (VRE) 1Bacillus subtilis ATCC6633 05Candida albicans ATCC10231 025

of action it was hypothesized that cytopiloyne protectsNOD mice from diabetes by a generalized suppression ofadaptive immunity To evaluate this hypothesis ovalbumin(Ova) was used as a T-cell dependent antigen to primeNOD mice which had already received cytopiloyne orPBS vehicle Ova priming boosted similar anti-Ova titersin cytopiloyne-treated mice and PBS-treated mice but adifference in immunoglobulin isotype was observed in thetwo groups Thus it was concluded that cytopiloyne is animmunomodulatory compound rather than an immunosup-pressive compound [50 53]

T2D is a chronic metabolic disease with serious com-plications resulting from defects in either insulin secretion

insulin action or both [124] A study by Ubillas et alshowed that the aqueous ethanol extract of the aerial partof B pilosa at 1 gkg body weight (BW) lowered bloodglucose in dbdb mice a T2D mouse model [51] Basedon a bioactivity-guided identification compounds 69 and50 were identified Further the mixture of the compounds(69 50) in a 2 3 ratio significantly decreased blood glucoseconcentration and reduced food intake on the second dayof treatment when administered at doses of 250mgkg twicea day to C5BLKs-dbdb mice When tested at 500mgkga more substantial drop in blood glucose level as wellas the stronger anorexic effect (food intake reduced from58 gmouseday to 25 gmouseday) was observed [51] Inthis study it was suggested that the blood glucose lower-ing effect of B pilosa was caused in part by the hungersuppressing effect of its polyynes [51] However the hungersuppressing effect of the ethanol extract of B pilosa wasnot found in the studies described below In another study[24] water extracts of B pilosa (BPWE) were used indiabetic dbdb mice aged 6ndash8 weeks with postprandialblood glucose levels of 350 to 400mgdL Like oral anti-diabetic glimepiride which stimulates insulin release onesingle dose of BPWE reduced blood glucose levels from374 to 144mgdL The antihyperglycemic effect of BPWEwas inversely correlated to an increase in serum insulinlevels suggesting that BPWE acts to lower blood glucose viaincreased insulin production However BPWE had differentinsulin secretion kinetics to glimepiride [24] One flaw incurrent anti-diabetics is their decreasing efficacy over timeThe authors investigated the long term anti-diabetic effectof BPWE in dbdb mice BPWE reduced blood glucoseincreased blood insulin improved glucose tolerance andreduced the percentage of glycosylated hemoglobin (HbA1c)Both long-term and one-time experiments strongly supportthe anti-diabetic action of BPWE [24] In sharp contrast toglimepiride BPWE protected against islet atrophy in mousepancreas The investigators further evaluated anti-diabeticproperties of 3 B pilosa varieties B pilosa L var radiate(BPR) Bpilosa L var pilosa (BPP) and B pilosa L varminor(BPM) in dbdb mice [8] One single oral dose (10 50 and250mgkg body weight) of BPR BPP or BPM crude extractsdecreased postprandial blood glucose levels in dbdb micefor up to four hours and the reduction of glucose levels inthe blood appeared to be dose-dependent Comparing thethree variants BPR extract resulted in a higher reduction inblood glucose levels when administered at the same dose as

42 Evidence-Based Complementary and Alternative Medicine

Table 20 Antifungal activity of B pilosa [92]

Partextract Concentration (ppm) Strain InhibitionCortiicum rolfsii Fusarium solani Fusarium oxysporum

Leaves

Essentialoils

100 857 plusmn 09 682 plusmn 0 745 plusmn 17250 960 plusmn 08 779 plusmn 18 879 plusmn 04

AqueousExtracts

100 446 plusmn 17 605 plusmn 21 716 plusmn 07250 942 plusmn 03 689 plusmn 07 824 plusmn 19

Flowers

Essentialoils

100 604 plusmn 09 892 plusmn 04 869 plusmn 05250 894 plusmn 12 980 plusmn 03 949 plusmn 06

AqueousExtracts

100 331 plusmn 11 714 plusmn 07 573 plusmn 22250 661 plusmn 14 912 plusmn 0 900 plusmn 07

Table 21 Antifungal activity of B pilosa root extracts [93]

Strain LC50 (mgmL)Acetoneextracts

Methanolextracts

Waterextracts

Aspergillus niger 014 006 007Aspergillus flavus 1091 658 0Penicillium notatum 005 005 005

the other two varieties In terms of serum insulin levels a doseof 50mgkg of each extract was used and the BPR extracttogether with the three polyynes significantly increased theserum insulin level in dbdb mice Long-term experiments(28-day treatment) were then conducted using diabetic micewith postprandial glucose levels from 370 to 420mgdL andglimepiride was used as positive controlThe range of dosagesapplied was from 10mgkg BW to 250mgkg BW Resultsshowed that the positive control as well as the crude extractsof the three varieties lowered the blood glucose levels indbdb mice However only BPR extract containing a higherpercentage of cytopiloyne (53) reduced blood glucose levelsand augmented blood insulin levelsmore than BPP andBPMThe percentage of glycosylated hemoglobin A1c (HbA1c) wasalso measured and found to be 79 plusmn 05 in mice aged 10ndash12 weeks and 66 plusmn 02 61 plusmn 03 and 62 plusmn 03 inthe blood of age-matchedmice following treatment with BPRcrude extract (50mgkg) glimepiride (1mgkg) and com-pound 53 (05mgkg) respectively [8] Among the polyynesfound in B pilosa cytopiloyne was the most effective againstT2D Hence cytopiloyne was used for further study on anti-diabetic action and mechanism [125] The data confirmedthat cytopiloyne reduced postprandial blood glucose lev-els increased blood insulin improved glucose tolerancesuppressed HbA1c level and protected pancreatic islets indbdb mice Nevertheless cytopiloyne failed to decreaseblood glucose in streptoztocin (STZ)-treated mice whose bcells were already destroyed Additionally cytopiloyne dose-dependently increased insulin secretion and expression in bcells as well as calcium influx diacylglycerol and activationof protein kinase C120572 Collectively the mechanistic studiessuggest that cytopiloyne treats T2D via regulation of insulin

production involving the calciumDAGPKC120572 cascade in bcells

The studies detailed above point to the conclusion thatcytopiloyne and related polyynes (compounds 69 and 49)are anti-diabetics in animal models The data uncover a newbiological action of polyynes It should be noted that likeall anti-diabetic drugs cytopiloyne failed to prevent or curediabetes completely but reduced diabetic complications [125]Intriguingly 34 polyynes have been found in B pilosa so farIt remains to be seen whether all the polyynes present in thisplant have anti-diabetic activities

34 Antioxidant Activity Free radicals can damage cellularcomponents via a series of chemical reactions [89] leadingto development and progression of cardiovascular diseasecancer neurodegenerative diseases and ageing [70] Freeradicals nitric oxide (NO) and superoxide anions can beproduced in macrophages to kill microbes However anexcessive generation of the free radicals under pathologicalconditions is associated with a wide range of illnessesPlants are known to be rich in antioxidant phytochemicalsChiang and colleagues evaluated the free radical scaveng-ing activity of crude extract fractions and compounds ofB pilosa using 11-diphenyl-2-picrylhydrazyl (DPPH) andhypoxanthinexanthine oxidase assays [70] Using DPPHand hypoxanthinexanthine oxidase assays they found thatthe B pilosa crude extract and the ethyl acetate butanoland water fractions had free radical scavenging activityNine compounds Heptyl- 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside (199) 3-O-Rabinobioside (124) Quercetin3-O-rutinoside (130) Chlorogenic acid (167) 34-Di-O-caffeoylquinic acid (169) 35-Di-O-caffeoylquinic acid (170)45-Di-O-caffeoylquinic acid (171) Jacein (119) and Cen-taurein (110) had DPPH radical scavenging activity [70]The IC

50values of the B pilosa crude extractfractions and

compounds are summarized in Tables 14 and 15 respectivelyMeasurement of free radical scavenging activities is one

way of assessing the antioxidant activities of B pilosa andits fractions and compounds It is interesting that the ethylacetate and butanol fractions are more active than the waterfraction and B pilosa crude extract [70] Of the secondarymetabolites only phenolic compounds 124 130 167 169 and171 showed significant DPPH-radical scavenging activities

Evidence-Based Complementary and Alternative Medicine 43

Further analysis of the structure-activity relationship of thecompounds suggested that substitution of the C3 hydroxylgroup with glycosides increased the activity approximately2-fold (for example in compounds 124 and 130) relative toquercetin (-OH in its C3) [70] Further modification of thestructures of the active compounds needs to be performed totest the effects of various substituents on activity The reasonwhy most of the antioxidant compounds contain phenolmoieties in their structure could be that the reduction-oxidation (redox) properties of phenols allow them to actas reducing agents singlet oxygen quenchers and hydrogendonors

A complementary study by Muchuweti and colleaguesdetermined phenolic content antioxidant activity and thephenolic profile of B pilosa methanol extract [89] Theyestimated that the phenolic content of the methanol extractof B pilosawas 11028plusmn22mgg [89] Vanillin hydroxyben-zaldehyde caffeic acid coumaric acid and ferulic acid werefound in this extractThe B pilosa extract also showed DPPHradical scavenging activity Furthermore the antioxidantactivity of the flavonoids found in B pilosa was correlatedwith its hepatoprotective effects through their inhibitionof NF-120581B activation which may lessen the oxidative stresscaused by the production of free radicals during liver injury[60]This activity might also be due to the anti-inflammatoryeffects of the aqueous extracts of B pilosa aerial parts on theinhibition of COX-2 and PGE

2production [88]

Essential oils from B pilosa flowers and leaves are alsoreported to possess antioxidant activity With the aim ofreplacing chemically synthesized additives Deba and col-leagues [92] worked on the antioxidant antibacterial andantifungal activities of essential oils and water extracts of Bpilosarsquos leaves and flowers Table 16 summarizes the resultsobtained from DPPH free radical scavenging assay

It can be inferred fromTable 16 that essential oils from theleaves possessed the highest activity It is reported elsewherethat monoterpenes present in essential oils such that of Bpilosa have protective effects and antioxidant properties [92]Beta-carotene bleaching method was also performed Leavesessential oils and aqueous extracts of the leaves and flowersshowed higher activity than the flower essential oils This isdue to the volatility of the flower essential oils The activityexhibited by the aqueous extracts was accounted to thepresence of phenolic compounds that are reported to donatea hydrogen atom to free radicals such that the propagation ofthe chain reaction during lipid oxidation is terminated [92]Overall essential oils and phenolics present in B pilosa canbe thought of as major antioxidant compounds

35 Immunomodulatory Activity B pilosa is thought to bean immunomodulatory plant and is reported to be effectivein the treatment of immune disorders such as allergy [75]arthritis [75] and T1D [48 50 53]

As pointed out in the discussion of its anti-diabeticactivities (Section 33) a combination of phytochemicalsmethod and T-cell activation assays was used to studyimmunomodulatory properties of B pilosa IFN-120574 is a keycytokine released by T and NK cells that mediates immune

cells and sustains immunity against pathogens Defects inIFN-120574 expression regulation and activation result in vulner-ability to diseases caused by bacteria and viruses [69] Anelegant study performed byChang and colleagues using IFN-120574 promoter-driven luciferase reporter construct in JurkatT cells showed that hot water crude extracts of B pilosaincreased IFN-120574 promoter activity two-fold [69] Out of thesubfractions of this extract the butanol fraction but not thewater or ethyl acetate fractions increased IFN-120574 promoteractivity six-fold Centaurein (110) and centaureidin (109)were identified from the butanol fraction and were stated tocause a four-fold increase in IFN-120574 promoter activity withEC50

values of 75 120583gmL and 09120583gmL respectively Themechanism of action of centaurein was determined usingtranscription factors such as AP-1 NFAT and NF120581B whichare reported to bind to IFN-120574 promoter and regulate IFN-120574transcription Unlike with the activity of the positive controlPHA centaurein caused a four-fold increase in NFAT a3-fold increase in NF120581B and had little if any effect onAP-1 enhancer activities (23-fold three-fold and ten-foldincreases respectively were seenwith PHA) [69]The authorsconcluded that centaureinmodulates IFN-120574 expression by theNFAT and NF120581B pathways The article only determined themechanism of action of centaurein Its aglycone centaureidinmay act through the samemechanism though this conclusionneeds to be further verified

B pilosa extract and its compounds are reported to inhibitdifferentiation of naıve CD4+ helper T (Th0) cells into Th1cells [49] Using theThcell differentiation assay as a screeningplatform 3 polyynes 2-120573-D-glucopyranosyloxy-1 -hydroxytrideca-57911-tetrayne (53) 3-120573-D-glucopyranosyl-1-hydroxy-6(E)- tetradecene-81012-triyne (69) and 2-120573-D-glucopyranosyloxy-1-hydroxy-5(E)- tridecene-7911-triyne(49) were discovered from B pilosa [49 53] The data showsthat cytopiloyne and other two polyynes suppressed thedifferentiation of type 1 helper T (Th1) cells and productionof Th1 cytokines and promoted that of type 2 helper T (Th2)cells and production of Th2 cytokines thus explaining theimmunomodulatory and anti-inflammatory effects of Bpilosa and its polyynes

Chang and colleagues were the first to report the effect ofthe butanol extract of B pilosa on the autoimmune diabetesand airway inflammation inmice [48] Imbalance in the levelsofTh1 andTh2 and of various cytokines leads to autoimmunediseases T1D and other autoimmune diseases (rheumatoidarthritis Crohnrsquos disease among others) are exacerbated byan increase Th1 levels (specifically CD4+ Th1 cells) whileTh2 cells antagonize this effect [19] Moreover Th2 cellsmediate asthma in ovalbumin-induced hypersensitivity inBALBc mice [48] In their work [48] Chang and colleaguesshowed that 10mgkg butanol extracts with 15 (ww)compound 69 and 11 (ww) compound 49 (Table 11)ameliorated the development of Th1-mediated diabetes inNOD mice through inhibition of 120573 cell death and leukocyteinfiltration At the same dosage the butanol extracts alsoexacerbated ovalbumin-induced pulmonary inflammationin BALBc mice with an increase in the infiltration ofeosinophils and mast cells into the airway of the mice [48]Despite the different outcomes both mouse models proved

44 Evidence-Based Complementary and Alternative Medicine

the concept that control over the Th1Th2 shift is associatedwith autoimmune diseases and that the B pilosa butanolextract can shift the differentiation of Th0 cells to Th2 cells[48 49]

An extended study presented by Chiang and colleaguesshowed that compound 53 modulates T-cell functions [50]Using CD4+ T cells from BALBc mice they demonstratedthat cytopiloyne decreased levels of IFN-120574 producing cells(Th1) by 122 (from 72 to 598) Since Th1 and Th2 celldifferentiation is antagonistic it was expected that compound53 increased the percentage of mouse IL-4 producing cells(Th2) by 72 (from 237 to 309) Subsequent assess-ment of effect of compound 53 on the modulation of thetranscription of IL-4 and IFN-120574 showed that cytopiloyne asexpected decreased the splenocyte levels of IFN-120574 mRNAand increasing that of IL-4 in a dose-dependent manner Inthe range of 01 to 3 120583gmL of compound 53 the effects werenot attributed to its cytotoxicity Consequently using 3120583gmLcytopiloyne for 72 and 96 hours the protein concentration ofIFN-120574 decreased to 186 and 444 respectively Under thesame conditions cytopiloyne increased IL-4 concentrationsto 1985 and 2470 (for 72 and 96 hours resp) Thismodulation of T-cell differentiation exhibited by compound53 was used to explain its anti-diabetic activity [50]

The anti-diabetic role of cytopiloyne was extensivelydiscussed above (Section 33 anti-diabetic activity) Themolecular basis of the regulation of cytokine expression bycytopiloyne has been described Cytopiloyne directly elevatedthe expression level of IL-4 via GATA-3 upregulation inT cells [53] However reduction of IFN-120574 expression in Tcells seemed to come from the indirect opposing effect IL-4cytokine because the expression level of T-bet was unaltered[53] In this way cytopiloyne skewedTh1 polarization into theTh2 state conferring protection against T1D in NOD miceAside frompolarization ofThcell differentiation cytopiloynealso activated the expression of Fas ligand in pancreaticb cells this increase leading to the partial depletion of Tcells and reduction of immune response in local areas suchas the pancreas Of note cytopiloyne also inhibited T-cellproliferation and activation By targeting T cells from threeimmunomodulatory actions cytopiloyne protects againstT1D and probably other Th1-mediated autoimmune diseases[19]

The phytochemical constituents of B pilosa exert theirfunctions on different immune cells to modulate immuneresponse It is possible that some of the compounds mayhave agonistic or antagonistic effects on immune responseImmune function of B pilosa may depend on its compo-sition and amount of compounds which could explain theapparently conflicting report of B pilosa butanol extractaggravating allergy in mice [48] while cellulosine-treatedextract ameliorated allergy [48 75] IFN-120574 promoter reporterassays and T-cell differentiation assays were used to isolate2 flavonoids [69] and 3 polyynes [49 50] as immunomod-ulatory compounds from the butanol fraction of B pilosaInterestingly the flavonoids promote IFN-120574 expression inNK and T cells In marked contrast the polyynes promoteIL-4 expression and indirectly inhibit IFN-120574 expression indifferentiating T cells Sensitivity appears to be the key to

identifying structure- and bioactivity-related phytochemicalsfrom medicinal plants

36 Antimalarial Activity The use of chemical drugs againstpathogens has resulted in drug-resistant mutants Examplesof drug resistance can be found in the species of thePlasmodium that cause malaria It is important to search fornew compounds to combat Plasmodium parasites [47] Astudy of the anti-malarial activity of the leaf extracts of Bpilosa using a combination of phytochemistry and bioassaysshowed that compound 49 (Table 11) showed activity againsta malaria parasite (P falciparum NF54 strain) with an IC

50

value of 60 120583gmL [77] In addition compound 49 isolatedfrom the aerial parts of B pilosa inhibited growth of the Pfalciparum FCR-3 strain with an IC

50value of 035 120583gmL

This compound was tested for its in vivo effect in miceinfected with P berghei NK-65 strain Results showed thatcompound 49 decreased the average parasitemia in the redblood cells by 207 (from 328 of that of the control to121) after an intravenous injection of 08mgkg BWday forfour days [47] Further studies addressing the anti-malarialmechanism underlying both polyynes and clinical studies areneeded

37 Antibacterial Activity Emergence of multiple antibiotic-resistantmicrobes is becoming a global threat to public healthand a challenge to disease treatment For instance penicillinis commonly used to combat a food-borne intracellularbacteriumListeria however penicillin-resistant bacteria havebeen discovered recently [69] Chang and coworkers iso-lated centaurein (110) and centaureidin (109) from B pilosaextract [69 70] Centaurein enhances expression of IFN-120574a key cytokine for macrophage activation and consequentlyenhances bactericidal activity in macrophages [69 70] Inagreement with observed in vitro effects centaurein wasreported to prevent and treat Listeria infection in C57BL6Jmice [91] Mechanistic studies confirmed that centaureinexerted antilisterial action via IFN-120574 expression in wild-typemice but not IFN-120574 knockout mice [91] Further in vitrostudies on centaurein showed that this compound increasedIFN-120574 expression by 13 (from 17 to 20) 20 (from 21to 41) and 11 (from 6 to 17) in CD4+ T cells CD8+Tcells and NK cells respectively That is to say there wasan increase in IFN-120574 producing immune cells As expectedcentaurein also enhanced the expression level of T-bet a keynuclear factor for IFN-120574 expression Consistently centaureinaugmented the serum IFN-120574 levels inC57BL6Jmice and thisaugmentation peaked 24 hours after compound injectionThe quantity of mouse serum IFN-120574 was sufficient to activatemacrophages in vitro and eradicated GFP-producing Listeriainside macrophages However the entry of Listeria intomacrophages was not affected by centaurein-treated mousesera Centaurein treatment at 20120583g per mouse rescued 30of the mice infected with a lethal dose of Listeria (2 times106 CFU) It is noteworthy that in the presence of ampicillin(5 120583gmouse) centaurein (20 120583gmouse) rescued 70 of themice suggesting an additive effect between ampicillin andcentaurein [91] Despite lower abundance centaureidin was

Evidence-Based Complementary and Alternative Medicine 45

30 times more active than centaurein in terms of IFN-120574production [91]

Aside from the indirect antibacterial action mentionedabove extract andor compounds of B pilosa also showeddirect bacteriostatic andor bactericidal action One studyreported that essential oils and leafflower extracts of Bpilosa could suppress the growth of gram positive and gramnegative bacteria as evidenced by zone of inhibition assaysIn this study antibacterial activity of the essential oils fromthe leaves and flowers of B pilosa was determined in anattempt to identify natural products as food preservatives forprevention of microbial multiplication and food oxidationThe essential oils and extracts of B pilosa leaves and flow-ers showed moderate but different extents of antibacterialactivity (Table 17) In general essential oils had higherantibacterial activity than crude extracts One explanation forthis could be that monoterpenes in the essential oils destroycellular integrity and subsequently inhibit the respirationand ion transport processes The presence of antibacterial120573-caryophyllene could be another explanation as reportedelsewhere [92] Another study reported that the methanoland acetone extract of B pilosa roots displayed antibacterialactivities against the bacteria listed in Table 18 [93] andmethanol extracts from the roots seemed to be the mosteffective

Another study indicated that the polyyne (R)-12-dihydroxytrideca-357911-pentayne (49) from this plantalso suppressed bacterial growth as shown by the minimuminhibitory concentration required to inhibit 50 bacterialgrowth (MIC

50) in Table 19 This compound was highly

effective against several Gram positive and Gram negativebacteria including the drug-resistant bacteria Staphylococcusaureus N315 (MRSA) and Enterococcus faecalis NCTC12201(VRE) [47] Strikingly compound 49 had a similar MIC

50

value to antibiotics (ampicillin tetracycline norfloxacin andamphotericin B) in most of the bacteria tested

Antibacterial activity of B pilosa extracts and compo-nents expressed as MIC

50and the mean zone of inhibition

is tabulated in Tables 17 18 and 19The zone of inhibition forAmpicilline (positive control) ranges from 153 plusmn 03mm to443 plusmn 02mm [92]

38 Antifungal Activity B pilosa has traditionally been usedto treat microbial infection Recently different parts of Bpilosa have been tested for antifungal activities Deba andcolleagues first evaluated the antifungal effect of the hotwater extracts of the B pilosa roots stems and leaves againstCorticium rolfsii Fusarium solani and Fusarium oxysporumThey discovered that C rolfsii was most suppressed bytreatment with B pilosa as its growth was reduced at almostall the tested doses followed by F oxysporum and F solani[99] However the fungicidal activities of the stems androots were greater than the leaves [99] Moreover the samegroup assessed the antifungal activity of the essential oilsand aqueous extracts from B pilosa flowers and leaves [92]They showed that the extracts and oils had antifungal activityagainst C rolfsii F solani and F oxysporum Essential oils

appeared to have better fungicidal activity thanwater extractsas summarized in Table 20

Another study by Ashafa and colleagues showed thatacetone methanol and water extracts of the B pilosa rootsshowed antifungal activities against Aspergillus niger Aflavus and Penicillium notatum using the agar dilutionmethod The results are tabulated in Table 21 [93] Negativecontrols showed 0 growth inhibitionThe methanol extractof the B pilosa roots at 10mgmL was also effective againstCandida albicans [93]Of note B pilosa obtained fromPapuaNew Guinea had no activity against A niger and C albicans[126] but the SouthAfrican (EasternCape) ecotype exhibitedmoderate activity against C albicans [93] This discrepancymay depend on extraction solvents extraction procedureassay techniques different plant parts and abundance ofactive compounds

B pilosa produces a variety of secondary metabolitessuch as flavonoids phenylacetylenes alkaloids sterols ter-penoids and tannis [92 93] However none of them havebeen confirmed as active compounds against fungi Furtherinvestigation of active compounds from B pilosa is necessaryto further understand the antifungal efficacy of this plant

39 Hypotensive and Vasodilatory Activities In early studiesDimo and colleagues used three rat models normotensiveWistar rats (NTR) salt-loading hypertensive rats (SLHR)and spontaneous hypertensive rats (SHR) to investigate thehypotensive effect of the methanol crude extract of B pilosaleaves [28 127] The extract lowered systolic blood pressurein hypertensive rats (SLHR and SHR) to a greater degreethan NTR [28 127] In addition a decrease in urinarysodium ions and an increase in urinary potassium ions wereobserved after treatment with the methanol extract of Bpilosa leaves although neither differences were statisticallysignificant [28 127] Taking the data together the studyproposed that B pilosa leaf extract reduced blood pressurevia vasodilation [28 127] The same group continued totest the antihypertensive effect of aqueous and methylenechloride extracts of B pilosa leaves in a hypertensive ratmodel [28 127] To establish a fructose-induced hypertensionmodel male Wistar rats were given 10 fructose solutionto drink ad libitum for three weeks In addition to freeaccess to 10 fructose the rats were treated with the aqueous(150mgkg) or methyl chloride (350mgkg) extracts of Bpilosa for additional three weeks [28 127] Both extracts ofB pilosa leaves had a hypotensive effect on rats Howeverneither extracts reversed the elevation of serum insulin infructose-fed rats Therefore B pilosa lowered blood pressureirrespective of insulin [28 127] To better understand thehypotensive mechanism the authors investigated the effectof a neutral extract of B pilosa (NBP) a mixture of methanolandmethylene chloride (1 1) extract after neutralization withNaOH and HCl on the heart and the blood pressure ofNTR and SHR [110] This study showed that an intravenousinjection of the NBP resulted in a biphasic reduction insystolic blood pressure In addition one intravenous dose ofthe extract at 10 20 and 30mgkg BW decreased systolicblood pressure in normal rats by 183 425 and 30

46 Evidence-Based Complementary and Alternative Medicine

respectively and the same doses reduced the blood pressurein hypertensive rats by 258 389 and 286 respectivelyOnly the highest dose (30mgkg) affected the force of thecontraction of the heart Atropine and propranolol were usedto interfere with the hypotensive action of the NBP Atropinereduced the initial phase of the hypotensive response in NBPand completely abolished the second phase of hypotensiveresponse in NBP In contrast propranolol increased thefirst hypotensive response but partially abolished the secondhypotensive response provoked by the NBP [110]This mech-anistic study suggested that B pilosa invokes the biphasichypotensive responses via targeting cardiac pump efficiencyduring the first phase and vasodilation at the second phase[110]

A further study was performed to investigate the relaxingeffect of a neutral extract of B pilosa (NBP) on rat aorta con-tractedwithKCl (60mM) andnorepinephrine (01mM) [94]Cumulative addition of NBP relaxed the rat aorta previouslycontracted by KCl in a dose-dependent manner The EC

50

value of theNBP for vasorelaxationwas 032mgmLThedataalso showed that the NBP reduced the contraction of aortapreviously contracted by KCl irrespective of the presence ofaortic endothelium [94]

Pretreatment with glibenclamide an ATP-dependent K+channel blocker did not considerably affect the relaxant effectof the NBP on KCl-induced contraction suggesting thatthe vasodilatory effect of B pilosa was not related to theopening of this ATP-dependent K+ channel [94] On theother hand in the presence of indomethacin or pyrilaminemaleate the relaxant response induced by the plant extractwas significantly inhibited at the lower concentrations Theplant extract was able to reduce the aorta resting tone inhibitthe KCl-induced contractions by 90 at 15mgmL and theCaCl2-induced contractions by 95 at 075mgmL These

results demonstrate that B pilosa can act as a vasodilatorprobably via acting as a calcium antagonist [94]

However no specific compound for the above activityhas been identified from B pilosa to date A bioactivity-guided identification approach may be adopted to identifythe active compounds in B pilosa that possess hypotensiveand vasodilatory effects and understand their mechanism ofaction

310 Wound Healing Activity B pilosa has been traditionallyused to treat tissue injury inCameroon Brazil andVenezuela[26] Hassan and colleagues investigated the wound healingpotential of B pilosa in Wistar rats [128] Mirroring thepositive control neomycin sulfate the ethanol extract of Bpilosa had faster wound closure than control rats 3 6 and 9days after topical application Histological examination alsorevealed better collagenation angiogenesis and organizationof wound tissue seven days after application Epithelializationand total healing time in B pilosa-treated rats were compara-ble to those of neomycin sulfate Together these data suggestthat B pilosa may be a viable alternative to neomycin lotionfor the treatment of wounds

In addition to studying the wound healing effect of Bpilosa on external ulcers Tan and colleagues also examined

the effect of methanol cyclohexane and methyl chlorideextracts of B pilosa on gastric ulcers in Wistar rats fedwith 1mL HClethanol gastric necrotizing solution (150mMHCl in 60 ethanol) and macroscopically visible lesionswere scored [26] Among the three extracts methylene chlo-ride extracts exhibited the highest activity showing 464inhibition of lesion formation at a dose of 500mgkg BWand complete inhibition at 750mgkg [26] The efficacy ofthe ethylene chloride extract was followed by that of themethanol extracts which had inhibition ranging from 304to 822 at concentrations of 500mgkg and 1000mgkgBW respectively [26] The cyclohexane extracts showed thelowest activity against gastric ulcers in rats with 13340 and 797 inhibition at 500 750 and 1000mgkg BWrespectively [26] To better understand the mode of actionof the methylene chloride extract of B pilosa rats werepretreated with indomethacin a COX-2 inhibitor involved inprostaglandin synthesis Pretreatment significantly reducedthe protection against HClethanol-induced ulcers to 313inhibition at 750mgkg BW suggesting a link between theantiulcerative activity of B pilosa and prostaglandin syn-thesis Unexpectedly the methylene chloride extract of Bpilosa showed little gastric mucosal protection against gastriclesions induced by 95 ethanol (1mL) [26] Absolute alcoholis known to cause mucosalsubmucosal tissue destruction viacellular necrosis and the release of tissue-derived mediators(histamine and leukotrieneC4)Thus these data imply thatBpilosa did not prevent the generation or the necrotic action ofthese mediators on the gastric microvasculature In additionpylorus ligation can increase gastric acid secretion withoutan alteration of mucosal histamine content The methylenechloride extract of B pilosa did not possess antisecretoryactivity On the contrary it was observed that increases in thedose of the extract led to elevated gastric juice acidity [26]Results with both absolute ethanol and pylorus ligation ratmodels suggested the possibility that the ineffectiveness of Bpilosa against gastric ulcers was due to lack of antihistaminicactivity in the plant In summary overall the data suggest thatB pilosa protects against HCLethanol-mediated ulcers viainhibition of prostaglandin biosynthesis

Previous phytochemical studies showed that a groupof flavonoids acyclichalcones are present in B pilosa[129 130] and the chalcones were proposed to have anti-ulcerative activity [131] Moreover nine hydroxychalcones 10were reported to possess gastric cytoprotective effects with24-dihydroxychalcone being the most active [132] Sincemethylene chloride extracts appear to be the most active Bpilosa extracts next the specific anti-ulcerative phytochemi-cals in the methylene chloride extracts of B pilosa and theirmodes of action needs to be probed

Despite the claims listed inTable 3 relatively few scientificstudies have been conducted in vitro and in vivo to address thetraditional ethnomedical uses of B pilosa Information aboutthe use of B pilosa as a botanical therapy recorded so far isfar from complete Studies conducted thus far only serve asa starting point for further investigation of B pilosa and theultimate efficacious use of the herb in clinical applications

Evidence-Based Complementary and Alternative Medicine 47

4 Toxicology

Despite its use as an ingredient in food for human con-sumption studies on systemic toxicity (eg acute subacutechronic and subchronic toxicities) of B pilosa in humansand animals are still inadequate and insufficient So faracute andor subchronic toxicities have been evaluated inrats and mice Oral acute and 28-day toxicities of waterextract of B pilosa leaves were evaluated in Wistar rats[133] An oral dose of water extract of B pilosa leaves at10 gkg BW showed no obvious mortality or changes inthe appearance in rats [134] The same extract at 08 gkgBWday once a day showed no obvious sub-chronic toxicityin rats over 28 days as measured by survival rate bodyweight and gross examination of organs [134] These dataare consistent with our data indicating that oral deliveryof the water extract of the B pilosa whole plant at 1 gkgBWday once a day is safe in rats over 28 days (unpublisheddata) Taken together these studies suggest that ingestion ofB pilosa aqueous extract at up to at 1 gkg BWday oncea day is highly safe in rats In addition the acute toxicityof aqueous and ethanol extracts of B pilosa in mice havebeen reported [134] Five- to six-week-old mice with weightsbetween 28 and 35 g received a peritoneal injection of bothextracts at the different doses The LD

50 the dose that causes

50 lethality of the aqueous and ethanol extracts in micewas 1230 gkg BW and 615 gkg BW respectively [134] Acomplete toxicological study has not been completed forhumans Furthermore the drug interactions of B pilosa withother drugs are unknown Further safety verification andclinical trials should be performed before B pilosa can beconsidered for medicinal use

5 Conclusions

B pilosa is an erect perennial plant with green leaves whiteor yellow flowers and tiny black seeds As it is distributedworldwide and is widely used as a folk remedy B pilosacan be thought of as an extraordinary source of food andmedicine However a comprehensive up-to-date review ofresearch on B pilosa has hitherto been unavailable In thisarticle scientific studies on B pilosa have been summarizedand critically discussed from the perspectives of botanyethnomedicine phytochemistry pharmacology and toxicol-ogy B pilosa is claimed to treat more than 40 disordersand 201 compounds have been identified from this plantThe medicinal utility of B pilosa and its modes of actionin relation to its known phytochemicals were discussedherein Polyynes flavonoids phenylpropanoids fatty acidsand phenolics are the primary bioactive compounds of Bpilosa and they have been reported to be effective in thetreatment of tumors inflammationimmune modulationdiabetes viruses microbes protozoans gastrointestinal dis-eases hypertension and cardiovascular diseases Cautionshould be exercised in the therapeutic use of B pilosa forhypoglycemia hypotension bleeding and allergy

Acknowledgments

The authors thank their laboratory members for construc-tive suggestions and the authors whose publications theycited for their contributions The authors also thank MsMiranda Loney of Academia Sinica for English editing of thispaper This work was supported by Grants 99-CDA-L11 and101S0010056 from Academia Sinica Taiwan Arlene P Bar-tolome is a recipient of MECO-TECO Sandwich ScholarshipProgram between Taiwan and the Philippines

References

[1] T Shen G H Li X N Wang and H X Lou ldquoThe genusCommiphora a review of its traditional uses phytochemistryand pharmacologyrdquo Journal of Ethnopharmacology vol 142 no2 pp 319ndash330

[2] C Long P Sauleau B David et al ldquoBioactive flavonoids ofTanacetum parthenium revisitedrdquo Phytochemistry vol 64 no2 pp 567ndash569 2003

[3] P O Karis and O Ryding Asteraceae Cladistics and Classifica-tion Bremer K Eds pp 559ndash569 Timber press Portland OreUSA 1994

[4] O N Pozharitskaya A N Shikov M N Makarova et alldquoAnti-inflammatory activity of a HPLC-fingerprinted aqueousinfusion of aerial part of Bidens tripartita LrdquoPhytomedicine vol17 no 6 pp 463ndash468 2010

[5] F Q Oliveira V Andrade-Neto A U Krettli and MG L Brandao ldquoNew evidences of antimalarial activity ofBidens pilosa roots extract correlated with polyacetylene andflavonoidsrdquo Journal of Ethnopharmacology vol 93 no 1 pp 39ndash42 2004

[6] Agriculture USDA ldquoPlants databaserdquo 2013in Natural Resources Conservation Servicehttpwwwnrcsusdagovwpsportalnrcssitenationalhome

[7] Y Zhou and X Z Yan ldquoExperimental study of qi deficiencysyndrome and Codonopsis pillosulae and Astragalus injectionon the immune response in micerdquo Zhong Xi Yi Jie He Za Zhivol 9 no 5 pp 286ndash262 1989

[8] S C Chien P H Young Y J Hsu et al ldquoAnti-diabeticproperties of three common Bidens pilosa variants in TaiwanrdquoPhytochemistry vol 70 no 10 pp 1246ndash1254 2009

[9] M J Alcaraz and M J Jimenez ldquoFlavonoids as anti-inflammatory agentsrdquo Fitoterapia vol 59 no 1 pp 25ndash38 1988

[10] FAO Agriculture Food and Nutrition for AfricamdashA ResourceBook for Teachers of Agriculture Publishing ManagementGroup FAO Information Division Rome Italy 1997

[11] M B Rokaya Z Munzbergova B Timsina and K R BhattaraildquoRheum australe D Don a review of its botany ethnobotanyphytochemistry and pharmacologyrdquo Journal of Ethnopharma-cology vol 141 no 3 pp 761ndash774 2012

[12] P H Young Y J Hsu and W C Yang ldquoBidens pilosa L andits medicinal userdquo in Recent Progress in Medicinal Plants DrugPlant II A S AwaadVK Singh and JNGovil Eds StandiumPress Houston Tex USA 2010

[13] C Ge ldquoCytologic study of Bidens bipinnata LrdquoZhongguo ZhongYao Za Zhi vol 15 no 2 pp 72ndash125 1990

[14] L C Chiang J S Chang C C Chen L T Ng and C CLin ldquoAnti-herpes simplex virus activity of Bidens pilosa and

48 Evidence-Based Complementary and Alternative Medicine

Houttuynia cordatardquoAmerican Journal of ChineseMedicine vol31 no 3 pp 355ndash362 2003

[15] N P Rybalchenko V A Prykhodko S S Nagorna et alldquoIn vitro antifungal activity of phenylheptatriyne from Bidenscernua L against yeastsrdquo Fitoterapia vol 81 no 5 pp 336ndash3382010

[16] K Redl W Breu B Davis and R Bauer ldquoAnti-inflammatoryactive polyacetylenes from Bidens campylothecardquo Planta Med-ica vol 60 no 1 pp 58ndash62 1994

[17] M Ayyanar and S Ignacimuthu ldquoTraditional knowledge ofKani tribals inKouthalai of Tirunelveli hills TamilNadu IndiardquoJournal of Ethnopharmacology vol 102 no 2 pp 246ndash255 2005

[18] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant intraditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[19] AKAbbas KMMurphy andA Sher ldquoFunctional diversity ofhelper T lymphocytesrdquo Nature vol 383 no 6603 pp 787ndash7931996

[20] J H Cano and G Volpato ldquoHerbal mixtures in the traditionalmedicine of Eastern Cubardquo Journal of Ethnopharmacology vol90 no 2-3 pp 293ndash316 2004

[21] J Namukobe J M Kasenene B T Kiremire et al ldquoTraditionalplants used for medicinal purposes by local communitiesaround the Northern sector of Kibale National Park UgandardquoJournal of Ethnopharmacology vol 136 no 1 pp 236ndash245 2011

[22] C Lans ldquoComparison of plants used for skin and stomachproblems in Trinidad and Tobago with Asian ethnomedicinerdquoJournal of Ethnobiology andEthnomedicine vol 3 article 3 2007

[23] S Dharmananda ldquoA Popular Remedy Ecapes Notice ofWesternPractitionersrdquo 2013 httpwwwitmonlineorgartsbidenshtm

[24] Y J Hsu T H Lee C L T Chang Y T Huang andW C YangldquoAnti-hyperglycemic effects and mechanism of Bidens pilosawater extractrdquo Journal of Ethnopharmacology vol 122 no 2 pp379ndash383 2009

[25] E Noumi F Houngue and D Lontsi ldquoTraditional medicinesin primary health care plants used for the treatment ofhypertension in Bafia Cameroonrdquo Fitoterapia vol 70 no 2 pp134ndash139 1999

[26] P V Tan T Dimo and E Dongo ldquoEffects of methanolcyclohexane and methylene chlo ride extracts of Bidens pilosaon various gastric ulcer models in ratsrdquo Journal of Ethnophar-macology vol 73 no 3 pp 415ndash421 2000

[27] R L C Pereira T Ibrahim L Lucchetti A J R Da Silva and VL G De Moraes ldquoImmunosuppressive and anti-inflammatoryeffects of methanolic extract and the polyacetylene isolatedfrom Bidens pilosa Lrdquo Immunopharmacology vol 43 no 1 pp31ndash37 1999

[28] T Dimo J Azay P V Tan et al ldquoEffects of the aqueous andmethylene chloride extracts of Bidens pilosa leaf on fructose-hypertensive ratsrdquo Journal of Ethnopharmacology vol 76 no 3pp 215ndash221 2001

[29] CWiartMedicinal Plants of Southeast Asia Pelanduk SelabgorDarul Ehsan Malaysia 2000

[30] F L Silva D C H Fischer J F Tavares M S Silva P FDe Athayde-Filho and J M Barbosa-Filho ldquoCompilation ofsecondary metabolites from Bidens pilosa LrdquoMolecules vol 16no 2 pp 1070ndash1102 2011

[31] A H Chen S R Lin and C H Hong ldquoPhytochemical studyon Bidens pilosa L varminorrdquo Huaxue pp 38ndash42 1975

[32] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[33] M C Zulueta M Tada and C Y Ragasa ldquoA diterpene fromBidens pilosardquo Phytochemistry vol 38 no 6 pp 1449ndash14501995

[34] J Wang H Yang Z W Lin and H D Sun ldquoFlavonoids fromBidens pilosa var radiatardquo Phytochemistry vol 46 no 7 pp1275ndash1278 1997

[35] A Zhao Q Zhao L Peng et al ldquoA new chalcone glycoside fromBidens pilosardquo Acta Botanica Yunnanica vol 26 no 1 pp 121ndash126

[36] P Gao Y Wu S Cui and Y Zou ldquoSynthesis and absoluteconfiguration of the 7-phenylhepta-4 6-diyne-1 2-diol isolatedfrom Bidens pilosardquo Synthesis no 13 pp 2131ndash2135 2011

[37] S Wang B Yang D Zhu D He and L Wang ldquoActivecomponents of Bidens pilosa Lrdquo Zhongcaoyao vol 36 pp 20ndash21

[38] F Bohlmann T Burkhardt and C Zdero Naturally OccuringAcetylenes Academic Press New York NY USA 1973

[39] T M Sarg A M Ateya N M Farrag and F A AbbasldquoConstituents and biological activity of Bidens pilosa L grownin EgyptrdquoActa Pharmaceutica Hungarica vol 61 no 6 pp 317ndash323 1991

[40] H A L Valdes and H P Rego ldquoBidens pilosa Linnerdquo RevistaCubana de Plantas Medicinales vol 6 pp 28ndash33

[41] F Bohlmann H Bornowski and K M Kleine ldquoNew polyynesfrom the tribe Heliantheaerdquo Chemische Berichte vol 97 pp2135ndash2138

[42] RWang Q XWu and Y P Shi ldquoPolyacetylenes and flavonoidsfrom the aerial parts ofBidens pilosardquo PlantaMedica vol 76 no9 pp 893ndash896 2010

[43] L W Wu Y M Chiang H C Chuang et al ldquoA novelpolyacetylene significantly inhibits angiogenesis and promotesapoptosis in human endothelial cells through activation of theCDK inhibitors and caspase-7rdquo Planta Medica vol 73 no 7 pp655ndash661 2007

[44] L W Wu Y M Chiang H C Chuang et al ldquoPolyacetylenesfunction as anti-angiogenic agentsrdquo Pharmaceutical Researchvol 21 no 11 pp 2112ndash2119 2004

[45] H L Yang S C Chen N W Chang et al ldquoProtection fromoxidative damage using Bidens pilosa extracts in normal humanerythrocytesrdquo Food and Chemical Toxicology vol 44 no 9 pp1513ndash1521 2006

[46] H Q Wang S J Lu H Li and Z H Yao ldquoEDTA-enhancedphytoremediation of lead contaminated soil by Bidens maxi-mowiczianardquo Journal of Environmental Sciences vol 19 no 12pp 1496ndash1499 2007

[47] S TobinagaMK SharmaWG L Aalbersberg et al ldquoIsolationand identification of a potent antimalarial and antibacterialpolyacetylene from Bidens pilosardquo Planta Medica vol 75 no 6pp 624ndash628 2009

[48] C L T Chang H K Kuo S L Chang et al ldquoThe distincteffects of a butanol fraction of Bidens pilosa plant extract onthe development of Th1-mediated diabetes and Th2-mediatedairway inflammation in micerdquo Journal of Biomedical Sciencevol 12 no 1 pp 79ndash89 2005

[49] S L Chang C L T Chang Y M Chiang et al ldquoPolyacetyleniccompounds and butanol fraction from Bidens pilosa can mod-ulate the differentiation of helper T cells and prevent autoim-mune diabetes in non-obese diabetic micerdquo Planta Medica vol70 no 11 pp 1045ndash1051 2004

Evidence-Based Complementary and Alternative Medicine 49

[50] Y M Chiang C L T Chang S L Chang W C Yang and LF Shyur ldquoCytopiloyne a novel polyacetylenic glucoside fromBidens pilosa functions as a T helper cell modulatorrdquo Journal ofEthnopharmacology vol 110 no 3 pp 532ndash538 2007

[51] R PUbillas CDMendez S D Jolad et al ldquoAntihyperglycemicacetylenic glucosides from Bidens pilosardquo Planta Medica vol66 no 1 pp 82ndash83 2000

[52] L Alvarez S Marquina M L Villarreal D Alonso E Arandaand G Delgado ldquoBioactive polyacetylenes from Bidens pilosardquoPlanta Medica vol 62 no 4 pp 355ndash357 1996

[53] C L T Chang S L Chang Y M Lee et al ldquoCytopiloyne apolyacetylenic glucoside prevents type 1 diabetes in nonobesediabetic micerdquo Journal of Immunology vol 178 no 11 pp 6984ndash6993 2007

[54] G Kusano A Kusano and Y Seyama Novel Hypoglycemicand Anti-Inflammatory Polyacetylenic Compounds Their Com-positions Bidens Plant Extract Fractions and CompositionsContaining the Plant or Fraction JPO Tokyo Japan 2004

[55] M G L Brandao A U Krettli L S R Soares C G CNery and H C Marinuzzi ldquoAntimalarial activity of extractsand fractions from Bidens pilosa and other Bidens species(Asteraceae) correlated with the presence of acetylene andflavonoid compoundsrdquo Journal of Ethnopharmacology vol 57no 2 pp 131ndash138 1997

[56] A U Krettli V F Andrade-Neto M D G L Brandao andW M S Ferrari ldquoThe search for new anti-malarial drugs fromplants used to treat fever and malaria or plants ramdomlyselected a reviewrdquoMemorias do Instituto Oswaldo Cruz vol 96no 8 pp 1033ndash1042 2001

[57] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[58] C K Wat R K Biswas E A Graham L Bohm G H NTowers and E R Waygood ldquoUltraviolet-mediated cytotoxicactivity of phenylheptatriyne from Bidens pilosa Lrdquo Journal ofNatural Products vol 42 no 1 pp 103ndash111 1979

[59] Y Sashida K Ogawa M Kitada H Karikome Y Mimakiand H Shimomura ldquoNew aurone glucosides and new phenyl-propanoid glucosides from Bidens pilosardquo Chemical and Phar-maceutical Bulletin vol 39 no 3 pp 709ndash711 1991

[60] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens pilosa L (TFB) on animal liver injury andliver fibrosisrdquo Journal of Ethnopharmacology vol 116 no 3 pp539ndash546 2008

[61] B Hoffmann and J Holzl ldquoAcylated compounds from Bidenspilosardquo Planta Medica vol 55 pp 108ndash109

[62] B Hoffmann and J Holzl ldquoA methylated chalcone glucosidefrom Bidens pilosardquo Phytochemistry vol 27 no 11 pp 3700ndash3701 1988

[63] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[64] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[65] B Hoffmann and J Holzl ldquoWeitere acylierte chalkone ausBidens pilosardquo Planta Medica vol 54 pp 450ndash451 1988

[66] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant in11traditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[67] V V Pham V V T K P V L Hoang and V K Phan ldquoFlavonoidcompounds from the plant Bidens pilosa L (Asteraceae)rdquo TapChi Duoc Hoc vol 50 pp 48ndash53

[68] S D Sarker B Bartholomew R J Nash and N Robinson ldquo5-O-methylhoslundin an unusual flavonoid from Bidens pilosa(Asteraceae)rdquo Biochemical Systematics and Ecology vol 28 no6 pp 591ndash593 2000

[69] S L Chang Y M Chiang C L T Chang et al ldquoFlavonoidscentaurein and centaureidin from Bidens pilosa stimulate IFN-120574 expressionrdquo Journal of Ethnopharmacology vol 112 no 2 pp232ndash236 2007

[70] Y M Chiang D Y Chuang S Y Wang Y H Kuo P WTsai and L F Shyur ldquoMetabolite profiling and chemopreventivebioactivity of plant extracts from Bidens pilosardquo Journal ofEthnopharmacology vol 95 no 2-3 pp 409ndash419 2004

[71] A Kusano Y Seyama E Usami et al ldquoStudies on the antioxi-dant active constituents of the dried powder from Bidens pilosaL var radiata SchrdquoNaturalMedicines vol 57 no 3 pp 100ndash1042003

[72] I Doleckova L Rarova J Gruz et al ldquoAnti-proliferative andanti-angiogenic effects of flavone eupatorin an active con-stituent of chloroform extract ofOrthosiphon stamineus leavesrdquoFitoterapia vol 83 no 6 pp 1000ndash1007

[73] K Bairwa R Kumar R J Sharma and R K Roy ldquoAn updatedreview on Bidens pilosa Lrdquo Der Pharma Chemica vol 2 no 3pp 325ndash337

[74] Q Xia Y Liu and Y Li ldquoDetermination of hyperoside indifferent parts and different species of Herba Bidens by RP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol 50pp 48ndash53

[75] M Horiuchi and Y Seyama ldquoImprovement of the antiinflam-matory and antiallergic activity of Bidens pilosa L var radiataSCHERFF treated with enzyme (Cellulosine)rdquo Journal of HealthScience vol 54 no 3 pp 294ndash301 2008

[76] M G L Brandao C G C Nery M A S Mamao and AU Krettli ldquoTwo methoxylated flavone glycosides from Bidenspilosardquo Phytochemistry vol 48 no 2 pp 397ndash400 1998

[77] P Kumari K Misra B S Sisodia et al ldquoA promising anticancerand antimalarial component from the leaves of Bidens pilosardquoPlanta Medica vol 75 no 1 pp 59ndash61 2009

[78] G Seelinger I Merfort U Wolfle and C M Schempp ldquoAnti-carcinogenic effects of the flavonoid luteolinrdquoMolecules vol 13no 10 pp 2628ndash2651 2008

[79] G Seelinger I Merfort and C M Schempp ldquoAnti-oxidantanti-inflammatory and anti-allergic activities of luteolinrdquoPlanta Medica vol 74 no 14 pp 1667ndash1677 2008

[80] C C Yit and N P Das ldquoCytotoxic effect of butein on humancolon adenocarcinoma cell proliferationrdquo Cancer Letters vol82 no 1 pp 65ndash72 1994

[81] J A Beutler E Hamel A J Vlietinck et al ldquoStructure-activityrequirements for flavone cytotoxicity and binding to tubulinrdquoJournal of Medicinal Chemistry vol 41 no 13 pp 2333ndash23381998

[82] M R Kviecinski K B Felipe T Schoenfelder et al ldquoStudy ofthe antitumor potential of Bidens pilosa (Asteraceae) used inBrazilian folkmedicinerdquo Journal of Ethnopharmacology vol 117no 1 pp 69ndash75 2008

[83] P Sundararajan A Dey A Smith A G Doss M Rajappanand S Natarajan ldquoStudies of anticancer and antipyretic activityof Bidens pilosawhole plantrdquo African Health Sciences vol 6 no1 pp 27ndash30 2006

[84] YMChiang C P Lo Y P Chen et al ldquoEthyl caffeate suppressesNF-120581B activation and its downstream inflammatory mediatorsiNOS COX-2 and PGE2 in vitro or in mouse skinrdquo BritishJournal of Pharmacology vol 146 no 3 pp 352ndash363 2005

50 Evidence-Based Complementary and Alternative Medicine

[85] J S Kim H J Lee M H Lee J Kim C Jin and J HRyu ldquoLuteolin inhibits LPS-stimulated inducible nitric oxidesynthase expression in BV-2 microglial cellsrdquo Planta Medicavol 72 no 1 pp 65ndash68 2006

[86] P A Ruiz and D Haller ldquoFunctional diversity of flavonoidsin the inhibition of the proinflammatory NF-120581B IRF and Aktsignaling pathways in murine intestinal epithelial cellsrdquo Journalof Nutrition vol 136 no 3 pp 664ndash671 2006

[87] A Xagorari A Papapetropoulos A Mauromatis MEconomou T Fotsis and C Roussos ldquoLuteolin inhibitsan endotoxin-stimulated phosphorylation cascade andproinflammatory cytokine production in macrophagesrdquoJournal of Pharmacology and Experimental Therapeutics vol296 no 1 pp 181ndash187 2001

[88] N Yoshida T Kanekura Y Higashi and T Kanzaki ldquoBidenspilosa suppresses interleukin-1120573-induced cyclooxygenase-2expression through the inhibition of mitogen activated proteinkinases phosphorylation in normal human dermal fibroblastsrdquoJournal of Dermatology vol 33 no 10 pp 676ndash683 2006

[89] M Muchuweti C Mupure A Ndhlala T Murenje and M AN Benhura ldquoScreening of antioxidant and radical scavengingactivity of Vigna ungiculata Bidens pilosa and Cleome gynan-drardquo American Journal of Food Technology vol 2 no 3 pp 161ndash168 2007

[90] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens bipinnata L against carbon tetrachloride-induced liver fibrosis in ratsrdquo Journal of Pharmacy and Pharma-cology vol 60 no 10 pp 1393ndash1402 2008

[91] S L Chang H H Yeh Y S Lin Y M Chiang T K Wuand W C Yang ldquoThe effect of centaurein on interferon-120574 expression and Listeria infection in micerdquo Toxicology andApplied Pharmacology vol 219 no 1 pp 54ndash61 2007

[92] F Deba T D Xuan M Yasuda and S Tawata ldquoChemical com-position and antioxidant antibacterial and antifungal activitiesof the essential oils from Bidens pilosa Linn var Radiatardquo FoodControl vol 19 no 4 pp 346ndash352 2008

[93] A O T Ashafa and A J Afolayan ldquoScreening the root extractsfrom Biden pilosa L var radiata (Asteraceae) for antimicrobialpotentialsrdquo Journal of Medicinal Plant Research vol 3 no 8 pp568ndash572 2009

[94] T B Nguelefack T Dimo E P Nguelefack Mbuyo P V TanS V Rakotonirina and A Kamanyi ldquoRelaxant effects of theneutral extract of the leaves of Bidens pilosa linn on isolated ratvascular smooth musclerdquo Phytotherapy Research vol 19 no 3pp 207ndash210 2005

[95] M T Grombone-Guaratini K L Silva-Brandao V N SolferiniJ Semir and J R Trigo ldquoSesquiterpene and polyacetyleneprofile of the Bidens pilosa complex (Asteraceae Heliantheae)from Southeast of Brazilrdquo Biochemical Systematics and Ecologyvol 33 no 5 pp 479ndash486 2005

[96] L L Lin C Y Wu H C Hsiu M T Wang and H ChuangldquoStudies on diabetes mellitus I The hypoglycemic activity ofphytosterin on alloxan diabetic ratsrdquo Taiwan Yi Xue Hui Za Zhivol 66 no 2 pp 58ndash65 1967

[97] M A N Benhura and I C Chitsiku ldquoThe extractable 120573-carotene content of Guku (Bidens pilosa) leaves after cookingdrying and storagerdquo International Journal of Food Science andTechnology vol 32 no 6 pp 495ndash500 1997

[98] J S Chang L C Chiang C C Chen L T Liu K C Wangand C C Lin ldquoAtileukemic activity of Bidens pilosa l varminor (blume) sherff andHouttuynia cordata thunbrdquo AmericanJournal of Chinese Medicine vol 29 no 2 pp 303ndash312 2001

[99] F Deba T D Xuan M Yasuda and S Tawata ldquoHerbicidal andfungicidal activities and identification of potential phytotoxinsfrom Bidens pilosa L var radiata Scherff research paperrdquoWeedBiology and Management vol 7 no 2 pp 77ndash83 2007

[100] J K Kumar and A K Sinha ldquoA new disubstituted acetylacetonefrom the leaves ofBidens pilosa LinnrdquoNatural Product Researchvol 17 no 1 pp 71ndash74 2003

[101] K Ogawa and Y Sashida ldquoCaffeoyl derivatives of a sugarlactone and its hydroxy acid from the leaves of Bidens pilosardquoPhytochemistry vol 31 no 10 pp 3657ndash3658 1992

[102] Q Xia Y Liu and Y Li ldquoDetermination of gallic acid fromdifferent species and different medical parts of Herba Bidens byRP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol24 no 03 pp 308ndash310

[103] W J Lee L F Wu W K Chen C J Wang and T HTseng ldquoInhibitory effect of luteolin on hepatocyte growth fac-torscatter factor-induced HepG2 cell invasion involving bothMAPKERKs and PI3K-Akt pathwaysrdquo Chemico-BiologicalInteractions vol 160 no 2 pp 123ndash133 2006

[104] J A Beutler J H Cardellina C M Lin E Hamel G MCragg andM R Boyd ldquoCentaureidin a cytotoxic flavone fromPolymnia fruticosa inhibits tubulin polymerizationrdquo BioorganicandMedicinal Chemistry Letters vol 3 no 4 pp 581ndash584 1993

[105] A Verma A Su A M Golin B OrsquoMarrah and J K AmorosaldquoThe lateral view a screening method for knee traumardquo Aca-demic Radiology vol 8 no 5 pp 392ndash397 2001

[106] J Corren M Lemay Y Lin L Rozga and R K RandolphldquoClinical and biochemical effects of a combination botanicalproduct (ClearGuard) for allergy a pilot randomized double-blind placebo-controlled trialrdquo Nutrition Journal vol 7 no 1article 20 2008

[107] M S Gachet J S Lecaro M Kaiser et al ldquoAssessment of anti-protozoal activity of plants traditionally used in Ecuador in thetreatment of leishmaniasisrdquo Journal of Ethnopharmacology vol128 no 1 pp 184ndash197 2010

[108] W C Yang M Ghiotto B Barbarat and D Olive ldquoThe role ofTec protein-tyrosine kinase in T cell signalingrdquo The Journal ofBiological Chemistry vol 274 no 2 pp 607ndash617 1999

[109] S Tewtrakul H Miyashiro N Nakamura et al ldquoHIV-1 inte-grase inhibitory substances from Coleus parvifoliusrdquo Phytother-apy Research vol 17 no 3 pp 232ndash239 2003

[110] T Dimo T B Nguelefack P V Tan et al ldquoPossible mechanismsof action of the neutral extract from Bidens pilosa L leaves onthe cardiovascular system of anaesthetized ratsrdquo PhytotherapyResearch vol 17 no 10 pp 1135ndash1139 2003

[111] T S C Li Chinese and Related North American Herbs CRCPress New York NY USA 2002

[112] H Wu H Chen X Hua Z Shi L Zhang and J ChenldquoClinical therapeutic effect of drug-separated moxibustion onchronic diarrhea and its immunologic mechanismsrdquo Journal ofTraditional Chinese Medicine vol 17 no 4 pp 253ndash258 1997

[113] S W Wright R R Harris J S Kerr et al ldquoSynthesis chemicaland biological properties of vinylogous hydroxamic acids dualinhibitors of 5-lipoxygenase and IL-1 biosynthesisrdquo Journal ofMedicinal Chemistry vol 35 no 22 pp 4061ndash4068 1992

[114] W R Almiron and M E Brewer ldquoClassification of immaturestage habitats of Culicidae (Diptera) collected in CordobaArgentinardquo Memorias do Instituto Oswaldo Cruz vol 91 no 1pp 1ndash9 1996

[115] N L Wang J Wang X S Yao and S Kitanaka ldquoTwo newmonoterpene glycosides and a new (+)-jasmololone glucoside

Evidence-Based Complementary and Alternative Medicine 51

from Bidens parvifloraWilldrdquo Journal of Asian Natural ProductsResearch vol 9 no 5 pp 473ndash479 2007

[116] P Champagnat ldquoRole of the terminal bud in the actionexercised by the cotyledon of Bidens pilosus L var radiatus onits axillary budrdquo Comptes Rendus des Seances et Memoires de laSociete de Biologie vol 145 no 17-18 pp 1374ndash1376 1951

[117] S F Nielsen S B Christensen G Cruciani A Kharazmiand T Liljefors ldquoAntileishmaniai chalcones statistical designsynthesis and three-dimensional quantitative structure-activityrelationship analysisrdquo Journal of Medicinal Chemistry vol 41no 24 pp 4819ndash4832 1998

[118] Y C Hwang J J H Chu P L Yang W Chen and M V YatesldquoRapid identification of inhibitors that interfere with poliovirusreplication using a cell-based assayrdquo Antiviral Research vol 77no 3 pp 232ndash236 2008

[119] V F Andrade-Neto M G L Brandao F Q Oliveira et alldquoAntimalarial activity of Bidens pilosa L (Asteraceae) ethanolextracts fromwild plants collected in various localities or plantscultivated in humus soilrdquo Phytotherapy Research vol 18 no 8pp 634ndash639 2004

[120] R J Marles and N R Farnsworth ldquoAnti-diabetic plants andtheir active constituentsrdquo Phytomedicine vol 2 no 2 pp 137ndash189

[121] M Habeck ldquoDiabetes treatments get sweet help from naturerdquoNature Medicine vol 9 no 10 p 1228 2003

[122] H W Lin G Y Han and S X Liao ldquoStudies on the activeconstituents of the Chinese traditional medicine Polygonatumodoratum (Mill) DrucerdquoActa Pharmaceutica Sinica vol 29 no3 pp 215ndash222 1994

[123] C L T Chang Y C Chen H M Chen N S Yang andW C Yang ldquoNatural cures for type 1 diabetes a reviewof phytochemicals biological actions and clinical potentialrdquoCurrent Medicinal Chemistry vol 20 no 7 pp 899ndash907 2013

[124] L Dey A S Attele and C S Yuan ldquoAlternative therapies fortype 2 diabetesrdquo Alternative Medicine Review vol 7 no 1 pp45ndash58 2002

[125] C L T ChangH Y Liu T F Kuo et al ldquoAnti-diabetic effect andmode of action of cytopiloynerdquo Evidence-Based Complementaryand Alternative Medicine vol 2013 Article ID 685642 13 pages2013

[126] M R Khan M Kihara and A D Omoloso ldquoAnti-microbialactivity of Bidens pilosa Bischofia javanica Elmerillia papuanaand Sigesbekia orientalisrdquo Fitoterapia vol 72 no 6 pp 662ndash6652001

[127] T Dimo T B Nguelefack P Kamtchouing E Dongo ARakotonirina and S V Rakotonirina ldquoHypotensive effects ofmethanol extract from Bidens pilosa Linn on hypertensive ratsrdquoComptes Rendus de lrsquoAcademie des Sciences Serie III vol 322no 4 pp 323ndash329 1999

[128] A K Hassan O Deogratius J F Nyafuono O Francis and OP Engeu ldquoWound healing potential of the ethanolic extracts ofBidens pilosa and Ocimum suaverdquo African Journal of Pharmacyand Pharmacology vol 5 no 2 pp 132ndash136 2011

[129] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[130] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[131] C Kandaswami and EMiddleton ldquoFree radical scavenging andantioxidant activity of plant flavonoidsrdquo Advances in Experi-mental Medicine and Biology vol 366 pp 351ndash376 1994

[132] K Yamamoto H Kakegawa H Ueda et al ldquoGastric cytopro-tective anti-ulcerogenic actions of hydroxychalcones in ratsrdquoPlanta Medica vol 58 no 5 pp 389ndash393 1992

[133] J O C Ezeonwumelu A K Julius C N Muhoho et alldquoBiochemical and histological studies of aqueous extract ofBidens pilosa leaves from Ugandan Rift valley in ratsrdquo BritishJournal of Pharmacology and Toxicology vol 2 no 6 pp 302ndash309

[134] L Frida S Rakotonirina A Rakotonirina and J P SavineauldquoIn vivo and in vitro effects of Bidens pilosa L (Asteraceae)leaf aqueous and ethanol extracts on primed-oestrogenized ratuterine musclerdquo African Journal of Traditional Complementaryand Alternative Medicines vol 5 no 1 pp 79ndash91 2008

52 Evidence-Based Complementary and Alternative Medicine

Composition Comments

1 Please provide valid postal code to the address if

possible

2 We made the highlighted change in Table 4 for the sake

of correctness Please check

3 Please check the correctness of the highlighted part in

Table 5

4 We considered the highlighted part in Table 11 as

reference Please check similar cases throughout

5 Please check the format of Table 18

6 We reformatted Table 20 Please check

7 We made the highlighted change in Table 4 for the sake

of consistency Please check similar cases throughout the

paper

8 We made the highlighted change(s) for the sake

of consistency Please check simila highlighted cases

throughout

9 We made the highlighted change in Table 9 for the sake

of clarity Please check similar cases through out

10 We made the highlighted change(s) for the sake of

correctness Please check

11 Comment on ref [66] This reference is a repetition

of [18] Please check similar cases throughout

Author(s) Name(s)

Author 1

Author 2

Author 3

6 Evidence-Based Complementary and Alternative Medicine

Table 4 Aliphatic natural products isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

1 Heneicosane CH3(CH2)19CH3 Aerial (Tanzania) [18]2 Docosane CH3(CH2)20CH3 Aerial (Tanzania) [18]3 Tricosane CH3(CH2)21CH3 Aerial (Tanzania) [18]4 Tetracosane CH3(CH2)22CH3 Aerial (Tanzania) [18]5 Pentacosane CH3(CH2)23CH3 Aerial (Tanzania) [18]6 Hexacosane CH3(CH2)24CH3 Aerial (Tanzania) [18]7 Heptacosane CH3(CH2)25CH3 Aerial (Tanzania) [18]

8 Octacosone CH3(CH2)26CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

9 Nonacosane CH3(CH2)27CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

10 Triacontane CH3(CH2)28CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

11 Hentriacontane CH3(CH2)29CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

12 Dotriacontane CH3(CH2)30CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

13 Tritriacontane CH3(CH2)31CH3

Aerial(Tanzania) notfound(Taiwan)

[18 31]

14 2-Butoxy-ethanol CH3(CH2)3OCH2CH2OH Whole (Taiwan) [32]15 Tetracosan-1-ol CH3(CH2)22CH2OH Aerial (Tanzania) [18]16 Hexacosan-1-ol CH3(CH2)24CH2OH Aerial (Tanzania) [18]17 1-Octacosanol CH3(CH2)26CH2OH Aerial (Tanzania) [18]

18 1-Hentriacontanol CH3(CH2)29CH2OHNot found(Taiwan) [31]

19 Tetradecanoic acid Myristic acid CH3(CH2)12CO2H Aerial (Tanzania) [18]20 Hexadecanoic acid Palmitic acid CH3(CH2)14CO2H Aerial (Tanzania) [18]21 Octadecanoic acid Stearic acid CH3(CH2)16CO2H Aerial (Tanzania) [18]22 Eicosanoic acid Arachidic acid CH3(CH2)18CO2H Aerial (Tanzania) [18]

23 Docosanoic acid Behenic acid CH3(CH2)20CO2HLeaves(Philippines) [33]

24 2-butenedioic acidO

OHO

HO Aerial (China) [34 35]

25 (Z)-9-Octadecenoicacid Oleic acid

O

OH Aerial (Tanzania) [18]

Evidence-Based Complementary and Alternative Medicine 7

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

26 (E)-9-Octadecenoicacid Elaidic acid

O

HOLeaves(Philippines) [33]

27 (ZZ)-912-Octadecadienoic acid

Linolic acid or Linoleicacid

O

OHAerial(Tanzania)whole (Taiwan)

[18 36]

28(ZZZ)-91215-Octadecatrienoicacid

120572-Linoleic acid

O

OH Whole (Taiwan) [32]

29(ZZ)-912-Octadecadienoic acidethyl ester

Ethyl linoleate O

O

Whole (Taiwan) [32]

30(ZZZ)-91215-Octadecatrienoicacid methyl ester

Methyl linolenate O

O

Whole (Taiwan) [32]

31(ZZZ)-91215-Octadecatrienoicacid ethyl ester

Ethyl linolenate O

O

Whole (Taiwan) [32]

32(Z)-9-Octadecenoicacid 2-butoxyethylester

2-Butoxyethyl oleate O

OO

Whole (Taiwan) [32]

33 2-Butoxyethyllinoleate O

OO Whole (Taiwan) [32]

34(ZZZ)-91215-Octadecatrienoicacid butoxyethyl ester

2-Butoxyethyl linoleate O

OO

Whole (Taiwan) [32]

3517E9E15E-Heptadecatetraene-1113-diyne

Heptadeca-2E8E10E16-tetraen-46-diyne

Not found(China) [37]

8 Evidence-Based Complementary and Alternative Medicine

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

36 111-Tridecadiene-3579-tetrayne

Roots (notstated) [38]

37 1-Tridecene-357911-pentayne Pentayneene

Leaves (notstated) andnot found(Egypt)

[38 39]

38 5-Tridecene-7911-triyne-3-ol

HO

not found(Egypt) [39]

39 21012-Tridecatriene-468-triyn-1-ol

OHPart not specified(Not stated) [40]

40 212-Tridecadiene-46810-tetrayn-1-ol

111-Tridecadiene-3579-

tetrayn-13-ol

OH Roots (Notstated) notfound(Egypt)

[38 39]

41 212-Tridecadiene-46810-tetraynal

111-Tridecadiene-3579-

tetrayn-13-al

O

Roots (Germany) [41]

42212-Tridecadiene-46810-tetrayn-1-ol1-acetate

111-Tridecadiene-3579-

tetrayn-13-acetateO

O

Roots (notstated) [38]

43(5E)-15-Tridecadiene-79-diyn-3413-triol OH

OH

HO

Aerial (China) [42]

44(6E12E)-3-oxo-tetradeca-612-dien-810-diyn-1-ol

OH

O

Aerial (China) [42]

45 (E)-5-Tridecene-7911-triyne-12-diol

12-Dihydroxy-5(E)-tridecene-7911-triyne

HO

OHWhole (Taiwan) [43]

46 (E)-6-Tetradecene-81012-triyne-13-diol

13-Dihydroxy-6(E)-tetradecene-81012-triyne

OH

OHWhole (Taiwan) [43ndash45]

47(2R3E11E)-311-Tridecadiene-579-triyne-12-diol

Safynol

OH

HO

Not found (Egyptand China) [37 39]

Evidence-Based Complementary and Alternative Medicine 9

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

4857911-Tridecatetrayne-12-diol

12-Dihydroxytrideca-57911-tetrayne

HOOH

Whole (Taiwan) [43 44 46]

49(R)-357911-Tridecapentayne-12-diol

(R)-12-Dihydroxytrideca-357911-pentayne

HO

OH

Aerial (Japan) [47]

50

(4E)-1-(Hydroxymethyl)-4-dodecene-6810-triyn-1-yl-120573-D-glucopyranoside

2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-

tridecene-7911-triyne

O

HO

H

OH

H

H

HOH

O

OH

HO

H Aerial (USA)whole (Taiwan)leaves (Taiwan)

[8 45 48ndash51]

51

(4E)-1-(2-Hydroxyethyl)-4-dodecene-6810-triyn-1-yl-120573-D-glucopyranoside

3-120573-D-Glucopyranosyloxy-1-hydroxy-6(E)-

tridecene-81012-triyne

OHO

H

OH

H

H

HOH

O

OH

OH

HAerial (USA andChina) whole(Taiwan) leaves(Taiwan)

[8 35 4245 48ndash51]

52

3-Hydroxy-6-tetradecene-81012-triynyl-120573-D-glucopyranoside

120573-D-Glucopyranosyloxy-

3-hydroxy-6E-tetradecence-81012-triyne

OH

O

OHO

H

OH

H

H

HOH

OH

HWhole (Mexico) [52]

53

1-(Hydroxymethyl)-46810-dodecatetrayne-1-yl-120573-D-glucopyranoside

Cytopiloyne2-120573-D-

Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne

O

HO

OHO

H

OH

H

H

HOH

OH

HWhole (Taiwan)leaves (Taiwan) [8 50 53]

54

2-O-D-Glucosyltrideca-11E-en-3579-tetrayn-12-diol

OH

O

O

OHH

HO

H

H

HHOH

OH

Leaves (Brazil) [27]

10 Evidence-Based Complementary and Alternative Medicine

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

55

(R)-1-(Hydroxymethyl)-246810-dodecapentayn-1-yl-a-D-glucopyranoside

2- 120573 -D-Glucopyranosyloxy-1-hydroxytrideca-357911-pentayne

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Chinaand Japan) [35 47]

56

1-[[(Carboxyacetyl)oxy]methyl]46810-dodecatetraynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Japan) [54]

57

(4E)-1-[[(Carboxyacetyl)oxy]-methyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHO

H

OH

Aerial (Japan) [54]

58

(4E)-1-[[(Carboxyacetyl)oxy]-ethyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

O

O

O

OHO

O

OHOH

H

HO

H

H

HHO

HOH

Aerial (Japan) [54]

59 (5E)-5-Heptene-13-diyn-1-yl-benzene

1-Phenylhepta-13-diyn-5en Whole (Taiwan) [32]

60 7-Phenyl-2(E)-heptene-46-diyn-1-ol

OHRoots (notstated) Aerial(China)

[38 42]

617-Phenyl-2(E)-heptene-46-diyn-1-ol-acetate

OO

Roots (notstated Brazil) [38 55 56]

62 7-Phenyl-46-heptadiyn-2-ol Pilosol A

OH

Whole (Taiwan)aerial (China) [42 57]

Evidence-Based Complementary and Alternative Medicine 11

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

63 7-Phenylhepta-46-diyn-12-diol HO

OH

Aerial (China) [42]

64 135-Heptatriyn-1-yl-benzene

1-Phenylhepta-135-triyne

Leaves (notstated) leaves oftissue culture(not stated)aerial (TanzaniaChina)Whole (Taiwan)roots (Brazil)

[18 34 3842 56ndash58]

65 7-Phenyl-246-heptatriyn-1-ol HO

Leaves (notstated) aerial(China)

[38 42]

667-Phenyl-246-heptatriyn-1-ol-acetate O

O Leaves (notstated) [38]

675-(2-Phenylethynyl)-2-thiophenemethanol

S OHAerial (China) [42]

685-(2-Phenylethynyl)-2120573-glucosylmethyl-thiophene

O

O

OH

H

OH

H

H

HOH H

O

HO

S

Aerial (China) [42]

69

3- 120573 -D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne

OH

O

OH

HOHO

H

H

HHOH

OH

Leaves(Cameroon) [28]

701-Phenyl-13-diyn-5-en-7-ol-acetate O

O

Leaves (Brazil) [27]

SN denotes serial number

the chloroform fraction was the most toxic with a halfmaximal inhibitory concentration (IC

50) of 97 plusmn 72 and 83 plusmn

52 120583gmL in NRU and MTT respectively [82] Kumari andcolleagues also reported the anti-cancer and anti-malarialactivities of B pilosa leaves [77] Based on a cytotoxicity-directed fractionation strategy they identified phenyl-135-heptatriene with IC

50values of 8plusmn001 049plusmn045 07plusmn001

and 10 plusmn 001 120583gmL against human oral liver colon andbreast cancer cell lines respectively However phenyl-135-heptatriyne showed lower activity against breast cancer celllines than the chloroform leaf extract which had an IC

50

value of 65 plusmn 001 120583gmL Moreover the positive controltaxol showed higher activity than phenyl-135-heptatriyne[77] Furthermore in vitro comet assays were performed toevaluate the toxicity of n-hexane chloroform and methanolextracts of B pilosa and its ethyl acetate acetone and waterfractions onHela andKB cellsThe ethyl acetate fraction fromthe methanol extract exhibited the highest activity with halfmaximal cytotoxic concentrations (CTC

50) of 9652120583gmL

and 5862120583gmL against Hela and KB cells respectivelyDespite the moderate toxicity these findings suggest thatthese B pilosa extractsfractions could be useful for future

12 Evidence-Based Complementary and Alternative Medicine

Table5Flavon

oids

isolated

from

Bpilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

71

2-[(34-Dihydroxyph

enyl)-

methylene]-

6-hydroxy-3(2H

)-benzofuranon

e

Sulfu

retin

O

OHO

OH

OH

Aeria

l(Ch

ina)

[35]

72

2-[(34-Dihydroxyph

enyl)-

methylene]-

67-dihydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-673101584041015840-

tetrahydroxy

maritimetin

O

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

73

2-[(34-Dihydroxyph

enyl)-

methylene]-6-

(120573-D

-glucopyrano

syloxy)-7-

hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-120573-D

-glucop

yranosyl-

673101584041015840-te

trahydroxy

maritimein

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japanand

China)aerial(Ch

ina)

[355960]

74

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

(120573-D

-glucopyrano

syloxy)-

6-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-7-O-120573-D

-glucop

yranosyl-

673101584041015840-

tetrahydroxy

O

OHO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japan)

[59]

75

6-[(6-O-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-acetyl-

120573-D

-glucopyrano

syl)

-673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OHO

H HO

H

H

HOH

HO

O

O

Leaves

(Not

stated)

aeria

l(Ch

ina)

[3559]

Evidence-Based Complementary and Alternative Medicine 13

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

76

6-[(36-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-

2-[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone(Z)-6-O

-(36-di-O

-acetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

bideno

sideA

O

O

HO

OH

OH

OHOH O

H

H

HOH

HO

O

O

O

Leaves

(China)

[60]

77

6-[(46-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(410158401015840610158401015840-

diacetyl-120573-D

-glucropyrano

syl)-

673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

HO

O

OO

Leaves

(not

stated)

aeria

l(Ch

ina)

[343561]

78

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

hydroxy-6-

[(246-tri-O

-acetyl-120573

-D-

glucop

yranosyl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(210158401015840410158401015840610158401015840-

triacetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

O

OOO

O

Leaves

(not

stated)

aeria

l(Ch

ina)

[3461]

79

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-6-

[(346-tri-O

-acetyl-120573

-D-glucopyrano

syl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(310158401015840410158401015840610158401015840-

triacetyl-120573

-D-glucrop

yranosyl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H OH

H

HOH

HO

OOO O

Aeria

l(Ch

ina)

[3435]

80

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-

6-[[6-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propenyl]-120573-

D-glucopyrano

syl]o

xy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-

p-coum

aroyl-

120573-D

-glucropyrano

syl)-

673101584041015840-te

trahydroxy

OH

O

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OLeaves

(Japan)

[59]

14 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

81

1-[2-(120573-D

-Glucopyrano

syloxy)-

4-hydroxyphenyl]-2-

hydroxy-3-(3-

hydroxyphenyl)-2-

prop

en-1-on

e

Chalcone120572

32101584041015840-

tetrahydroxy-

21015840-O

-120573-D

-glucop

yranosyl

HO

O

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[35]

821-(24-Dihydroxyph

enyl)-

3-(34-dihydroxyph

enyl)-

2-prop

en-1-on

eBu

tein

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

833-(34-D

ihydroxyph

enyl)-

1-(234-trihydroxyph

enyl)-

2-prop

en-1-on

eOkanin

O

HO

HO

OH

OH

OH

Leaves

(China)

[60]

84

3-(34-D

ihydroxyph

enyl)-

1-[3-(120573-D

-glucop

yranosyloxy)-

24-dihydroxyphenyl]-2-

prop

en-1-on

e

Okanin

31015840-O

-120573-D

-glucosid

e

O

HO

OH

OH

OH

OO

HO

H

HO

H

H

HOH

H

OH

Leaves

(Germany)

flowers(Germany)

[61ndash63]

85

3-(34-D

ihydroxyph

enyl)-

1-[4-(120573-D

-glucop

yranosyloxy)-

23-dihydroxyphenyl]-

2-prop

en-1-on

e

Okanin

41015840-O

-120573-D

-glucosid

e

OOH

OH

OH

HOO

OHO

H

HO

H

H

HOH

H

OH

Flow

ers(Germany)

leaves

(Japan)

[5963]

Evidence-Based Complementary and Alternative Medicine 15

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

86Okanin41015840-O

-120573-D

-(610158401015840-

O-acetylglucosid

e)

O

O

OH

OH

OH

OHO

H

HO

H

H

HOH

H

HO

O

O

Flow

ers(Germany)

[63]

87

1-[4-[(46-di-O

-Acetyl-

120573-D

-glucopyrano

syl)-oxy]-

23-dihydroxyphenyl]-3-

(34-dihydroxyph

enyl)-2-

prop

en-1-on

e

Okanin

41015840-O

-120573-D

-(410158401015840610158401015840-diacetyl)-

glucop

yranoside

O

O

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Aeria

l(Ch

ina)

[34]

88Okanin41015840-O

-120573-D

-(210158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[64]

89Okanin41015840-O

-120573-D

-(310158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

O

OAe

rial(Ch

ina)

[34]

16 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

90

1-[23-Dihydroxy-4-

[[6-O-[3-(4-hydroxyph

enyl)-

1-oxo-2-prop

enyl]-120573-D

-glucop

yranosyl]oxy]-ph

enyl]-

3-(34-dihydroxyph

enyl)-

2-prop

en-1on

e

Okanin

41015840-O

-120573-D

-(610158401015840-tr

ans-

p-coum

aroyl)-

glucoside

O

O

OH

OH

OH

OHOH

HO

H

H

HOH

H

HO

OO

OH

Leaves

(Germany)

[64]

91Okanin41015840-O

-120573-D

-(410158401015840-acetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Leaves

(Germany)

[65]

92

Okanin

41015840-O

-120573-D

-(210158401015840410158401015840-diacetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[65]

Evidence-Based Complementary and Alternative Medicine 17

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

93

Okanin

41015840-O

-120573-D

-(310158401015840410158401015840-diacetyl-610158401015840-

trans-p-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

OH

O O

Leaves

(Germany)

[65]

94

Okanin

41015840-O

-[120573-D

-glucopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

OOH

OH

OH

HO

OH

HO

HHO

H

HOH

HO

OH

OH

HO

HHO

H

HOH

HO

Flow

er(G

ermany)

[63]

95Okanin

3101584041015840-di-O

-120573-D

-glucosid

e

OOH

OH

OH

O OO

H HO

HHOH

HOH

HHO

OHO

H

HO

H

H

HOH

H

OH

Flow

er(G

ermany)

[63]

18 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

96

1-[3-(120573-D

-Glucopyrano

syloxy)-

24-dihydroxyphenyl]-

3-(3-hydroxy)-

4-metho

xyph

enyl)-

2-prop

en-1-on

e

Okanin

4-methylether-

31015840-O

-120573-D

-glucop

yranoside

OOH

O

OH

HO

O

H HO

HHOH

HOH

H

OHO

Leaves

(Germany)aerial

(China)

[3562]

97

Okanin4-methylether-

3101584041015840-di-O

-120573-(41015840101584061015840101584041015840101584010158406101584010158401015840-

tetraacetyl)-

glucop

yranoside

OOH

OOH

O

HH

OH

HO

HOH

H

OO

O

Ac

Ac

AcAc

OO

H

HO

H

H

HOH

H

O

Aeria

l(Ch

ina)

[42]

98Ch

alcone210158404101584061015840-tr

imetho

xy-

4-O-D

-glucopyrano

syl-

dihydro

Not

foun

dLeaves

(China)

[60]

992-(34-D

ihydroxyph

enyl)-23-

dihydro-78

-dihydroxy-

4H-1-

benzop

yran-4-one

Okanin

isoOO

OH

HO

OH

OH

Leaves

(China)

[60]

100

2-(34-D

ihydroxyph

enyl)-

23-dihydro-8-hydroxy-

7[(246-tr

i-O-acetyl-120573

-D-

glucop

yranosyl)oxy]-4H

-1-b

enzopyran-4-on

eOO

OH

OH

OH

OH

O

HHO

H

HO

HO

OAc

Ac

Ac

Aeria

l(Ch

ina)

[34]

Evidence-Based Complementary and Alternative Medicine 19

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

101

57-Dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

OO

OH

HO

OH

Aeria

l(TanzaniaC

hina)

[4266]

102

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

7-O-glucopyrano

side

OO

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Tanzania)

[66]

103

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-4H

-1-benzop

yran-4-one

Luteolin

OO

HO

OH

OH

OH

Aeria

l(TanzaniaC

hina

Vietnam)

[343542

6667]

104

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5hydroxy-4H-1-

benzop

yran-4-one

Luteolin

7-O-120573-D

-glucop

yranoside

OOOH

OH

OH

OO

HO

H

HO

H

H

HOH

OH H

Aeria

l(Tanzania)

[66]

105

57-Dim

etho

xy-6-

(5-m

etho

xy-6-m

ethyl-

4-oxo-4H

-pyran-3-yl)-

2-ph

enyl-4H-1-

benzop

yran-4-one

5-O-M

ethylhosludin

O OOO

O

O

O

Aeria

l(Ugand

a)[68]

20 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

106

3-(120573-D

-Glucopyrano

syloxy)-

57-dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Astragalin

Kaem

pferol-3-O

-120573-D

-glucopyrano

side

HO

OH

O O

OH

O

O

HO

HHO

H

H

HOH

OH

H

Aeria

l(Ch

ina)

[35]

107

Kaem

pferol3-

(23-di-E

-p-cou

maroyl-

120572-L-rhamno

pyrano

side)

Not

foun

dAe

rial(Vietnam)

[67]

108

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5-hydroxy-36-dimetho

xy-

4H-1-

benzop

yran-4-one

Axillarosid

eO O

OH

OH

OO

OH

OO

HO

H

HO

H

H

HOH

H

OH

Aeria

l(Ch

ina)

[42]

109

57-Dihydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaureidin

O

O

O

O

O

HO

OH

OH

Who

le(Taiwan)

[69]

110

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaurein

O O

OO

O OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan)w

hole

(Taiwan)

[69ndash

71]

Evidence-Based Complementary and Alternative Medicine 21

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

111

5-Hydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-

67-dimetho

xy-4H-

chromen-4-one

Eupatorin

iso

OO O

OH

O

O OH

Not

foun

d(C

hina)

[3772]

112

2-(34-D

imetho

xyph

enyl)-7-

(120573-D

-glucopyrano

syloxy)-35-

dihydroxy-

8-metho

xy-4H-1-

benzop

yran-4-one

O O

OH

O O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

113

7-(120573-D

-Glucopyrano

syloxy)-

5hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-38-

dimehtoxy-4H-1-

benzop

yran-4-one

O O

O

OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

114

Isorhamnetin

3-[O

-120572-

L-rham

nopyrano

syl-

(1-2)-120573-D

-glucop

yranoside]

Not

foun

dAe

rial(Vietnam)

[67]

115

7-[(6-deoxy-120572-L-

Manno

pyrano

syl)o

xy]-

3-120573-D

-glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-4H

-1-benzop

yran-4-one

Luteoside

O O

OH

O

OH

OH

OH

HOH

H

H HO

HO

HO

OH

HO

H

HO

OH

OH

HH

Aeria

l(Ch

ina)

[42]

22 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

116

Luteolin

3-O-120573-D

-Glucopyrano

side

O O

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

HO

Aeria

l(Tanzania)

[66]

117

57-Dihydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Quercetagetin

3631015840-tr

imethylether

OOO

O

O OH

HO

OH

Aeria

l(Ch

ina)

[42]

118

6-((2S3S4S5S)-

Tetrahydro-345-tr

ihydroxy-

6-(hydroxymethyl)-2H

-pyran-2-yloxy)-5-hydroxy-

2-(4-hydroxy-3-

metho

xyph

enyl)-

37-dimetho

xy-4H-

chromen-4-one

Quercetagetin

3731015840-

trim

ethylether-6-O

-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[42]

119

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

JaceinQ

uercetin

3631015840-

trim

ethyl

ether-7-O-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan

China)

who

le(Taiwan)

[425470]

120

2-(34-D

ihydroxyph

enyl)-

357-trihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

OO

HO

OH

OH

OH

OH

Aeria

l(Ch

ina)leaves

(China)who

le(C

hina)

[356074]

Evidence-Based Complementary and Alternative Medicine 23

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

121

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-galactopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-4-one

Quercetin

3-O-120573-D

-galactosidehyperin

hyperosid

e

O O

HO

OH

OH

OH

OOHH

HOH

H

H HO

HO

OH

Aeria

l(TanzaniaJapan)

Leaves

(China)Who

le(C

hina)

[376066

717475]

122

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-glucopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

3-O-120573-

D-glucopyrano

side

O O

HO

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

Aeria

l(TanzaniaC

hina

Japan)L

eaves(Japan)

[355966

75]

123

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-

4-oxo-4H

-1-benzop

yran-3-yl-

120573-D

-glucopyrano

siduron

icacid

Quercetin

3-O-120573-D

-glucuron

opyranoside

O O

HO

OH

OH

OH

OHOH

HOH

H

H HO

HO

OHO

Aeria

l(TanzaniaJapan)

[6671]

124

3-[[6-O-(6-

Deoxy-120572-L-m

anno

pyrano

syl)-

120573-D

-galactopyrano

syl]o

xy]-

2-(34-dihydroxyph

enyl)-57-

dihydroxy-4H

-1-benzop

yran-4-one

Quercetin

3-O-rob

inob

iosid

e

O O

HO

OH

OH

OH

O

HOHH OH

HO

H HH

OO

OHH

HOH

H

H HO

HO

Aeria

l(Japan)W

hole

(Taiwan)

[7071]

24 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

125

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-

3-metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

OH

Roots(Brazil)

[55676]

126

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-

120573-D

-glucopyrano

syl]o

xy]-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120572-

L-rham

nopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

O

HHO HHO

HO H

HH

ORo

ots(Brazil)

[5676]

127

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-120573-D

-glucop

yranosyl]oxy]-5-

hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-3-metho

xy-

4H-1-

benzop

yran-4-one

Quercetin

341015840-

dimethyl

ether-7-O-rutinoside

O OOH

OH

O

O

OH O

HOH

H

HHO

H

O

HHO HHO

HO H

HH

O

HO

Aeria

l(Ch

ina)

[3435]

128

2-(34-D

ihydroxyph

enyl)-3-

(120573-D

-glucofurano

syloxy)-57-

dihydroxy-4H

-1-benzop

yran-

4-on

e

Isoq

uercitrin

O OOH

OH

OHO

H

OHH

OH

HO

H O HOH

OH

Aeria

l(Japan

China)

[3571]

Evidence-Based Complementary and Alternative Medicine 25

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

129

Quercetin

3-O-120573-D

-galactop

yranoside

O

OH

HO

OH OH

O

O

HO

OH

OHOH

OAe

rial(no

tstated)

[18]

130

Quercetin

3-O-rutinoside

O

O

R

OO

OH

OHO

H

O

OH

OH

H3C

H3C

H3C

OH

HO

Who

le(Taiwan)

[70]

26 Evidence-Based Complementary and Alternative Medicine

Table 6 Terpenoids isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(Country) References

131371111-Tetramethylbicyclo[810]undeca-

26-dieneBicyclogermacrene Leaves

(Brazil) [95]

13241111-Trimethyl-8-

methylenebicyclo[720]undec-4-ene

E-Caryophyllene Leaves(Brazil) [95]

1331-Methyl-5-methylene-8-

(1-methylethyl)-16-cyclodecadiene

Germacrene D Leaves(Brazil) [95]

1344-(15-Dimethyl-4-hexen-1-ylidene)-1-methyl-cyclohexene

Z-120574-Bisabolene Leaves(Brazil) [95]

135Decahydro-114-trimethyl-7-methylene-1H-cycloprop[e]-

azulene120573-Gurjunene Leaves

(Brazil) [95]

136 2669-Tetramethyl-148-cycloundecatriene

120572-Humulene120572-caryophyllene

Leaves(Brazil) [95]

13712344a568a-Octahydro-1-isopropyl-7-methyl-4-methylenenaphthalene

120575-Muurolene Leaves(Brazil) [95]

138

12344a568a-Octahydro-4a8-dimethyl-2-(1-methylethylidene)-

naphthalene

Selina-37(11)-diene

Leaves(Brazil) [95]

139(2E7R11R)-371115-

Tetramethyl-2-hexadecen-1-ol

Phytol

HO

Whole(Taiwan) [57]

Evidence-Based Complementary and Alternative Medicine 27

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

140 371115-Tetramethyl-2-hexadecenoic acid Phytanic acid

O

OH

Whole(Taiwan) [57]

141371115-Tetramethyl-2-

hexadecenyl ester-heptanoicacid

Phythylheptanoate O O

Leaves(Not stated) [33]

142

(3S10R13R)-17-((2R5R)-5-Ethyl-6-methylheptan-2-yl)-

2347891011121314151617-tetradecahydro-

1013-dimethyl-1H-cyclopenta[a]phenanthren-

3-ol

Campestrol

HO

Aerial(Tanzania) [18]

143 Not found Phytosterin-B Not foundNot found(TaiwanEgypt)

[39 96]

144 Stigmast-5-en-3-ol 120573-sitosterol

HOH

H

H

Aerial(Tanzania)

whole(Taiwan)

[18 31 57]

145

1314151617-Tetradecahydro-1013-

dimethyl-1H-cyclopenta[a]phenanthren-

3-yloxy)-tetrahydro-6-(hydroxymethyl)-2H-pyran-345-triol

120573-Sitosterolglucoside

H

H

HOH

HO

HHO

H

HOHH O

OH Not found(Egypt) [39]

146 5120572-Stigmasta-7-en-3120573-ol

HO

Whole(Taiwan) [57]

147 5120572-Stigmasta-722t-dien-3120573-ol

HO

Whole(Taiwan) [57]

148 Stigmasta-522-dien-3-ol Stigmasterol

HOH

H

H

Not found(Taiwan)aerial

(Tanzania)leaves (notstated)whole

(Taiwan)

[18 31 3357]

28 Evidence-Based Complementary and Alternative Medicine

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

149 Lup-20(29)-en-3-ol Lupeol

H

HH

HOH

Not found(Egypt) [39]

150 Lup-20(29)-en-3-ol acetate Lupeol acetate

H

HH

O

OH

Not found(Egypt) [39]

151 Olean-12-en-3-ol 120573-amyrin

H

HOH

Not found(Egypt) [39]

152 5913-Trimethyl-242526-trinoroleanan-3-ol Friedelan-3120573-ol

HO

Aerial(Tanzania) [18]

153 5913-Trimethyl-242526-trinoroleanan-3-one

Friedelinfriedelan-3-one

O

Aerial(Tanzania) [18]

1542610151923-

Hexamethyl-2610141822-tetracosahexaene

Squalene

Aerial(Tanzania)Leaves (Notstated)Whole(Taiwan)

[18 33 57]

155 120573120573-Carotene 120573-Carotene Leaves (Notstated) [97]

Evidence-Based Complementary and Alternative Medicine 29

studies [83] Hot water extracts of B pilosa varminor Sheriffwere also assessed for its antileukemic effects on leukemiccell lines L1210 U937 K562 Raji and P3HR1 using XTT-based colorimetric assays The extract inhibited the five celllines with IC

50values ranging from 145120583gmL to 586120583gmL

L1210 K562 Raji and P3HR1 were more sensitive to B pilosaextract with IC

50values below 200120583gmL [98]

Consistent with the antitumor activities of B pilosaextracts and fractions some of its phytochemicals alsoshowed anticancer activity as outlined in Table 11 Amongthem luteolin (103) a well-studied flavonoid with multiplebioactivities was more effective against tumor cell prolifera-tion than its derivatives with IC

50values ranging from 3 120583M

to 50120583M in cells and 5 to 10mgkg in animals Luteolin wasalso found to fight cancer as a food additive at concentrationsof 50 to 200 ppm [78] and prevent skin cancer [78] and cancer3invasion [103] Lee and colleagues reported that luteolinprevents cancer by inhibiting cell adhesion and invasion[103] Significant inhibition concentration was reported to be5 120583M and complete inhibition concentration was reported tobe 40120583M Moreover luteolin was reported to inhibit hepa-tocyte growth factor (HGF)-induced cell scattering as wellas cytoskeleton changes such as filopodia and lamellipodiawhichwas determined using phase-contrast and fluorescencemicroscopy Furthermore luteolin also inhibited the HGF-induced phosphorylation of c-Met ERK 12 and Akt aswell as the MAPKERK and P13K-Akt pathways [78 103]Other mechanisms underlying the anticancer activities ofluteolin are the inhibition of topoisomerase I and II whichinhibits cell replication and DNA repair thus promotingapoptosis regulation of PI-3-KinaseAktMAPKERKJNKactivation of apoptosis in the mitochondrial pathway byactivating caspase 9 and caspase 3 which were found inmalignant cells but not in normal human peripheral bloodmononuclear cells death receptor-induced apoptosis and acell-cycle arrest mechanism inhibition of fatty acid synthasewhich is upregulated in many cancer cells and sensitizationto chemotherapywhereby luteolin increases the susceptibilityof cancer cells to chemotherapy [78] Butein (82) is anotherflavonoid that showed a cytotoxic effect on human colonadenocarcinoma cell proliferation with a reported IC

50value

of 175 120583M Butein at 2 120583M affected the incorporation of[14C]-labeled leucine thymidine and uridine which cancause the inhibition of DNA RNA and protein synthesis ofhuman colon cancer cells Moreover butein also exhibitednoncompetitive inhibition of 1-chloro-24-dinitrobenzene(CDNB) in glutathione S-transferase (GST) activity Tumorresistance was correlated with high levels of GST thus buteininhibited proliferation of cancer cells [80] Another flavonoidpresent in B pilosa centaureidin (109) also showed anti-cancer activity in B lymphoma cells Centaureidin isolatedfrom Polymnia fruticosa inhibited tubulin polymerization invitro and induced mitotic figure formation in CA46 Burkittlymphoma cells Using turbidimetric assay the IC

50value of

centaureidin in inhibition of mitosis was 3120583M [104] Cyto-toxicity of centaureidin was further analyzed using AmericanNational Cancer Institute (NCI) 60 human tumor cell linesThe cytotoxicity potency of centaureidin expressed as GI

50

(50 growth inhibition in the NCI tumor line panel) was024120583M [81] These data mark centaureidin as a promisingantimitotic agent for tumor therapy

In addition to anti-tumor flavones polyynes found in B 4pilosa have also been shown to possess anti-tumor proper-ties Based on a bioactivity-directed isolation approach Wuand colleagues identified two polyyne aglycones from theethyl acetate fraction of B pilosa [44] 12-Dihydroxytrideca-57911-tetrayne (48) and 13-Dihydroxy-6(E)- tetradecene-81012-triyne (46) exhibited significant anticell prolifera-tion activity in primary human umbilical vein endotheliumcells (HUVEC) with IC

50values of 125 120583M and 173 120583M

respectively They also decreased angiogenesis and pro-moted apoptosis in human endothelial cells Their anti-angiogenic and cytotoxic effects correlated with activationof the CDK inhibitors and caspase-7 [44] In addition 12-Dihyroxy-5(E)-tridecene-7911-triyne (45) showed antian-giogenic effects in HUVECs with an IC

50value of 124 120583M as

evidenced by a decrease in the tube formation and migrationof HUVECs [43] The IC

50value of compound 45 in the

inhibition of basic fibroblast growth factor-induced HUVECgrowth was 282120583M However it had higher IC

50values

than those for lung carcinoma cells and keratinocytes Thiscompound could also inhibit cell proliferation of HUVECslung carcinoma A549 cells and HACAT keratinocytes Themechanism by which compound 45 inhibits HUVEC growthand angiogenesis is complicated and includes decreasing theexpression of cell cycle regulators (CDK4 cyclins D1 and Aretinoblastoma (Rb) and vascular endothelial growth factorreceptor 1) caspase-mediated activation of CDK inhibitorsp21 (Cip1) and p27 (Kip) upregulation of Fas ligand expres-sion downregulation of Bcl-2 expression and activation ofcaspase-7 and poly (ADP-ribose) polymerase [43]

32 Anti-Inflammatory Activity B pilosa is commonly usedto treat inflammatory disorders The anti-inflammatory phy-tochemicals present in B pilosa are listed in Table 11 5Cyclooxygenase-2 (COX-2) is a physiologically importantenzyme that converts arachidonic acid to prostaglandin(PGE2) Its expression is induced by a wide variety of external

stimuli indicating its involvement in inflammatory diseasesand it is used as an inflammatory marker [88] Yoshida andcolleagues studied the effects of the aqueous extracts of Bpilosa aerial parts in the production of COX-2 and PGE

2as

well as on the activation of mitogen activated protein kinases(MAPKs) in normal human dermal fibroblasts (HDFs) inresponse to inflammatory cytokine IL-1120573 This work showedthat IL-1120573 activated MAPKs such as ERK12 p38 and JNKto different extents and induced COX-2 expression TheCOX-2 expression in HDFs was regulated mainly by p38following IL-1120573 stimulation Consistently the p38 inhibitorSB203580 blocked this expression Using this cell platformB pilosa extracts were tested for inhibition of inflammationThe extract dose-dependently suppressed the activation ofp38 and JNK and moderately suppressed ERK12 as wellas suppressing COX-2 expression and PGE

2production

30 Evidence-Based Complementary and Alternative Medicine

Table7Ph

enylprop

anoids

isolatedfro

mB

pilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

156

3-(4-H

ydroxyph

enyl)-2-prop

enoica

cid

p-Cou

maricacid

OHO

OH

Who

le(Ja

pan)

[99]

157

2-Metho

xy-4-(2-prop

en-1-yl)-ph

enol

Eugeno

lOOH

Leaves

androots(Japan)

[99]

158

3-(4-H

ydroxy-3-m

etho

xyph

enyl)-

2-prop

enoica

cid

Ferulic

acid

O

OH

O

HO

Who

le(Ja

pan)

[99]

159

3-(34-D

ihydroxyph

enyl)-2-prop

enoic

acid

Caffeicacid

O

HO

OH

OH

Who

leandaeria

l(Japan)

[7599]

160

3-Prop

yl-3[(245-trim

etho

xyph

enyl)-

metho

xy]-24-pentanedione

3-Prop

yl-3-(245-

trim

etho

xy)benzyloxy-

pentan-24-dione

O

O

O

O

O

O

Leaves

(India)

[100]

161

3-(34-D

ihydroxyph

enyl)-2-

prop

enoica

cidethyleste

rCa

ffeateethyl

O

O

OH

OH

Not

foun

d(Taiwan)

who

le(Taiwan)

[434584]

Evidence-Based Complementary and Alternative Medicine 31Ta

ble7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

162

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

d-Erythron

icacid

2-O-caffeoyl-2-C

-methyl

O

HO

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

163

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

2-O-caffeoyl-2-C

-methyl

O

O

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

164

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-24-

dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

3-O-caffeoyl-2-C

-methyl

O

O

HO

HO

O

O

OH

OH

Leaves

(Japan)

[101]

165

4-(A

cetyloxy)-3-[[3-(34-

dihydroxyphenyl)-

1-oxo-2-propen-1-y

l-]oxy]-2-hydroxy-

2-methyl-b

utanoica

cid

O

HO

OH

O OHO

HO

OO

Not

foun

d(Ja

pan)

[73]

166

3-(34-D

ihydroxyph

enyl)-tetrahydro-4-

hydroxy-4-

methyl-5

-oxo-3-fu

ranyl

ester-2-prop

enoica

cid

3-O-C

affeoyl-2-C

-methyl-

D-erythrono

-14-la

cton

eHO

HO

O

OO

OOH

Leaves

(Japan)

[101]

167

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]-oxy]-145-

trihydroxy-cyclohexanecarboxylicacid

Chlorogenica

cid

O

HO

HO

OH

O

OH

O

OH

HO

Aeria

l(Japan)w

hole

(Taiwan)

[707175]

32 Evidence-Based Complementary and Alternative Medicine

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

168

4-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]oxy]-

135-tr

ihydroxy-cyclohexanecarboxylic

acid

4-O-C

affeoylqu

inicacid

OHO

HO

OH

OH

O

O

OH

OH

Aeria

l(Japan)

[71]

169

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-15-

dihydroxy-cyclo

hexanecarboxylicacid

34-Di-O

-caffeoylqu

inic

acid

HO

HO

O

O

O

O

OH

OH

HO

OH O

H

O

Aeria

l(Japan)W

hole

(Taiwan)

[45497071]

170

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-14-dihydroxy-

cyclo

hexanecarboxylicacid

35-Di-O

-caffeoylqu

inic

acid

HO

HO

O O

HO

O

HO

OH OO

OHOH

Aeria

l(Japan)w

hole

(Taiwan)

[45497071]

Evidence-Based Complementary and Alternative Medicine 33

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

171

34-bis[[(2E

)-3-(34-D

ihydroxyph

enyl)-1-

oxo-2-prop

en-1-

yl]oxo]-15

-dihydroxy-

cyclo

hexanecarboxylicacid

45-Di-O

-caffeoylqu

inic

acid

OHO

OH

HO

O

O

OH

OH

O

OH

HO

OWho

le(Taiwan)

[454970]

172

3-[4-[[-6-O-[3-(4-H

ydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]-phenyl]-2-prop

enoica

cid

120573-D

-p-C

oumaricacid4-O

-(6-O

-p-caoum

aroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HOH

HO

O

O

Leaves

(Japan)

[59]

173

3-[4-[[2-O

-Acetyl-6

-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]ph

enyl]2-propeno

icacid

120573-D

-p-C

oumaricacid4-O

-(2-O

-acetyl-6

-O-p-

caou

maroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HO

HO

O

O

Leaves

(Japan)aerial

(China)

[3459]

174

67-Dihydroxy-2-chrom

enon

eEsculetin

cicho

rigenin

OHO HO

O

Not

foun

d(Egypt)

[39]

34 Evidence-Based Complementary and Alternative Medicine

Table 8 Aromatic compounds isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

175 12-Benzenediol PyrocatechinHO

HOWhole(Japan) [99]

176 4-Ethyl-12-benzenediol Pyrocatechol

OH

OH

Whole(Japan) [99]

177 Dimethoxyphenol26-dimethoxyphenol

O

HO

O

Roots (Japan) [99]

178 4-Ethenyl-2-methoxy-phenol p-Vinylguaiacol

O

OHWhole(Japan) [99]

179 2-Hydroxy-6-methylbenzaldehyde

6-Methyl-salicylaldehyde O

OH

Whole(Japan) [99]

180 Benzene-ethanol 2-Phenyl-ethanolHO

Whole(Japan) [57]

181 4-Hydroxy-3-methoxy-benzaldehyde Vanillin O

O

HOAerial(Japan) [99]

182 3-Hydroxy-4-methoxy-benzaldehyde Vanillin iso

OO

OH

Leaves(Japan) [99]

183 4-Hydroxybenzoic acid p-Hydroxybenzoicacid

O

HOOH Whole

(Japan) [99]

184 2-Hydroxybenzoic acid Salicylic acidO

HO

HO

Stem androots (Japan) [99]

185 34-Dihydroxybenzoicacid Protocatechuic acid

O

HOOH

OH

Whole(Japan) [99]

1864-Hydroxy-

3methoxybenzoicacid

Vanillic acid

OOH

O

OH

Aerial(Uganda)

Roots (Japan)[68 99]

Evidence-Based Complementary and Alternative Medicine 35

Table 8 Continued

S N IUPAC names Common names Structure Plant part(country) References

187 345-Trihydroxybenzoicacid Gallic acid

O

OHHO

HO

HO

Whole(China) [102]

[88] This work supports the use of B pilosa as an anti-inflammatory agent however no compounds responsible forthe anti-inflammatory activity of B pilosa were identified

A further study also reported the anti-inflammatoryactivity as well as the antiallergic activity of B pilosa [75] Inthis study dried powder of the aerial part of B pilosa whichhad been pretreated with the enzyme cellulosine was usedfor further tests The results showed that oral administrationof the cellulosine-treated B pilosa lowered the level of serumIgE in mice 10 days after immunization with DNP (24-dintrophenyl)-Ascaris as an antigen This treatment alsoreduced dye exudation in skin induced by passive cutaneousanaphylaxis and production of inflammatory mediators his-tamine and substance P in rats [75] Phytochemical analysisshowed that cellulosine treatment increased the percentage ofcaffeic acid and flavonoids This study suggests that B pilosaand its phenolics have anti-inflammatory functions

Phenolics and polyynes are major anti-inflammatoryphytochemicals present in B pilosa (Table 11) Unsurpris-6ingly phenolics such as luteolin (103) and ethyl caffeate(161) that are major constituents of B pilosa have alsobeen reported to possess anti-inflammatory activity Lute-olin was reported to exhibit anti-inflammatory activity inmacrophages Xagorari and colleagues showed that luteolininhibited the release of inflammatory cytokines TNF-120572 andinterleukin-6 in RAW 2647 cells following LPS stimulation[87] It inhibited TNF-120572 production with an IC

50value

of 1120583M The underlying anti-inflammatory mechanism ofluteolin was reported to be the inactivation of Akt andNF-120581B activation [87] In addition luteolin was reportedto confer anti-inflammatory activity through inhibition ofLPS-stimulated iNOS expression in BV-2 microglial cellsIt inhibited LPS-activated microglia in a dose-dependentmanner with an IC

50value of 69 120583MMoreover immunoblot

and RT-PCR data proved that luteolin suppressed I120581B-120572degradation and iNOS expression in LPS-activated microglia[85] Kim and colleagues stated that luteolin may havebeneficial effects on inflammatory neural diseases throughinhibition of iNOS expression [85] A related study revealedthat luteolin decreased the transcriptional activity of NF-120581BRelA via partial inhibition of TNF-mediated NF-120581B DNAbinding activity Luteolin also inhibited Akt phosphorylationand induced degradation of a transcription factor interferonregulatory factor (IRF) [86]

Chiang and colleagues showed that ethyl caffeate(161) significantly inhibited NO production in mousemacrophages RAW 2647 cells [84] Based on MTT assays

they concluded that this inhibition was not due to thecytotoxicity of ethyl caffeate The IC

50value of ethyl caffeate

in the inhibition of NO production was 55 120583gmL slightlylower than curcumin (positive control) which has anIC50

value of 65120583gmL They demonstrated that ethylcaffeate exerted anti-inflammatory activity via the reducedtranscription and translation of iNOS (inducible nitric oxidesynthase) in RAW 2467 cells In addition this compoundalso suppressed COX-2 expression in RAW 2467 cellsand MCF-7 cells The in vivo anti-inflammatory effectof ethyl caffeate was verified by testing in TPA-treatedmouse skin Like celecoxib the positive control ethylcaffeate significantly abolished COX-2 expression in adose-dependent manner Ethyl caffeate at 1mg200120583Lsite(24mM) inhibited COX-2 expression at a level comparableto celecoxib at 1mg200120583Lsite (13mM) Remarkably ethylcaffeate at 48mM (2mg200120583Lsite) was more effectivethan celecoxib at 131mM (10mg200120583Lsite) Moreoverthis compound inhibited the activation of nuclear factor-120581B(NF-120581B) by LPS via the prevention of NF-120581B binding toDNA [84] In addition 3 ethyl caffeate analogs (ethyl 34-dihydroxyhydrocinnamate ethyl cinnamate and catechol)also showed different degrees of NF-120581B binding to DNA aslisted in Table 12 Ethyl cinnamate lacks a catechol moietywhich results in ineffective inhibition of NF-120581B binding toDNA [84]

Pereira and colleagues assessed the anti- 7inflammatory and immunomodulatory activities ofB pilosa methanol extract as well as one polyyne2-O-120573-glucosyltrideca-11(E)-en-3 579-tetrayn-12-diol(54) in T lymphocytes and a zymosan-induced arthritismouse model [27] They first examined the in vitro effect of 8the B pilosa extract and compound 54 on cell proliferation ofhuman T cells stimulated with 5120583gmL phytohemagglutinin(PHA) or 100 nM 12-O-tetradecanoyl phorbol-13-acetate(TPA) plus 15 120583M ionomycin and on cell proliferation ofmouse T cells stimulated with 5 120583gmL concanavalin A(Con A) The data demonstrated that both methanol extractand compound 54 suppressed T-cell proliferation in adose-dependent manner The estimated IC

50values of the B

pilosa extract against human T cells stimulated with 5120583gmLPHA and 100 nM TPA plus 15120583M ionomycin were 125 and25 120583gmL respectively In comparison with the methanolextract compound 54 showed 10-fold more inhibition ofhuman T-cell proliferation with an estimated IC

50value of

15 120583gmL Accordingly the B pilosa extract and compound54 dose-dependently suppressed mouse T-cell proliferation

36 Evidence-Based Complementary and Alternative Medicine

Table 9 Porphyrins isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

188

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(15S16S)-10-ethenyl-5-ethyl-1161820-tetrahydro-6111522-tetramethyl-1820-dioxo-15H-912-imino-212-

metheno-471714-dinitrilopyrano[43-

b]azacyclononadecine-16-propanoic

acid

AristophyllC

NH N

HNN

O

O OOOPhytyl

O

Leaves(Taiwan) [100]

189

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -20b-(methoxycarbonyl)-9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-dinitrilo-12-

dioxeto-[310158404101584034]-cyclo-pent[1 2]azacyclononadecine-

19-propanoic acid

Bidenphytin A

NH N

HNN

OPhytylO O O

O O

Leaves(Taiwan) [100]

190

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -2a-

hydroxy-20b-(methoxycarbonyl)-

9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-

dinitrilo-12-dioxeto[310158404101584034]-cyclo-pent[12-b]-

azacyclononadecine-19-propanoic acid

Bidenphytin B

NH N

HNN

OPhytylO O O

O O

OH

Leaves(Taiwan) [100]

191

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO O

HOOO

Leaves(Taiwan) [100]

192

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

Evidence-Based Complementary and Alternative Medicine 37

Table 9 Continued

S N IUPAC names Common names Structure Plant part(country) References

193

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

Leaves(Taiwan) [100]

194

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

195

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3S4S21R)-9-ethenyl-14-ethyl-21-(methoxycarbonyl)-

481318-tetramethyl-20-oxo-3-phorbinepropanoic acid

Pheophytin A

NH N

HNN

PhytylO

O

OO O

Leaves(Taiwan) [100]

with estimated IC50values of 30 and 25 120583gmL respectively

Taken together the data indicate that the B pilosa extractand compound 54 act on human and mouse T cells Totest the in vivo effect of the B pilosa extract and compound54 a zymosan-induced arthritis mouse model was usedThis model was established from B10ASgSnJ mice with aninjection of zymosan (015mg) The zymosan-injected mice9received an intraperitoneal injection of the B pilosa extract(1 5 or 10mg) at one dose a day for 5 days Popliteal lymphnode (PLN) weight was monitored to check the developmentof arthritis The results revealed that 10mg of the methanolextract of B pilosa extract could significantly diminishinflammation as evidenced by PLN weight [27] This worksuggests that B pilosa (and compound 54) can suppressimmune response and inflammation

33 Antidiabetic Activity Anti-diabetic agents are primarilydeveloped from plants and other natural resources [8 24 4951 53] B pilosa is one of 1200 plant species that have beeninvestigated for antidiabetic activity [120 121]B pilosa is usedas an anti-diabetic herb in America Africa and Asia [51 120

122] Many studies have indicated that B pilosa could treattype 1 diabetes (T1D) and type 2 diabetes (T2D) in animals

Etiologically speaking T1D is caused by autoimmune-mediated destruction of pancreatic 120573 cells leading to insulindeficiency hyperglycemia and complications Currentlythere is no cure for T1D Polarization ofTh cell differentiationcontrols the development of T1D Suppression of Th1 celldifferentiation and promotion of Th2 cell differentiationameliorate T1D [123] One study showed that the butanolfraction of B pilosa inhibited T-cell proliferation decreasedTh1 cells and cytokines and increased Th2 cells andcytokines leading to prevention of T1D in nonobesediabetic (NOD) mice [49] Based on a bioactivity-directedisolation strategy 3 polyynes 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne (53) also known ascytopiloyne 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D- Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne (50) were identifiedfrom B pilosa [49 53] The IC

50value of the butanol fraction

was 200120583gmL This inhibition was reported to be partiallyattributed to cytotoxicity because the butanol fraction at

38 Evidence-Based Complementary and Alternative Medicine

Table 10 Other compounds isolated from B pilosa [30]

SN IUPAC names Common names Structure Plant part(country) References

19637-Dihydro-137-

trimethyl-1H-purine-26-dione

CaffeineN

NN

NO

O

Aerial(Uganda) [68]

197

1-((2R4S5R)-Tetrahydro-4-hydroxy-5-(hydroxymethyl)furan-2-yl)-5-

methylpyrimidine-24(1H3H)-dione

ThymidineHO

HO

ON

O

NHO Not found

(China) [37]

198 1-(2-Thienyl)-ethanone 2-Acetyl-thiophene

S

O

Roots(Germany) [41]

199

(2R3S4S5S)-2-(Heptan-2-yloxy)-tetrahydro-6-

((tetrahydro-34-dihydroxy-5-

(hydroxymethyl)furan-2-yloxy)methyl)-

2H-pyran-345-triol

Heptanyl2-O-120573-xylofuranosyl-

(1rarr 6)-120573-glucopyranoside

O

OOH

OH

HO

OHO

H

HO

H

H

HOHH

O

Whole(Taiwan) [70]

200

2-[(3R7R11R)-3-Hydroxy-371115-

tetramethylhexadecyl]-356-trimethyl-

25-cyclohexadiene-14-dione

120572-Tocopherylquinone

OO

HO

Whole(Taiwan) [32]

2017-O-(410158401015840610158401015840-Diacetyl)-

120573-D-glucopyranoside

Not found Leaves(China) [60]

180 120583gmL could cause 50 death of Th1 cells Moreoverthis study suggested that the butanol fraction may preventdiabetes in NOD mice in vivo via downregulation of Th1cells or upregulation Th2 cells that have effects which areantagonistic of those of Th1 cells [49] This was proven byintraperitoneal injection of the butanol fraction at a doseof 3mgkg BW 3 times a week to NOD mice from 4 to 27weeks This dosage resulted in lower incidence of diabetes(33) At a dose of 10mgkg the butanol fraction of Bpilosa totally eliminated (0) the initiation of the diseaseTo further support this result assessment of IgG2a and IgEproduction was performed in the serum of NOD mice Asin vivo results obtained from intracellular cytokine stainingexperiments were not very conclusive levels of IgG2a and IgE

were measured since Th1 cytokine IFN120574 and Th2 cytokineIL-4 favor the production of IgG2a and IgE respectively Asexpected high levels of IgE and some decline in the levels ofIgG2a were observed in the serum Profiling of the butanolextract revealed five compounds 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne(50) 45-Di-O-caffeoyl- quinic acid 35-Di-O-caffeoylquinicacid and 34-Di-O-caffeoylquinic acid Only the first twocompounds showed similar effects on the prevention ofdiabetes in NOD mice as the B pilosa butanol fractionMoreover compound 50 showed greater activity thancompound 69 in terms of enhancement (by 34 comparedto 8) of differentiation of Th0 to Th2 at 15 120583gmL (both

Evidence-Based Complementary and Alternative Medicine 39

Table 11 Chemical constituents of B pilosa and their biological activities

SN Name Classification Molecularformula Biological activities

109 Centaureidin [91] Flavonoid C18H16O8Anti-listerial [69 91]

Cytotoxic [10]

110 Centaurein [91] Flavonoid C24H26O13

Anti-listerial [69 91]Cytotoxic [10]Anti-viral [105]

103 Luteolin [106] Flavonoid C15H10O6

Anti-viral [107 108]Cytotoxic [77]

Anti-inflammatory [109]Anti-allergic [109]

82 Butein [110] Flavonoid C15H12O5Anti-leishmanial [111]

Cytotoxic [78]48 12-Dihydroxytrideca-57911-tetrayne [112] Polyyne C13H12O2 Anti-angiogeneic [112]46 13-Dihyroxy-6(E)-tetradecene-81012-triyne [112] Polyyne C14H16O2 Anti-angiogeneic [112]

45 12-Dihyroxy-5(E)-tridecene-7911-triyne [113] Polyyne C13H14O2Anti-angiogeneic [113]Anti-proliferative [113]

64 1-Phenylhepta-135-triyne [114] Polyyne C13H8

Anti-microbial [115]Anti-malarial [3]Cytotoxic [3]Antifungal [15]

27 Linoleic acid [79] Fatty acid C18H32O2Anti-viral (100)Cytotoxic [17]

161 Ethyl caffeate [84] Phenylpropanoid C11H12O4 Anti-inflammatory [84]

54 2-O-120573-Glucosyltrideca-11(E)-en-3579-tetrayn-12-diol [26] Polyyne C19H20O7Immunosuppressive andAnti-inflammatory [26]

53 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne [50] Polyyne C19H22O7Anti-diabetic [116]

Anti-inflammatory [50]

69 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne [28] Polyyne C20H26O7Anti-diabetic [28]

Anti-inflammatory [94]

50 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne [28] Polyyne C19H24O7

Anti-diabetic [28]Anti-inflammatory [94]

Anti-malarial andantibacterial [47]

129 Quercetin 3-O-120573-D-galactopyranoside [18] Flavonoid C21H20O12 Anti-inflammatory [117]

170 35-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

171 45-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

169 34-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

126 Quercetin 331015840-dimethyl ether7-O-120572-L-rhamnopyranosyl-(1rarr 6)-120573-D-glucopyranoside [118] Flavonoid C21H20O11 Anti-malarial [119]

125 Quercetin 331015840-dimethyl ether-7-O-120573-D-glucopyranoside [118] Flavonoid C21H20O12 Anti-malarial [119]70 1-Phenyl-13-diyn-5-en-7-ol-acetate [27] Polyyne C15H12O2 Anti-malarial [27]199 Heptanyl 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside [70] Miscellaneous C18H44O10 Antioxidant [70]124 3-O-Rabinobioside [70] Saccharide C27H38O15 Antioxidant [70]130 Quercetin 3-O-rutinoside [70] Flavonoid C27H38O15 Antioxidant [70]167 Chlorogenic acid [70] Phenolic C16H26O9 Antioxidant [70]119 Jacein [70] Flavonoid C24H26O13 Antioxidant [70]

49 (R)-12-dihydroxytrideca-357911-pentayne [47] Polyyne C13H8O2Anti-malarial andAntibacterial [47]

40 Evidence-Based Complementary and Alternative Medicine

Table 12 Structure and activity relationship studies of ethyl caffeateusing in vitro NF-120581BDNA binding assays [84]

Compound Concentration(120583M)

NF-120581BDNA binding

Ethyl caffeate 50 100inhibition

Ethyl 34-dihydroxyhydrocinnamate 100 100inhibition

Catechol 400 100inhibition

Ethyl cinnamate 400 Noinhibition

Table 13 Apoptosis in cocultures of T cells and pancreatic 120573 cells[53]

Cellmedium apoptosis andnecrosis

CD4+ T cellscontrol medium lt4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 2

CD4+ T cellswith cytopiloyne-treated 120573 cells 18CD4+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 7

CD8+ T cellscontrol medium 4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 4

CD8+ T cellswith cytopiloyne-treated 120573 cells 4CD8+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 4

Table 14 Radical scavenging activities of B pilosa extracts [70]

Extractscontrol DPPH assayIC50 (120583gmL)

NBThypoxanthinesuperoxide assayIC50 (120583gmL)

Quercetin 198 15Ascorbic acid 634 Not determined120572-tocopherol 897 Not determinedEthyl acetateextract 1383 597

Butanol extract 1669 114Water extract gt100 gt100

compounds) and inhibition (by 40 compared to 10) ofdifferentiation toTh1 at the same concentration [49]

Among the three polyynes found in B pilosa cytopiloyne(53) had the most potent anti-T1D activity [53] To test the invivo effect of cytopiloyne NODmice received intraperitonealor intramuscular injection of cytopiloyne at 25120583gkg BW3 times per week Twelve-week-old NOD mice started todevelop T1D and 70 of NOD mice aged 23 weeks andover developed T1D Remarkably 12- to 30-week-old NODmice treated with cytopiloyne showed normal levels of bloodglucose (lt200mgdL) and insulin (1-2 ngmL) Consistent

Table 15 Radical scavenging activity of secondarymetabolites fromB pilosa [70]

Metabolite (Table 11)control DPPH assay IC50 (120583gmL)199 Not determined124 53130 68167 105169 33171 38119 Not determined110 Not determinedQuercetin 256Caffeic acid 890

Table 16 Antioxidant activity of the essential oils and water extractsfrom B pilosa [92]

Extract IC50 (120583gmL)Leaf essential oils 47Flower essential oils 50Leaf extract 61Flower extract 172

with T1D incidence cytopiloyne delayed and reduced theinvasion of CD4+ T cells into the pancreatic islets [53]

In vitro study showed that cytopiloyne (53) inhibited thedifferentiation of naıveTh (Th0) cells (ie CD4+ T cells) intoTh1 cells and promoted differentiation of Th0 cells into Th2cells [50] The in vitro data are consistent with the in vivoresults indicating that cytopiloyne reducedTh1differentiationand increased Th2 differentiation as shown by intracellularcytokine staining and FACS analysis [53] In line with theskewing of Th differentiation the level of serum IFN-120574 andIgG2c decreased while that of serum IL-4 and serum IgEincreased compared to the negative controls (PBS-treatedmice) Cytopiloyne also enhanced the expression of GATA-3 a master gene for Th2 cell differentiation but not theexpression of T-bet a master gene forTh1 cell differentiationfurther supporting its role in skewingTh differentiation [53]

Also importantly cytopiloyne partially depleted CD4+rather than CD8+ T cells in NOD mice [53] As shown inTable 13 coculture assays showed that the depletion of CD4+T cells was mediated through the induction of Fas ligandexpression on pancreatic islet cells by cytopiloyne leading toapoptosis of infiltrating CD4+ T cells in the pancreas via theFas and Fas ligand pathways However cytopiloyne did notinduce the expression of TNF-120572 in pancreatic islet cells andthus had no effect on CD8+ T cells [53]

In addition Chang and colleagues showed that cytopi-loyne dose-dependently inhibited T-cell proliferation stimu-lated by IL-2 plus Con A or anti-CD3 antibody using [3H]thymidine incorporation assay [53]

Overall the mechanism of action of cytopiloyne andprobably its derivatives in T1D includes inhibition of T-cellproliferation skewing of Th cell differentiation and partialdepletion of Th cells Due to the anti-diabetic mechanisms

Evidence-Based Complementary and Alternative Medicine 41

Table 17 Antibacterial activity of essential oils and flower extracts from B pilosa [92]

Strain Mean zone of inhibition (mm)Leaf essential oil Flower essential oil Leaf extract Flower extract

Micrococcus flavus 127 plusmn 03 87 plusmn 03 102 plusmn 02 108 plusmn 03Bacillus subtilis 173 plusmn 19 117 plusmn 02 109 plusmn 02 103 plusmn 02Bacillus cereus 190 plusmn 14 112 plusmn 03 118 plusmn 04 185 plusmn 10Bacillus pumilus 123 plusmn 07 108 plusmn 02 105 plusmn 04 77 plusmn 02Escherichia coli 137 plusmn 04 203 plusmn 07 102 plusmn 11 140 plusmn 13Pseudomonas ovalis 125 plusmn 08 137 plusmn 15 102 plusmn 06 125 plusmn 06

Table 18 Antibacterial activity of root extracts from B pilosa [93]

Strain MIC50 (mgmL)Methanol extract Acetone extract

Bacillus cereus 10 mdashEscherichia coli 5 5Klebsilla pneumonia 5 10Micrococcus kristinae 10 mdashPseudomonas aeruginosa 10 10Staphylococcus aureus 5 10Sraphylococcus epidermidis 5 5Serratia marcescens 10 mdashShigelea flexneri 10 mdashStreptococcus faecalis 10 mdash

Table 19 Antibacterial activity of B pilosa of compound 29 [47]

Strain MIC50 (120583gmL)Escherichia coli NIHJ 1Escherichia coli ATCC25922 1Klebsiella pneumoniae ATCC700603 128Serratia marcescens ATCC13880 16Pseudomonas aeruginosa ATCC27853 8Staphylococcus aureus FDA209P 05Staphylococcus aureus ATCC29213 025Staphylococcus aureus N315 (MRSA) 05Enterococcus faecalis ATCC29212 2Enterococcus faecalis NCTC12201 (VRE) 1Bacillus subtilis ATCC6633 05Candida albicans ATCC10231 025

of action it was hypothesized that cytopiloyne protectsNOD mice from diabetes by a generalized suppression ofadaptive immunity To evaluate this hypothesis ovalbumin(Ova) was used as a T-cell dependent antigen to primeNOD mice which had already received cytopiloyne orPBS vehicle Ova priming boosted similar anti-Ova titersin cytopiloyne-treated mice and PBS-treated mice but adifference in immunoglobulin isotype was observed in thetwo groups Thus it was concluded that cytopiloyne is animmunomodulatory compound rather than an immunosup-pressive compound [50 53]

T2D is a chronic metabolic disease with serious com-plications resulting from defects in either insulin secretion

insulin action or both [124] A study by Ubillas et alshowed that the aqueous ethanol extract of the aerial partof B pilosa at 1 gkg body weight (BW) lowered bloodglucose in dbdb mice a T2D mouse model [51] Basedon a bioactivity-guided identification compounds 69 and50 were identified Further the mixture of the compounds(69 50) in a 2 3 ratio significantly decreased blood glucoseconcentration and reduced food intake on the second dayof treatment when administered at doses of 250mgkg twicea day to C5BLKs-dbdb mice When tested at 500mgkga more substantial drop in blood glucose level as wellas the stronger anorexic effect (food intake reduced from58 gmouseday to 25 gmouseday) was observed [51] Inthis study it was suggested that the blood glucose lower-ing effect of B pilosa was caused in part by the hungersuppressing effect of its polyynes [51] However the hungersuppressing effect of the ethanol extract of B pilosa wasnot found in the studies described below In another study[24] water extracts of B pilosa (BPWE) were used indiabetic dbdb mice aged 6ndash8 weeks with postprandialblood glucose levels of 350 to 400mgdL Like oral anti-diabetic glimepiride which stimulates insulin release onesingle dose of BPWE reduced blood glucose levels from374 to 144mgdL The antihyperglycemic effect of BPWEwas inversely correlated to an increase in serum insulinlevels suggesting that BPWE acts to lower blood glucose viaincreased insulin production However BPWE had differentinsulin secretion kinetics to glimepiride [24] One flaw incurrent anti-diabetics is their decreasing efficacy over timeThe authors investigated the long term anti-diabetic effectof BPWE in dbdb mice BPWE reduced blood glucoseincreased blood insulin improved glucose tolerance andreduced the percentage of glycosylated hemoglobin (HbA1c)Both long-term and one-time experiments strongly supportthe anti-diabetic action of BPWE [24] In sharp contrast toglimepiride BPWE protected against islet atrophy in mousepancreas The investigators further evaluated anti-diabeticproperties of 3 B pilosa varieties B pilosa L var radiate(BPR) Bpilosa L var pilosa (BPP) and B pilosa L varminor(BPM) in dbdb mice [8] One single oral dose (10 50 and250mgkg body weight) of BPR BPP or BPM crude extractsdecreased postprandial blood glucose levels in dbdb micefor up to four hours and the reduction of glucose levels inthe blood appeared to be dose-dependent Comparing thethree variants BPR extract resulted in a higher reduction inblood glucose levels when administered at the same dose as

42 Evidence-Based Complementary and Alternative Medicine

Table 20 Antifungal activity of B pilosa [92]

Partextract Concentration (ppm) Strain InhibitionCortiicum rolfsii Fusarium solani Fusarium oxysporum

Leaves

Essentialoils

100 857 plusmn 09 682 plusmn 0 745 plusmn 17250 960 plusmn 08 779 plusmn 18 879 plusmn 04

AqueousExtracts

100 446 plusmn 17 605 plusmn 21 716 plusmn 07250 942 plusmn 03 689 plusmn 07 824 plusmn 19

Flowers

Essentialoils

100 604 plusmn 09 892 plusmn 04 869 plusmn 05250 894 plusmn 12 980 plusmn 03 949 plusmn 06

AqueousExtracts

100 331 plusmn 11 714 plusmn 07 573 plusmn 22250 661 plusmn 14 912 plusmn 0 900 plusmn 07

Table 21 Antifungal activity of B pilosa root extracts [93]

Strain LC50 (mgmL)Acetoneextracts

Methanolextracts

Waterextracts

Aspergillus niger 014 006 007Aspergillus flavus 1091 658 0Penicillium notatum 005 005 005

the other two varieties In terms of serum insulin levels a doseof 50mgkg of each extract was used and the BPR extracttogether with the three polyynes significantly increased theserum insulin level in dbdb mice Long-term experiments(28-day treatment) were then conducted using diabetic micewith postprandial glucose levels from 370 to 420mgdL andglimepiride was used as positive controlThe range of dosagesapplied was from 10mgkg BW to 250mgkg BW Resultsshowed that the positive control as well as the crude extractsof the three varieties lowered the blood glucose levels indbdb mice However only BPR extract containing a higherpercentage of cytopiloyne (53) reduced blood glucose levelsand augmented blood insulin levelsmore than BPP andBPMThe percentage of glycosylated hemoglobin A1c (HbA1c) wasalso measured and found to be 79 plusmn 05 in mice aged 10ndash12 weeks and 66 plusmn 02 61 plusmn 03 and 62 plusmn 03 inthe blood of age-matchedmice following treatment with BPRcrude extract (50mgkg) glimepiride (1mgkg) and com-pound 53 (05mgkg) respectively [8] Among the polyynesfound in B pilosa cytopiloyne was the most effective againstT2D Hence cytopiloyne was used for further study on anti-diabetic action and mechanism [125] The data confirmedthat cytopiloyne reduced postprandial blood glucose lev-els increased blood insulin improved glucose tolerancesuppressed HbA1c level and protected pancreatic islets indbdb mice Nevertheless cytopiloyne failed to decreaseblood glucose in streptoztocin (STZ)-treated mice whose bcells were already destroyed Additionally cytopiloyne dose-dependently increased insulin secretion and expression in bcells as well as calcium influx diacylglycerol and activationof protein kinase C120572 Collectively the mechanistic studiessuggest that cytopiloyne treats T2D via regulation of insulin

production involving the calciumDAGPKC120572 cascade in bcells

The studies detailed above point to the conclusion thatcytopiloyne and related polyynes (compounds 69 and 49)are anti-diabetics in animal models The data uncover a newbiological action of polyynes It should be noted that likeall anti-diabetic drugs cytopiloyne failed to prevent or curediabetes completely but reduced diabetic complications [125]Intriguingly 34 polyynes have been found in B pilosa so farIt remains to be seen whether all the polyynes present in thisplant have anti-diabetic activities

34 Antioxidant Activity Free radicals can damage cellularcomponents via a series of chemical reactions [89] leadingto development and progression of cardiovascular diseasecancer neurodegenerative diseases and ageing [70] Freeradicals nitric oxide (NO) and superoxide anions can beproduced in macrophages to kill microbes However anexcessive generation of the free radicals under pathologicalconditions is associated with a wide range of illnessesPlants are known to be rich in antioxidant phytochemicalsChiang and colleagues evaluated the free radical scaveng-ing activity of crude extract fractions and compounds ofB pilosa using 11-diphenyl-2-picrylhydrazyl (DPPH) andhypoxanthinexanthine oxidase assays [70] Using DPPHand hypoxanthinexanthine oxidase assays they found thatthe B pilosa crude extract and the ethyl acetate butanoland water fractions had free radical scavenging activityNine compounds Heptyl- 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside (199) 3-O-Rabinobioside (124) Quercetin3-O-rutinoside (130) Chlorogenic acid (167) 34-Di-O-caffeoylquinic acid (169) 35-Di-O-caffeoylquinic acid (170)45-Di-O-caffeoylquinic acid (171) Jacein (119) and Cen-taurein (110) had DPPH radical scavenging activity [70]The IC

50values of the B pilosa crude extractfractions and

compounds are summarized in Tables 14 and 15 respectivelyMeasurement of free radical scavenging activities is one

way of assessing the antioxidant activities of B pilosa andits fractions and compounds It is interesting that the ethylacetate and butanol fractions are more active than the waterfraction and B pilosa crude extract [70] Of the secondarymetabolites only phenolic compounds 124 130 167 169 and171 showed significant DPPH-radical scavenging activities

Evidence-Based Complementary and Alternative Medicine 43

Further analysis of the structure-activity relationship of thecompounds suggested that substitution of the C3 hydroxylgroup with glycosides increased the activity approximately2-fold (for example in compounds 124 and 130) relative toquercetin (-OH in its C3) [70] Further modification of thestructures of the active compounds needs to be performed totest the effects of various substituents on activity The reasonwhy most of the antioxidant compounds contain phenolmoieties in their structure could be that the reduction-oxidation (redox) properties of phenols allow them to actas reducing agents singlet oxygen quenchers and hydrogendonors

A complementary study by Muchuweti and colleaguesdetermined phenolic content antioxidant activity and thephenolic profile of B pilosa methanol extract [89] Theyestimated that the phenolic content of the methanol extractof B pilosawas 11028plusmn22mgg [89] Vanillin hydroxyben-zaldehyde caffeic acid coumaric acid and ferulic acid werefound in this extractThe B pilosa extract also showed DPPHradical scavenging activity Furthermore the antioxidantactivity of the flavonoids found in B pilosa was correlatedwith its hepatoprotective effects through their inhibitionof NF-120581B activation which may lessen the oxidative stresscaused by the production of free radicals during liver injury[60]This activity might also be due to the anti-inflammatoryeffects of the aqueous extracts of B pilosa aerial parts on theinhibition of COX-2 and PGE

2production [88]

Essential oils from B pilosa flowers and leaves are alsoreported to possess antioxidant activity With the aim ofreplacing chemically synthesized additives Deba and col-leagues [92] worked on the antioxidant antibacterial andantifungal activities of essential oils and water extracts of Bpilosarsquos leaves and flowers Table 16 summarizes the resultsobtained from DPPH free radical scavenging assay

It can be inferred fromTable 16 that essential oils from theleaves possessed the highest activity It is reported elsewherethat monoterpenes present in essential oils such that of Bpilosa have protective effects and antioxidant properties [92]Beta-carotene bleaching method was also performed Leavesessential oils and aqueous extracts of the leaves and flowersshowed higher activity than the flower essential oils This isdue to the volatility of the flower essential oils The activityexhibited by the aqueous extracts was accounted to thepresence of phenolic compounds that are reported to donatea hydrogen atom to free radicals such that the propagation ofthe chain reaction during lipid oxidation is terminated [92]Overall essential oils and phenolics present in B pilosa canbe thought of as major antioxidant compounds

35 Immunomodulatory Activity B pilosa is thought to bean immunomodulatory plant and is reported to be effectivein the treatment of immune disorders such as allergy [75]arthritis [75] and T1D [48 50 53]

As pointed out in the discussion of its anti-diabeticactivities (Section 33) a combination of phytochemicalsmethod and T-cell activation assays was used to studyimmunomodulatory properties of B pilosa IFN-120574 is a keycytokine released by T and NK cells that mediates immune

cells and sustains immunity against pathogens Defects inIFN-120574 expression regulation and activation result in vulner-ability to diseases caused by bacteria and viruses [69] Anelegant study performed byChang and colleagues using IFN-120574 promoter-driven luciferase reporter construct in JurkatT cells showed that hot water crude extracts of B pilosaincreased IFN-120574 promoter activity two-fold [69] Out of thesubfractions of this extract the butanol fraction but not thewater or ethyl acetate fractions increased IFN-120574 promoteractivity six-fold Centaurein (110) and centaureidin (109)were identified from the butanol fraction and were stated tocause a four-fold increase in IFN-120574 promoter activity withEC50

values of 75 120583gmL and 09120583gmL respectively Themechanism of action of centaurein was determined usingtranscription factors such as AP-1 NFAT and NF120581B whichare reported to bind to IFN-120574 promoter and regulate IFN-120574transcription Unlike with the activity of the positive controlPHA centaurein caused a four-fold increase in NFAT a3-fold increase in NF120581B and had little if any effect onAP-1 enhancer activities (23-fold three-fold and ten-foldincreases respectively were seenwith PHA) [69]The authorsconcluded that centaureinmodulates IFN-120574 expression by theNFAT and NF120581B pathways The article only determined themechanism of action of centaurein Its aglycone centaureidinmay act through the samemechanism though this conclusionneeds to be further verified

B pilosa extract and its compounds are reported to inhibitdifferentiation of naıve CD4+ helper T (Th0) cells into Th1cells [49] Using theThcell differentiation assay as a screeningplatform 3 polyynes 2-120573-D-glucopyranosyloxy-1 -hydroxytrideca-57911-tetrayne (53) 3-120573-D-glucopyranosyl-1-hydroxy-6(E)- tetradecene-81012-triyne (69) and 2-120573-D-glucopyranosyloxy-1-hydroxy-5(E)- tridecene-7911-triyne(49) were discovered from B pilosa [49 53] The data showsthat cytopiloyne and other two polyynes suppressed thedifferentiation of type 1 helper T (Th1) cells and productionof Th1 cytokines and promoted that of type 2 helper T (Th2)cells and production of Th2 cytokines thus explaining theimmunomodulatory and anti-inflammatory effects of Bpilosa and its polyynes

Chang and colleagues were the first to report the effect ofthe butanol extract of B pilosa on the autoimmune diabetesand airway inflammation inmice [48] Imbalance in the levelsofTh1 andTh2 and of various cytokines leads to autoimmunediseases T1D and other autoimmune diseases (rheumatoidarthritis Crohnrsquos disease among others) are exacerbated byan increase Th1 levels (specifically CD4+ Th1 cells) whileTh2 cells antagonize this effect [19] Moreover Th2 cellsmediate asthma in ovalbumin-induced hypersensitivity inBALBc mice [48] In their work [48] Chang and colleaguesshowed that 10mgkg butanol extracts with 15 (ww)compound 69 and 11 (ww) compound 49 (Table 11)ameliorated the development of Th1-mediated diabetes inNOD mice through inhibition of 120573 cell death and leukocyteinfiltration At the same dosage the butanol extracts alsoexacerbated ovalbumin-induced pulmonary inflammationin BALBc mice with an increase in the infiltration ofeosinophils and mast cells into the airway of the mice [48]Despite the different outcomes both mouse models proved

44 Evidence-Based Complementary and Alternative Medicine

the concept that control over the Th1Th2 shift is associatedwith autoimmune diseases and that the B pilosa butanolextract can shift the differentiation of Th0 cells to Th2 cells[48 49]

An extended study presented by Chiang and colleaguesshowed that compound 53 modulates T-cell functions [50]Using CD4+ T cells from BALBc mice they demonstratedthat cytopiloyne decreased levels of IFN-120574 producing cells(Th1) by 122 (from 72 to 598) Since Th1 and Th2 celldifferentiation is antagonistic it was expected that compound53 increased the percentage of mouse IL-4 producing cells(Th2) by 72 (from 237 to 309) Subsequent assess-ment of effect of compound 53 on the modulation of thetranscription of IL-4 and IFN-120574 showed that cytopiloyne asexpected decreased the splenocyte levels of IFN-120574 mRNAand increasing that of IL-4 in a dose-dependent manner Inthe range of 01 to 3 120583gmL of compound 53 the effects werenot attributed to its cytotoxicity Consequently using 3120583gmLcytopiloyne for 72 and 96 hours the protein concentration ofIFN-120574 decreased to 186 and 444 respectively Under thesame conditions cytopiloyne increased IL-4 concentrationsto 1985 and 2470 (for 72 and 96 hours resp) Thismodulation of T-cell differentiation exhibited by compound53 was used to explain its anti-diabetic activity [50]

The anti-diabetic role of cytopiloyne was extensivelydiscussed above (Section 33 anti-diabetic activity) Themolecular basis of the regulation of cytokine expression bycytopiloyne has been described Cytopiloyne directly elevatedthe expression level of IL-4 via GATA-3 upregulation inT cells [53] However reduction of IFN-120574 expression in Tcells seemed to come from the indirect opposing effect IL-4cytokine because the expression level of T-bet was unaltered[53] In this way cytopiloyne skewedTh1 polarization into theTh2 state conferring protection against T1D in NOD miceAside frompolarization ofThcell differentiation cytopiloynealso activated the expression of Fas ligand in pancreaticb cells this increase leading to the partial depletion of Tcells and reduction of immune response in local areas suchas the pancreas Of note cytopiloyne also inhibited T-cellproliferation and activation By targeting T cells from threeimmunomodulatory actions cytopiloyne protects againstT1D and probably other Th1-mediated autoimmune diseases[19]

The phytochemical constituents of B pilosa exert theirfunctions on different immune cells to modulate immuneresponse It is possible that some of the compounds mayhave agonistic or antagonistic effects on immune responseImmune function of B pilosa may depend on its compo-sition and amount of compounds which could explain theapparently conflicting report of B pilosa butanol extractaggravating allergy in mice [48] while cellulosine-treatedextract ameliorated allergy [48 75] IFN-120574 promoter reporterassays and T-cell differentiation assays were used to isolate2 flavonoids [69] and 3 polyynes [49 50] as immunomod-ulatory compounds from the butanol fraction of B pilosaInterestingly the flavonoids promote IFN-120574 expression inNK and T cells In marked contrast the polyynes promoteIL-4 expression and indirectly inhibit IFN-120574 expression indifferentiating T cells Sensitivity appears to be the key to

identifying structure- and bioactivity-related phytochemicalsfrom medicinal plants

36 Antimalarial Activity The use of chemical drugs againstpathogens has resulted in drug-resistant mutants Examplesof drug resistance can be found in the species of thePlasmodium that cause malaria It is important to search fornew compounds to combat Plasmodium parasites [47] Astudy of the anti-malarial activity of the leaf extracts of Bpilosa using a combination of phytochemistry and bioassaysshowed that compound 49 (Table 11) showed activity againsta malaria parasite (P falciparum NF54 strain) with an IC

50

value of 60 120583gmL [77] In addition compound 49 isolatedfrom the aerial parts of B pilosa inhibited growth of the Pfalciparum FCR-3 strain with an IC

50value of 035 120583gmL

This compound was tested for its in vivo effect in miceinfected with P berghei NK-65 strain Results showed thatcompound 49 decreased the average parasitemia in the redblood cells by 207 (from 328 of that of the control to121) after an intravenous injection of 08mgkg BWday forfour days [47] Further studies addressing the anti-malarialmechanism underlying both polyynes and clinical studies areneeded

37 Antibacterial Activity Emergence of multiple antibiotic-resistantmicrobes is becoming a global threat to public healthand a challenge to disease treatment For instance penicillinis commonly used to combat a food-borne intracellularbacteriumListeria however penicillin-resistant bacteria havebeen discovered recently [69] Chang and coworkers iso-lated centaurein (110) and centaureidin (109) from B pilosaextract [69 70] Centaurein enhances expression of IFN-120574a key cytokine for macrophage activation and consequentlyenhances bactericidal activity in macrophages [69 70] Inagreement with observed in vitro effects centaurein wasreported to prevent and treat Listeria infection in C57BL6Jmice [91] Mechanistic studies confirmed that centaureinexerted antilisterial action via IFN-120574 expression in wild-typemice but not IFN-120574 knockout mice [91] Further in vitrostudies on centaurein showed that this compound increasedIFN-120574 expression by 13 (from 17 to 20) 20 (from 21to 41) and 11 (from 6 to 17) in CD4+ T cells CD8+Tcells and NK cells respectively That is to say there wasan increase in IFN-120574 producing immune cells As expectedcentaurein also enhanced the expression level of T-bet a keynuclear factor for IFN-120574 expression Consistently centaureinaugmented the serum IFN-120574 levels inC57BL6Jmice and thisaugmentation peaked 24 hours after compound injectionThe quantity of mouse serum IFN-120574 was sufficient to activatemacrophages in vitro and eradicated GFP-producing Listeriainside macrophages However the entry of Listeria intomacrophages was not affected by centaurein-treated mousesera Centaurein treatment at 20120583g per mouse rescued 30of the mice infected with a lethal dose of Listeria (2 times106 CFU) It is noteworthy that in the presence of ampicillin(5 120583gmouse) centaurein (20 120583gmouse) rescued 70 of themice suggesting an additive effect between ampicillin andcentaurein [91] Despite lower abundance centaureidin was

Evidence-Based Complementary and Alternative Medicine 45

30 times more active than centaurein in terms of IFN-120574production [91]

Aside from the indirect antibacterial action mentionedabove extract andor compounds of B pilosa also showeddirect bacteriostatic andor bactericidal action One studyreported that essential oils and leafflower extracts of Bpilosa could suppress the growth of gram positive and gramnegative bacteria as evidenced by zone of inhibition assaysIn this study antibacterial activity of the essential oils fromthe leaves and flowers of B pilosa was determined in anattempt to identify natural products as food preservatives forprevention of microbial multiplication and food oxidationThe essential oils and extracts of B pilosa leaves and flow-ers showed moderate but different extents of antibacterialactivity (Table 17) In general essential oils had higherantibacterial activity than crude extracts One explanation forthis could be that monoterpenes in the essential oils destroycellular integrity and subsequently inhibit the respirationand ion transport processes The presence of antibacterial120573-caryophyllene could be another explanation as reportedelsewhere [92] Another study reported that the methanoland acetone extract of B pilosa roots displayed antibacterialactivities against the bacteria listed in Table 18 [93] andmethanol extracts from the roots seemed to be the mosteffective

Another study indicated that the polyyne (R)-12-dihydroxytrideca-357911-pentayne (49) from this plantalso suppressed bacterial growth as shown by the minimuminhibitory concentration required to inhibit 50 bacterialgrowth (MIC

50) in Table 19 This compound was highly

effective against several Gram positive and Gram negativebacteria including the drug-resistant bacteria Staphylococcusaureus N315 (MRSA) and Enterococcus faecalis NCTC12201(VRE) [47] Strikingly compound 49 had a similar MIC

50

value to antibiotics (ampicillin tetracycline norfloxacin andamphotericin B) in most of the bacteria tested

Antibacterial activity of B pilosa extracts and compo-nents expressed as MIC

50and the mean zone of inhibition

is tabulated in Tables 17 18 and 19The zone of inhibition forAmpicilline (positive control) ranges from 153 plusmn 03mm to443 plusmn 02mm [92]

38 Antifungal Activity B pilosa has traditionally been usedto treat microbial infection Recently different parts of Bpilosa have been tested for antifungal activities Deba andcolleagues first evaluated the antifungal effect of the hotwater extracts of the B pilosa roots stems and leaves againstCorticium rolfsii Fusarium solani and Fusarium oxysporumThey discovered that C rolfsii was most suppressed bytreatment with B pilosa as its growth was reduced at almostall the tested doses followed by F oxysporum and F solani[99] However the fungicidal activities of the stems androots were greater than the leaves [99] Moreover the samegroup assessed the antifungal activity of the essential oilsand aqueous extracts from B pilosa flowers and leaves [92]They showed that the extracts and oils had antifungal activityagainst C rolfsii F solani and F oxysporum Essential oils

appeared to have better fungicidal activity thanwater extractsas summarized in Table 20

Another study by Ashafa and colleagues showed thatacetone methanol and water extracts of the B pilosa rootsshowed antifungal activities against Aspergillus niger Aflavus and Penicillium notatum using the agar dilutionmethod The results are tabulated in Table 21 [93] Negativecontrols showed 0 growth inhibitionThe methanol extractof the B pilosa roots at 10mgmL was also effective againstCandida albicans [93]Of note B pilosa obtained fromPapuaNew Guinea had no activity against A niger and C albicans[126] but the SouthAfrican (EasternCape) ecotype exhibitedmoderate activity against C albicans [93] This discrepancymay depend on extraction solvents extraction procedureassay techniques different plant parts and abundance ofactive compounds

B pilosa produces a variety of secondary metabolitessuch as flavonoids phenylacetylenes alkaloids sterols ter-penoids and tannis [92 93] However none of them havebeen confirmed as active compounds against fungi Furtherinvestigation of active compounds from B pilosa is necessaryto further understand the antifungal efficacy of this plant

39 Hypotensive and Vasodilatory Activities In early studiesDimo and colleagues used three rat models normotensiveWistar rats (NTR) salt-loading hypertensive rats (SLHR)and spontaneous hypertensive rats (SHR) to investigate thehypotensive effect of the methanol crude extract of B pilosaleaves [28 127] The extract lowered systolic blood pressurein hypertensive rats (SLHR and SHR) to a greater degreethan NTR [28 127] In addition a decrease in urinarysodium ions and an increase in urinary potassium ions wereobserved after treatment with the methanol extract of Bpilosa leaves although neither differences were statisticallysignificant [28 127] Taking the data together the studyproposed that B pilosa leaf extract reduced blood pressurevia vasodilation [28 127] The same group continued totest the antihypertensive effect of aqueous and methylenechloride extracts of B pilosa leaves in a hypertensive ratmodel [28 127] To establish a fructose-induced hypertensionmodel male Wistar rats were given 10 fructose solutionto drink ad libitum for three weeks In addition to freeaccess to 10 fructose the rats were treated with the aqueous(150mgkg) or methyl chloride (350mgkg) extracts of Bpilosa for additional three weeks [28 127] Both extracts ofB pilosa leaves had a hypotensive effect on rats Howeverneither extracts reversed the elevation of serum insulin infructose-fed rats Therefore B pilosa lowered blood pressureirrespective of insulin [28 127] To better understand thehypotensive mechanism the authors investigated the effectof a neutral extract of B pilosa (NBP) a mixture of methanolandmethylene chloride (1 1) extract after neutralization withNaOH and HCl on the heart and the blood pressure ofNTR and SHR [110] This study showed that an intravenousinjection of the NBP resulted in a biphasic reduction insystolic blood pressure In addition one intravenous dose ofthe extract at 10 20 and 30mgkg BW decreased systolicblood pressure in normal rats by 183 425 and 30

46 Evidence-Based Complementary and Alternative Medicine

respectively and the same doses reduced the blood pressurein hypertensive rats by 258 389 and 286 respectivelyOnly the highest dose (30mgkg) affected the force of thecontraction of the heart Atropine and propranolol were usedto interfere with the hypotensive action of the NBP Atropinereduced the initial phase of the hypotensive response in NBPand completely abolished the second phase of hypotensiveresponse in NBP In contrast propranolol increased thefirst hypotensive response but partially abolished the secondhypotensive response provoked by the NBP [110]This mech-anistic study suggested that B pilosa invokes the biphasichypotensive responses via targeting cardiac pump efficiencyduring the first phase and vasodilation at the second phase[110]

A further study was performed to investigate the relaxingeffect of a neutral extract of B pilosa (NBP) on rat aorta con-tractedwithKCl (60mM) andnorepinephrine (01mM) [94]Cumulative addition of NBP relaxed the rat aorta previouslycontracted by KCl in a dose-dependent manner The EC

50

value of theNBP for vasorelaxationwas 032mgmLThedataalso showed that the NBP reduced the contraction of aortapreviously contracted by KCl irrespective of the presence ofaortic endothelium [94]

Pretreatment with glibenclamide an ATP-dependent K+channel blocker did not considerably affect the relaxant effectof the NBP on KCl-induced contraction suggesting thatthe vasodilatory effect of B pilosa was not related to theopening of this ATP-dependent K+ channel [94] On theother hand in the presence of indomethacin or pyrilaminemaleate the relaxant response induced by the plant extractwas significantly inhibited at the lower concentrations Theplant extract was able to reduce the aorta resting tone inhibitthe KCl-induced contractions by 90 at 15mgmL and theCaCl2-induced contractions by 95 at 075mgmL These

results demonstrate that B pilosa can act as a vasodilatorprobably via acting as a calcium antagonist [94]

However no specific compound for the above activityhas been identified from B pilosa to date A bioactivity-guided identification approach may be adopted to identifythe active compounds in B pilosa that possess hypotensiveand vasodilatory effects and understand their mechanism ofaction

310 Wound Healing Activity B pilosa has been traditionallyused to treat tissue injury inCameroon Brazil andVenezuela[26] Hassan and colleagues investigated the wound healingpotential of B pilosa in Wistar rats [128] Mirroring thepositive control neomycin sulfate the ethanol extract of Bpilosa had faster wound closure than control rats 3 6 and 9days after topical application Histological examination alsorevealed better collagenation angiogenesis and organizationof wound tissue seven days after application Epithelializationand total healing time in B pilosa-treated rats were compara-ble to those of neomycin sulfate Together these data suggestthat B pilosa may be a viable alternative to neomycin lotionfor the treatment of wounds

In addition to studying the wound healing effect of Bpilosa on external ulcers Tan and colleagues also examined

the effect of methanol cyclohexane and methyl chlorideextracts of B pilosa on gastric ulcers in Wistar rats fedwith 1mL HClethanol gastric necrotizing solution (150mMHCl in 60 ethanol) and macroscopically visible lesionswere scored [26] Among the three extracts methylene chlo-ride extracts exhibited the highest activity showing 464inhibition of lesion formation at a dose of 500mgkg BWand complete inhibition at 750mgkg [26] The efficacy ofthe ethylene chloride extract was followed by that of themethanol extracts which had inhibition ranging from 304to 822 at concentrations of 500mgkg and 1000mgkgBW respectively [26] The cyclohexane extracts showed thelowest activity against gastric ulcers in rats with 13340 and 797 inhibition at 500 750 and 1000mgkg BWrespectively [26] To better understand the mode of actionof the methylene chloride extract of B pilosa rats werepretreated with indomethacin a COX-2 inhibitor involved inprostaglandin synthesis Pretreatment significantly reducedthe protection against HClethanol-induced ulcers to 313inhibition at 750mgkg BW suggesting a link between theantiulcerative activity of B pilosa and prostaglandin syn-thesis Unexpectedly the methylene chloride extract of Bpilosa showed little gastric mucosal protection against gastriclesions induced by 95 ethanol (1mL) [26] Absolute alcoholis known to cause mucosalsubmucosal tissue destruction viacellular necrosis and the release of tissue-derived mediators(histamine and leukotrieneC4)Thus these data imply thatBpilosa did not prevent the generation or the necrotic action ofthese mediators on the gastric microvasculature In additionpylorus ligation can increase gastric acid secretion withoutan alteration of mucosal histamine content The methylenechloride extract of B pilosa did not possess antisecretoryactivity On the contrary it was observed that increases in thedose of the extract led to elevated gastric juice acidity [26]Results with both absolute ethanol and pylorus ligation ratmodels suggested the possibility that the ineffectiveness of Bpilosa against gastric ulcers was due to lack of antihistaminicactivity in the plant In summary overall the data suggest thatB pilosa protects against HCLethanol-mediated ulcers viainhibition of prostaglandin biosynthesis

Previous phytochemical studies showed that a groupof flavonoids acyclichalcones are present in B pilosa[129 130] and the chalcones were proposed to have anti-ulcerative activity [131] Moreover nine hydroxychalcones 10were reported to possess gastric cytoprotective effects with24-dihydroxychalcone being the most active [132] Sincemethylene chloride extracts appear to be the most active Bpilosa extracts next the specific anti-ulcerative phytochemi-cals in the methylene chloride extracts of B pilosa and theirmodes of action needs to be probed

Despite the claims listed inTable 3 relatively few scientificstudies have been conducted in vitro and in vivo to address thetraditional ethnomedical uses of B pilosa Information aboutthe use of B pilosa as a botanical therapy recorded so far isfar from complete Studies conducted thus far only serve asa starting point for further investigation of B pilosa and theultimate efficacious use of the herb in clinical applications

Evidence-Based Complementary and Alternative Medicine 47

4 Toxicology

Despite its use as an ingredient in food for human con-sumption studies on systemic toxicity (eg acute subacutechronic and subchronic toxicities) of B pilosa in humansand animals are still inadequate and insufficient So faracute andor subchronic toxicities have been evaluated inrats and mice Oral acute and 28-day toxicities of waterextract of B pilosa leaves were evaluated in Wistar rats[133] An oral dose of water extract of B pilosa leaves at10 gkg BW showed no obvious mortality or changes inthe appearance in rats [134] The same extract at 08 gkgBWday once a day showed no obvious sub-chronic toxicityin rats over 28 days as measured by survival rate bodyweight and gross examination of organs [134] These dataare consistent with our data indicating that oral deliveryof the water extract of the B pilosa whole plant at 1 gkgBWday once a day is safe in rats over 28 days (unpublisheddata) Taken together these studies suggest that ingestion ofB pilosa aqueous extract at up to at 1 gkg BWday oncea day is highly safe in rats In addition the acute toxicityof aqueous and ethanol extracts of B pilosa in mice havebeen reported [134] Five- to six-week-old mice with weightsbetween 28 and 35 g received a peritoneal injection of bothextracts at the different doses The LD

50 the dose that causes

50 lethality of the aqueous and ethanol extracts in micewas 1230 gkg BW and 615 gkg BW respectively [134] Acomplete toxicological study has not been completed forhumans Furthermore the drug interactions of B pilosa withother drugs are unknown Further safety verification andclinical trials should be performed before B pilosa can beconsidered for medicinal use

5 Conclusions

B pilosa is an erect perennial plant with green leaves whiteor yellow flowers and tiny black seeds As it is distributedworldwide and is widely used as a folk remedy B pilosacan be thought of as an extraordinary source of food andmedicine However a comprehensive up-to-date review ofresearch on B pilosa has hitherto been unavailable In thisarticle scientific studies on B pilosa have been summarizedand critically discussed from the perspectives of botanyethnomedicine phytochemistry pharmacology and toxicol-ogy B pilosa is claimed to treat more than 40 disordersand 201 compounds have been identified from this plantThe medicinal utility of B pilosa and its modes of actionin relation to its known phytochemicals were discussedherein Polyynes flavonoids phenylpropanoids fatty acidsand phenolics are the primary bioactive compounds of Bpilosa and they have been reported to be effective in thetreatment of tumors inflammationimmune modulationdiabetes viruses microbes protozoans gastrointestinal dis-eases hypertension and cardiovascular diseases Cautionshould be exercised in the therapeutic use of B pilosa forhypoglycemia hypotension bleeding and allergy

Acknowledgments

The authors thank their laboratory members for construc-tive suggestions and the authors whose publications theycited for their contributions The authors also thank MsMiranda Loney of Academia Sinica for English editing of thispaper This work was supported by Grants 99-CDA-L11 and101S0010056 from Academia Sinica Taiwan Arlene P Bar-tolome is a recipient of MECO-TECO Sandwich ScholarshipProgram between Taiwan and the Philippines

References

[1] T Shen G H Li X N Wang and H X Lou ldquoThe genusCommiphora a review of its traditional uses phytochemistryand pharmacologyrdquo Journal of Ethnopharmacology vol 142 no2 pp 319ndash330

[2] C Long P Sauleau B David et al ldquoBioactive flavonoids ofTanacetum parthenium revisitedrdquo Phytochemistry vol 64 no2 pp 567ndash569 2003

[3] P O Karis and O Ryding Asteraceae Cladistics and Classifica-tion Bremer K Eds pp 559ndash569 Timber press Portland OreUSA 1994

[4] O N Pozharitskaya A N Shikov M N Makarova et alldquoAnti-inflammatory activity of a HPLC-fingerprinted aqueousinfusion of aerial part of Bidens tripartita LrdquoPhytomedicine vol17 no 6 pp 463ndash468 2010

[5] F Q Oliveira V Andrade-Neto A U Krettli and MG L Brandao ldquoNew evidences of antimalarial activity ofBidens pilosa roots extract correlated with polyacetylene andflavonoidsrdquo Journal of Ethnopharmacology vol 93 no 1 pp 39ndash42 2004

[6] Agriculture USDA ldquoPlants databaserdquo 2013in Natural Resources Conservation Servicehttpwwwnrcsusdagovwpsportalnrcssitenationalhome

[7] Y Zhou and X Z Yan ldquoExperimental study of qi deficiencysyndrome and Codonopsis pillosulae and Astragalus injectionon the immune response in micerdquo Zhong Xi Yi Jie He Za Zhivol 9 no 5 pp 286ndash262 1989

[8] S C Chien P H Young Y J Hsu et al ldquoAnti-diabeticproperties of three common Bidens pilosa variants in TaiwanrdquoPhytochemistry vol 70 no 10 pp 1246ndash1254 2009

[9] M J Alcaraz and M J Jimenez ldquoFlavonoids as anti-inflammatory agentsrdquo Fitoterapia vol 59 no 1 pp 25ndash38 1988

[10] FAO Agriculture Food and Nutrition for AfricamdashA ResourceBook for Teachers of Agriculture Publishing ManagementGroup FAO Information Division Rome Italy 1997

[11] M B Rokaya Z Munzbergova B Timsina and K R BhattaraildquoRheum australe D Don a review of its botany ethnobotanyphytochemistry and pharmacologyrdquo Journal of Ethnopharma-cology vol 141 no 3 pp 761ndash774 2012

[12] P H Young Y J Hsu and W C Yang ldquoBidens pilosa L andits medicinal userdquo in Recent Progress in Medicinal Plants DrugPlant II A S AwaadVK Singh and JNGovil Eds StandiumPress Houston Tex USA 2010

[13] C Ge ldquoCytologic study of Bidens bipinnata LrdquoZhongguo ZhongYao Za Zhi vol 15 no 2 pp 72ndash125 1990

[14] L C Chiang J S Chang C C Chen L T Ng and C CLin ldquoAnti-herpes simplex virus activity of Bidens pilosa and

48 Evidence-Based Complementary and Alternative Medicine

Houttuynia cordatardquoAmerican Journal of ChineseMedicine vol31 no 3 pp 355ndash362 2003

[15] N P Rybalchenko V A Prykhodko S S Nagorna et alldquoIn vitro antifungal activity of phenylheptatriyne from Bidenscernua L against yeastsrdquo Fitoterapia vol 81 no 5 pp 336ndash3382010

[16] K Redl W Breu B Davis and R Bauer ldquoAnti-inflammatoryactive polyacetylenes from Bidens campylothecardquo Planta Med-ica vol 60 no 1 pp 58ndash62 1994

[17] M Ayyanar and S Ignacimuthu ldquoTraditional knowledge ofKani tribals inKouthalai of Tirunelveli hills TamilNadu IndiardquoJournal of Ethnopharmacology vol 102 no 2 pp 246ndash255 2005

[18] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant intraditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[19] AKAbbas KMMurphy andA Sher ldquoFunctional diversity ofhelper T lymphocytesrdquo Nature vol 383 no 6603 pp 787ndash7931996

[20] J H Cano and G Volpato ldquoHerbal mixtures in the traditionalmedicine of Eastern Cubardquo Journal of Ethnopharmacology vol90 no 2-3 pp 293ndash316 2004

[21] J Namukobe J M Kasenene B T Kiremire et al ldquoTraditionalplants used for medicinal purposes by local communitiesaround the Northern sector of Kibale National Park UgandardquoJournal of Ethnopharmacology vol 136 no 1 pp 236ndash245 2011

[22] C Lans ldquoComparison of plants used for skin and stomachproblems in Trinidad and Tobago with Asian ethnomedicinerdquoJournal of Ethnobiology andEthnomedicine vol 3 article 3 2007

[23] S Dharmananda ldquoA Popular Remedy Ecapes Notice ofWesternPractitionersrdquo 2013 httpwwwitmonlineorgartsbidenshtm

[24] Y J Hsu T H Lee C L T Chang Y T Huang andW C YangldquoAnti-hyperglycemic effects and mechanism of Bidens pilosawater extractrdquo Journal of Ethnopharmacology vol 122 no 2 pp379ndash383 2009

[25] E Noumi F Houngue and D Lontsi ldquoTraditional medicinesin primary health care plants used for the treatment ofhypertension in Bafia Cameroonrdquo Fitoterapia vol 70 no 2 pp134ndash139 1999

[26] P V Tan T Dimo and E Dongo ldquoEffects of methanolcyclohexane and methylene chlo ride extracts of Bidens pilosaon various gastric ulcer models in ratsrdquo Journal of Ethnophar-macology vol 73 no 3 pp 415ndash421 2000

[27] R L C Pereira T Ibrahim L Lucchetti A J R Da Silva and VL G De Moraes ldquoImmunosuppressive and anti-inflammatoryeffects of methanolic extract and the polyacetylene isolatedfrom Bidens pilosa Lrdquo Immunopharmacology vol 43 no 1 pp31ndash37 1999

[28] T Dimo J Azay P V Tan et al ldquoEffects of the aqueous andmethylene chloride extracts of Bidens pilosa leaf on fructose-hypertensive ratsrdquo Journal of Ethnopharmacology vol 76 no 3pp 215ndash221 2001

[29] CWiartMedicinal Plants of Southeast Asia Pelanduk SelabgorDarul Ehsan Malaysia 2000

[30] F L Silva D C H Fischer J F Tavares M S Silva P FDe Athayde-Filho and J M Barbosa-Filho ldquoCompilation ofsecondary metabolites from Bidens pilosa LrdquoMolecules vol 16no 2 pp 1070ndash1102 2011

[31] A H Chen S R Lin and C H Hong ldquoPhytochemical studyon Bidens pilosa L varminorrdquo Huaxue pp 38ndash42 1975

[32] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[33] M C Zulueta M Tada and C Y Ragasa ldquoA diterpene fromBidens pilosardquo Phytochemistry vol 38 no 6 pp 1449ndash14501995

[34] J Wang H Yang Z W Lin and H D Sun ldquoFlavonoids fromBidens pilosa var radiatardquo Phytochemistry vol 46 no 7 pp1275ndash1278 1997

[35] A Zhao Q Zhao L Peng et al ldquoA new chalcone glycoside fromBidens pilosardquo Acta Botanica Yunnanica vol 26 no 1 pp 121ndash126

[36] P Gao Y Wu S Cui and Y Zou ldquoSynthesis and absoluteconfiguration of the 7-phenylhepta-4 6-diyne-1 2-diol isolatedfrom Bidens pilosardquo Synthesis no 13 pp 2131ndash2135 2011

[37] S Wang B Yang D Zhu D He and L Wang ldquoActivecomponents of Bidens pilosa Lrdquo Zhongcaoyao vol 36 pp 20ndash21

[38] F Bohlmann T Burkhardt and C Zdero Naturally OccuringAcetylenes Academic Press New York NY USA 1973

[39] T M Sarg A M Ateya N M Farrag and F A AbbasldquoConstituents and biological activity of Bidens pilosa L grownin EgyptrdquoActa Pharmaceutica Hungarica vol 61 no 6 pp 317ndash323 1991

[40] H A L Valdes and H P Rego ldquoBidens pilosa Linnerdquo RevistaCubana de Plantas Medicinales vol 6 pp 28ndash33

[41] F Bohlmann H Bornowski and K M Kleine ldquoNew polyynesfrom the tribe Heliantheaerdquo Chemische Berichte vol 97 pp2135ndash2138

[42] RWang Q XWu and Y P Shi ldquoPolyacetylenes and flavonoidsfrom the aerial parts ofBidens pilosardquo PlantaMedica vol 76 no9 pp 893ndash896 2010

[43] L W Wu Y M Chiang H C Chuang et al ldquoA novelpolyacetylene significantly inhibits angiogenesis and promotesapoptosis in human endothelial cells through activation of theCDK inhibitors and caspase-7rdquo Planta Medica vol 73 no 7 pp655ndash661 2007

[44] L W Wu Y M Chiang H C Chuang et al ldquoPolyacetylenesfunction as anti-angiogenic agentsrdquo Pharmaceutical Researchvol 21 no 11 pp 2112ndash2119 2004

[45] H L Yang S C Chen N W Chang et al ldquoProtection fromoxidative damage using Bidens pilosa extracts in normal humanerythrocytesrdquo Food and Chemical Toxicology vol 44 no 9 pp1513ndash1521 2006

[46] H Q Wang S J Lu H Li and Z H Yao ldquoEDTA-enhancedphytoremediation of lead contaminated soil by Bidens maxi-mowiczianardquo Journal of Environmental Sciences vol 19 no 12pp 1496ndash1499 2007

[47] S TobinagaMK SharmaWG L Aalbersberg et al ldquoIsolationand identification of a potent antimalarial and antibacterialpolyacetylene from Bidens pilosardquo Planta Medica vol 75 no 6pp 624ndash628 2009

[48] C L T Chang H K Kuo S L Chang et al ldquoThe distincteffects of a butanol fraction of Bidens pilosa plant extract onthe development of Th1-mediated diabetes and Th2-mediatedairway inflammation in micerdquo Journal of Biomedical Sciencevol 12 no 1 pp 79ndash89 2005

[49] S L Chang C L T Chang Y M Chiang et al ldquoPolyacetyleniccompounds and butanol fraction from Bidens pilosa can mod-ulate the differentiation of helper T cells and prevent autoim-mune diabetes in non-obese diabetic micerdquo Planta Medica vol70 no 11 pp 1045ndash1051 2004

Evidence-Based Complementary and Alternative Medicine 49

[50] Y M Chiang C L T Chang S L Chang W C Yang and LF Shyur ldquoCytopiloyne a novel polyacetylenic glucoside fromBidens pilosa functions as a T helper cell modulatorrdquo Journal ofEthnopharmacology vol 110 no 3 pp 532ndash538 2007

[51] R PUbillas CDMendez S D Jolad et al ldquoAntihyperglycemicacetylenic glucosides from Bidens pilosardquo Planta Medica vol66 no 1 pp 82ndash83 2000

[52] L Alvarez S Marquina M L Villarreal D Alonso E Arandaand G Delgado ldquoBioactive polyacetylenes from Bidens pilosardquoPlanta Medica vol 62 no 4 pp 355ndash357 1996

[53] C L T Chang S L Chang Y M Lee et al ldquoCytopiloyne apolyacetylenic glucoside prevents type 1 diabetes in nonobesediabetic micerdquo Journal of Immunology vol 178 no 11 pp 6984ndash6993 2007

[54] G Kusano A Kusano and Y Seyama Novel Hypoglycemicand Anti-Inflammatory Polyacetylenic Compounds Their Com-positions Bidens Plant Extract Fractions and CompositionsContaining the Plant or Fraction JPO Tokyo Japan 2004

[55] M G L Brandao A U Krettli L S R Soares C G CNery and H C Marinuzzi ldquoAntimalarial activity of extractsand fractions from Bidens pilosa and other Bidens species(Asteraceae) correlated with the presence of acetylene andflavonoid compoundsrdquo Journal of Ethnopharmacology vol 57no 2 pp 131ndash138 1997

[56] A U Krettli V F Andrade-Neto M D G L Brandao andW M S Ferrari ldquoThe search for new anti-malarial drugs fromplants used to treat fever and malaria or plants ramdomlyselected a reviewrdquoMemorias do Instituto Oswaldo Cruz vol 96no 8 pp 1033ndash1042 2001

[57] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[58] C K Wat R K Biswas E A Graham L Bohm G H NTowers and E R Waygood ldquoUltraviolet-mediated cytotoxicactivity of phenylheptatriyne from Bidens pilosa Lrdquo Journal ofNatural Products vol 42 no 1 pp 103ndash111 1979

[59] Y Sashida K Ogawa M Kitada H Karikome Y Mimakiand H Shimomura ldquoNew aurone glucosides and new phenyl-propanoid glucosides from Bidens pilosardquo Chemical and Phar-maceutical Bulletin vol 39 no 3 pp 709ndash711 1991

[60] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens pilosa L (TFB) on animal liver injury andliver fibrosisrdquo Journal of Ethnopharmacology vol 116 no 3 pp539ndash546 2008

[61] B Hoffmann and J Holzl ldquoAcylated compounds from Bidenspilosardquo Planta Medica vol 55 pp 108ndash109

[62] B Hoffmann and J Holzl ldquoA methylated chalcone glucosidefrom Bidens pilosardquo Phytochemistry vol 27 no 11 pp 3700ndash3701 1988

[63] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[64] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[65] B Hoffmann and J Holzl ldquoWeitere acylierte chalkone ausBidens pilosardquo Planta Medica vol 54 pp 450ndash451 1988

[66] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant in11traditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[67] V V Pham V V T K P V L Hoang and V K Phan ldquoFlavonoidcompounds from the plant Bidens pilosa L (Asteraceae)rdquo TapChi Duoc Hoc vol 50 pp 48ndash53

[68] S D Sarker B Bartholomew R J Nash and N Robinson ldquo5-O-methylhoslundin an unusual flavonoid from Bidens pilosa(Asteraceae)rdquo Biochemical Systematics and Ecology vol 28 no6 pp 591ndash593 2000

[69] S L Chang Y M Chiang C L T Chang et al ldquoFlavonoidscentaurein and centaureidin from Bidens pilosa stimulate IFN-120574 expressionrdquo Journal of Ethnopharmacology vol 112 no 2 pp232ndash236 2007

[70] Y M Chiang D Y Chuang S Y Wang Y H Kuo P WTsai and L F Shyur ldquoMetabolite profiling and chemopreventivebioactivity of plant extracts from Bidens pilosardquo Journal ofEthnopharmacology vol 95 no 2-3 pp 409ndash419 2004

[71] A Kusano Y Seyama E Usami et al ldquoStudies on the antioxi-dant active constituents of the dried powder from Bidens pilosaL var radiata SchrdquoNaturalMedicines vol 57 no 3 pp 100ndash1042003

[72] I Doleckova L Rarova J Gruz et al ldquoAnti-proliferative andanti-angiogenic effects of flavone eupatorin an active con-stituent of chloroform extract ofOrthosiphon stamineus leavesrdquoFitoterapia vol 83 no 6 pp 1000ndash1007

[73] K Bairwa R Kumar R J Sharma and R K Roy ldquoAn updatedreview on Bidens pilosa Lrdquo Der Pharma Chemica vol 2 no 3pp 325ndash337

[74] Q Xia Y Liu and Y Li ldquoDetermination of hyperoside indifferent parts and different species of Herba Bidens by RP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol 50pp 48ndash53

[75] M Horiuchi and Y Seyama ldquoImprovement of the antiinflam-matory and antiallergic activity of Bidens pilosa L var radiataSCHERFF treated with enzyme (Cellulosine)rdquo Journal of HealthScience vol 54 no 3 pp 294ndash301 2008

[76] M G L Brandao C G C Nery M A S Mamao and AU Krettli ldquoTwo methoxylated flavone glycosides from Bidenspilosardquo Phytochemistry vol 48 no 2 pp 397ndash400 1998

[77] P Kumari K Misra B S Sisodia et al ldquoA promising anticancerand antimalarial component from the leaves of Bidens pilosardquoPlanta Medica vol 75 no 1 pp 59ndash61 2009

[78] G Seelinger I Merfort U Wolfle and C M Schempp ldquoAnti-carcinogenic effects of the flavonoid luteolinrdquoMolecules vol 13no 10 pp 2628ndash2651 2008

[79] G Seelinger I Merfort and C M Schempp ldquoAnti-oxidantanti-inflammatory and anti-allergic activities of luteolinrdquoPlanta Medica vol 74 no 14 pp 1667ndash1677 2008

[80] C C Yit and N P Das ldquoCytotoxic effect of butein on humancolon adenocarcinoma cell proliferationrdquo Cancer Letters vol82 no 1 pp 65ndash72 1994

[81] J A Beutler E Hamel A J Vlietinck et al ldquoStructure-activityrequirements for flavone cytotoxicity and binding to tubulinrdquoJournal of Medicinal Chemistry vol 41 no 13 pp 2333ndash23381998

[82] M R Kviecinski K B Felipe T Schoenfelder et al ldquoStudy ofthe antitumor potential of Bidens pilosa (Asteraceae) used inBrazilian folkmedicinerdquo Journal of Ethnopharmacology vol 117no 1 pp 69ndash75 2008

[83] P Sundararajan A Dey A Smith A G Doss M Rajappanand S Natarajan ldquoStudies of anticancer and antipyretic activityof Bidens pilosawhole plantrdquo African Health Sciences vol 6 no1 pp 27ndash30 2006

[84] YMChiang C P Lo Y P Chen et al ldquoEthyl caffeate suppressesNF-120581B activation and its downstream inflammatory mediatorsiNOS COX-2 and PGE2 in vitro or in mouse skinrdquo BritishJournal of Pharmacology vol 146 no 3 pp 352ndash363 2005

50 Evidence-Based Complementary and Alternative Medicine

[85] J S Kim H J Lee M H Lee J Kim C Jin and J HRyu ldquoLuteolin inhibits LPS-stimulated inducible nitric oxidesynthase expression in BV-2 microglial cellsrdquo Planta Medicavol 72 no 1 pp 65ndash68 2006

[86] P A Ruiz and D Haller ldquoFunctional diversity of flavonoidsin the inhibition of the proinflammatory NF-120581B IRF and Aktsignaling pathways in murine intestinal epithelial cellsrdquo Journalof Nutrition vol 136 no 3 pp 664ndash671 2006

[87] A Xagorari A Papapetropoulos A Mauromatis MEconomou T Fotsis and C Roussos ldquoLuteolin inhibitsan endotoxin-stimulated phosphorylation cascade andproinflammatory cytokine production in macrophagesrdquoJournal of Pharmacology and Experimental Therapeutics vol296 no 1 pp 181ndash187 2001

[88] N Yoshida T Kanekura Y Higashi and T Kanzaki ldquoBidenspilosa suppresses interleukin-1120573-induced cyclooxygenase-2expression through the inhibition of mitogen activated proteinkinases phosphorylation in normal human dermal fibroblastsrdquoJournal of Dermatology vol 33 no 10 pp 676ndash683 2006

[89] M Muchuweti C Mupure A Ndhlala T Murenje and M AN Benhura ldquoScreening of antioxidant and radical scavengingactivity of Vigna ungiculata Bidens pilosa and Cleome gynan-drardquo American Journal of Food Technology vol 2 no 3 pp 161ndash168 2007

[90] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens bipinnata L against carbon tetrachloride-induced liver fibrosis in ratsrdquo Journal of Pharmacy and Pharma-cology vol 60 no 10 pp 1393ndash1402 2008

[91] S L Chang H H Yeh Y S Lin Y M Chiang T K Wuand W C Yang ldquoThe effect of centaurein on interferon-120574 expression and Listeria infection in micerdquo Toxicology andApplied Pharmacology vol 219 no 1 pp 54ndash61 2007

[92] F Deba T D Xuan M Yasuda and S Tawata ldquoChemical com-position and antioxidant antibacterial and antifungal activitiesof the essential oils from Bidens pilosa Linn var Radiatardquo FoodControl vol 19 no 4 pp 346ndash352 2008

[93] A O T Ashafa and A J Afolayan ldquoScreening the root extractsfrom Biden pilosa L var radiata (Asteraceae) for antimicrobialpotentialsrdquo Journal of Medicinal Plant Research vol 3 no 8 pp568ndash572 2009

[94] T B Nguelefack T Dimo E P Nguelefack Mbuyo P V TanS V Rakotonirina and A Kamanyi ldquoRelaxant effects of theneutral extract of the leaves of Bidens pilosa linn on isolated ratvascular smooth musclerdquo Phytotherapy Research vol 19 no 3pp 207ndash210 2005

[95] M T Grombone-Guaratini K L Silva-Brandao V N SolferiniJ Semir and J R Trigo ldquoSesquiterpene and polyacetyleneprofile of the Bidens pilosa complex (Asteraceae Heliantheae)from Southeast of Brazilrdquo Biochemical Systematics and Ecologyvol 33 no 5 pp 479ndash486 2005

[96] L L Lin C Y Wu H C Hsiu M T Wang and H ChuangldquoStudies on diabetes mellitus I The hypoglycemic activity ofphytosterin on alloxan diabetic ratsrdquo Taiwan Yi Xue Hui Za Zhivol 66 no 2 pp 58ndash65 1967

[97] M A N Benhura and I C Chitsiku ldquoThe extractable 120573-carotene content of Guku (Bidens pilosa) leaves after cookingdrying and storagerdquo International Journal of Food Science andTechnology vol 32 no 6 pp 495ndash500 1997

[98] J S Chang L C Chiang C C Chen L T Liu K C Wangand C C Lin ldquoAtileukemic activity of Bidens pilosa l varminor (blume) sherff andHouttuynia cordata thunbrdquo AmericanJournal of Chinese Medicine vol 29 no 2 pp 303ndash312 2001

[99] F Deba T D Xuan M Yasuda and S Tawata ldquoHerbicidal andfungicidal activities and identification of potential phytotoxinsfrom Bidens pilosa L var radiata Scherff research paperrdquoWeedBiology and Management vol 7 no 2 pp 77ndash83 2007

[100] J K Kumar and A K Sinha ldquoA new disubstituted acetylacetonefrom the leaves ofBidens pilosa LinnrdquoNatural Product Researchvol 17 no 1 pp 71ndash74 2003

[101] K Ogawa and Y Sashida ldquoCaffeoyl derivatives of a sugarlactone and its hydroxy acid from the leaves of Bidens pilosardquoPhytochemistry vol 31 no 10 pp 3657ndash3658 1992

[102] Q Xia Y Liu and Y Li ldquoDetermination of gallic acid fromdifferent species and different medical parts of Herba Bidens byRP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol24 no 03 pp 308ndash310

[103] W J Lee L F Wu W K Chen C J Wang and T HTseng ldquoInhibitory effect of luteolin on hepatocyte growth fac-torscatter factor-induced HepG2 cell invasion involving bothMAPKERKs and PI3K-Akt pathwaysrdquo Chemico-BiologicalInteractions vol 160 no 2 pp 123ndash133 2006

[104] J A Beutler J H Cardellina C M Lin E Hamel G MCragg andM R Boyd ldquoCentaureidin a cytotoxic flavone fromPolymnia fruticosa inhibits tubulin polymerizationrdquo BioorganicandMedicinal Chemistry Letters vol 3 no 4 pp 581ndash584 1993

[105] A Verma A Su A M Golin B OrsquoMarrah and J K AmorosaldquoThe lateral view a screening method for knee traumardquo Aca-demic Radiology vol 8 no 5 pp 392ndash397 2001

[106] J Corren M Lemay Y Lin L Rozga and R K RandolphldquoClinical and biochemical effects of a combination botanicalproduct (ClearGuard) for allergy a pilot randomized double-blind placebo-controlled trialrdquo Nutrition Journal vol 7 no 1article 20 2008

[107] M S Gachet J S Lecaro M Kaiser et al ldquoAssessment of anti-protozoal activity of plants traditionally used in Ecuador in thetreatment of leishmaniasisrdquo Journal of Ethnopharmacology vol128 no 1 pp 184ndash197 2010

[108] W C Yang M Ghiotto B Barbarat and D Olive ldquoThe role ofTec protein-tyrosine kinase in T cell signalingrdquo The Journal ofBiological Chemistry vol 274 no 2 pp 607ndash617 1999

[109] S Tewtrakul H Miyashiro N Nakamura et al ldquoHIV-1 inte-grase inhibitory substances from Coleus parvifoliusrdquo Phytother-apy Research vol 17 no 3 pp 232ndash239 2003

[110] T Dimo T B Nguelefack P V Tan et al ldquoPossible mechanismsof action of the neutral extract from Bidens pilosa L leaves onthe cardiovascular system of anaesthetized ratsrdquo PhytotherapyResearch vol 17 no 10 pp 1135ndash1139 2003

[111] T S C Li Chinese and Related North American Herbs CRCPress New York NY USA 2002

[112] H Wu H Chen X Hua Z Shi L Zhang and J ChenldquoClinical therapeutic effect of drug-separated moxibustion onchronic diarrhea and its immunologic mechanismsrdquo Journal ofTraditional Chinese Medicine vol 17 no 4 pp 253ndash258 1997

[113] S W Wright R R Harris J S Kerr et al ldquoSynthesis chemicaland biological properties of vinylogous hydroxamic acids dualinhibitors of 5-lipoxygenase and IL-1 biosynthesisrdquo Journal ofMedicinal Chemistry vol 35 no 22 pp 4061ndash4068 1992

[114] W R Almiron and M E Brewer ldquoClassification of immaturestage habitats of Culicidae (Diptera) collected in CordobaArgentinardquo Memorias do Instituto Oswaldo Cruz vol 91 no 1pp 1ndash9 1996

[115] N L Wang J Wang X S Yao and S Kitanaka ldquoTwo newmonoterpene glycosides and a new (+)-jasmololone glucoside

Evidence-Based Complementary and Alternative Medicine 51

from Bidens parvifloraWilldrdquo Journal of Asian Natural ProductsResearch vol 9 no 5 pp 473ndash479 2007

[116] P Champagnat ldquoRole of the terminal bud in the actionexercised by the cotyledon of Bidens pilosus L var radiatus onits axillary budrdquo Comptes Rendus des Seances et Memoires de laSociete de Biologie vol 145 no 17-18 pp 1374ndash1376 1951

[117] S F Nielsen S B Christensen G Cruciani A Kharazmiand T Liljefors ldquoAntileishmaniai chalcones statistical designsynthesis and three-dimensional quantitative structure-activityrelationship analysisrdquo Journal of Medicinal Chemistry vol 41no 24 pp 4819ndash4832 1998

[118] Y C Hwang J J H Chu P L Yang W Chen and M V YatesldquoRapid identification of inhibitors that interfere with poliovirusreplication using a cell-based assayrdquo Antiviral Research vol 77no 3 pp 232ndash236 2008

[119] V F Andrade-Neto M G L Brandao F Q Oliveira et alldquoAntimalarial activity of Bidens pilosa L (Asteraceae) ethanolextracts fromwild plants collected in various localities or plantscultivated in humus soilrdquo Phytotherapy Research vol 18 no 8pp 634ndash639 2004

[120] R J Marles and N R Farnsworth ldquoAnti-diabetic plants andtheir active constituentsrdquo Phytomedicine vol 2 no 2 pp 137ndash189

[121] M Habeck ldquoDiabetes treatments get sweet help from naturerdquoNature Medicine vol 9 no 10 p 1228 2003

[122] H W Lin G Y Han and S X Liao ldquoStudies on the activeconstituents of the Chinese traditional medicine Polygonatumodoratum (Mill) DrucerdquoActa Pharmaceutica Sinica vol 29 no3 pp 215ndash222 1994

[123] C L T Chang Y C Chen H M Chen N S Yang andW C Yang ldquoNatural cures for type 1 diabetes a reviewof phytochemicals biological actions and clinical potentialrdquoCurrent Medicinal Chemistry vol 20 no 7 pp 899ndash907 2013

[124] L Dey A S Attele and C S Yuan ldquoAlternative therapies fortype 2 diabetesrdquo Alternative Medicine Review vol 7 no 1 pp45ndash58 2002

[125] C L T ChangH Y Liu T F Kuo et al ldquoAnti-diabetic effect andmode of action of cytopiloynerdquo Evidence-Based Complementaryand Alternative Medicine vol 2013 Article ID 685642 13 pages2013

[126] M R Khan M Kihara and A D Omoloso ldquoAnti-microbialactivity of Bidens pilosa Bischofia javanica Elmerillia papuanaand Sigesbekia orientalisrdquo Fitoterapia vol 72 no 6 pp 662ndash6652001

[127] T Dimo T B Nguelefack P Kamtchouing E Dongo ARakotonirina and S V Rakotonirina ldquoHypotensive effects ofmethanol extract from Bidens pilosa Linn on hypertensive ratsrdquoComptes Rendus de lrsquoAcademie des Sciences Serie III vol 322no 4 pp 323ndash329 1999

[128] A K Hassan O Deogratius J F Nyafuono O Francis and OP Engeu ldquoWound healing potential of the ethanolic extracts ofBidens pilosa and Ocimum suaverdquo African Journal of Pharmacyand Pharmacology vol 5 no 2 pp 132ndash136 2011

[129] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[130] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[131] C Kandaswami and EMiddleton ldquoFree radical scavenging andantioxidant activity of plant flavonoidsrdquo Advances in Experi-mental Medicine and Biology vol 366 pp 351ndash376 1994

[132] K Yamamoto H Kakegawa H Ueda et al ldquoGastric cytopro-tective anti-ulcerogenic actions of hydroxychalcones in ratsrdquoPlanta Medica vol 58 no 5 pp 389ndash393 1992

[133] J O C Ezeonwumelu A K Julius C N Muhoho et alldquoBiochemical and histological studies of aqueous extract ofBidens pilosa leaves from Ugandan Rift valley in ratsrdquo BritishJournal of Pharmacology and Toxicology vol 2 no 6 pp 302ndash309

[134] L Frida S Rakotonirina A Rakotonirina and J P SavineauldquoIn vivo and in vitro effects of Bidens pilosa L (Asteraceae)leaf aqueous and ethanol extracts on primed-oestrogenized ratuterine musclerdquo African Journal of Traditional Complementaryand Alternative Medicines vol 5 no 1 pp 79ndash91 2008

52 Evidence-Based Complementary and Alternative Medicine

Composition Comments

1 Please provide valid postal code to the address if

possible

2 We made the highlighted change in Table 4 for the sake

of correctness Please check

3 Please check the correctness of the highlighted part in

Table 5

4 We considered the highlighted part in Table 11 as

reference Please check similar cases throughout

5 Please check the format of Table 18

6 We reformatted Table 20 Please check

7 We made the highlighted change in Table 4 for the sake

of consistency Please check similar cases throughout the

paper

8 We made the highlighted change(s) for the sake

of consistency Please check simila highlighted cases

throughout

9 We made the highlighted change in Table 9 for the sake

of clarity Please check similar cases through out

10 We made the highlighted change(s) for the sake of

correctness Please check

11 Comment on ref [66] This reference is a repetition

of [18] Please check similar cases throughout

Author(s) Name(s)

Author 1

Author 2

Author 3

Evidence-Based Complementary and Alternative Medicine 7

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

26 (E)-9-Octadecenoicacid Elaidic acid

O

HOLeaves(Philippines) [33]

27 (ZZ)-912-Octadecadienoic acid

Linolic acid or Linoleicacid

O

OHAerial(Tanzania)whole (Taiwan)

[18 36]

28(ZZZ)-91215-Octadecatrienoicacid

120572-Linoleic acid

O

OH Whole (Taiwan) [32]

29(ZZ)-912-Octadecadienoic acidethyl ester

Ethyl linoleate O

O

Whole (Taiwan) [32]

30(ZZZ)-91215-Octadecatrienoicacid methyl ester

Methyl linolenate O

O

Whole (Taiwan) [32]

31(ZZZ)-91215-Octadecatrienoicacid ethyl ester

Ethyl linolenate O

O

Whole (Taiwan) [32]

32(Z)-9-Octadecenoicacid 2-butoxyethylester

2-Butoxyethyl oleate O

OO

Whole (Taiwan) [32]

33 2-Butoxyethyllinoleate O

OO Whole (Taiwan) [32]

34(ZZZ)-91215-Octadecatrienoicacid butoxyethyl ester

2-Butoxyethyl linoleate O

OO

Whole (Taiwan) [32]

3517E9E15E-Heptadecatetraene-1113-diyne

Heptadeca-2E8E10E16-tetraen-46-diyne

Not found(China) [37]

8 Evidence-Based Complementary and Alternative Medicine

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

36 111-Tridecadiene-3579-tetrayne

Roots (notstated) [38]

37 1-Tridecene-357911-pentayne Pentayneene

Leaves (notstated) andnot found(Egypt)

[38 39]

38 5-Tridecene-7911-triyne-3-ol

HO

not found(Egypt) [39]

39 21012-Tridecatriene-468-triyn-1-ol

OHPart not specified(Not stated) [40]

40 212-Tridecadiene-46810-tetrayn-1-ol

111-Tridecadiene-3579-

tetrayn-13-ol

OH Roots (Notstated) notfound(Egypt)

[38 39]

41 212-Tridecadiene-46810-tetraynal

111-Tridecadiene-3579-

tetrayn-13-al

O

Roots (Germany) [41]

42212-Tridecadiene-46810-tetrayn-1-ol1-acetate

111-Tridecadiene-3579-

tetrayn-13-acetateO

O

Roots (notstated) [38]

43(5E)-15-Tridecadiene-79-diyn-3413-triol OH

OH

HO

Aerial (China) [42]

44(6E12E)-3-oxo-tetradeca-612-dien-810-diyn-1-ol

OH

O

Aerial (China) [42]

45 (E)-5-Tridecene-7911-triyne-12-diol

12-Dihydroxy-5(E)-tridecene-7911-triyne

HO

OHWhole (Taiwan) [43]

46 (E)-6-Tetradecene-81012-triyne-13-diol

13-Dihydroxy-6(E)-tetradecene-81012-triyne

OH

OHWhole (Taiwan) [43ndash45]

47(2R3E11E)-311-Tridecadiene-579-triyne-12-diol

Safynol

OH

HO

Not found (Egyptand China) [37 39]

Evidence-Based Complementary and Alternative Medicine 9

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

4857911-Tridecatetrayne-12-diol

12-Dihydroxytrideca-57911-tetrayne

HOOH

Whole (Taiwan) [43 44 46]

49(R)-357911-Tridecapentayne-12-diol

(R)-12-Dihydroxytrideca-357911-pentayne

HO

OH

Aerial (Japan) [47]

50

(4E)-1-(Hydroxymethyl)-4-dodecene-6810-triyn-1-yl-120573-D-glucopyranoside

2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-

tridecene-7911-triyne

O

HO

H

OH

H

H

HOH

O

OH

HO

H Aerial (USA)whole (Taiwan)leaves (Taiwan)

[8 45 48ndash51]

51

(4E)-1-(2-Hydroxyethyl)-4-dodecene-6810-triyn-1-yl-120573-D-glucopyranoside

3-120573-D-Glucopyranosyloxy-1-hydroxy-6(E)-

tridecene-81012-triyne

OHO

H

OH

H

H

HOH

O

OH

OH

HAerial (USA andChina) whole(Taiwan) leaves(Taiwan)

[8 35 4245 48ndash51]

52

3-Hydroxy-6-tetradecene-81012-triynyl-120573-D-glucopyranoside

120573-D-Glucopyranosyloxy-

3-hydroxy-6E-tetradecence-81012-triyne

OH

O

OHO

H

OH

H

H

HOH

OH

HWhole (Mexico) [52]

53

1-(Hydroxymethyl)-46810-dodecatetrayne-1-yl-120573-D-glucopyranoside

Cytopiloyne2-120573-D-

Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne

O

HO

OHO

H

OH

H

H

HOH

OH

HWhole (Taiwan)leaves (Taiwan) [8 50 53]

54

2-O-D-Glucosyltrideca-11E-en-3579-tetrayn-12-diol

OH

O

O

OHH

HO

H

H

HHOH

OH

Leaves (Brazil) [27]

10 Evidence-Based Complementary and Alternative Medicine

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

55

(R)-1-(Hydroxymethyl)-246810-dodecapentayn-1-yl-a-D-glucopyranoside

2- 120573 -D-Glucopyranosyloxy-1-hydroxytrideca-357911-pentayne

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Chinaand Japan) [35 47]

56

1-[[(Carboxyacetyl)oxy]methyl]46810-dodecatetraynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Japan) [54]

57

(4E)-1-[[(Carboxyacetyl)oxy]-methyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHO

H

OH

Aerial (Japan) [54]

58

(4E)-1-[[(Carboxyacetyl)oxy]-ethyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

O

O

O

OHO

O

OHOH

H

HO

H

H

HHO

HOH

Aerial (Japan) [54]

59 (5E)-5-Heptene-13-diyn-1-yl-benzene

1-Phenylhepta-13-diyn-5en Whole (Taiwan) [32]

60 7-Phenyl-2(E)-heptene-46-diyn-1-ol

OHRoots (notstated) Aerial(China)

[38 42]

617-Phenyl-2(E)-heptene-46-diyn-1-ol-acetate

OO

Roots (notstated Brazil) [38 55 56]

62 7-Phenyl-46-heptadiyn-2-ol Pilosol A

OH

Whole (Taiwan)aerial (China) [42 57]

Evidence-Based Complementary and Alternative Medicine 11

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

63 7-Phenylhepta-46-diyn-12-diol HO

OH

Aerial (China) [42]

64 135-Heptatriyn-1-yl-benzene

1-Phenylhepta-135-triyne

Leaves (notstated) leaves oftissue culture(not stated)aerial (TanzaniaChina)Whole (Taiwan)roots (Brazil)

[18 34 3842 56ndash58]

65 7-Phenyl-246-heptatriyn-1-ol HO

Leaves (notstated) aerial(China)

[38 42]

667-Phenyl-246-heptatriyn-1-ol-acetate O

O Leaves (notstated) [38]

675-(2-Phenylethynyl)-2-thiophenemethanol

S OHAerial (China) [42]

685-(2-Phenylethynyl)-2120573-glucosylmethyl-thiophene

O

O

OH

H

OH

H

H

HOH H

O

HO

S

Aerial (China) [42]

69

3- 120573 -D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne

OH

O

OH

HOHO

H

H

HHOH

OH

Leaves(Cameroon) [28]

701-Phenyl-13-diyn-5-en-7-ol-acetate O

O

Leaves (Brazil) [27]

SN denotes serial number

the chloroform fraction was the most toxic with a halfmaximal inhibitory concentration (IC

50) of 97 plusmn 72 and 83 plusmn

52 120583gmL in NRU and MTT respectively [82] Kumari andcolleagues also reported the anti-cancer and anti-malarialactivities of B pilosa leaves [77] Based on a cytotoxicity-directed fractionation strategy they identified phenyl-135-heptatriene with IC

50values of 8plusmn001 049plusmn045 07plusmn001

and 10 plusmn 001 120583gmL against human oral liver colon andbreast cancer cell lines respectively However phenyl-135-heptatriyne showed lower activity against breast cancer celllines than the chloroform leaf extract which had an IC

50

value of 65 plusmn 001 120583gmL Moreover the positive controltaxol showed higher activity than phenyl-135-heptatriyne[77] Furthermore in vitro comet assays were performed toevaluate the toxicity of n-hexane chloroform and methanolextracts of B pilosa and its ethyl acetate acetone and waterfractions onHela andKB cellsThe ethyl acetate fraction fromthe methanol extract exhibited the highest activity with halfmaximal cytotoxic concentrations (CTC

50) of 9652120583gmL

and 5862120583gmL against Hela and KB cells respectivelyDespite the moderate toxicity these findings suggest thatthese B pilosa extractsfractions could be useful for future

12 Evidence-Based Complementary and Alternative Medicine

Table5Flavon

oids

isolated

from

Bpilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

71

2-[(34-Dihydroxyph

enyl)-

methylene]-

6-hydroxy-3(2H

)-benzofuranon

e

Sulfu

retin

O

OHO

OH

OH

Aeria

l(Ch

ina)

[35]

72

2-[(34-Dihydroxyph

enyl)-

methylene]-

67-dihydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-673101584041015840-

tetrahydroxy

maritimetin

O

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

73

2-[(34-Dihydroxyph

enyl)-

methylene]-6-

(120573-D

-glucopyrano

syloxy)-7-

hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-120573-D

-glucop

yranosyl-

673101584041015840-te

trahydroxy

maritimein

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japanand

China)aerial(Ch

ina)

[355960]

74

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

(120573-D

-glucopyrano

syloxy)-

6-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-7-O-120573-D

-glucop

yranosyl-

673101584041015840-

tetrahydroxy

O

OHO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japan)

[59]

75

6-[(6-O-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-acetyl-

120573-D

-glucopyrano

syl)

-673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OHO

H HO

H

H

HOH

HO

O

O

Leaves

(Not

stated)

aeria

l(Ch

ina)

[3559]

Evidence-Based Complementary and Alternative Medicine 13

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

76

6-[(36-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-

2-[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone(Z)-6-O

-(36-di-O

-acetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

bideno

sideA

O

O

HO

OH

OH

OHOH O

H

H

HOH

HO

O

O

O

Leaves

(China)

[60]

77

6-[(46-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(410158401015840610158401015840-

diacetyl-120573-D

-glucropyrano

syl)-

673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

HO

O

OO

Leaves

(not

stated)

aeria

l(Ch

ina)

[343561]

78

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

hydroxy-6-

[(246-tri-O

-acetyl-120573

-D-

glucop

yranosyl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(210158401015840410158401015840610158401015840-

triacetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

O

OOO

O

Leaves

(not

stated)

aeria

l(Ch

ina)

[3461]

79

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-6-

[(346-tri-O

-acetyl-120573

-D-glucopyrano

syl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(310158401015840410158401015840610158401015840-

triacetyl-120573

-D-glucrop

yranosyl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H OH

H

HOH

HO

OOO O

Aeria

l(Ch

ina)

[3435]

80

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-

6-[[6-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propenyl]-120573-

D-glucopyrano

syl]o

xy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-

p-coum

aroyl-

120573-D

-glucropyrano

syl)-

673101584041015840-te

trahydroxy

OH

O

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OLeaves

(Japan)

[59]

14 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

81

1-[2-(120573-D

-Glucopyrano

syloxy)-

4-hydroxyphenyl]-2-

hydroxy-3-(3-

hydroxyphenyl)-2-

prop

en-1-on

e

Chalcone120572

32101584041015840-

tetrahydroxy-

21015840-O

-120573-D

-glucop

yranosyl

HO

O

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[35]

821-(24-Dihydroxyph

enyl)-

3-(34-dihydroxyph

enyl)-

2-prop

en-1-on

eBu

tein

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

833-(34-D

ihydroxyph

enyl)-

1-(234-trihydroxyph

enyl)-

2-prop

en-1-on

eOkanin

O

HO

HO

OH

OH

OH

Leaves

(China)

[60]

84

3-(34-D

ihydroxyph

enyl)-

1-[3-(120573-D

-glucop

yranosyloxy)-

24-dihydroxyphenyl]-2-

prop

en-1-on

e

Okanin

31015840-O

-120573-D

-glucosid

e

O

HO

OH

OH

OH

OO

HO

H

HO

H

H

HOH

H

OH

Leaves

(Germany)

flowers(Germany)

[61ndash63]

85

3-(34-D

ihydroxyph

enyl)-

1-[4-(120573-D

-glucop

yranosyloxy)-

23-dihydroxyphenyl]-

2-prop

en-1-on

e

Okanin

41015840-O

-120573-D

-glucosid

e

OOH

OH

OH

HOO

OHO

H

HO

H

H

HOH

H

OH

Flow

ers(Germany)

leaves

(Japan)

[5963]

Evidence-Based Complementary and Alternative Medicine 15

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

86Okanin41015840-O

-120573-D

-(610158401015840-

O-acetylglucosid

e)

O

O

OH

OH

OH

OHO

H

HO

H

H

HOH

H

HO

O

O

Flow

ers(Germany)

[63]

87

1-[4-[(46-di-O

-Acetyl-

120573-D

-glucopyrano

syl)-oxy]-

23-dihydroxyphenyl]-3-

(34-dihydroxyph

enyl)-2-

prop

en-1-on

e

Okanin

41015840-O

-120573-D

-(410158401015840610158401015840-diacetyl)-

glucop

yranoside

O

O

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Aeria

l(Ch

ina)

[34]

88Okanin41015840-O

-120573-D

-(210158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[64]

89Okanin41015840-O

-120573-D

-(310158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

O

OAe

rial(Ch

ina)

[34]

16 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

90

1-[23-Dihydroxy-4-

[[6-O-[3-(4-hydroxyph

enyl)-

1-oxo-2-prop

enyl]-120573-D

-glucop

yranosyl]oxy]-ph

enyl]-

3-(34-dihydroxyph

enyl)-

2-prop

en-1on

e

Okanin

41015840-O

-120573-D

-(610158401015840-tr

ans-

p-coum

aroyl)-

glucoside

O

O

OH

OH

OH

OHOH

HO

H

H

HOH

H

HO

OO

OH

Leaves

(Germany)

[64]

91Okanin41015840-O

-120573-D

-(410158401015840-acetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Leaves

(Germany)

[65]

92

Okanin

41015840-O

-120573-D

-(210158401015840410158401015840-diacetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[65]

Evidence-Based Complementary and Alternative Medicine 17

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

93

Okanin

41015840-O

-120573-D

-(310158401015840410158401015840-diacetyl-610158401015840-

trans-p-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

OH

O O

Leaves

(Germany)

[65]

94

Okanin

41015840-O

-[120573-D

-glucopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

OOH

OH

OH

HO

OH

HO

HHO

H

HOH

HO

OH

OH

HO

HHO

H

HOH

HO

Flow

er(G

ermany)

[63]

95Okanin

3101584041015840-di-O

-120573-D

-glucosid

e

OOH

OH

OH

O OO

H HO

HHOH

HOH

HHO

OHO

H

HO

H

H

HOH

H

OH

Flow

er(G

ermany)

[63]

18 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

96

1-[3-(120573-D

-Glucopyrano

syloxy)-

24-dihydroxyphenyl]-

3-(3-hydroxy)-

4-metho

xyph

enyl)-

2-prop

en-1-on

e

Okanin

4-methylether-

31015840-O

-120573-D

-glucop

yranoside

OOH

O

OH

HO

O

H HO

HHOH

HOH

H

OHO

Leaves

(Germany)aerial

(China)

[3562]

97

Okanin4-methylether-

3101584041015840-di-O

-120573-(41015840101584061015840101584041015840101584010158406101584010158401015840-

tetraacetyl)-

glucop

yranoside

OOH

OOH

O

HH

OH

HO

HOH

H

OO

O

Ac

Ac

AcAc

OO

H

HO

H

H

HOH

H

O

Aeria

l(Ch

ina)

[42]

98Ch

alcone210158404101584061015840-tr

imetho

xy-

4-O-D

-glucopyrano

syl-

dihydro

Not

foun

dLeaves

(China)

[60]

992-(34-D

ihydroxyph

enyl)-23-

dihydro-78

-dihydroxy-

4H-1-

benzop

yran-4-one

Okanin

isoOO

OH

HO

OH

OH

Leaves

(China)

[60]

100

2-(34-D

ihydroxyph

enyl)-

23-dihydro-8-hydroxy-

7[(246-tr

i-O-acetyl-120573

-D-

glucop

yranosyl)oxy]-4H

-1-b

enzopyran-4-on

eOO

OH

OH

OH

OH

O

HHO

H

HO

HO

OAc

Ac

Ac

Aeria

l(Ch

ina)

[34]

Evidence-Based Complementary and Alternative Medicine 19

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

101

57-Dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

OO

OH

HO

OH

Aeria

l(TanzaniaC

hina)

[4266]

102

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

7-O-glucopyrano

side

OO

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Tanzania)

[66]

103

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-4H

-1-benzop

yran-4-one

Luteolin

OO

HO

OH

OH

OH

Aeria

l(TanzaniaC

hina

Vietnam)

[343542

6667]

104

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5hydroxy-4H-1-

benzop

yran-4-one

Luteolin

7-O-120573-D

-glucop

yranoside

OOOH

OH

OH

OO

HO

H

HO

H

H

HOH

OH H

Aeria

l(Tanzania)

[66]

105

57-Dim

etho

xy-6-

(5-m

etho

xy-6-m

ethyl-

4-oxo-4H

-pyran-3-yl)-

2-ph

enyl-4H-1-

benzop

yran-4-one

5-O-M

ethylhosludin

O OOO

O

O

O

Aeria

l(Ugand

a)[68]

20 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

106

3-(120573-D

-Glucopyrano

syloxy)-

57-dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Astragalin

Kaem

pferol-3-O

-120573-D

-glucopyrano

side

HO

OH

O O

OH

O

O

HO

HHO

H

H

HOH

OH

H

Aeria

l(Ch

ina)

[35]

107

Kaem

pferol3-

(23-di-E

-p-cou

maroyl-

120572-L-rhamno

pyrano

side)

Not

foun

dAe

rial(Vietnam)

[67]

108

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5-hydroxy-36-dimetho

xy-

4H-1-

benzop

yran-4-one

Axillarosid

eO O

OH

OH

OO

OH

OO

HO

H

HO

H

H

HOH

H

OH

Aeria

l(Ch

ina)

[42]

109

57-Dihydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaureidin

O

O

O

O

O

HO

OH

OH

Who

le(Taiwan)

[69]

110

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaurein

O O

OO

O OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan)w

hole

(Taiwan)

[69ndash

71]

Evidence-Based Complementary and Alternative Medicine 21

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

111

5-Hydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-

67-dimetho

xy-4H-

chromen-4-one

Eupatorin

iso

OO O

OH

O

O OH

Not

foun

d(C

hina)

[3772]

112

2-(34-D

imetho

xyph

enyl)-7-

(120573-D

-glucopyrano

syloxy)-35-

dihydroxy-

8-metho

xy-4H-1-

benzop

yran-4-one

O O

OH

O O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

113

7-(120573-D

-Glucopyrano

syloxy)-

5hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-38-

dimehtoxy-4H-1-

benzop

yran-4-one

O O

O

OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

114

Isorhamnetin

3-[O

-120572-

L-rham

nopyrano

syl-

(1-2)-120573-D

-glucop

yranoside]

Not

foun

dAe

rial(Vietnam)

[67]

115

7-[(6-deoxy-120572-L-

Manno

pyrano

syl)o

xy]-

3-120573-D

-glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-4H

-1-benzop

yran-4-one

Luteoside

O O

OH

O

OH

OH

OH

HOH

H

H HO

HO

HO

OH

HO

H

HO

OH

OH

HH

Aeria

l(Ch

ina)

[42]

22 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

116

Luteolin

3-O-120573-D

-Glucopyrano

side

O O

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

HO

Aeria

l(Tanzania)

[66]

117

57-Dihydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Quercetagetin

3631015840-tr

imethylether

OOO

O

O OH

HO

OH

Aeria

l(Ch

ina)

[42]

118

6-((2S3S4S5S)-

Tetrahydro-345-tr

ihydroxy-

6-(hydroxymethyl)-2H

-pyran-2-yloxy)-5-hydroxy-

2-(4-hydroxy-3-

metho

xyph

enyl)-

37-dimetho

xy-4H-

chromen-4-one

Quercetagetin

3731015840-

trim

ethylether-6-O

-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[42]

119

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

JaceinQ

uercetin

3631015840-

trim

ethyl

ether-7-O-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan

China)

who

le(Taiwan)

[425470]

120

2-(34-D

ihydroxyph

enyl)-

357-trihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

OO

HO

OH

OH

OH

OH

Aeria

l(Ch

ina)leaves

(China)who

le(C

hina)

[356074]

Evidence-Based Complementary and Alternative Medicine 23

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

121

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-galactopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-4-one

Quercetin

3-O-120573-D

-galactosidehyperin

hyperosid

e

O O

HO

OH

OH

OH

OOHH

HOH

H

H HO

HO

OH

Aeria

l(TanzaniaJapan)

Leaves

(China)Who

le(C

hina)

[376066

717475]

122

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-glucopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

3-O-120573-

D-glucopyrano

side

O O

HO

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

Aeria

l(TanzaniaC

hina

Japan)L

eaves(Japan)

[355966

75]

123

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-

4-oxo-4H

-1-benzop

yran-3-yl-

120573-D

-glucopyrano

siduron

icacid

Quercetin

3-O-120573-D

-glucuron

opyranoside

O O

HO

OH

OH

OH

OHOH

HOH

H

H HO

HO

OHO

Aeria

l(TanzaniaJapan)

[6671]

124

3-[[6-O-(6-

Deoxy-120572-L-m

anno

pyrano

syl)-

120573-D

-galactopyrano

syl]o

xy]-

2-(34-dihydroxyph

enyl)-57-

dihydroxy-4H

-1-benzop

yran-4-one

Quercetin

3-O-rob

inob

iosid

e

O O

HO

OH

OH

OH

O

HOHH OH

HO

H HH

OO

OHH

HOH

H

H HO

HO

Aeria

l(Japan)W

hole

(Taiwan)

[7071]

24 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

125

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-

3-metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

OH

Roots(Brazil)

[55676]

126

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-

120573-D

-glucopyrano

syl]o

xy]-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120572-

L-rham

nopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

O

HHO HHO

HO H

HH

ORo

ots(Brazil)

[5676]

127

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-120573-D

-glucop

yranosyl]oxy]-5-

hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-3-metho

xy-

4H-1-

benzop

yran-4-one

Quercetin

341015840-

dimethyl

ether-7-O-rutinoside

O OOH

OH

O

O

OH O

HOH

H

HHO

H

O

HHO HHO

HO H

HH

O

HO

Aeria

l(Ch

ina)

[3435]

128

2-(34-D

ihydroxyph

enyl)-3-

(120573-D

-glucofurano

syloxy)-57-

dihydroxy-4H

-1-benzop

yran-

4-on

e

Isoq

uercitrin

O OOH

OH

OHO

H

OHH

OH

HO

H O HOH

OH

Aeria

l(Japan

China)

[3571]

Evidence-Based Complementary and Alternative Medicine 25

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

129

Quercetin

3-O-120573-D

-galactop

yranoside

O

OH

HO

OH OH

O

O

HO

OH

OHOH

OAe

rial(no

tstated)

[18]

130

Quercetin

3-O-rutinoside

O

O

R

OO

OH

OHO

H

O

OH

OH

H3C

H3C

H3C

OH

HO

Who

le(Taiwan)

[70]

26 Evidence-Based Complementary and Alternative Medicine

Table 6 Terpenoids isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(Country) References

131371111-Tetramethylbicyclo[810]undeca-

26-dieneBicyclogermacrene Leaves

(Brazil) [95]

13241111-Trimethyl-8-

methylenebicyclo[720]undec-4-ene

E-Caryophyllene Leaves(Brazil) [95]

1331-Methyl-5-methylene-8-

(1-methylethyl)-16-cyclodecadiene

Germacrene D Leaves(Brazil) [95]

1344-(15-Dimethyl-4-hexen-1-ylidene)-1-methyl-cyclohexene

Z-120574-Bisabolene Leaves(Brazil) [95]

135Decahydro-114-trimethyl-7-methylene-1H-cycloprop[e]-

azulene120573-Gurjunene Leaves

(Brazil) [95]

136 2669-Tetramethyl-148-cycloundecatriene

120572-Humulene120572-caryophyllene

Leaves(Brazil) [95]

13712344a568a-Octahydro-1-isopropyl-7-methyl-4-methylenenaphthalene

120575-Muurolene Leaves(Brazil) [95]

138

12344a568a-Octahydro-4a8-dimethyl-2-(1-methylethylidene)-

naphthalene

Selina-37(11)-diene

Leaves(Brazil) [95]

139(2E7R11R)-371115-

Tetramethyl-2-hexadecen-1-ol

Phytol

HO

Whole(Taiwan) [57]

Evidence-Based Complementary and Alternative Medicine 27

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

140 371115-Tetramethyl-2-hexadecenoic acid Phytanic acid

O

OH

Whole(Taiwan) [57]

141371115-Tetramethyl-2-

hexadecenyl ester-heptanoicacid

Phythylheptanoate O O

Leaves(Not stated) [33]

142

(3S10R13R)-17-((2R5R)-5-Ethyl-6-methylheptan-2-yl)-

2347891011121314151617-tetradecahydro-

1013-dimethyl-1H-cyclopenta[a]phenanthren-

3-ol

Campestrol

HO

Aerial(Tanzania) [18]

143 Not found Phytosterin-B Not foundNot found(TaiwanEgypt)

[39 96]

144 Stigmast-5-en-3-ol 120573-sitosterol

HOH

H

H

Aerial(Tanzania)

whole(Taiwan)

[18 31 57]

145

1314151617-Tetradecahydro-1013-

dimethyl-1H-cyclopenta[a]phenanthren-

3-yloxy)-tetrahydro-6-(hydroxymethyl)-2H-pyran-345-triol

120573-Sitosterolglucoside

H

H

HOH

HO

HHO

H

HOHH O

OH Not found(Egypt) [39]

146 5120572-Stigmasta-7-en-3120573-ol

HO

Whole(Taiwan) [57]

147 5120572-Stigmasta-722t-dien-3120573-ol

HO

Whole(Taiwan) [57]

148 Stigmasta-522-dien-3-ol Stigmasterol

HOH

H

H

Not found(Taiwan)aerial

(Tanzania)leaves (notstated)whole

(Taiwan)

[18 31 3357]

28 Evidence-Based Complementary and Alternative Medicine

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

149 Lup-20(29)-en-3-ol Lupeol

H

HH

HOH

Not found(Egypt) [39]

150 Lup-20(29)-en-3-ol acetate Lupeol acetate

H

HH

O

OH

Not found(Egypt) [39]

151 Olean-12-en-3-ol 120573-amyrin

H

HOH

Not found(Egypt) [39]

152 5913-Trimethyl-242526-trinoroleanan-3-ol Friedelan-3120573-ol

HO

Aerial(Tanzania) [18]

153 5913-Trimethyl-242526-trinoroleanan-3-one

Friedelinfriedelan-3-one

O

Aerial(Tanzania) [18]

1542610151923-

Hexamethyl-2610141822-tetracosahexaene

Squalene

Aerial(Tanzania)Leaves (Notstated)Whole(Taiwan)

[18 33 57]

155 120573120573-Carotene 120573-Carotene Leaves (Notstated) [97]

Evidence-Based Complementary and Alternative Medicine 29

studies [83] Hot water extracts of B pilosa varminor Sheriffwere also assessed for its antileukemic effects on leukemiccell lines L1210 U937 K562 Raji and P3HR1 using XTT-based colorimetric assays The extract inhibited the five celllines with IC

50values ranging from 145120583gmL to 586120583gmL

L1210 K562 Raji and P3HR1 were more sensitive to B pilosaextract with IC

50values below 200120583gmL [98]

Consistent with the antitumor activities of B pilosaextracts and fractions some of its phytochemicals alsoshowed anticancer activity as outlined in Table 11 Amongthem luteolin (103) a well-studied flavonoid with multiplebioactivities was more effective against tumor cell prolifera-tion than its derivatives with IC

50values ranging from 3 120583M

to 50120583M in cells and 5 to 10mgkg in animals Luteolin wasalso found to fight cancer as a food additive at concentrationsof 50 to 200 ppm [78] and prevent skin cancer [78] and cancer3invasion [103] Lee and colleagues reported that luteolinprevents cancer by inhibiting cell adhesion and invasion[103] Significant inhibition concentration was reported to be5 120583M and complete inhibition concentration was reported tobe 40120583M Moreover luteolin was reported to inhibit hepa-tocyte growth factor (HGF)-induced cell scattering as wellas cytoskeleton changes such as filopodia and lamellipodiawhichwas determined using phase-contrast and fluorescencemicroscopy Furthermore luteolin also inhibited the HGF-induced phosphorylation of c-Met ERK 12 and Akt aswell as the MAPKERK and P13K-Akt pathways [78 103]Other mechanisms underlying the anticancer activities ofluteolin are the inhibition of topoisomerase I and II whichinhibits cell replication and DNA repair thus promotingapoptosis regulation of PI-3-KinaseAktMAPKERKJNKactivation of apoptosis in the mitochondrial pathway byactivating caspase 9 and caspase 3 which were found inmalignant cells but not in normal human peripheral bloodmononuclear cells death receptor-induced apoptosis and acell-cycle arrest mechanism inhibition of fatty acid synthasewhich is upregulated in many cancer cells and sensitizationto chemotherapywhereby luteolin increases the susceptibilityof cancer cells to chemotherapy [78] Butein (82) is anotherflavonoid that showed a cytotoxic effect on human colonadenocarcinoma cell proliferation with a reported IC

50value

of 175 120583M Butein at 2 120583M affected the incorporation of[14C]-labeled leucine thymidine and uridine which cancause the inhibition of DNA RNA and protein synthesis ofhuman colon cancer cells Moreover butein also exhibitednoncompetitive inhibition of 1-chloro-24-dinitrobenzene(CDNB) in glutathione S-transferase (GST) activity Tumorresistance was correlated with high levels of GST thus buteininhibited proliferation of cancer cells [80] Another flavonoidpresent in B pilosa centaureidin (109) also showed anti-cancer activity in B lymphoma cells Centaureidin isolatedfrom Polymnia fruticosa inhibited tubulin polymerization invitro and induced mitotic figure formation in CA46 Burkittlymphoma cells Using turbidimetric assay the IC

50value of

centaureidin in inhibition of mitosis was 3120583M [104] Cyto-toxicity of centaureidin was further analyzed using AmericanNational Cancer Institute (NCI) 60 human tumor cell linesThe cytotoxicity potency of centaureidin expressed as GI

50

(50 growth inhibition in the NCI tumor line panel) was024120583M [81] These data mark centaureidin as a promisingantimitotic agent for tumor therapy

In addition to anti-tumor flavones polyynes found in B 4pilosa have also been shown to possess anti-tumor proper-ties Based on a bioactivity-directed isolation approach Wuand colleagues identified two polyyne aglycones from theethyl acetate fraction of B pilosa [44] 12-Dihydroxytrideca-57911-tetrayne (48) and 13-Dihydroxy-6(E)- tetradecene-81012-triyne (46) exhibited significant anticell prolifera-tion activity in primary human umbilical vein endotheliumcells (HUVEC) with IC

50values of 125 120583M and 173 120583M

respectively They also decreased angiogenesis and pro-moted apoptosis in human endothelial cells Their anti-angiogenic and cytotoxic effects correlated with activationof the CDK inhibitors and caspase-7 [44] In addition 12-Dihyroxy-5(E)-tridecene-7911-triyne (45) showed antian-giogenic effects in HUVECs with an IC

50value of 124 120583M as

evidenced by a decrease in the tube formation and migrationof HUVECs [43] The IC

50value of compound 45 in the

inhibition of basic fibroblast growth factor-induced HUVECgrowth was 282120583M However it had higher IC

50values

than those for lung carcinoma cells and keratinocytes Thiscompound could also inhibit cell proliferation of HUVECslung carcinoma A549 cells and HACAT keratinocytes Themechanism by which compound 45 inhibits HUVEC growthand angiogenesis is complicated and includes decreasing theexpression of cell cycle regulators (CDK4 cyclins D1 and Aretinoblastoma (Rb) and vascular endothelial growth factorreceptor 1) caspase-mediated activation of CDK inhibitorsp21 (Cip1) and p27 (Kip) upregulation of Fas ligand expres-sion downregulation of Bcl-2 expression and activation ofcaspase-7 and poly (ADP-ribose) polymerase [43]

32 Anti-Inflammatory Activity B pilosa is commonly usedto treat inflammatory disorders The anti-inflammatory phy-tochemicals present in B pilosa are listed in Table 11 5Cyclooxygenase-2 (COX-2) is a physiologically importantenzyme that converts arachidonic acid to prostaglandin(PGE2) Its expression is induced by a wide variety of external

stimuli indicating its involvement in inflammatory diseasesand it is used as an inflammatory marker [88] Yoshida andcolleagues studied the effects of the aqueous extracts of Bpilosa aerial parts in the production of COX-2 and PGE

2as

well as on the activation of mitogen activated protein kinases(MAPKs) in normal human dermal fibroblasts (HDFs) inresponse to inflammatory cytokine IL-1120573 This work showedthat IL-1120573 activated MAPKs such as ERK12 p38 and JNKto different extents and induced COX-2 expression TheCOX-2 expression in HDFs was regulated mainly by p38following IL-1120573 stimulation Consistently the p38 inhibitorSB203580 blocked this expression Using this cell platformB pilosa extracts were tested for inhibition of inflammationThe extract dose-dependently suppressed the activation ofp38 and JNK and moderately suppressed ERK12 as wellas suppressing COX-2 expression and PGE

2production

30 Evidence-Based Complementary and Alternative Medicine

Table7Ph

enylprop

anoids

isolatedfro

mB

pilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

156

3-(4-H

ydroxyph

enyl)-2-prop

enoica

cid

p-Cou

maricacid

OHO

OH

Who

le(Ja

pan)

[99]

157

2-Metho

xy-4-(2-prop

en-1-yl)-ph

enol

Eugeno

lOOH

Leaves

androots(Japan)

[99]

158

3-(4-H

ydroxy-3-m

etho

xyph

enyl)-

2-prop

enoica

cid

Ferulic

acid

O

OH

O

HO

Who

le(Ja

pan)

[99]

159

3-(34-D

ihydroxyph

enyl)-2-prop

enoic

acid

Caffeicacid

O

HO

OH

OH

Who

leandaeria

l(Japan)

[7599]

160

3-Prop

yl-3[(245-trim

etho

xyph

enyl)-

metho

xy]-24-pentanedione

3-Prop

yl-3-(245-

trim

etho

xy)benzyloxy-

pentan-24-dione

O

O

O

O

O

O

Leaves

(India)

[100]

161

3-(34-D

ihydroxyph

enyl)-2-

prop

enoica

cidethyleste

rCa

ffeateethyl

O

O

OH

OH

Not

foun

d(Taiwan)

who

le(Taiwan)

[434584]

Evidence-Based Complementary and Alternative Medicine 31Ta

ble7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

162

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

d-Erythron

icacid

2-O-caffeoyl-2-C

-methyl

O

HO

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

163

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

2-O-caffeoyl-2-C

-methyl

O

O

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

164

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-24-

dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

3-O-caffeoyl-2-C

-methyl

O

O

HO

HO

O

O

OH

OH

Leaves

(Japan)

[101]

165

4-(A

cetyloxy)-3-[[3-(34-

dihydroxyphenyl)-

1-oxo-2-propen-1-y

l-]oxy]-2-hydroxy-

2-methyl-b

utanoica

cid

O

HO

OH

O OHO

HO

OO

Not

foun

d(Ja

pan)

[73]

166

3-(34-D

ihydroxyph

enyl)-tetrahydro-4-

hydroxy-4-

methyl-5

-oxo-3-fu

ranyl

ester-2-prop

enoica

cid

3-O-C

affeoyl-2-C

-methyl-

D-erythrono

-14-la

cton

eHO

HO

O

OO

OOH

Leaves

(Japan)

[101]

167

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]-oxy]-145-

trihydroxy-cyclohexanecarboxylicacid

Chlorogenica

cid

O

HO

HO

OH

O

OH

O

OH

HO

Aeria

l(Japan)w

hole

(Taiwan)

[707175]

32 Evidence-Based Complementary and Alternative Medicine

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

168

4-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]oxy]-

135-tr

ihydroxy-cyclohexanecarboxylic

acid

4-O-C

affeoylqu

inicacid

OHO

HO

OH

OH

O

O

OH

OH

Aeria

l(Japan)

[71]

169

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-15-

dihydroxy-cyclo

hexanecarboxylicacid

34-Di-O

-caffeoylqu

inic

acid

HO

HO

O

O

O

O

OH

OH

HO

OH O

H

O

Aeria

l(Japan)W

hole

(Taiwan)

[45497071]

170

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-14-dihydroxy-

cyclo

hexanecarboxylicacid

35-Di-O

-caffeoylqu

inic

acid

HO

HO

O O

HO

O

HO

OH OO

OHOH

Aeria

l(Japan)w

hole

(Taiwan)

[45497071]

Evidence-Based Complementary and Alternative Medicine 33

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

171

34-bis[[(2E

)-3-(34-D

ihydroxyph

enyl)-1-

oxo-2-prop

en-1-

yl]oxo]-15

-dihydroxy-

cyclo

hexanecarboxylicacid

45-Di-O

-caffeoylqu

inic

acid

OHO

OH

HO

O

O

OH

OH

O

OH

HO

OWho

le(Taiwan)

[454970]

172

3-[4-[[-6-O-[3-(4-H

ydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]-phenyl]-2-prop

enoica

cid

120573-D

-p-C

oumaricacid4-O

-(6-O

-p-caoum

aroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HOH

HO

O

O

Leaves

(Japan)

[59]

173

3-[4-[[2-O

-Acetyl-6

-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]ph

enyl]2-propeno

icacid

120573-D

-p-C

oumaricacid4-O

-(2-O

-acetyl-6

-O-p-

caou

maroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HO

HO

O

O

Leaves

(Japan)aerial

(China)

[3459]

174

67-Dihydroxy-2-chrom

enon

eEsculetin

cicho

rigenin

OHO HO

O

Not

foun

d(Egypt)

[39]

34 Evidence-Based Complementary and Alternative Medicine

Table 8 Aromatic compounds isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

175 12-Benzenediol PyrocatechinHO

HOWhole(Japan) [99]

176 4-Ethyl-12-benzenediol Pyrocatechol

OH

OH

Whole(Japan) [99]

177 Dimethoxyphenol26-dimethoxyphenol

O

HO

O

Roots (Japan) [99]

178 4-Ethenyl-2-methoxy-phenol p-Vinylguaiacol

O

OHWhole(Japan) [99]

179 2-Hydroxy-6-methylbenzaldehyde

6-Methyl-salicylaldehyde O

OH

Whole(Japan) [99]

180 Benzene-ethanol 2-Phenyl-ethanolHO

Whole(Japan) [57]

181 4-Hydroxy-3-methoxy-benzaldehyde Vanillin O

O

HOAerial(Japan) [99]

182 3-Hydroxy-4-methoxy-benzaldehyde Vanillin iso

OO

OH

Leaves(Japan) [99]

183 4-Hydroxybenzoic acid p-Hydroxybenzoicacid

O

HOOH Whole

(Japan) [99]

184 2-Hydroxybenzoic acid Salicylic acidO

HO

HO

Stem androots (Japan) [99]

185 34-Dihydroxybenzoicacid Protocatechuic acid

O

HOOH

OH

Whole(Japan) [99]

1864-Hydroxy-

3methoxybenzoicacid

Vanillic acid

OOH

O

OH

Aerial(Uganda)

Roots (Japan)[68 99]

Evidence-Based Complementary and Alternative Medicine 35

Table 8 Continued

S N IUPAC names Common names Structure Plant part(country) References

187 345-Trihydroxybenzoicacid Gallic acid

O

OHHO

HO

HO

Whole(China) [102]

[88] This work supports the use of B pilosa as an anti-inflammatory agent however no compounds responsible forthe anti-inflammatory activity of B pilosa were identified

A further study also reported the anti-inflammatoryactivity as well as the antiallergic activity of B pilosa [75] Inthis study dried powder of the aerial part of B pilosa whichhad been pretreated with the enzyme cellulosine was usedfor further tests The results showed that oral administrationof the cellulosine-treated B pilosa lowered the level of serumIgE in mice 10 days after immunization with DNP (24-dintrophenyl)-Ascaris as an antigen This treatment alsoreduced dye exudation in skin induced by passive cutaneousanaphylaxis and production of inflammatory mediators his-tamine and substance P in rats [75] Phytochemical analysisshowed that cellulosine treatment increased the percentage ofcaffeic acid and flavonoids This study suggests that B pilosaand its phenolics have anti-inflammatory functions

Phenolics and polyynes are major anti-inflammatoryphytochemicals present in B pilosa (Table 11) Unsurpris-6ingly phenolics such as luteolin (103) and ethyl caffeate(161) that are major constituents of B pilosa have alsobeen reported to possess anti-inflammatory activity Lute-olin was reported to exhibit anti-inflammatory activity inmacrophages Xagorari and colleagues showed that luteolininhibited the release of inflammatory cytokines TNF-120572 andinterleukin-6 in RAW 2647 cells following LPS stimulation[87] It inhibited TNF-120572 production with an IC

50value

of 1120583M The underlying anti-inflammatory mechanism ofluteolin was reported to be the inactivation of Akt andNF-120581B activation [87] In addition luteolin was reportedto confer anti-inflammatory activity through inhibition ofLPS-stimulated iNOS expression in BV-2 microglial cellsIt inhibited LPS-activated microglia in a dose-dependentmanner with an IC

50value of 69 120583MMoreover immunoblot

and RT-PCR data proved that luteolin suppressed I120581B-120572degradation and iNOS expression in LPS-activated microglia[85] Kim and colleagues stated that luteolin may havebeneficial effects on inflammatory neural diseases throughinhibition of iNOS expression [85] A related study revealedthat luteolin decreased the transcriptional activity of NF-120581BRelA via partial inhibition of TNF-mediated NF-120581B DNAbinding activity Luteolin also inhibited Akt phosphorylationand induced degradation of a transcription factor interferonregulatory factor (IRF) [86]

Chiang and colleagues showed that ethyl caffeate(161) significantly inhibited NO production in mousemacrophages RAW 2647 cells [84] Based on MTT assays

they concluded that this inhibition was not due to thecytotoxicity of ethyl caffeate The IC

50value of ethyl caffeate

in the inhibition of NO production was 55 120583gmL slightlylower than curcumin (positive control) which has anIC50

value of 65120583gmL They demonstrated that ethylcaffeate exerted anti-inflammatory activity via the reducedtranscription and translation of iNOS (inducible nitric oxidesynthase) in RAW 2467 cells In addition this compoundalso suppressed COX-2 expression in RAW 2467 cellsand MCF-7 cells The in vivo anti-inflammatory effectof ethyl caffeate was verified by testing in TPA-treatedmouse skin Like celecoxib the positive control ethylcaffeate significantly abolished COX-2 expression in adose-dependent manner Ethyl caffeate at 1mg200120583Lsite(24mM) inhibited COX-2 expression at a level comparableto celecoxib at 1mg200120583Lsite (13mM) Remarkably ethylcaffeate at 48mM (2mg200120583Lsite) was more effectivethan celecoxib at 131mM (10mg200120583Lsite) Moreoverthis compound inhibited the activation of nuclear factor-120581B(NF-120581B) by LPS via the prevention of NF-120581B binding toDNA [84] In addition 3 ethyl caffeate analogs (ethyl 34-dihydroxyhydrocinnamate ethyl cinnamate and catechol)also showed different degrees of NF-120581B binding to DNA aslisted in Table 12 Ethyl cinnamate lacks a catechol moietywhich results in ineffective inhibition of NF-120581B binding toDNA [84]

Pereira and colleagues assessed the anti- 7inflammatory and immunomodulatory activities ofB pilosa methanol extract as well as one polyyne2-O-120573-glucosyltrideca-11(E)-en-3 579-tetrayn-12-diol(54) in T lymphocytes and a zymosan-induced arthritismouse model [27] They first examined the in vitro effect of 8the B pilosa extract and compound 54 on cell proliferation ofhuman T cells stimulated with 5120583gmL phytohemagglutinin(PHA) or 100 nM 12-O-tetradecanoyl phorbol-13-acetate(TPA) plus 15 120583M ionomycin and on cell proliferation ofmouse T cells stimulated with 5 120583gmL concanavalin A(Con A) The data demonstrated that both methanol extractand compound 54 suppressed T-cell proliferation in adose-dependent manner The estimated IC

50values of the B

pilosa extract against human T cells stimulated with 5120583gmLPHA and 100 nM TPA plus 15120583M ionomycin were 125 and25 120583gmL respectively In comparison with the methanolextract compound 54 showed 10-fold more inhibition ofhuman T-cell proliferation with an estimated IC

50value of

15 120583gmL Accordingly the B pilosa extract and compound54 dose-dependently suppressed mouse T-cell proliferation

36 Evidence-Based Complementary and Alternative Medicine

Table 9 Porphyrins isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

188

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(15S16S)-10-ethenyl-5-ethyl-1161820-tetrahydro-6111522-tetramethyl-1820-dioxo-15H-912-imino-212-

metheno-471714-dinitrilopyrano[43-

b]azacyclononadecine-16-propanoic

acid

AristophyllC

NH N

HNN

O

O OOOPhytyl

O

Leaves(Taiwan) [100]

189

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -20b-(methoxycarbonyl)-9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-dinitrilo-12-

dioxeto-[310158404101584034]-cyclo-pent[1 2]azacyclononadecine-

19-propanoic acid

Bidenphytin A

NH N

HNN

OPhytylO O O

O O

Leaves(Taiwan) [100]

190

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -2a-

hydroxy-20b-(methoxycarbonyl)-

9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-

dinitrilo-12-dioxeto[310158404101584034]-cyclo-pent[12-b]-

azacyclononadecine-19-propanoic acid

Bidenphytin B

NH N

HNN

OPhytylO O O

O O

OH

Leaves(Taiwan) [100]

191

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO O

HOOO

Leaves(Taiwan) [100]

192

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

Evidence-Based Complementary and Alternative Medicine 37

Table 9 Continued

S N IUPAC names Common names Structure Plant part(country) References

193

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

Leaves(Taiwan) [100]

194

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

195

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3S4S21R)-9-ethenyl-14-ethyl-21-(methoxycarbonyl)-

481318-tetramethyl-20-oxo-3-phorbinepropanoic acid

Pheophytin A

NH N

HNN

PhytylO

O

OO O

Leaves(Taiwan) [100]

with estimated IC50values of 30 and 25 120583gmL respectively

Taken together the data indicate that the B pilosa extractand compound 54 act on human and mouse T cells Totest the in vivo effect of the B pilosa extract and compound54 a zymosan-induced arthritis mouse model was usedThis model was established from B10ASgSnJ mice with aninjection of zymosan (015mg) The zymosan-injected mice9received an intraperitoneal injection of the B pilosa extract(1 5 or 10mg) at one dose a day for 5 days Popliteal lymphnode (PLN) weight was monitored to check the developmentof arthritis The results revealed that 10mg of the methanolextract of B pilosa extract could significantly diminishinflammation as evidenced by PLN weight [27] This worksuggests that B pilosa (and compound 54) can suppressimmune response and inflammation

33 Antidiabetic Activity Anti-diabetic agents are primarilydeveloped from plants and other natural resources [8 24 4951 53] B pilosa is one of 1200 plant species that have beeninvestigated for antidiabetic activity [120 121]B pilosa is usedas an anti-diabetic herb in America Africa and Asia [51 120

122] Many studies have indicated that B pilosa could treattype 1 diabetes (T1D) and type 2 diabetes (T2D) in animals

Etiologically speaking T1D is caused by autoimmune-mediated destruction of pancreatic 120573 cells leading to insulindeficiency hyperglycemia and complications Currentlythere is no cure for T1D Polarization ofTh cell differentiationcontrols the development of T1D Suppression of Th1 celldifferentiation and promotion of Th2 cell differentiationameliorate T1D [123] One study showed that the butanolfraction of B pilosa inhibited T-cell proliferation decreasedTh1 cells and cytokines and increased Th2 cells andcytokines leading to prevention of T1D in nonobesediabetic (NOD) mice [49] Based on a bioactivity-directedisolation strategy 3 polyynes 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne (53) also known ascytopiloyne 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D- Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne (50) were identifiedfrom B pilosa [49 53] The IC

50value of the butanol fraction

was 200120583gmL This inhibition was reported to be partiallyattributed to cytotoxicity because the butanol fraction at

38 Evidence-Based Complementary and Alternative Medicine

Table 10 Other compounds isolated from B pilosa [30]

SN IUPAC names Common names Structure Plant part(country) References

19637-Dihydro-137-

trimethyl-1H-purine-26-dione

CaffeineN

NN

NO

O

Aerial(Uganda) [68]

197

1-((2R4S5R)-Tetrahydro-4-hydroxy-5-(hydroxymethyl)furan-2-yl)-5-

methylpyrimidine-24(1H3H)-dione

ThymidineHO

HO

ON

O

NHO Not found

(China) [37]

198 1-(2-Thienyl)-ethanone 2-Acetyl-thiophene

S

O

Roots(Germany) [41]

199

(2R3S4S5S)-2-(Heptan-2-yloxy)-tetrahydro-6-

((tetrahydro-34-dihydroxy-5-

(hydroxymethyl)furan-2-yloxy)methyl)-

2H-pyran-345-triol

Heptanyl2-O-120573-xylofuranosyl-

(1rarr 6)-120573-glucopyranoside

O

OOH

OH

HO

OHO

H

HO

H

H

HOHH

O

Whole(Taiwan) [70]

200

2-[(3R7R11R)-3-Hydroxy-371115-

tetramethylhexadecyl]-356-trimethyl-

25-cyclohexadiene-14-dione

120572-Tocopherylquinone

OO

HO

Whole(Taiwan) [32]

2017-O-(410158401015840610158401015840-Diacetyl)-

120573-D-glucopyranoside

Not found Leaves(China) [60]

180 120583gmL could cause 50 death of Th1 cells Moreoverthis study suggested that the butanol fraction may preventdiabetes in NOD mice in vivo via downregulation of Th1cells or upregulation Th2 cells that have effects which areantagonistic of those of Th1 cells [49] This was proven byintraperitoneal injection of the butanol fraction at a doseof 3mgkg BW 3 times a week to NOD mice from 4 to 27weeks This dosage resulted in lower incidence of diabetes(33) At a dose of 10mgkg the butanol fraction of Bpilosa totally eliminated (0) the initiation of the diseaseTo further support this result assessment of IgG2a and IgEproduction was performed in the serum of NOD mice Asin vivo results obtained from intracellular cytokine stainingexperiments were not very conclusive levels of IgG2a and IgE

were measured since Th1 cytokine IFN120574 and Th2 cytokineIL-4 favor the production of IgG2a and IgE respectively Asexpected high levels of IgE and some decline in the levels ofIgG2a were observed in the serum Profiling of the butanolextract revealed five compounds 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne(50) 45-Di-O-caffeoyl- quinic acid 35-Di-O-caffeoylquinicacid and 34-Di-O-caffeoylquinic acid Only the first twocompounds showed similar effects on the prevention ofdiabetes in NOD mice as the B pilosa butanol fractionMoreover compound 50 showed greater activity thancompound 69 in terms of enhancement (by 34 comparedto 8) of differentiation of Th0 to Th2 at 15 120583gmL (both

Evidence-Based Complementary and Alternative Medicine 39

Table 11 Chemical constituents of B pilosa and their biological activities

SN Name Classification Molecularformula Biological activities

109 Centaureidin [91] Flavonoid C18H16O8Anti-listerial [69 91]

Cytotoxic [10]

110 Centaurein [91] Flavonoid C24H26O13

Anti-listerial [69 91]Cytotoxic [10]Anti-viral [105]

103 Luteolin [106] Flavonoid C15H10O6

Anti-viral [107 108]Cytotoxic [77]

Anti-inflammatory [109]Anti-allergic [109]

82 Butein [110] Flavonoid C15H12O5Anti-leishmanial [111]

Cytotoxic [78]48 12-Dihydroxytrideca-57911-tetrayne [112] Polyyne C13H12O2 Anti-angiogeneic [112]46 13-Dihyroxy-6(E)-tetradecene-81012-triyne [112] Polyyne C14H16O2 Anti-angiogeneic [112]

45 12-Dihyroxy-5(E)-tridecene-7911-triyne [113] Polyyne C13H14O2Anti-angiogeneic [113]Anti-proliferative [113]

64 1-Phenylhepta-135-triyne [114] Polyyne C13H8

Anti-microbial [115]Anti-malarial [3]Cytotoxic [3]Antifungal [15]

27 Linoleic acid [79] Fatty acid C18H32O2Anti-viral (100)Cytotoxic [17]

161 Ethyl caffeate [84] Phenylpropanoid C11H12O4 Anti-inflammatory [84]

54 2-O-120573-Glucosyltrideca-11(E)-en-3579-tetrayn-12-diol [26] Polyyne C19H20O7Immunosuppressive andAnti-inflammatory [26]

53 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne [50] Polyyne C19H22O7Anti-diabetic [116]

Anti-inflammatory [50]

69 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne [28] Polyyne C20H26O7Anti-diabetic [28]

Anti-inflammatory [94]

50 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne [28] Polyyne C19H24O7

Anti-diabetic [28]Anti-inflammatory [94]

Anti-malarial andantibacterial [47]

129 Quercetin 3-O-120573-D-galactopyranoside [18] Flavonoid C21H20O12 Anti-inflammatory [117]

170 35-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

171 45-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

169 34-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

126 Quercetin 331015840-dimethyl ether7-O-120572-L-rhamnopyranosyl-(1rarr 6)-120573-D-glucopyranoside [118] Flavonoid C21H20O11 Anti-malarial [119]

125 Quercetin 331015840-dimethyl ether-7-O-120573-D-glucopyranoside [118] Flavonoid C21H20O12 Anti-malarial [119]70 1-Phenyl-13-diyn-5-en-7-ol-acetate [27] Polyyne C15H12O2 Anti-malarial [27]199 Heptanyl 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside [70] Miscellaneous C18H44O10 Antioxidant [70]124 3-O-Rabinobioside [70] Saccharide C27H38O15 Antioxidant [70]130 Quercetin 3-O-rutinoside [70] Flavonoid C27H38O15 Antioxidant [70]167 Chlorogenic acid [70] Phenolic C16H26O9 Antioxidant [70]119 Jacein [70] Flavonoid C24H26O13 Antioxidant [70]

49 (R)-12-dihydroxytrideca-357911-pentayne [47] Polyyne C13H8O2Anti-malarial andAntibacterial [47]

40 Evidence-Based Complementary and Alternative Medicine

Table 12 Structure and activity relationship studies of ethyl caffeateusing in vitro NF-120581BDNA binding assays [84]

Compound Concentration(120583M)

NF-120581BDNA binding

Ethyl caffeate 50 100inhibition

Ethyl 34-dihydroxyhydrocinnamate 100 100inhibition

Catechol 400 100inhibition

Ethyl cinnamate 400 Noinhibition

Table 13 Apoptosis in cocultures of T cells and pancreatic 120573 cells[53]

Cellmedium apoptosis andnecrosis

CD4+ T cellscontrol medium lt4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 2

CD4+ T cellswith cytopiloyne-treated 120573 cells 18CD4+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 7

CD8+ T cellscontrol medium 4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 4

CD8+ T cellswith cytopiloyne-treated 120573 cells 4CD8+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 4

Table 14 Radical scavenging activities of B pilosa extracts [70]

Extractscontrol DPPH assayIC50 (120583gmL)

NBThypoxanthinesuperoxide assayIC50 (120583gmL)

Quercetin 198 15Ascorbic acid 634 Not determined120572-tocopherol 897 Not determinedEthyl acetateextract 1383 597

Butanol extract 1669 114Water extract gt100 gt100

compounds) and inhibition (by 40 compared to 10) ofdifferentiation toTh1 at the same concentration [49]

Among the three polyynes found in B pilosa cytopiloyne(53) had the most potent anti-T1D activity [53] To test the invivo effect of cytopiloyne NODmice received intraperitonealor intramuscular injection of cytopiloyne at 25120583gkg BW3 times per week Twelve-week-old NOD mice started todevelop T1D and 70 of NOD mice aged 23 weeks andover developed T1D Remarkably 12- to 30-week-old NODmice treated with cytopiloyne showed normal levels of bloodglucose (lt200mgdL) and insulin (1-2 ngmL) Consistent

Table 15 Radical scavenging activity of secondarymetabolites fromB pilosa [70]

Metabolite (Table 11)control DPPH assay IC50 (120583gmL)199 Not determined124 53130 68167 105169 33171 38119 Not determined110 Not determinedQuercetin 256Caffeic acid 890

Table 16 Antioxidant activity of the essential oils and water extractsfrom B pilosa [92]

Extract IC50 (120583gmL)Leaf essential oils 47Flower essential oils 50Leaf extract 61Flower extract 172

with T1D incidence cytopiloyne delayed and reduced theinvasion of CD4+ T cells into the pancreatic islets [53]

In vitro study showed that cytopiloyne (53) inhibited thedifferentiation of naıveTh (Th0) cells (ie CD4+ T cells) intoTh1 cells and promoted differentiation of Th0 cells into Th2cells [50] The in vitro data are consistent with the in vivoresults indicating that cytopiloyne reducedTh1differentiationand increased Th2 differentiation as shown by intracellularcytokine staining and FACS analysis [53] In line with theskewing of Th differentiation the level of serum IFN-120574 andIgG2c decreased while that of serum IL-4 and serum IgEincreased compared to the negative controls (PBS-treatedmice) Cytopiloyne also enhanced the expression of GATA-3 a master gene for Th2 cell differentiation but not theexpression of T-bet a master gene forTh1 cell differentiationfurther supporting its role in skewingTh differentiation [53]

Also importantly cytopiloyne partially depleted CD4+rather than CD8+ T cells in NOD mice [53] As shown inTable 13 coculture assays showed that the depletion of CD4+T cells was mediated through the induction of Fas ligandexpression on pancreatic islet cells by cytopiloyne leading toapoptosis of infiltrating CD4+ T cells in the pancreas via theFas and Fas ligand pathways However cytopiloyne did notinduce the expression of TNF-120572 in pancreatic islet cells andthus had no effect on CD8+ T cells [53]

In addition Chang and colleagues showed that cytopi-loyne dose-dependently inhibited T-cell proliferation stimu-lated by IL-2 plus Con A or anti-CD3 antibody using [3H]thymidine incorporation assay [53]

Overall the mechanism of action of cytopiloyne andprobably its derivatives in T1D includes inhibition of T-cellproliferation skewing of Th cell differentiation and partialdepletion of Th cells Due to the anti-diabetic mechanisms

Evidence-Based Complementary and Alternative Medicine 41

Table 17 Antibacterial activity of essential oils and flower extracts from B pilosa [92]

Strain Mean zone of inhibition (mm)Leaf essential oil Flower essential oil Leaf extract Flower extract

Micrococcus flavus 127 plusmn 03 87 plusmn 03 102 plusmn 02 108 plusmn 03Bacillus subtilis 173 plusmn 19 117 plusmn 02 109 plusmn 02 103 plusmn 02Bacillus cereus 190 plusmn 14 112 plusmn 03 118 plusmn 04 185 plusmn 10Bacillus pumilus 123 plusmn 07 108 plusmn 02 105 plusmn 04 77 plusmn 02Escherichia coli 137 plusmn 04 203 plusmn 07 102 plusmn 11 140 plusmn 13Pseudomonas ovalis 125 plusmn 08 137 plusmn 15 102 plusmn 06 125 plusmn 06

Table 18 Antibacterial activity of root extracts from B pilosa [93]

Strain MIC50 (mgmL)Methanol extract Acetone extract

Bacillus cereus 10 mdashEscherichia coli 5 5Klebsilla pneumonia 5 10Micrococcus kristinae 10 mdashPseudomonas aeruginosa 10 10Staphylococcus aureus 5 10Sraphylococcus epidermidis 5 5Serratia marcescens 10 mdashShigelea flexneri 10 mdashStreptococcus faecalis 10 mdash

Table 19 Antibacterial activity of B pilosa of compound 29 [47]

Strain MIC50 (120583gmL)Escherichia coli NIHJ 1Escherichia coli ATCC25922 1Klebsiella pneumoniae ATCC700603 128Serratia marcescens ATCC13880 16Pseudomonas aeruginosa ATCC27853 8Staphylococcus aureus FDA209P 05Staphylococcus aureus ATCC29213 025Staphylococcus aureus N315 (MRSA) 05Enterococcus faecalis ATCC29212 2Enterococcus faecalis NCTC12201 (VRE) 1Bacillus subtilis ATCC6633 05Candida albicans ATCC10231 025

of action it was hypothesized that cytopiloyne protectsNOD mice from diabetes by a generalized suppression ofadaptive immunity To evaluate this hypothesis ovalbumin(Ova) was used as a T-cell dependent antigen to primeNOD mice which had already received cytopiloyne orPBS vehicle Ova priming boosted similar anti-Ova titersin cytopiloyne-treated mice and PBS-treated mice but adifference in immunoglobulin isotype was observed in thetwo groups Thus it was concluded that cytopiloyne is animmunomodulatory compound rather than an immunosup-pressive compound [50 53]

T2D is a chronic metabolic disease with serious com-plications resulting from defects in either insulin secretion

insulin action or both [124] A study by Ubillas et alshowed that the aqueous ethanol extract of the aerial partof B pilosa at 1 gkg body weight (BW) lowered bloodglucose in dbdb mice a T2D mouse model [51] Basedon a bioactivity-guided identification compounds 69 and50 were identified Further the mixture of the compounds(69 50) in a 2 3 ratio significantly decreased blood glucoseconcentration and reduced food intake on the second dayof treatment when administered at doses of 250mgkg twicea day to C5BLKs-dbdb mice When tested at 500mgkga more substantial drop in blood glucose level as wellas the stronger anorexic effect (food intake reduced from58 gmouseday to 25 gmouseday) was observed [51] Inthis study it was suggested that the blood glucose lower-ing effect of B pilosa was caused in part by the hungersuppressing effect of its polyynes [51] However the hungersuppressing effect of the ethanol extract of B pilosa wasnot found in the studies described below In another study[24] water extracts of B pilosa (BPWE) were used indiabetic dbdb mice aged 6ndash8 weeks with postprandialblood glucose levels of 350 to 400mgdL Like oral anti-diabetic glimepiride which stimulates insulin release onesingle dose of BPWE reduced blood glucose levels from374 to 144mgdL The antihyperglycemic effect of BPWEwas inversely correlated to an increase in serum insulinlevels suggesting that BPWE acts to lower blood glucose viaincreased insulin production However BPWE had differentinsulin secretion kinetics to glimepiride [24] One flaw incurrent anti-diabetics is their decreasing efficacy over timeThe authors investigated the long term anti-diabetic effectof BPWE in dbdb mice BPWE reduced blood glucoseincreased blood insulin improved glucose tolerance andreduced the percentage of glycosylated hemoglobin (HbA1c)Both long-term and one-time experiments strongly supportthe anti-diabetic action of BPWE [24] In sharp contrast toglimepiride BPWE protected against islet atrophy in mousepancreas The investigators further evaluated anti-diabeticproperties of 3 B pilosa varieties B pilosa L var radiate(BPR) Bpilosa L var pilosa (BPP) and B pilosa L varminor(BPM) in dbdb mice [8] One single oral dose (10 50 and250mgkg body weight) of BPR BPP or BPM crude extractsdecreased postprandial blood glucose levels in dbdb micefor up to four hours and the reduction of glucose levels inthe blood appeared to be dose-dependent Comparing thethree variants BPR extract resulted in a higher reduction inblood glucose levels when administered at the same dose as

42 Evidence-Based Complementary and Alternative Medicine

Table 20 Antifungal activity of B pilosa [92]

Partextract Concentration (ppm) Strain InhibitionCortiicum rolfsii Fusarium solani Fusarium oxysporum

Leaves

Essentialoils

100 857 plusmn 09 682 plusmn 0 745 plusmn 17250 960 plusmn 08 779 plusmn 18 879 plusmn 04

AqueousExtracts

100 446 plusmn 17 605 plusmn 21 716 plusmn 07250 942 plusmn 03 689 plusmn 07 824 plusmn 19

Flowers

Essentialoils

100 604 plusmn 09 892 plusmn 04 869 plusmn 05250 894 plusmn 12 980 plusmn 03 949 plusmn 06

AqueousExtracts

100 331 plusmn 11 714 plusmn 07 573 plusmn 22250 661 plusmn 14 912 plusmn 0 900 plusmn 07

Table 21 Antifungal activity of B pilosa root extracts [93]

Strain LC50 (mgmL)Acetoneextracts

Methanolextracts

Waterextracts

Aspergillus niger 014 006 007Aspergillus flavus 1091 658 0Penicillium notatum 005 005 005

the other two varieties In terms of serum insulin levels a doseof 50mgkg of each extract was used and the BPR extracttogether with the three polyynes significantly increased theserum insulin level in dbdb mice Long-term experiments(28-day treatment) were then conducted using diabetic micewith postprandial glucose levels from 370 to 420mgdL andglimepiride was used as positive controlThe range of dosagesapplied was from 10mgkg BW to 250mgkg BW Resultsshowed that the positive control as well as the crude extractsof the three varieties lowered the blood glucose levels indbdb mice However only BPR extract containing a higherpercentage of cytopiloyne (53) reduced blood glucose levelsand augmented blood insulin levelsmore than BPP andBPMThe percentage of glycosylated hemoglobin A1c (HbA1c) wasalso measured and found to be 79 plusmn 05 in mice aged 10ndash12 weeks and 66 plusmn 02 61 plusmn 03 and 62 plusmn 03 inthe blood of age-matchedmice following treatment with BPRcrude extract (50mgkg) glimepiride (1mgkg) and com-pound 53 (05mgkg) respectively [8] Among the polyynesfound in B pilosa cytopiloyne was the most effective againstT2D Hence cytopiloyne was used for further study on anti-diabetic action and mechanism [125] The data confirmedthat cytopiloyne reduced postprandial blood glucose lev-els increased blood insulin improved glucose tolerancesuppressed HbA1c level and protected pancreatic islets indbdb mice Nevertheless cytopiloyne failed to decreaseblood glucose in streptoztocin (STZ)-treated mice whose bcells were already destroyed Additionally cytopiloyne dose-dependently increased insulin secretion and expression in bcells as well as calcium influx diacylglycerol and activationof protein kinase C120572 Collectively the mechanistic studiessuggest that cytopiloyne treats T2D via regulation of insulin

production involving the calciumDAGPKC120572 cascade in bcells

The studies detailed above point to the conclusion thatcytopiloyne and related polyynes (compounds 69 and 49)are anti-diabetics in animal models The data uncover a newbiological action of polyynes It should be noted that likeall anti-diabetic drugs cytopiloyne failed to prevent or curediabetes completely but reduced diabetic complications [125]Intriguingly 34 polyynes have been found in B pilosa so farIt remains to be seen whether all the polyynes present in thisplant have anti-diabetic activities

34 Antioxidant Activity Free radicals can damage cellularcomponents via a series of chemical reactions [89] leadingto development and progression of cardiovascular diseasecancer neurodegenerative diseases and ageing [70] Freeradicals nitric oxide (NO) and superoxide anions can beproduced in macrophages to kill microbes However anexcessive generation of the free radicals under pathologicalconditions is associated with a wide range of illnessesPlants are known to be rich in antioxidant phytochemicalsChiang and colleagues evaluated the free radical scaveng-ing activity of crude extract fractions and compounds ofB pilosa using 11-diphenyl-2-picrylhydrazyl (DPPH) andhypoxanthinexanthine oxidase assays [70] Using DPPHand hypoxanthinexanthine oxidase assays they found thatthe B pilosa crude extract and the ethyl acetate butanoland water fractions had free radical scavenging activityNine compounds Heptyl- 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside (199) 3-O-Rabinobioside (124) Quercetin3-O-rutinoside (130) Chlorogenic acid (167) 34-Di-O-caffeoylquinic acid (169) 35-Di-O-caffeoylquinic acid (170)45-Di-O-caffeoylquinic acid (171) Jacein (119) and Cen-taurein (110) had DPPH radical scavenging activity [70]The IC

50values of the B pilosa crude extractfractions and

compounds are summarized in Tables 14 and 15 respectivelyMeasurement of free radical scavenging activities is one

way of assessing the antioxidant activities of B pilosa andits fractions and compounds It is interesting that the ethylacetate and butanol fractions are more active than the waterfraction and B pilosa crude extract [70] Of the secondarymetabolites only phenolic compounds 124 130 167 169 and171 showed significant DPPH-radical scavenging activities

Evidence-Based Complementary and Alternative Medicine 43

Further analysis of the structure-activity relationship of thecompounds suggested that substitution of the C3 hydroxylgroup with glycosides increased the activity approximately2-fold (for example in compounds 124 and 130) relative toquercetin (-OH in its C3) [70] Further modification of thestructures of the active compounds needs to be performed totest the effects of various substituents on activity The reasonwhy most of the antioxidant compounds contain phenolmoieties in their structure could be that the reduction-oxidation (redox) properties of phenols allow them to actas reducing agents singlet oxygen quenchers and hydrogendonors

A complementary study by Muchuweti and colleaguesdetermined phenolic content antioxidant activity and thephenolic profile of B pilosa methanol extract [89] Theyestimated that the phenolic content of the methanol extractof B pilosawas 11028plusmn22mgg [89] Vanillin hydroxyben-zaldehyde caffeic acid coumaric acid and ferulic acid werefound in this extractThe B pilosa extract also showed DPPHradical scavenging activity Furthermore the antioxidantactivity of the flavonoids found in B pilosa was correlatedwith its hepatoprotective effects through their inhibitionof NF-120581B activation which may lessen the oxidative stresscaused by the production of free radicals during liver injury[60]This activity might also be due to the anti-inflammatoryeffects of the aqueous extracts of B pilosa aerial parts on theinhibition of COX-2 and PGE

2production [88]

Essential oils from B pilosa flowers and leaves are alsoreported to possess antioxidant activity With the aim ofreplacing chemically synthesized additives Deba and col-leagues [92] worked on the antioxidant antibacterial andantifungal activities of essential oils and water extracts of Bpilosarsquos leaves and flowers Table 16 summarizes the resultsobtained from DPPH free radical scavenging assay

It can be inferred fromTable 16 that essential oils from theleaves possessed the highest activity It is reported elsewherethat monoterpenes present in essential oils such that of Bpilosa have protective effects and antioxidant properties [92]Beta-carotene bleaching method was also performed Leavesessential oils and aqueous extracts of the leaves and flowersshowed higher activity than the flower essential oils This isdue to the volatility of the flower essential oils The activityexhibited by the aqueous extracts was accounted to thepresence of phenolic compounds that are reported to donatea hydrogen atom to free radicals such that the propagation ofthe chain reaction during lipid oxidation is terminated [92]Overall essential oils and phenolics present in B pilosa canbe thought of as major antioxidant compounds

35 Immunomodulatory Activity B pilosa is thought to bean immunomodulatory plant and is reported to be effectivein the treatment of immune disorders such as allergy [75]arthritis [75] and T1D [48 50 53]

As pointed out in the discussion of its anti-diabeticactivities (Section 33) a combination of phytochemicalsmethod and T-cell activation assays was used to studyimmunomodulatory properties of B pilosa IFN-120574 is a keycytokine released by T and NK cells that mediates immune

cells and sustains immunity against pathogens Defects inIFN-120574 expression regulation and activation result in vulner-ability to diseases caused by bacteria and viruses [69] Anelegant study performed byChang and colleagues using IFN-120574 promoter-driven luciferase reporter construct in JurkatT cells showed that hot water crude extracts of B pilosaincreased IFN-120574 promoter activity two-fold [69] Out of thesubfractions of this extract the butanol fraction but not thewater or ethyl acetate fractions increased IFN-120574 promoteractivity six-fold Centaurein (110) and centaureidin (109)were identified from the butanol fraction and were stated tocause a four-fold increase in IFN-120574 promoter activity withEC50

values of 75 120583gmL and 09120583gmL respectively Themechanism of action of centaurein was determined usingtranscription factors such as AP-1 NFAT and NF120581B whichare reported to bind to IFN-120574 promoter and regulate IFN-120574transcription Unlike with the activity of the positive controlPHA centaurein caused a four-fold increase in NFAT a3-fold increase in NF120581B and had little if any effect onAP-1 enhancer activities (23-fold three-fold and ten-foldincreases respectively were seenwith PHA) [69]The authorsconcluded that centaureinmodulates IFN-120574 expression by theNFAT and NF120581B pathways The article only determined themechanism of action of centaurein Its aglycone centaureidinmay act through the samemechanism though this conclusionneeds to be further verified

B pilosa extract and its compounds are reported to inhibitdifferentiation of naıve CD4+ helper T (Th0) cells into Th1cells [49] Using theThcell differentiation assay as a screeningplatform 3 polyynes 2-120573-D-glucopyranosyloxy-1 -hydroxytrideca-57911-tetrayne (53) 3-120573-D-glucopyranosyl-1-hydroxy-6(E)- tetradecene-81012-triyne (69) and 2-120573-D-glucopyranosyloxy-1-hydroxy-5(E)- tridecene-7911-triyne(49) were discovered from B pilosa [49 53] The data showsthat cytopiloyne and other two polyynes suppressed thedifferentiation of type 1 helper T (Th1) cells and productionof Th1 cytokines and promoted that of type 2 helper T (Th2)cells and production of Th2 cytokines thus explaining theimmunomodulatory and anti-inflammatory effects of Bpilosa and its polyynes

Chang and colleagues were the first to report the effect ofthe butanol extract of B pilosa on the autoimmune diabetesand airway inflammation inmice [48] Imbalance in the levelsofTh1 andTh2 and of various cytokines leads to autoimmunediseases T1D and other autoimmune diseases (rheumatoidarthritis Crohnrsquos disease among others) are exacerbated byan increase Th1 levels (specifically CD4+ Th1 cells) whileTh2 cells antagonize this effect [19] Moreover Th2 cellsmediate asthma in ovalbumin-induced hypersensitivity inBALBc mice [48] In their work [48] Chang and colleaguesshowed that 10mgkg butanol extracts with 15 (ww)compound 69 and 11 (ww) compound 49 (Table 11)ameliorated the development of Th1-mediated diabetes inNOD mice through inhibition of 120573 cell death and leukocyteinfiltration At the same dosage the butanol extracts alsoexacerbated ovalbumin-induced pulmonary inflammationin BALBc mice with an increase in the infiltration ofeosinophils and mast cells into the airway of the mice [48]Despite the different outcomes both mouse models proved

44 Evidence-Based Complementary and Alternative Medicine

the concept that control over the Th1Th2 shift is associatedwith autoimmune diseases and that the B pilosa butanolextract can shift the differentiation of Th0 cells to Th2 cells[48 49]

An extended study presented by Chiang and colleaguesshowed that compound 53 modulates T-cell functions [50]Using CD4+ T cells from BALBc mice they demonstratedthat cytopiloyne decreased levels of IFN-120574 producing cells(Th1) by 122 (from 72 to 598) Since Th1 and Th2 celldifferentiation is antagonistic it was expected that compound53 increased the percentage of mouse IL-4 producing cells(Th2) by 72 (from 237 to 309) Subsequent assess-ment of effect of compound 53 on the modulation of thetranscription of IL-4 and IFN-120574 showed that cytopiloyne asexpected decreased the splenocyte levels of IFN-120574 mRNAand increasing that of IL-4 in a dose-dependent manner Inthe range of 01 to 3 120583gmL of compound 53 the effects werenot attributed to its cytotoxicity Consequently using 3120583gmLcytopiloyne for 72 and 96 hours the protein concentration ofIFN-120574 decreased to 186 and 444 respectively Under thesame conditions cytopiloyne increased IL-4 concentrationsto 1985 and 2470 (for 72 and 96 hours resp) Thismodulation of T-cell differentiation exhibited by compound53 was used to explain its anti-diabetic activity [50]

The anti-diabetic role of cytopiloyne was extensivelydiscussed above (Section 33 anti-diabetic activity) Themolecular basis of the regulation of cytokine expression bycytopiloyne has been described Cytopiloyne directly elevatedthe expression level of IL-4 via GATA-3 upregulation inT cells [53] However reduction of IFN-120574 expression in Tcells seemed to come from the indirect opposing effect IL-4cytokine because the expression level of T-bet was unaltered[53] In this way cytopiloyne skewedTh1 polarization into theTh2 state conferring protection against T1D in NOD miceAside frompolarization ofThcell differentiation cytopiloynealso activated the expression of Fas ligand in pancreaticb cells this increase leading to the partial depletion of Tcells and reduction of immune response in local areas suchas the pancreas Of note cytopiloyne also inhibited T-cellproliferation and activation By targeting T cells from threeimmunomodulatory actions cytopiloyne protects againstT1D and probably other Th1-mediated autoimmune diseases[19]

The phytochemical constituents of B pilosa exert theirfunctions on different immune cells to modulate immuneresponse It is possible that some of the compounds mayhave agonistic or antagonistic effects on immune responseImmune function of B pilosa may depend on its compo-sition and amount of compounds which could explain theapparently conflicting report of B pilosa butanol extractaggravating allergy in mice [48] while cellulosine-treatedextract ameliorated allergy [48 75] IFN-120574 promoter reporterassays and T-cell differentiation assays were used to isolate2 flavonoids [69] and 3 polyynes [49 50] as immunomod-ulatory compounds from the butanol fraction of B pilosaInterestingly the flavonoids promote IFN-120574 expression inNK and T cells In marked contrast the polyynes promoteIL-4 expression and indirectly inhibit IFN-120574 expression indifferentiating T cells Sensitivity appears to be the key to

identifying structure- and bioactivity-related phytochemicalsfrom medicinal plants

36 Antimalarial Activity The use of chemical drugs againstpathogens has resulted in drug-resistant mutants Examplesof drug resistance can be found in the species of thePlasmodium that cause malaria It is important to search fornew compounds to combat Plasmodium parasites [47] Astudy of the anti-malarial activity of the leaf extracts of Bpilosa using a combination of phytochemistry and bioassaysshowed that compound 49 (Table 11) showed activity againsta malaria parasite (P falciparum NF54 strain) with an IC

50

value of 60 120583gmL [77] In addition compound 49 isolatedfrom the aerial parts of B pilosa inhibited growth of the Pfalciparum FCR-3 strain with an IC

50value of 035 120583gmL

This compound was tested for its in vivo effect in miceinfected with P berghei NK-65 strain Results showed thatcompound 49 decreased the average parasitemia in the redblood cells by 207 (from 328 of that of the control to121) after an intravenous injection of 08mgkg BWday forfour days [47] Further studies addressing the anti-malarialmechanism underlying both polyynes and clinical studies areneeded

37 Antibacterial Activity Emergence of multiple antibiotic-resistantmicrobes is becoming a global threat to public healthand a challenge to disease treatment For instance penicillinis commonly used to combat a food-borne intracellularbacteriumListeria however penicillin-resistant bacteria havebeen discovered recently [69] Chang and coworkers iso-lated centaurein (110) and centaureidin (109) from B pilosaextract [69 70] Centaurein enhances expression of IFN-120574a key cytokine for macrophage activation and consequentlyenhances bactericidal activity in macrophages [69 70] Inagreement with observed in vitro effects centaurein wasreported to prevent and treat Listeria infection in C57BL6Jmice [91] Mechanistic studies confirmed that centaureinexerted antilisterial action via IFN-120574 expression in wild-typemice but not IFN-120574 knockout mice [91] Further in vitrostudies on centaurein showed that this compound increasedIFN-120574 expression by 13 (from 17 to 20) 20 (from 21to 41) and 11 (from 6 to 17) in CD4+ T cells CD8+Tcells and NK cells respectively That is to say there wasan increase in IFN-120574 producing immune cells As expectedcentaurein also enhanced the expression level of T-bet a keynuclear factor for IFN-120574 expression Consistently centaureinaugmented the serum IFN-120574 levels inC57BL6Jmice and thisaugmentation peaked 24 hours after compound injectionThe quantity of mouse serum IFN-120574 was sufficient to activatemacrophages in vitro and eradicated GFP-producing Listeriainside macrophages However the entry of Listeria intomacrophages was not affected by centaurein-treated mousesera Centaurein treatment at 20120583g per mouse rescued 30of the mice infected with a lethal dose of Listeria (2 times106 CFU) It is noteworthy that in the presence of ampicillin(5 120583gmouse) centaurein (20 120583gmouse) rescued 70 of themice suggesting an additive effect between ampicillin andcentaurein [91] Despite lower abundance centaureidin was

Evidence-Based Complementary and Alternative Medicine 45

30 times more active than centaurein in terms of IFN-120574production [91]

Aside from the indirect antibacterial action mentionedabove extract andor compounds of B pilosa also showeddirect bacteriostatic andor bactericidal action One studyreported that essential oils and leafflower extracts of Bpilosa could suppress the growth of gram positive and gramnegative bacteria as evidenced by zone of inhibition assaysIn this study antibacterial activity of the essential oils fromthe leaves and flowers of B pilosa was determined in anattempt to identify natural products as food preservatives forprevention of microbial multiplication and food oxidationThe essential oils and extracts of B pilosa leaves and flow-ers showed moderate but different extents of antibacterialactivity (Table 17) In general essential oils had higherantibacterial activity than crude extracts One explanation forthis could be that monoterpenes in the essential oils destroycellular integrity and subsequently inhibit the respirationand ion transport processes The presence of antibacterial120573-caryophyllene could be another explanation as reportedelsewhere [92] Another study reported that the methanoland acetone extract of B pilosa roots displayed antibacterialactivities against the bacteria listed in Table 18 [93] andmethanol extracts from the roots seemed to be the mosteffective

Another study indicated that the polyyne (R)-12-dihydroxytrideca-357911-pentayne (49) from this plantalso suppressed bacterial growth as shown by the minimuminhibitory concentration required to inhibit 50 bacterialgrowth (MIC

50) in Table 19 This compound was highly

effective against several Gram positive and Gram negativebacteria including the drug-resistant bacteria Staphylococcusaureus N315 (MRSA) and Enterococcus faecalis NCTC12201(VRE) [47] Strikingly compound 49 had a similar MIC

50

value to antibiotics (ampicillin tetracycline norfloxacin andamphotericin B) in most of the bacteria tested

Antibacterial activity of B pilosa extracts and compo-nents expressed as MIC

50and the mean zone of inhibition

is tabulated in Tables 17 18 and 19The zone of inhibition forAmpicilline (positive control) ranges from 153 plusmn 03mm to443 plusmn 02mm [92]

38 Antifungal Activity B pilosa has traditionally been usedto treat microbial infection Recently different parts of Bpilosa have been tested for antifungal activities Deba andcolleagues first evaluated the antifungal effect of the hotwater extracts of the B pilosa roots stems and leaves againstCorticium rolfsii Fusarium solani and Fusarium oxysporumThey discovered that C rolfsii was most suppressed bytreatment with B pilosa as its growth was reduced at almostall the tested doses followed by F oxysporum and F solani[99] However the fungicidal activities of the stems androots were greater than the leaves [99] Moreover the samegroup assessed the antifungal activity of the essential oilsand aqueous extracts from B pilosa flowers and leaves [92]They showed that the extracts and oils had antifungal activityagainst C rolfsii F solani and F oxysporum Essential oils

appeared to have better fungicidal activity thanwater extractsas summarized in Table 20

Another study by Ashafa and colleagues showed thatacetone methanol and water extracts of the B pilosa rootsshowed antifungal activities against Aspergillus niger Aflavus and Penicillium notatum using the agar dilutionmethod The results are tabulated in Table 21 [93] Negativecontrols showed 0 growth inhibitionThe methanol extractof the B pilosa roots at 10mgmL was also effective againstCandida albicans [93]Of note B pilosa obtained fromPapuaNew Guinea had no activity against A niger and C albicans[126] but the SouthAfrican (EasternCape) ecotype exhibitedmoderate activity against C albicans [93] This discrepancymay depend on extraction solvents extraction procedureassay techniques different plant parts and abundance ofactive compounds

B pilosa produces a variety of secondary metabolitessuch as flavonoids phenylacetylenes alkaloids sterols ter-penoids and tannis [92 93] However none of them havebeen confirmed as active compounds against fungi Furtherinvestigation of active compounds from B pilosa is necessaryto further understand the antifungal efficacy of this plant

39 Hypotensive and Vasodilatory Activities In early studiesDimo and colleagues used three rat models normotensiveWistar rats (NTR) salt-loading hypertensive rats (SLHR)and spontaneous hypertensive rats (SHR) to investigate thehypotensive effect of the methanol crude extract of B pilosaleaves [28 127] The extract lowered systolic blood pressurein hypertensive rats (SLHR and SHR) to a greater degreethan NTR [28 127] In addition a decrease in urinarysodium ions and an increase in urinary potassium ions wereobserved after treatment with the methanol extract of Bpilosa leaves although neither differences were statisticallysignificant [28 127] Taking the data together the studyproposed that B pilosa leaf extract reduced blood pressurevia vasodilation [28 127] The same group continued totest the antihypertensive effect of aqueous and methylenechloride extracts of B pilosa leaves in a hypertensive ratmodel [28 127] To establish a fructose-induced hypertensionmodel male Wistar rats were given 10 fructose solutionto drink ad libitum for three weeks In addition to freeaccess to 10 fructose the rats were treated with the aqueous(150mgkg) or methyl chloride (350mgkg) extracts of Bpilosa for additional three weeks [28 127] Both extracts ofB pilosa leaves had a hypotensive effect on rats Howeverneither extracts reversed the elevation of serum insulin infructose-fed rats Therefore B pilosa lowered blood pressureirrespective of insulin [28 127] To better understand thehypotensive mechanism the authors investigated the effectof a neutral extract of B pilosa (NBP) a mixture of methanolandmethylene chloride (1 1) extract after neutralization withNaOH and HCl on the heart and the blood pressure ofNTR and SHR [110] This study showed that an intravenousinjection of the NBP resulted in a biphasic reduction insystolic blood pressure In addition one intravenous dose ofthe extract at 10 20 and 30mgkg BW decreased systolicblood pressure in normal rats by 183 425 and 30

46 Evidence-Based Complementary and Alternative Medicine

respectively and the same doses reduced the blood pressurein hypertensive rats by 258 389 and 286 respectivelyOnly the highest dose (30mgkg) affected the force of thecontraction of the heart Atropine and propranolol were usedto interfere with the hypotensive action of the NBP Atropinereduced the initial phase of the hypotensive response in NBPand completely abolished the second phase of hypotensiveresponse in NBP In contrast propranolol increased thefirst hypotensive response but partially abolished the secondhypotensive response provoked by the NBP [110]This mech-anistic study suggested that B pilosa invokes the biphasichypotensive responses via targeting cardiac pump efficiencyduring the first phase and vasodilation at the second phase[110]

A further study was performed to investigate the relaxingeffect of a neutral extract of B pilosa (NBP) on rat aorta con-tractedwithKCl (60mM) andnorepinephrine (01mM) [94]Cumulative addition of NBP relaxed the rat aorta previouslycontracted by KCl in a dose-dependent manner The EC

50

value of theNBP for vasorelaxationwas 032mgmLThedataalso showed that the NBP reduced the contraction of aortapreviously contracted by KCl irrespective of the presence ofaortic endothelium [94]

Pretreatment with glibenclamide an ATP-dependent K+channel blocker did not considerably affect the relaxant effectof the NBP on KCl-induced contraction suggesting thatthe vasodilatory effect of B pilosa was not related to theopening of this ATP-dependent K+ channel [94] On theother hand in the presence of indomethacin or pyrilaminemaleate the relaxant response induced by the plant extractwas significantly inhibited at the lower concentrations Theplant extract was able to reduce the aorta resting tone inhibitthe KCl-induced contractions by 90 at 15mgmL and theCaCl2-induced contractions by 95 at 075mgmL These

results demonstrate that B pilosa can act as a vasodilatorprobably via acting as a calcium antagonist [94]

However no specific compound for the above activityhas been identified from B pilosa to date A bioactivity-guided identification approach may be adopted to identifythe active compounds in B pilosa that possess hypotensiveand vasodilatory effects and understand their mechanism ofaction

310 Wound Healing Activity B pilosa has been traditionallyused to treat tissue injury inCameroon Brazil andVenezuela[26] Hassan and colleagues investigated the wound healingpotential of B pilosa in Wistar rats [128] Mirroring thepositive control neomycin sulfate the ethanol extract of Bpilosa had faster wound closure than control rats 3 6 and 9days after topical application Histological examination alsorevealed better collagenation angiogenesis and organizationof wound tissue seven days after application Epithelializationand total healing time in B pilosa-treated rats were compara-ble to those of neomycin sulfate Together these data suggestthat B pilosa may be a viable alternative to neomycin lotionfor the treatment of wounds

In addition to studying the wound healing effect of Bpilosa on external ulcers Tan and colleagues also examined

the effect of methanol cyclohexane and methyl chlorideextracts of B pilosa on gastric ulcers in Wistar rats fedwith 1mL HClethanol gastric necrotizing solution (150mMHCl in 60 ethanol) and macroscopically visible lesionswere scored [26] Among the three extracts methylene chlo-ride extracts exhibited the highest activity showing 464inhibition of lesion formation at a dose of 500mgkg BWand complete inhibition at 750mgkg [26] The efficacy ofthe ethylene chloride extract was followed by that of themethanol extracts which had inhibition ranging from 304to 822 at concentrations of 500mgkg and 1000mgkgBW respectively [26] The cyclohexane extracts showed thelowest activity against gastric ulcers in rats with 13340 and 797 inhibition at 500 750 and 1000mgkg BWrespectively [26] To better understand the mode of actionof the methylene chloride extract of B pilosa rats werepretreated with indomethacin a COX-2 inhibitor involved inprostaglandin synthesis Pretreatment significantly reducedthe protection against HClethanol-induced ulcers to 313inhibition at 750mgkg BW suggesting a link between theantiulcerative activity of B pilosa and prostaglandin syn-thesis Unexpectedly the methylene chloride extract of Bpilosa showed little gastric mucosal protection against gastriclesions induced by 95 ethanol (1mL) [26] Absolute alcoholis known to cause mucosalsubmucosal tissue destruction viacellular necrosis and the release of tissue-derived mediators(histamine and leukotrieneC4)Thus these data imply thatBpilosa did not prevent the generation or the necrotic action ofthese mediators on the gastric microvasculature In additionpylorus ligation can increase gastric acid secretion withoutan alteration of mucosal histamine content The methylenechloride extract of B pilosa did not possess antisecretoryactivity On the contrary it was observed that increases in thedose of the extract led to elevated gastric juice acidity [26]Results with both absolute ethanol and pylorus ligation ratmodels suggested the possibility that the ineffectiveness of Bpilosa against gastric ulcers was due to lack of antihistaminicactivity in the plant In summary overall the data suggest thatB pilosa protects against HCLethanol-mediated ulcers viainhibition of prostaglandin biosynthesis

Previous phytochemical studies showed that a groupof flavonoids acyclichalcones are present in B pilosa[129 130] and the chalcones were proposed to have anti-ulcerative activity [131] Moreover nine hydroxychalcones 10were reported to possess gastric cytoprotective effects with24-dihydroxychalcone being the most active [132] Sincemethylene chloride extracts appear to be the most active Bpilosa extracts next the specific anti-ulcerative phytochemi-cals in the methylene chloride extracts of B pilosa and theirmodes of action needs to be probed

Despite the claims listed inTable 3 relatively few scientificstudies have been conducted in vitro and in vivo to address thetraditional ethnomedical uses of B pilosa Information aboutthe use of B pilosa as a botanical therapy recorded so far isfar from complete Studies conducted thus far only serve asa starting point for further investigation of B pilosa and theultimate efficacious use of the herb in clinical applications

Evidence-Based Complementary and Alternative Medicine 47

4 Toxicology

Despite its use as an ingredient in food for human con-sumption studies on systemic toxicity (eg acute subacutechronic and subchronic toxicities) of B pilosa in humansand animals are still inadequate and insufficient So faracute andor subchronic toxicities have been evaluated inrats and mice Oral acute and 28-day toxicities of waterextract of B pilosa leaves were evaluated in Wistar rats[133] An oral dose of water extract of B pilosa leaves at10 gkg BW showed no obvious mortality or changes inthe appearance in rats [134] The same extract at 08 gkgBWday once a day showed no obvious sub-chronic toxicityin rats over 28 days as measured by survival rate bodyweight and gross examination of organs [134] These dataare consistent with our data indicating that oral deliveryof the water extract of the B pilosa whole plant at 1 gkgBWday once a day is safe in rats over 28 days (unpublisheddata) Taken together these studies suggest that ingestion ofB pilosa aqueous extract at up to at 1 gkg BWday oncea day is highly safe in rats In addition the acute toxicityof aqueous and ethanol extracts of B pilosa in mice havebeen reported [134] Five- to six-week-old mice with weightsbetween 28 and 35 g received a peritoneal injection of bothextracts at the different doses The LD

50 the dose that causes

50 lethality of the aqueous and ethanol extracts in micewas 1230 gkg BW and 615 gkg BW respectively [134] Acomplete toxicological study has not been completed forhumans Furthermore the drug interactions of B pilosa withother drugs are unknown Further safety verification andclinical trials should be performed before B pilosa can beconsidered for medicinal use

5 Conclusions

B pilosa is an erect perennial plant with green leaves whiteor yellow flowers and tiny black seeds As it is distributedworldwide and is widely used as a folk remedy B pilosacan be thought of as an extraordinary source of food andmedicine However a comprehensive up-to-date review ofresearch on B pilosa has hitherto been unavailable In thisarticle scientific studies on B pilosa have been summarizedand critically discussed from the perspectives of botanyethnomedicine phytochemistry pharmacology and toxicol-ogy B pilosa is claimed to treat more than 40 disordersand 201 compounds have been identified from this plantThe medicinal utility of B pilosa and its modes of actionin relation to its known phytochemicals were discussedherein Polyynes flavonoids phenylpropanoids fatty acidsand phenolics are the primary bioactive compounds of Bpilosa and they have been reported to be effective in thetreatment of tumors inflammationimmune modulationdiabetes viruses microbes protozoans gastrointestinal dis-eases hypertension and cardiovascular diseases Cautionshould be exercised in the therapeutic use of B pilosa forhypoglycemia hypotension bleeding and allergy

Acknowledgments

The authors thank their laboratory members for construc-tive suggestions and the authors whose publications theycited for their contributions The authors also thank MsMiranda Loney of Academia Sinica for English editing of thispaper This work was supported by Grants 99-CDA-L11 and101S0010056 from Academia Sinica Taiwan Arlene P Bar-tolome is a recipient of MECO-TECO Sandwich ScholarshipProgram between Taiwan and the Philippines

References

[1] T Shen G H Li X N Wang and H X Lou ldquoThe genusCommiphora a review of its traditional uses phytochemistryand pharmacologyrdquo Journal of Ethnopharmacology vol 142 no2 pp 319ndash330

[2] C Long P Sauleau B David et al ldquoBioactive flavonoids ofTanacetum parthenium revisitedrdquo Phytochemistry vol 64 no2 pp 567ndash569 2003

[3] P O Karis and O Ryding Asteraceae Cladistics and Classifica-tion Bremer K Eds pp 559ndash569 Timber press Portland OreUSA 1994

[4] O N Pozharitskaya A N Shikov M N Makarova et alldquoAnti-inflammatory activity of a HPLC-fingerprinted aqueousinfusion of aerial part of Bidens tripartita LrdquoPhytomedicine vol17 no 6 pp 463ndash468 2010

[5] F Q Oliveira V Andrade-Neto A U Krettli and MG L Brandao ldquoNew evidences of antimalarial activity ofBidens pilosa roots extract correlated with polyacetylene andflavonoidsrdquo Journal of Ethnopharmacology vol 93 no 1 pp 39ndash42 2004

[6] Agriculture USDA ldquoPlants databaserdquo 2013in Natural Resources Conservation Servicehttpwwwnrcsusdagovwpsportalnrcssitenationalhome

[7] Y Zhou and X Z Yan ldquoExperimental study of qi deficiencysyndrome and Codonopsis pillosulae and Astragalus injectionon the immune response in micerdquo Zhong Xi Yi Jie He Za Zhivol 9 no 5 pp 286ndash262 1989

[8] S C Chien P H Young Y J Hsu et al ldquoAnti-diabeticproperties of three common Bidens pilosa variants in TaiwanrdquoPhytochemistry vol 70 no 10 pp 1246ndash1254 2009

[9] M J Alcaraz and M J Jimenez ldquoFlavonoids as anti-inflammatory agentsrdquo Fitoterapia vol 59 no 1 pp 25ndash38 1988

[10] FAO Agriculture Food and Nutrition for AfricamdashA ResourceBook for Teachers of Agriculture Publishing ManagementGroup FAO Information Division Rome Italy 1997

[11] M B Rokaya Z Munzbergova B Timsina and K R BhattaraildquoRheum australe D Don a review of its botany ethnobotanyphytochemistry and pharmacologyrdquo Journal of Ethnopharma-cology vol 141 no 3 pp 761ndash774 2012

[12] P H Young Y J Hsu and W C Yang ldquoBidens pilosa L andits medicinal userdquo in Recent Progress in Medicinal Plants DrugPlant II A S AwaadVK Singh and JNGovil Eds StandiumPress Houston Tex USA 2010

[13] C Ge ldquoCytologic study of Bidens bipinnata LrdquoZhongguo ZhongYao Za Zhi vol 15 no 2 pp 72ndash125 1990

[14] L C Chiang J S Chang C C Chen L T Ng and C CLin ldquoAnti-herpes simplex virus activity of Bidens pilosa and

48 Evidence-Based Complementary and Alternative Medicine

Houttuynia cordatardquoAmerican Journal of ChineseMedicine vol31 no 3 pp 355ndash362 2003

[15] N P Rybalchenko V A Prykhodko S S Nagorna et alldquoIn vitro antifungal activity of phenylheptatriyne from Bidenscernua L against yeastsrdquo Fitoterapia vol 81 no 5 pp 336ndash3382010

[16] K Redl W Breu B Davis and R Bauer ldquoAnti-inflammatoryactive polyacetylenes from Bidens campylothecardquo Planta Med-ica vol 60 no 1 pp 58ndash62 1994

[17] M Ayyanar and S Ignacimuthu ldquoTraditional knowledge ofKani tribals inKouthalai of Tirunelveli hills TamilNadu IndiardquoJournal of Ethnopharmacology vol 102 no 2 pp 246ndash255 2005

[18] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant intraditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[19] AKAbbas KMMurphy andA Sher ldquoFunctional diversity ofhelper T lymphocytesrdquo Nature vol 383 no 6603 pp 787ndash7931996

[20] J H Cano and G Volpato ldquoHerbal mixtures in the traditionalmedicine of Eastern Cubardquo Journal of Ethnopharmacology vol90 no 2-3 pp 293ndash316 2004

[21] J Namukobe J M Kasenene B T Kiremire et al ldquoTraditionalplants used for medicinal purposes by local communitiesaround the Northern sector of Kibale National Park UgandardquoJournal of Ethnopharmacology vol 136 no 1 pp 236ndash245 2011

[22] C Lans ldquoComparison of plants used for skin and stomachproblems in Trinidad and Tobago with Asian ethnomedicinerdquoJournal of Ethnobiology andEthnomedicine vol 3 article 3 2007

[23] S Dharmananda ldquoA Popular Remedy Ecapes Notice ofWesternPractitionersrdquo 2013 httpwwwitmonlineorgartsbidenshtm

[24] Y J Hsu T H Lee C L T Chang Y T Huang andW C YangldquoAnti-hyperglycemic effects and mechanism of Bidens pilosawater extractrdquo Journal of Ethnopharmacology vol 122 no 2 pp379ndash383 2009

[25] E Noumi F Houngue and D Lontsi ldquoTraditional medicinesin primary health care plants used for the treatment ofhypertension in Bafia Cameroonrdquo Fitoterapia vol 70 no 2 pp134ndash139 1999

[26] P V Tan T Dimo and E Dongo ldquoEffects of methanolcyclohexane and methylene chlo ride extracts of Bidens pilosaon various gastric ulcer models in ratsrdquo Journal of Ethnophar-macology vol 73 no 3 pp 415ndash421 2000

[27] R L C Pereira T Ibrahim L Lucchetti A J R Da Silva and VL G De Moraes ldquoImmunosuppressive and anti-inflammatoryeffects of methanolic extract and the polyacetylene isolatedfrom Bidens pilosa Lrdquo Immunopharmacology vol 43 no 1 pp31ndash37 1999

[28] T Dimo J Azay P V Tan et al ldquoEffects of the aqueous andmethylene chloride extracts of Bidens pilosa leaf on fructose-hypertensive ratsrdquo Journal of Ethnopharmacology vol 76 no 3pp 215ndash221 2001

[29] CWiartMedicinal Plants of Southeast Asia Pelanduk SelabgorDarul Ehsan Malaysia 2000

[30] F L Silva D C H Fischer J F Tavares M S Silva P FDe Athayde-Filho and J M Barbosa-Filho ldquoCompilation ofsecondary metabolites from Bidens pilosa LrdquoMolecules vol 16no 2 pp 1070ndash1102 2011

[31] A H Chen S R Lin and C H Hong ldquoPhytochemical studyon Bidens pilosa L varminorrdquo Huaxue pp 38ndash42 1975

[32] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[33] M C Zulueta M Tada and C Y Ragasa ldquoA diterpene fromBidens pilosardquo Phytochemistry vol 38 no 6 pp 1449ndash14501995

[34] J Wang H Yang Z W Lin and H D Sun ldquoFlavonoids fromBidens pilosa var radiatardquo Phytochemistry vol 46 no 7 pp1275ndash1278 1997

[35] A Zhao Q Zhao L Peng et al ldquoA new chalcone glycoside fromBidens pilosardquo Acta Botanica Yunnanica vol 26 no 1 pp 121ndash126

[36] P Gao Y Wu S Cui and Y Zou ldquoSynthesis and absoluteconfiguration of the 7-phenylhepta-4 6-diyne-1 2-diol isolatedfrom Bidens pilosardquo Synthesis no 13 pp 2131ndash2135 2011

[37] S Wang B Yang D Zhu D He and L Wang ldquoActivecomponents of Bidens pilosa Lrdquo Zhongcaoyao vol 36 pp 20ndash21

[38] F Bohlmann T Burkhardt and C Zdero Naturally OccuringAcetylenes Academic Press New York NY USA 1973

[39] T M Sarg A M Ateya N M Farrag and F A AbbasldquoConstituents and biological activity of Bidens pilosa L grownin EgyptrdquoActa Pharmaceutica Hungarica vol 61 no 6 pp 317ndash323 1991

[40] H A L Valdes and H P Rego ldquoBidens pilosa Linnerdquo RevistaCubana de Plantas Medicinales vol 6 pp 28ndash33

[41] F Bohlmann H Bornowski and K M Kleine ldquoNew polyynesfrom the tribe Heliantheaerdquo Chemische Berichte vol 97 pp2135ndash2138

[42] RWang Q XWu and Y P Shi ldquoPolyacetylenes and flavonoidsfrom the aerial parts ofBidens pilosardquo PlantaMedica vol 76 no9 pp 893ndash896 2010

[43] L W Wu Y M Chiang H C Chuang et al ldquoA novelpolyacetylene significantly inhibits angiogenesis and promotesapoptosis in human endothelial cells through activation of theCDK inhibitors and caspase-7rdquo Planta Medica vol 73 no 7 pp655ndash661 2007

[44] L W Wu Y M Chiang H C Chuang et al ldquoPolyacetylenesfunction as anti-angiogenic agentsrdquo Pharmaceutical Researchvol 21 no 11 pp 2112ndash2119 2004

[45] H L Yang S C Chen N W Chang et al ldquoProtection fromoxidative damage using Bidens pilosa extracts in normal humanerythrocytesrdquo Food and Chemical Toxicology vol 44 no 9 pp1513ndash1521 2006

[46] H Q Wang S J Lu H Li and Z H Yao ldquoEDTA-enhancedphytoremediation of lead contaminated soil by Bidens maxi-mowiczianardquo Journal of Environmental Sciences vol 19 no 12pp 1496ndash1499 2007

[47] S TobinagaMK SharmaWG L Aalbersberg et al ldquoIsolationand identification of a potent antimalarial and antibacterialpolyacetylene from Bidens pilosardquo Planta Medica vol 75 no 6pp 624ndash628 2009

[48] C L T Chang H K Kuo S L Chang et al ldquoThe distincteffects of a butanol fraction of Bidens pilosa plant extract onthe development of Th1-mediated diabetes and Th2-mediatedairway inflammation in micerdquo Journal of Biomedical Sciencevol 12 no 1 pp 79ndash89 2005

[49] S L Chang C L T Chang Y M Chiang et al ldquoPolyacetyleniccompounds and butanol fraction from Bidens pilosa can mod-ulate the differentiation of helper T cells and prevent autoim-mune diabetes in non-obese diabetic micerdquo Planta Medica vol70 no 11 pp 1045ndash1051 2004

Evidence-Based Complementary and Alternative Medicine 49

[50] Y M Chiang C L T Chang S L Chang W C Yang and LF Shyur ldquoCytopiloyne a novel polyacetylenic glucoside fromBidens pilosa functions as a T helper cell modulatorrdquo Journal ofEthnopharmacology vol 110 no 3 pp 532ndash538 2007

[51] R PUbillas CDMendez S D Jolad et al ldquoAntihyperglycemicacetylenic glucosides from Bidens pilosardquo Planta Medica vol66 no 1 pp 82ndash83 2000

[52] L Alvarez S Marquina M L Villarreal D Alonso E Arandaand G Delgado ldquoBioactive polyacetylenes from Bidens pilosardquoPlanta Medica vol 62 no 4 pp 355ndash357 1996

[53] C L T Chang S L Chang Y M Lee et al ldquoCytopiloyne apolyacetylenic glucoside prevents type 1 diabetes in nonobesediabetic micerdquo Journal of Immunology vol 178 no 11 pp 6984ndash6993 2007

[54] G Kusano A Kusano and Y Seyama Novel Hypoglycemicand Anti-Inflammatory Polyacetylenic Compounds Their Com-positions Bidens Plant Extract Fractions and CompositionsContaining the Plant or Fraction JPO Tokyo Japan 2004

[55] M G L Brandao A U Krettli L S R Soares C G CNery and H C Marinuzzi ldquoAntimalarial activity of extractsand fractions from Bidens pilosa and other Bidens species(Asteraceae) correlated with the presence of acetylene andflavonoid compoundsrdquo Journal of Ethnopharmacology vol 57no 2 pp 131ndash138 1997

[56] A U Krettli V F Andrade-Neto M D G L Brandao andW M S Ferrari ldquoThe search for new anti-malarial drugs fromplants used to treat fever and malaria or plants ramdomlyselected a reviewrdquoMemorias do Instituto Oswaldo Cruz vol 96no 8 pp 1033ndash1042 2001

[57] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[58] C K Wat R K Biswas E A Graham L Bohm G H NTowers and E R Waygood ldquoUltraviolet-mediated cytotoxicactivity of phenylheptatriyne from Bidens pilosa Lrdquo Journal ofNatural Products vol 42 no 1 pp 103ndash111 1979

[59] Y Sashida K Ogawa M Kitada H Karikome Y Mimakiand H Shimomura ldquoNew aurone glucosides and new phenyl-propanoid glucosides from Bidens pilosardquo Chemical and Phar-maceutical Bulletin vol 39 no 3 pp 709ndash711 1991

[60] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens pilosa L (TFB) on animal liver injury andliver fibrosisrdquo Journal of Ethnopharmacology vol 116 no 3 pp539ndash546 2008

[61] B Hoffmann and J Holzl ldquoAcylated compounds from Bidenspilosardquo Planta Medica vol 55 pp 108ndash109

[62] B Hoffmann and J Holzl ldquoA methylated chalcone glucosidefrom Bidens pilosardquo Phytochemistry vol 27 no 11 pp 3700ndash3701 1988

[63] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[64] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[65] B Hoffmann and J Holzl ldquoWeitere acylierte chalkone ausBidens pilosardquo Planta Medica vol 54 pp 450ndash451 1988

[66] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant in11traditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[67] V V Pham V V T K P V L Hoang and V K Phan ldquoFlavonoidcompounds from the plant Bidens pilosa L (Asteraceae)rdquo TapChi Duoc Hoc vol 50 pp 48ndash53

[68] S D Sarker B Bartholomew R J Nash and N Robinson ldquo5-O-methylhoslundin an unusual flavonoid from Bidens pilosa(Asteraceae)rdquo Biochemical Systematics and Ecology vol 28 no6 pp 591ndash593 2000

[69] S L Chang Y M Chiang C L T Chang et al ldquoFlavonoidscentaurein and centaureidin from Bidens pilosa stimulate IFN-120574 expressionrdquo Journal of Ethnopharmacology vol 112 no 2 pp232ndash236 2007

[70] Y M Chiang D Y Chuang S Y Wang Y H Kuo P WTsai and L F Shyur ldquoMetabolite profiling and chemopreventivebioactivity of plant extracts from Bidens pilosardquo Journal ofEthnopharmacology vol 95 no 2-3 pp 409ndash419 2004

[71] A Kusano Y Seyama E Usami et al ldquoStudies on the antioxi-dant active constituents of the dried powder from Bidens pilosaL var radiata SchrdquoNaturalMedicines vol 57 no 3 pp 100ndash1042003

[72] I Doleckova L Rarova J Gruz et al ldquoAnti-proliferative andanti-angiogenic effects of flavone eupatorin an active con-stituent of chloroform extract ofOrthosiphon stamineus leavesrdquoFitoterapia vol 83 no 6 pp 1000ndash1007

[73] K Bairwa R Kumar R J Sharma and R K Roy ldquoAn updatedreview on Bidens pilosa Lrdquo Der Pharma Chemica vol 2 no 3pp 325ndash337

[74] Q Xia Y Liu and Y Li ldquoDetermination of hyperoside indifferent parts and different species of Herba Bidens by RP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol 50pp 48ndash53

[75] M Horiuchi and Y Seyama ldquoImprovement of the antiinflam-matory and antiallergic activity of Bidens pilosa L var radiataSCHERFF treated with enzyme (Cellulosine)rdquo Journal of HealthScience vol 54 no 3 pp 294ndash301 2008

[76] M G L Brandao C G C Nery M A S Mamao and AU Krettli ldquoTwo methoxylated flavone glycosides from Bidenspilosardquo Phytochemistry vol 48 no 2 pp 397ndash400 1998

[77] P Kumari K Misra B S Sisodia et al ldquoA promising anticancerand antimalarial component from the leaves of Bidens pilosardquoPlanta Medica vol 75 no 1 pp 59ndash61 2009

[78] G Seelinger I Merfort U Wolfle and C M Schempp ldquoAnti-carcinogenic effects of the flavonoid luteolinrdquoMolecules vol 13no 10 pp 2628ndash2651 2008

[79] G Seelinger I Merfort and C M Schempp ldquoAnti-oxidantanti-inflammatory and anti-allergic activities of luteolinrdquoPlanta Medica vol 74 no 14 pp 1667ndash1677 2008

[80] C C Yit and N P Das ldquoCytotoxic effect of butein on humancolon adenocarcinoma cell proliferationrdquo Cancer Letters vol82 no 1 pp 65ndash72 1994

[81] J A Beutler E Hamel A J Vlietinck et al ldquoStructure-activityrequirements for flavone cytotoxicity and binding to tubulinrdquoJournal of Medicinal Chemistry vol 41 no 13 pp 2333ndash23381998

[82] M R Kviecinski K B Felipe T Schoenfelder et al ldquoStudy ofthe antitumor potential of Bidens pilosa (Asteraceae) used inBrazilian folkmedicinerdquo Journal of Ethnopharmacology vol 117no 1 pp 69ndash75 2008

[83] P Sundararajan A Dey A Smith A G Doss M Rajappanand S Natarajan ldquoStudies of anticancer and antipyretic activityof Bidens pilosawhole plantrdquo African Health Sciences vol 6 no1 pp 27ndash30 2006

[84] YMChiang C P Lo Y P Chen et al ldquoEthyl caffeate suppressesNF-120581B activation and its downstream inflammatory mediatorsiNOS COX-2 and PGE2 in vitro or in mouse skinrdquo BritishJournal of Pharmacology vol 146 no 3 pp 352ndash363 2005

50 Evidence-Based Complementary and Alternative Medicine

[85] J S Kim H J Lee M H Lee J Kim C Jin and J HRyu ldquoLuteolin inhibits LPS-stimulated inducible nitric oxidesynthase expression in BV-2 microglial cellsrdquo Planta Medicavol 72 no 1 pp 65ndash68 2006

[86] P A Ruiz and D Haller ldquoFunctional diversity of flavonoidsin the inhibition of the proinflammatory NF-120581B IRF and Aktsignaling pathways in murine intestinal epithelial cellsrdquo Journalof Nutrition vol 136 no 3 pp 664ndash671 2006

[87] A Xagorari A Papapetropoulos A Mauromatis MEconomou T Fotsis and C Roussos ldquoLuteolin inhibitsan endotoxin-stimulated phosphorylation cascade andproinflammatory cytokine production in macrophagesrdquoJournal of Pharmacology and Experimental Therapeutics vol296 no 1 pp 181ndash187 2001

[88] N Yoshida T Kanekura Y Higashi and T Kanzaki ldquoBidenspilosa suppresses interleukin-1120573-induced cyclooxygenase-2expression through the inhibition of mitogen activated proteinkinases phosphorylation in normal human dermal fibroblastsrdquoJournal of Dermatology vol 33 no 10 pp 676ndash683 2006

[89] M Muchuweti C Mupure A Ndhlala T Murenje and M AN Benhura ldquoScreening of antioxidant and radical scavengingactivity of Vigna ungiculata Bidens pilosa and Cleome gynan-drardquo American Journal of Food Technology vol 2 no 3 pp 161ndash168 2007

[90] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens bipinnata L against carbon tetrachloride-induced liver fibrosis in ratsrdquo Journal of Pharmacy and Pharma-cology vol 60 no 10 pp 1393ndash1402 2008

[91] S L Chang H H Yeh Y S Lin Y M Chiang T K Wuand W C Yang ldquoThe effect of centaurein on interferon-120574 expression and Listeria infection in micerdquo Toxicology andApplied Pharmacology vol 219 no 1 pp 54ndash61 2007

[92] F Deba T D Xuan M Yasuda and S Tawata ldquoChemical com-position and antioxidant antibacterial and antifungal activitiesof the essential oils from Bidens pilosa Linn var Radiatardquo FoodControl vol 19 no 4 pp 346ndash352 2008

[93] A O T Ashafa and A J Afolayan ldquoScreening the root extractsfrom Biden pilosa L var radiata (Asteraceae) for antimicrobialpotentialsrdquo Journal of Medicinal Plant Research vol 3 no 8 pp568ndash572 2009

[94] T B Nguelefack T Dimo E P Nguelefack Mbuyo P V TanS V Rakotonirina and A Kamanyi ldquoRelaxant effects of theneutral extract of the leaves of Bidens pilosa linn on isolated ratvascular smooth musclerdquo Phytotherapy Research vol 19 no 3pp 207ndash210 2005

[95] M T Grombone-Guaratini K L Silva-Brandao V N SolferiniJ Semir and J R Trigo ldquoSesquiterpene and polyacetyleneprofile of the Bidens pilosa complex (Asteraceae Heliantheae)from Southeast of Brazilrdquo Biochemical Systematics and Ecologyvol 33 no 5 pp 479ndash486 2005

[96] L L Lin C Y Wu H C Hsiu M T Wang and H ChuangldquoStudies on diabetes mellitus I The hypoglycemic activity ofphytosterin on alloxan diabetic ratsrdquo Taiwan Yi Xue Hui Za Zhivol 66 no 2 pp 58ndash65 1967

[97] M A N Benhura and I C Chitsiku ldquoThe extractable 120573-carotene content of Guku (Bidens pilosa) leaves after cookingdrying and storagerdquo International Journal of Food Science andTechnology vol 32 no 6 pp 495ndash500 1997

[98] J S Chang L C Chiang C C Chen L T Liu K C Wangand C C Lin ldquoAtileukemic activity of Bidens pilosa l varminor (blume) sherff andHouttuynia cordata thunbrdquo AmericanJournal of Chinese Medicine vol 29 no 2 pp 303ndash312 2001

[99] F Deba T D Xuan M Yasuda and S Tawata ldquoHerbicidal andfungicidal activities and identification of potential phytotoxinsfrom Bidens pilosa L var radiata Scherff research paperrdquoWeedBiology and Management vol 7 no 2 pp 77ndash83 2007

[100] J K Kumar and A K Sinha ldquoA new disubstituted acetylacetonefrom the leaves ofBidens pilosa LinnrdquoNatural Product Researchvol 17 no 1 pp 71ndash74 2003

[101] K Ogawa and Y Sashida ldquoCaffeoyl derivatives of a sugarlactone and its hydroxy acid from the leaves of Bidens pilosardquoPhytochemistry vol 31 no 10 pp 3657ndash3658 1992

[102] Q Xia Y Liu and Y Li ldquoDetermination of gallic acid fromdifferent species and different medical parts of Herba Bidens byRP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol24 no 03 pp 308ndash310

[103] W J Lee L F Wu W K Chen C J Wang and T HTseng ldquoInhibitory effect of luteolin on hepatocyte growth fac-torscatter factor-induced HepG2 cell invasion involving bothMAPKERKs and PI3K-Akt pathwaysrdquo Chemico-BiologicalInteractions vol 160 no 2 pp 123ndash133 2006

[104] J A Beutler J H Cardellina C M Lin E Hamel G MCragg andM R Boyd ldquoCentaureidin a cytotoxic flavone fromPolymnia fruticosa inhibits tubulin polymerizationrdquo BioorganicandMedicinal Chemistry Letters vol 3 no 4 pp 581ndash584 1993

[105] A Verma A Su A M Golin B OrsquoMarrah and J K AmorosaldquoThe lateral view a screening method for knee traumardquo Aca-demic Radiology vol 8 no 5 pp 392ndash397 2001

[106] J Corren M Lemay Y Lin L Rozga and R K RandolphldquoClinical and biochemical effects of a combination botanicalproduct (ClearGuard) for allergy a pilot randomized double-blind placebo-controlled trialrdquo Nutrition Journal vol 7 no 1article 20 2008

[107] M S Gachet J S Lecaro M Kaiser et al ldquoAssessment of anti-protozoal activity of plants traditionally used in Ecuador in thetreatment of leishmaniasisrdquo Journal of Ethnopharmacology vol128 no 1 pp 184ndash197 2010

[108] W C Yang M Ghiotto B Barbarat and D Olive ldquoThe role ofTec protein-tyrosine kinase in T cell signalingrdquo The Journal ofBiological Chemistry vol 274 no 2 pp 607ndash617 1999

[109] S Tewtrakul H Miyashiro N Nakamura et al ldquoHIV-1 inte-grase inhibitory substances from Coleus parvifoliusrdquo Phytother-apy Research vol 17 no 3 pp 232ndash239 2003

[110] T Dimo T B Nguelefack P V Tan et al ldquoPossible mechanismsof action of the neutral extract from Bidens pilosa L leaves onthe cardiovascular system of anaesthetized ratsrdquo PhytotherapyResearch vol 17 no 10 pp 1135ndash1139 2003

[111] T S C Li Chinese and Related North American Herbs CRCPress New York NY USA 2002

[112] H Wu H Chen X Hua Z Shi L Zhang and J ChenldquoClinical therapeutic effect of drug-separated moxibustion onchronic diarrhea and its immunologic mechanismsrdquo Journal ofTraditional Chinese Medicine vol 17 no 4 pp 253ndash258 1997

[113] S W Wright R R Harris J S Kerr et al ldquoSynthesis chemicaland biological properties of vinylogous hydroxamic acids dualinhibitors of 5-lipoxygenase and IL-1 biosynthesisrdquo Journal ofMedicinal Chemistry vol 35 no 22 pp 4061ndash4068 1992

[114] W R Almiron and M E Brewer ldquoClassification of immaturestage habitats of Culicidae (Diptera) collected in CordobaArgentinardquo Memorias do Instituto Oswaldo Cruz vol 91 no 1pp 1ndash9 1996

[115] N L Wang J Wang X S Yao and S Kitanaka ldquoTwo newmonoterpene glycosides and a new (+)-jasmololone glucoside

Evidence-Based Complementary and Alternative Medicine 51

from Bidens parvifloraWilldrdquo Journal of Asian Natural ProductsResearch vol 9 no 5 pp 473ndash479 2007

[116] P Champagnat ldquoRole of the terminal bud in the actionexercised by the cotyledon of Bidens pilosus L var radiatus onits axillary budrdquo Comptes Rendus des Seances et Memoires de laSociete de Biologie vol 145 no 17-18 pp 1374ndash1376 1951

[117] S F Nielsen S B Christensen G Cruciani A Kharazmiand T Liljefors ldquoAntileishmaniai chalcones statistical designsynthesis and three-dimensional quantitative structure-activityrelationship analysisrdquo Journal of Medicinal Chemistry vol 41no 24 pp 4819ndash4832 1998

[118] Y C Hwang J J H Chu P L Yang W Chen and M V YatesldquoRapid identification of inhibitors that interfere with poliovirusreplication using a cell-based assayrdquo Antiviral Research vol 77no 3 pp 232ndash236 2008

[119] V F Andrade-Neto M G L Brandao F Q Oliveira et alldquoAntimalarial activity of Bidens pilosa L (Asteraceae) ethanolextracts fromwild plants collected in various localities or plantscultivated in humus soilrdquo Phytotherapy Research vol 18 no 8pp 634ndash639 2004

[120] R J Marles and N R Farnsworth ldquoAnti-diabetic plants andtheir active constituentsrdquo Phytomedicine vol 2 no 2 pp 137ndash189

[121] M Habeck ldquoDiabetes treatments get sweet help from naturerdquoNature Medicine vol 9 no 10 p 1228 2003

[122] H W Lin G Y Han and S X Liao ldquoStudies on the activeconstituents of the Chinese traditional medicine Polygonatumodoratum (Mill) DrucerdquoActa Pharmaceutica Sinica vol 29 no3 pp 215ndash222 1994

[123] C L T Chang Y C Chen H M Chen N S Yang andW C Yang ldquoNatural cures for type 1 diabetes a reviewof phytochemicals biological actions and clinical potentialrdquoCurrent Medicinal Chemistry vol 20 no 7 pp 899ndash907 2013

[124] L Dey A S Attele and C S Yuan ldquoAlternative therapies fortype 2 diabetesrdquo Alternative Medicine Review vol 7 no 1 pp45ndash58 2002

[125] C L T ChangH Y Liu T F Kuo et al ldquoAnti-diabetic effect andmode of action of cytopiloynerdquo Evidence-Based Complementaryand Alternative Medicine vol 2013 Article ID 685642 13 pages2013

[126] M R Khan M Kihara and A D Omoloso ldquoAnti-microbialactivity of Bidens pilosa Bischofia javanica Elmerillia papuanaand Sigesbekia orientalisrdquo Fitoterapia vol 72 no 6 pp 662ndash6652001

[127] T Dimo T B Nguelefack P Kamtchouing E Dongo ARakotonirina and S V Rakotonirina ldquoHypotensive effects ofmethanol extract from Bidens pilosa Linn on hypertensive ratsrdquoComptes Rendus de lrsquoAcademie des Sciences Serie III vol 322no 4 pp 323ndash329 1999

[128] A K Hassan O Deogratius J F Nyafuono O Francis and OP Engeu ldquoWound healing potential of the ethanolic extracts ofBidens pilosa and Ocimum suaverdquo African Journal of Pharmacyand Pharmacology vol 5 no 2 pp 132ndash136 2011

[129] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[130] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[131] C Kandaswami and EMiddleton ldquoFree radical scavenging andantioxidant activity of plant flavonoidsrdquo Advances in Experi-mental Medicine and Biology vol 366 pp 351ndash376 1994

[132] K Yamamoto H Kakegawa H Ueda et al ldquoGastric cytopro-tective anti-ulcerogenic actions of hydroxychalcones in ratsrdquoPlanta Medica vol 58 no 5 pp 389ndash393 1992

[133] J O C Ezeonwumelu A K Julius C N Muhoho et alldquoBiochemical and histological studies of aqueous extract ofBidens pilosa leaves from Ugandan Rift valley in ratsrdquo BritishJournal of Pharmacology and Toxicology vol 2 no 6 pp 302ndash309

[134] L Frida S Rakotonirina A Rakotonirina and J P SavineauldquoIn vivo and in vitro effects of Bidens pilosa L (Asteraceae)leaf aqueous and ethanol extracts on primed-oestrogenized ratuterine musclerdquo African Journal of Traditional Complementaryand Alternative Medicines vol 5 no 1 pp 79ndash91 2008

52 Evidence-Based Complementary and Alternative Medicine

Composition Comments

1 Please provide valid postal code to the address if

possible

2 We made the highlighted change in Table 4 for the sake

of correctness Please check

3 Please check the correctness of the highlighted part in

Table 5

4 We considered the highlighted part in Table 11 as

reference Please check similar cases throughout

5 Please check the format of Table 18

6 We reformatted Table 20 Please check

7 We made the highlighted change in Table 4 for the sake

of consistency Please check similar cases throughout the

paper

8 We made the highlighted change(s) for the sake

of consistency Please check simila highlighted cases

throughout

9 We made the highlighted change in Table 9 for the sake

of clarity Please check similar cases through out

10 We made the highlighted change(s) for the sake of

correctness Please check

11 Comment on ref [66] This reference is a repetition

of [18] Please check similar cases throughout

Author(s) Name(s)

Author 1

Author 2

Author 3

8 Evidence-Based Complementary and Alternative Medicine

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

36 111-Tridecadiene-3579-tetrayne

Roots (notstated) [38]

37 1-Tridecene-357911-pentayne Pentayneene

Leaves (notstated) andnot found(Egypt)

[38 39]

38 5-Tridecene-7911-triyne-3-ol

HO

not found(Egypt) [39]

39 21012-Tridecatriene-468-triyn-1-ol

OHPart not specified(Not stated) [40]

40 212-Tridecadiene-46810-tetrayn-1-ol

111-Tridecadiene-3579-

tetrayn-13-ol

OH Roots (Notstated) notfound(Egypt)

[38 39]

41 212-Tridecadiene-46810-tetraynal

111-Tridecadiene-3579-

tetrayn-13-al

O

Roots (Germany) [41]

42212-Tridecadiene-46810-tetrayn-1-ol1-acetate

111-Tridecadiene-3579-

tetrayn-13-acetateO

O

Roots (notstated) [38]

43(5E)-15-Tridecadiene-79-diyn-3413-triol OH

OH

HO

Aerial (China) [42]

44(6E12E)-3-oxo-tetradeca-612-dien-810-diyn-1-ol

OH

O

Aerial (China) [42]

45 (E)-5-Tridecene-7911-triyne-12-diol

12-Dihydroxy-5(E)-tridecene-7911-triyne

HO

OHWhole (Taiwan) [43]

46 (E)-6-Tetradecene-81012-triyne-13-diol

13-Dihydroxy-6(E)-tetradecene-81012-triyne

OH

OHWhole (Taiwan) [43ndash45]

47(2R3E11E)-311-Tridecadiene-579-triyne-12-diol

Safynol

OH

HO

Not found (Egyptand China) [37 39]

Evidence-Based Complementary and Alternative Medicine 9

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

4857911-Tridecatetrayne-12-diol

12-Dihydroxytrideca-57911-tetrayne

HOOH

Whole (Taiwan) [43 44 46]

49(R)-357911-Tridecapentayne-12-diol

(R)-12-Dihydroxytrideca-357911-pentayne

HO

OH

Aerial (Japan) [47]

50

(4E)-1-(Hydroxymethyl)-4-dodecene-6810-triyn-1-yl-120573-D-glucopyranoside

2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-

tridecene-7911-triyne

O

HO

H

OH

H

H

HOH

O

OH

HO

H Aerial (USA)whole (Taiwan)leaves (Taiwan)

[8 45 48ndash51]

51

(4E)-1-(2-Hydroxyethyl)-4-dodecene-6810-triyn-1-yl-120573-D-glucopyranoside

3-120573-D-Glucopyranosyloxy-1-hydroxy-6(E)-

tridecene-81012-triyne

OHO

H

OH

H

H

HOH

O

OH

OH

HAerial (USA andChina) whole(Taiwan) leaves(Taiwan)

[8 35 4245 48ndash51]

52

3-Hydroxy-6-tetradecene-81012-triynyl-120573-D-glucopyranoside

120573-D-Glucopyranosyloxy-

3-hydroxy-6E-tetradecence-81012-triyne

OH

O

OHO

H

OH

H

H

HOH

OH

HWhole (Mexico) [52]

53

1-(Hydroxymethyl)-46810-dodecatetrayne-1-yl-120573-D-glucopyranoside

Cytopiloyne2-120573-D-

Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne

O

HO

OHO

H

OH

H

H

HOH

OH

HWhole (Taiwan)leaves (Taiwan) [8 50 53]

54

2-O-D-Glucosyltrideca-11E-en-3579-tetrayn-12-diol

OH

O

O

OHH

HO

H

H

HHOH

OH

Leaves (Brazil) [27]

10 Evidence-Based Complementary and Alternative Medicine

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

55

(R)-1-(Hydroxymethyl)-246810-dodecapentayn-1-yl-a-D-glucopyranoside

2- 120573 -D-Glucopyranosyloxy-1-hydroxytrideca-357911-pentayne

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Chinaand Japan) [35 47]

56

1-[[(Carboxyacetyl)oxy]methyl]46810-dodecatetraynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Japan) [54]

57

(4E)-1-[[(Carboxyacetyl)oxy]-methyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHO

H

OH

Aerial (Japan) [54]

58

(4E)-1-[[(Carboxyacetyl)oxy]-ethyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

O

O

O

OHO

O

OHOH

H

HO

H

H

HHO

HOH

Aerial (Japan) [54]

59 (5E)-5-Heptene-13-diyn-1-yl-benzene

1-Phenylhepta-13-diyn-5en Whole (Taiwan) [32]

60 7-Phenyl-2(E)-heptene-46-diyn-1-ol

OHRoots (notstated) Aerial(China)

[38 42]

617-Phenyl-2(E)-heptene-46-diyn-1-ol-acetate

OO

Roots (notstated Brazil) [38 55 56]

62 7-Phenyl-46-heptadiyn-2-ol Pilosol A

OH

Whole (Taiwan)aerial (China) [42 57]

Evidence-Based Complementary and Alternative Medicine 11

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

63 7-Phenylhepta-46-diyn-12-diol HO

OH

Aerial (China) [42]

64 135-Heptatriyn-1-yl-benzene

1-Phenylhepta-135-triyne

Leaves (notstated) leaves oftissue culture(not stated)aerial (TanzaniaChina)Whole (Taiwan)roots (Brazil)

[18 34 3842 56ndash58]

65 7-Phenyl-246-heptatriyn-1-ol HO

Leaves (notstated) aerial(China)

[38 42]

667-Phenyl-246-heptatriyn-1-ol-acetate O

O Leaves (notstated) [38]

675-(2-Phenylethynyl)-2-thiophenemethanol

S OHAerial (China) [42]

685-(2-Phenylethynyl)-2120573-glucosylmethyl-thiophene

O

O

OH

H

OH

H

H

HOH H

O

HO

S

Aerial (China) [42]

69

3- 120573 -D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne

OH

O

OH

HOHO

H

H

HHOH

OH

Leaves(Cameroon) [28]

701-Phenyl-13-diyn-5-en-7-ol-acetate O

O

Leaves (Brazil) [27]

SN denotes serial number

the chloroform fraction was the most toxic with a halfmaximal inhibitory concentration (IC

50) of 97 plusmn 72 and 83 plusmn

52 120583gmL in NRU and MTT respectively [82] Kumari andcolleagues also reported the anti-cancer and anti-malarialactivities of B pilosa leaves [77] Based on a cytotoxicity-directed fractionation strategy they identified phenyl-135-heptatriene with IC

50values of 8plusmn001 049plusmn045 07plusmn001

and 10 plusmn 001 120583gmL against human oral liver colon andbreast cancer cell lines respectively However phenyl-135-heptatriyne showed lower activity against breast cancer celllines than the chloroform leaf extract which had an IC

50

value of 65 plusmn 001 120583gmL Moreover the positive controltaxol showed higher activity than phenyl-135-heptatriyne[77] Furthermore in vitro comet assays were performed toevaluate the toxicity of n-hexane chloroform and methanolextracts of B pilosa and its ethyl acetate acetone and waterfractions onHela andKB cellsThe ethyl acetate fraction fromthe methanol extract exhibited the highest activity with halfmaximal cytotoxic concentrations (CTC

50) of 9652120583gmL

and 5862120583gmL against Hela and KB cells respectivelyDespite the moderate toxicity these findings suggest thatthese B pilosa extractsfractions could be useful for future

12 Evidence-Based Complementary and Alternative Medicine

Table5Flavon

oids

isolated

from

Bpilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

71

2-[(34-Dihydroxyph

enyl)-

methylene]-

6-hydroxy-3(2H

)-benzofuranon

e

Sulfu

retin

O

OHO

OH

OH

Aeria

l(Ch

ina)

[35]

72

2-[(34-Dihydroxyph

enyl)-

methylene]-

67-dihydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-673101584041015840-

tetrahydroxy

maritimetin

O

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

73

2-[(34-Dihydroxyph

enyl)-

methylene]-6-

(120573-D

-glucopyrano

syloxy)-7-

hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-120573-D

-glucop

yranosyl-

673101584041015840-te

trahydroxy

maritimein

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japanand

China)aerial(Ch

ina)

[355960]

74

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

(120573-D

-glucopyrano

syloxy)-

6-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-7-O-120573-D

-glucop

yranosyl-

673101584041015840-

tetrahydroxy

O

OHO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japan)

[59]

75

6-[(6-O-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-acetyl-

120573-D

-glucopyrano

syl)

-673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OHO

H HO

H

H

HOH

HO

O

O

Leaves

(Not

stated)

aeria

l(Ch

ina)

[3559]

Evidence-Based Complementary and Alternative Medicine 13

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

76

6-[(36-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-

2-[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone(Z)-6-O

-(36-di-O

-acetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

bideno

sideA

O

O

HO

OH

OH

OHOH O

H

H

HOH

HO

O

O

O

Leaves

(China)

[60]

77

6-[(46-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(410158401015840610158401015840-

diacetyl-120573-D

-glucropyrano

syl)-

673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

HO

O

OO

Leaves

(not

stated)

aeria

l(Ch

ina)

[343561]

78

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

hydroxy-6-

[(246-tri-O

-acetyl-120573

-D-

glucop

yranosyl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(210158401015840410158401015840610158401015840-

triacetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

O

OOO

O

Leaves

(not

stated)

aeria

l(Ch

ina)

[3461]

79

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-6-

[(346-tri-O

-acetyl-120573

-D-glucopyrano

syl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(310158401015840410158401015840610158401015840-

triacetyl-120573

-D-glucrop

yranosyl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H OH

H

HOH

HO

OOO O

Aeria

l(Ch

ina)

[3435]

80

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-

6-[[6-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propenyl]-120573-

D-glucopyrano

syl]o

xy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-

p-coum

aroyl-

120573-D

-glucropyrano

syl)-

673101584041015840-te

trahydroxy

OH

O

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OLeaves

(Japan)

[59]

14 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

81

1-[2-(120573-D

-Glucopyrano

syloxy)-

4-hydroxyphenyl]-2-

hydroxy-3-(3-

hydroxyphenyl)-2-

prop

en-1-on

e

Chalcone120572

32101584041015840-

tetrahydroxy-

21015840-O

-120573-D

-glucop

yranosyl

HO

O

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[35]

821-(24-Dihydroxyph

enyl)-

3-(34-dihydroxyph

enyl)-

2-prop

en-1-on

eBu

tein

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

833-(34-D

ihydroxyph

enyl)-

1-(234-trihydroxyph

enyl)-

2-prop

en-1-on

eOkanin

O

HO

HO

OH

OH

OH

Leaves

(China)

[60]

84

3-(34-D

ihydroxyph

enyl)-

1-[3-(120573-D

-glucop

yranosyloxy)-

24-dihydroxyphenyl]-2-

prop

en-1-on

e

Okanin

31015840-O

-120573-D

-glucosid

e

O

HO

OH

OH

OH

OO

HO

H

HO

H

H

HOH

H

OH

Leaves

(Germany)

flowers(Germany)

[61ndash63]

85

3-(34-D

ihydroxyph

enyl)-

1-[4-(120573-D

-glucop

yranosyloxy)-

23-dihydroxyphenyl]-

2-prop

en-1-on

e

Okanin

41015840-O

-120573-D

-glucosid

e

OOH

OH

OH

HOO

OHO

H

HO

H

H

HOH

H

OH

Flow

ers(Germany)

leaves

(Japan)

[5963]

Evidence-Based Complementary and Alternative Medicine 15

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

86Okanin41015840-O

-120573-D

-(610158401015840-

O-acetylglucosid

e)

O

O

OH

OH

OH

OHO

H

HO

H

H

HOH

H

HO

O

O

Flow

ers(Germany)

[63]

87

1-[4-[(46-di-O

-Acetyl-

120573-D

-glucopyrano

syl)-oxy]-

23-dihydroxyphenyl]-3-

(34-dihydroxyph

enyl)-2-

prop

en-1-on

e

Okanin

41015840-O

-120573-D

-(410158401015840610158401015840-diacetyl)-

glucop

yranoside

O

O

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Aeria

l(Ch

ina)

[34]

88Okanin41015840-O

-120573-D

-(210158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[64]

89Okanin41015840-O

-120573-D

-(310158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

O

OAe

rial(Ch

ina)

[34]

16 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

90

1-[23-Dihydroxy-4-

[[6-O-[3-(4-hydroxyph

enyl)-

1-oxo-2-prop

enyl]-120573-D

-glucop

yranosyl]oxy]-ph

enyl]-

3-(34-dihydroxyph

enyl)-

2-prop

en-1on

e

Okanin

41015840-O

-120573-D

-(610158401015840-tr

ans-

p-coum

aroyl)-

glucoside

O

O

OH

OH

OH

OHOH

HO

H

H

HOH

H

HO

OO

OH

Leaves

(Germany)

[64]

91Okanin41015840-O

-120573-D

-(410158401015840-acetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Leaves

(Germany)

[65]

92

Okanin

41015840-O

-120573-D

-(210158401015840410158401015840-diacetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[65]

Evidence-Based Complementary and Alternative Medicine 17

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

93

Okanin

41015840-O

-120573-D

-(310158401015840410158401015840-diacetyl-610158401015840-

trans-p-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

OH

O O

Leaves

(Germany)

[65]

94

Okanin

41015840-O

-[120573-D

-glucopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

OOH

OH

OH

HO

OH

HO

HHO

H

HOH

HO

OH

OH

HO

HHO

H

HOH

HO

Flow

er(G

ermany)

[63]

95Okanin

3101584041015840-di-O

-120573-D

-glucosid

e

OOH

OH

OH

O OO

H HO

HHOH

HOH

HHO

OHO

H

HO

H

H

HOH

H

OH

Flow

er(G

ermany)

[63]

18 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

96

1-[3-(120573-D

-Glucopyrano

syloxy)-

24-dihydroxyphenyl]-

3-(3-hydroxy)-

4-metho

xyph

enyl)-

2-prop

en-1-on

e

Okanin

4-methylether-

31015840-O

-120573-D

-glucop

yranoside

OOH

O

OH

HO

O

H HO

HHOH

HOH

H

OHO

Leaves

(Germany)aerial

(China)

[3562]

97

Okanin4-methylether-

3101584041015840-di-O

-120573-(41015840101584061015840101584041015840101584010158406101584010158401015840-

tetraacetyl)-

glucop

yranoside

OOH

OOH

O

HH

OH

HO

HOH

H

OO

O

Ac

Ac

AcAc

OO

H

HO

H

H

HOH

H

O

Aeria

l(Ch

ina)

[42]

98Ch

alcone210158404101584061015840-tr

imetho

xy-

4-O-D

-glucopyrano

syl-

dihydro

Not

foun

dLeaves

(China)

[60]

992-(34-D

ihydroxyph

enyl)-23-

dihydro-78

-dihydroxy-

4H-1-

benzop

yran-4-one

Okanin

isoOO

OH

HO

OH

OH

Leaves

(China)

[60]

100

2-(34-D

ihydroxyph

enyl)-

23-dihydro-8-hydroxy-

7[(246-tr

i-O-acetyl-120573

-D-

glucop

yranosyl)oxy]-4H

-1-b

enzopyran-4-on

eOO

OH

OH

OH

OH

O

HHO

H

HO

HO

OAc

Ac

Ac

Aeria

l(Ch

ina)

[34]

Evidence-Based Complementary and Alternative Medicine 19

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

101

57-Dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

OO

OH

HO

OH

Aeria

l(TanzaniaC

hina)

[4266]

102

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

7-O-glucopyrano

side

OO

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Tanzania)

[66]

103

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-4H

-1-benzop

yran-4-one

Luteolin

OO

HO

OH

OH

OH

Aeria

l(TanzaniaC

hina

Vietnam)

[343542

6667]

104

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5hydroxy-4H-1-

benzop

yran-4-one

Luteolin

7-O-120573-D

-glucop

yranoside

OOOH

OH

OH

OO

HO

H

HO

H

H

HOH

OH H

Aeria

l(Tanzania)

[66]

105

57-Dim

etho

xy-6-

(5-m

etho

xy-6-m

ethyl-

4-oxo-4H

-pyran-3-yl)-

2-ph

enyl-4H-1-

benzop

yran-4-one

5-O-M

ethylhosludin

O OOO

O

O

O

Aeria

l(Ugand

a)[68]

20 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

106

3-(120573-D

-Glucopyrano

syloxy)-

57-dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Astragalin

Kaem

pferol-3-O

-120573-D

-glucopyrano

side

HO

OH

O O

OH

O

O

HO

HHO

H

H

HOH

OH

H

Aeria

l(Ch

ina)

[35]

107

Kaem

pferol3-

(23-di-E

-p-cou

maroyl-

120572-L-rhamno

pyrano

side)

Not

foun

dAe

rial(Vietnam)

[67]

108

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5-hydroxy-36-dimetho

xy-

4H-1-

benzop

yran-4-one

Axillarosid

eO O

OH

OH

OO

OH

OO

HO

H

HO

H

H

HOH

H

OH

Aeria

l(Ch

ina)

[42]

109

57-Dihydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaureidin

O

O

O

O

O

HO

OH

OH

Who

le(Taiwan)

[69]

110

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaurein

O O

OO

O OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan)w

hole

(Taiwan)

[69ndash

71]

Evidence-Based Complementary and Alternative Medicine 21

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

111

5-Hydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-

67-dimetho

xy-4H-

chromen-4-one

Eupatorin

iso

OO O

OH

O

O OH

Not

foun

d(C

hina)

[3772]

112

2-(34-D

imetho

xyph

enyl)-7-

(120573-D

-glucopyrano

syloxy)-35-

dihydroxy-

8-metho

xy-4H-1-

benzop

yran-4-one

O O

OH

O O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

113

7-(120573-D

-Glucopyrano

syloxy)-

5hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-38-

dimehtoxy-4H-1-

benzop

yran-4-one

O O

O

OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

114

Isorhamnetin

3-[O

-120572-

L-rham

nopyrano

syl-

(1-2)-120573-D

-glucop

yranoside]

Not

foun

dAe

rial(Vietnam)

[67]

115

7-[(6-deoxy-120572-L-

Manno

pyrano

syl)o

xy]-

3-120573-D

-glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-4H

-1-benzop

yran-4-one

Luteoside

O O

OH

O

OH

OH

OH

HOH

H

H HO

HO

HO

OH

HO

H

HO

OH

OH

HH

Aeria

l(Ch

ina)

[42]

22 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

116

Luteolin

3-O-120573-D

-Glucopyrano

side

O O

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

HO

Aeria

l(Tanzania)

[66]

117

57-Dihydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Quercetagetin

3631015840-tr

imethylether

OOO

O

O OH

HO

OH

Aeria

l(Ch

ina)

[42]

118

6-((2S3S4S5S)-

Tetrahydro-345-tr

ihydroxy-

6-(hydroxymethyl)-2H

-pyran-2-yloxy)-5-hydroxy-

2-(4-hydroxy-3-

metho

xyph

enyl)-

37-dimetho

xy-4H-

chromen-4-one

Quercetagetin

3731015840-

trim

ethylether-6-O

-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[42]

119

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

JaceinQ

uercetin

3631015840-

trim

ethyl

ether-7-O-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan

China)

who

le(Taiwan)

[425470]

120

2-(34-D

ihydroxyph

enyl)-

357-trihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

OO

HO

OH

OH

OH

OH

Aeria

l(Ch

ina)leaves

(China)who

le(C

hina)

[356074]

Evidence-Based Complementary and Alternative Medicine 23

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

121

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-galactopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-4-one

Quercetin

3-O-120573-D

-galactosidehyperin

hyperosid

e

O O

HO

OH

OH

OH

OOHH

HOH

H

H HO

HO

OH

Aeria

l(TanzaniaJapan)

Leaves

(China)Who

le(C

hina)

[376066

717475]

122

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-glucopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

3-O-120573-

D-glucopyrano

side

O O

HO

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

Aeria

l(TanzaniaC

hina

Japan)L

eaves(Japan)

[355966

75]

123

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-

4-oxo-4H

-1-benzop

yran-3-yl-

120573-D

-glucopyrano

siduron

icacid

Quercetin

3-O-120573-D

-glucuron

opyranoside

O O

HO

OH

OH

OH

OHOH

HOH

H

H HO

HO

OHO

Aeria

l(TanzaniaJapan)

[6671]

124

3-[[6-O-(6-

Deoxy-120572-L-m

anno

pyrano

syl)-

120573-D

-galactopyrano

syl]o

xy]-

2-(34-dihydroxyph

enyl)-57-

dihydroxy-4H

-1-benzop

yran-4-one

Quercetin

3-O-rob

inob

iosid

e

O O

HO

OH

OH

OH

O

HOHH OH

HO

H HH

OO

OHH

HOH

H

H HO

HO

Aeria

l(Japan)W

hole

(Taiwan)

[7071]

24 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

125

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-

3-metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

OH

Roots(Brazil)

[55676]

126

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-

120573-D

-glucopyrano

syl]o

xy]-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120572-

L-rham

nopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

O

HHO HHO

HO H

HH

ORo

ots(Brazil)

[5676]

127

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-120573-D

-glucop

yranosyl]oxy]-5-

hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-3-metho

xy-

4H-1-

benzop

yran-4-one

Quercetin

341015840-

dimethyl

ether-7-O-rutinoside

O OOH

OH

O

O

OH O

HOH

H

HHO

H

O

HHO HHO

HO H

HH

O

HO

Aeria

l(Ch

ina)

[3435]

128

2-(34-D

ihydroxyph

enyl)-3-

(120573-D

-glucofurano

syloxy)-57-

dihydroxy-4H

-1-benzop

yran-

4-on

e

Isoq

uercitrin

O OOH

OH

OHO

H

OHH

OH

HO

H O HOH

OH

Aeria

l(Japan

China)

[3571]

Evidence-Based Complementary and Alternative Medicine 25

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

129

Quercetin

3-O-120573-D

-galactop

yranoside

O

OH

HO

OH OH

O

O

HO

OH

OHOH

OAe

rial(no

tstated)

[18]

130

Quercetin

3-O-rutinoside

O

O

R

OO

OH

OHO

H

O

OH

OH

H3C

H3C

H3C

OH

HO

Who

le(Taiwan)

[70]

26 Evidence-Based Complementary and Alternative Medicine

Table 6 Terpenoids isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(Country) References

131371111-Tetramethylbicyclo[810]undeca-

26-dieneBicyclogermacrene Leaves

(Brazil) [95]

13241111-Trimethyl-8-

methylenebicyclo[720]undec-4-ene

E-Caryophyllene Leaves(Brazil) [95]

1331-Methyl-5-methylene-8-

(1-methylethyl)-16-cyclodecadiene

Germacrene D Leaves(Brazil) [95]

1344-(15-Dimethyl-4-hexen-1-ylidene)-1-methyl-cyclohexene

Z-120574-Bisabolene Leaves(Brazil) [95]

135Decahydro-114-trimethyl-7-methylene-1H-cycloprop[e]-

azulene120573-Gurjunene Leaves

(Brazil) [95]

136 2669-Tetramethyl-148-cycloundecatriene

120572-Humulene120572-caryophyllene

Leaves(Brazil) [95]

13712344a568a-Octahydro-1-isopropyl-7-methyl-4-methylenenaphthalene

120575-Muurolene Leaves(Brazil) [95]

138

12344a568a-Octahydro-4a8-dimethyl-2-(1-methylethylidene)-

naphthalene

Selina-37(11)-diene

Leaves(Brazil) [95]

139(2E7R11R)-371115-

Tetramethyl-2-hexadecen-1-ol

Phytol

HO

Whole(Taiwan) [57]

Evidence-Based Complementary and Alternative Medicine 27

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

140 371115-Tetramethyl-2-hexadecenoic acid Phytanic acid

O

OH

Whole(Taiwan) [57]

141371115-Tetramethyl-2-

hexadecenyl ester-heptanoicacid

Phythylheptanoate O O

Leaves(Not stated) [33]

142

(3S10R13R)-17-((2R5R)-5-Ethyl-6-methylheptan-2-yl)-

2347891011121314151617-tetradecahydro-

1013-dimethyl-1H-cyclopenta[a]phenanthren-

3-ol

Campestrol

HO

Aerial(Tanzania) [18]

143 Not found Phytosterin-B Not foundNot found(TaiwanEgypt)

[39 96]

144 Stigmast-5-en-3-ol 120573-sitosterol

HOH

H

H

Aerial(Tanzania)

whole(Taiwan)

[18 31 57]

145

1314151617-Tetradecahydro-1013-

dimethyl-1H-cyclopenta[a]phenanthren-

3-yloxy)-tetrahydro-6-(hydroxymethyl)-2H-pyran-345-triol

120573-Sitosterolglucoside

H

H

HOH

HO

HHO

H

HOHH O

OH Not found(Egypt) [39]

146 5120572-Stigmasta-7-en-3120573-ol

HO

Whole(Taiwan) [57]

147 5120572-Stigmasta-722t-dien-3120573-ol

HO

Whole(Taiwan) [57]

148 Stigmasta-522-dien-3-ol Stigmasterol

HOH

H

H

Not found(Taiwan)aerial

(Tanzania)leaves (notstated)whole

(Taiwan)

[18 31 3357]

28 Evidence-Based Complementary and Alternative Medicine

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

149 Lup-20(29)-en-3-ol Lupeol

H

HH

HOH

Not found(Egypt) [39]

150 Lup-20(29)-en-3-ol acetate Lupeol acetate

H

HH

O

OH

Not found(Egypt) [39]

151 Olean-12-en-3-ol 120573-amyrin

H

HOH

Not found(Egypt) [39]

152 5913-Trimethyl-242526-trinoroleanan-3-ol Friedelan-3120573-ol

HO

Aerial(Tanzania) [18]

153 5913-Trimethyl-242526-trinoroleanan-3-one

Friedelinfriedelan-3-one

O

Aerial(Tanzania) [18]

1542610151923-

Hexamethyl-2610141822-tetracosahexaene

Squalene

Aerial(Tanzania)Leaves (Notstated)Whole(Taiwan)

[18 33 57]

155 120573120573-Carotene 120573-Carotene Leaves (Notstated) [97]

Evidence-Based Complementary and Alternative Medicine 29

studies [83] Hot water extracts of B pilosa varminor Sheriffwere also assessed for its antileukemic effects on leukemiccell lines L1210 U937 K562 Raji and P3HR1 using XTT-based colorimetric assays The extract inhibited the five celllines with IC

50values ranging from 145120583gmL to 586120583gmL

L1210 K562 Raji and P3HR1 were more sensitive to B pilosaextract with IC

50values below 200120583gmL [98]

Consistent with the antitumor activities of B pilosaextracts and fractions some of its phytochemicals alsoshowed anticancer activity as outlined in Table 11 Amongthem luteolin (103) a well-studied flavonoid with multiplebioactivities was more effective against tumor cell prolifera-tion than its derivatives with IC

50values ranging from 3 120583M

to 50120583M in cells and 5 to 10mgkg in animals Luteolin wasalso found to fight cancer as a food additive at concentrationsof 50 to 200 ppm [78] and prevent skin cancer [78] and cancer3invasion [103] Lee and colleagues reported that luteolinprevents cancer by inhibiting cell adhesion and invasion[103] Significant inhibition concentration was reported to be5 120583M and complete inhibition concentration was reported tobe 40120583M Moreover luteolin was reported to inhibit hepa-tocyte growth factor (HGF)-induced cell scattering as wellas cytoskeleton changes such as filopodia and lamellipodiawhichwas determined using phase-contrast and fluorescencemicroscopy Furthermore luteolin also inhibited the HGF-induced phosphorylation of c-Met ERK 12 and Akt aswell as the MAPKERK and P13K-Akt pathways [78 103]Other mechanisms underlying the anticancer activities ofluteolin are the inhibition of topoisomerase I and II whichinhibits cell replication and DNA repair thus promotingapoptosis regulation of PI-3-KinaseAktMAPKERKJNKactivation of apoptosis in the mitochondrial pathway byactivating caspase 9 and caspase 3 which were found inmalignant cells but not in normal human peripheral bloodmononuclear cells death receptor-induced apoptosis and acell-cycle arrest mechanism inhibition of fatty acid synthasewhich is upregulated in many cancer cells and sensitizationto chemotherapywhereby luteolin increases the susceptibilityof cancer cells to chemotherapy [78] Butein (82) is anotherflavonoid that showed a cytotoxic effect on human colonadenocarcinoma cell proliferation with a reported IC

50value

of 175 120583M Butein at 2 120583M affected the incorporation of[14C]-labeled leucine thymidine and uridine which cancause the inhibition of DNA RNA and protein synthesis ofhuman colon cancer cells Moreover butein also exhibitednoncompetitive inhibition of 1-chloro-24-dinitrobenzene(CDNB) in glutathione S-transferase (GST) activity Tumorresistance was correlated with high levels of GST thus buteininhibited proliferation of cancer cells [80] Another flavonoidpresent in B pilosa centaureidin (109) also showed anti-cancer activity in B lymphoma cells Centaureidin isolatedfrom Polymnia fruticosa inhibited tubulin polymerization invitro and induced mitotic figure formation in CA46 Burkittlymphoma cells Using turbidimetric assay the IC

50value of

centaureidin in inhibition of mitosis was 3120583M [104] Cyto-toxicity of centaureidin was further analyzed using AmericanNational Cancer Institute (NCI) 60 human tumor cell linesThe cytotoxicity potency of centaureidin expressed as GI

50

(50 growth inhibition in the NCI tumor line panel) was024120583M [81] These data mark centaureidin as a promisingantimitotic agent for tumor therapy

In addition to anti-tumor flavones polyynes found in B 4pilosa have also been shown to possess anti-tumor proper-ties Based on a bioactivity-directed isolation approach Wuand colleagues identified two polyyne aglycones from theethyl acetate fraction of B pilosa [44] 12-Dihydroxytrideca-57911-tetrayne (48) and 13-Dihydroxy-6(E)- tetradecene-81012-triyne (46) exhibited significant anticell prolifera-tion activity in primary human umbilical vein endotheliumcells (HUVEC) with IC

50values of 125 120583M and 173 120583M

respectively They also decreased angiogenesis and pro-moted apoptosis in human endothelial cells Their anti-angiogenic and cytotoxic effects correlated with activationof the CDK inhibitors and caspase-7 [44] In addition 12-Dihyroxy-5(E)-tridecene-7911-triyne (45) showed antian-giogenic effects in HUVECs with an IC

50value of 124 120583M as

evidenced by a decrease in the tube formation and migrationof HUVECs [43] The IC

50value of compound 45 in the

inhibition of basic fibroblast growth factor-induced HUVECgrowth was 282120583M However it had higher IC

50values

than those for lung carcinoma cells and keratinocytes Thiscompound could also inhibit cell proliferation of HUVECslung carcinoma A549 cells and HACAT keratinocytes Themechanism by which compound 45 inhibits HUVEC growthand angiogenesis is complicated and includes decreasing theexpression of cell cycle regulators (CDK4 cyclins D1 and Aretinoblastoma (Rb) and vascular endothelial growth factorreceptor 1) caspase-mediated activation of CDK inhibitorsp21 (Cip1) and p27 (Kip) upregulation of Fas ligand expres-sion downregulation of Bcl-2 expression and activation ofcaspase-7 and poly (ADP-ribose) polymerase [43]

32 Anti-Inflammatory Activity B pilosa is commonly usedto treat inflammatory disorders The anti-inflammatory phy-tochemicals present in B pilosa are listed in Table 11 5Cyclooxygenase-2 (COX-2) is a physiologically importantenzyme that converts arachidonic acid to prostaglandin(PGE2) Its expression is induced by a wide variety of external

stimuli indicating its involvement in inflammatory diseasesand it is used as an inflammatory marker [88] Yoshida andcolleagues studied the effects of the aqueous extracts of Bpilosa aerial parts in the production of COX-2 and PGE

2as

well as on the activation of mitogen activated protein kinases(MAPKs) in normal human dermal fibroblasts (HDFs) inresponse to inflammatory cytokine IL-1120573 This work showedthat IL-1120573 activated MAPKs such as ERK12 p38 and JNKto different extents and induced COX-2 expression TheCOX-2 expression in HDFs was regulated mainly by p38following IL-1120573 stimulation Consistently the p38 inhibitorSB203580 blocked this expression Using this cell platformB pilosa extracts were tested for inhibition of inflammationThe extract dose-dependently suppressed the activation ofp38 and JNK and moderately suppressed ERK12 as wellas suppressing COX-2 expression and PGE

2production

30 Evidence-Based Complementary and Alternative Medicine

Table7Ph

enylprop

anoids

isolatedfro

mB

pilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

156

3-(4-H

ydroxyph

enyl)-2-prop

enoica

cid

p-Cou

maricacid

OHO

OH

Who

le(Ja

pan)

[99]

157

2-Metho

xy-4-(2-prop

en-1-yl)-ph

enol

Eugeno

lOOH

Leaves

androots(Japan)

[99]

158

3-(4-H

ydroxy-3-m

etho

xyph

enyl)-

2-prop

enoica

cid

Ferulic

acid

O

OH

O

HO

Who

le(Ja

pan)

[99]

159

3-(34-D

ihydroxyph

enyl)-2-prop

enoic

acid

Caffeicacid

O

HO

OH

OH

Who

leandaeria

l(Japan)

[7599]

160

3-Prop

yl-3[(245-trim

etho

xyph

enyl)-

metho

xy]-24-pentanedione

3-Prop

yl-3-(245-

trim

etho

xy)benzyloxy-

pentan-24-dione

O

O

O

O

O

O

Leaves

(India)

[100]

161

3-(34-D

ihydroxyph

enyl)-2-

prop

enoica

cidethyleste

rCa

ffeateethyl

O

O

OH

OH

Not

foun

d(Taiwan)

who

le(Taiwan)

[434584]

Evidence-Based Complementary and Alternative Medicine 31Ta

ble7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

162

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

d-Erythron

icacid

2-O-caffeoyl-2-C

-methyl

O

HO

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

163

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

2-O-caffeoyl-2-C

-methyl

O

O

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

164

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-24-

dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

3-O-caffeoyl-2-C

-methyl

O

O

HO

HO

O

O

OH

OH

Leaves

(Japan)

[101]

165

4-(A

cetyloxy)-3-[[3-(34-

dihydroxyphenyl)-

1-oxo-2-propen-1-y

l-]oxy]-2-hydroxy-

2-methyl-b

utanoica

cid

O

HO

OH

O OHO

HO

OO

Not

foun

d(Ja

pan)

[73]

166

3-(34-D

ihydroxyph

enyl)-tetrahydro-4-

hydroxy-4-

methyl-5

-oxo-3-fu

ranyl

ester-2-prop

enoica

cid

3-O-C

affeoyl-2-C

-methyl-

D-erythrono

-14-la

cton

eHO

HO

O

OO

OOH

Leaves

(Japan)

[101]

167

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]-oxy]-145-

trihydroxy-cyclohexanecarboxylicacid

Chlorogenica

cid

O

HO

HO

OH

O

OH

O

OH

HO

Aeria

l(Japan)w

hole

(Taiwan)

[707175]

32 Evidence-Based Complementary and Alternative Medicine

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

168

4-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]oxy]-

135-tr

ihydroxy-cyclohexanecarboxylic

acid

4-O-C

affeoylqu

inicacid

OHO

HO

OH

OH

O

O

OH

OH

Aeria

l(Japan)

[71]

169

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-15-

dihydroxy-cyclo

hexanecarboxylicacid

34-Di-O

-caffeoylqu

inic

acid

HO

HO

O

O

O

O

OH

OH

HO

OH O

H

O

Aeria

l(Japan)W

hole

(Taiwan)

[45497071]

170

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-14-dihydroxy-

cyclo

hexanecarboxylicacid

35-Di-O

-caffeoylqu

inic

acid

HO

HO

O O

HO

O

HO

OH OO

OHOH

Aeria

l(Japan)w

hole

(Taiwan)

[45497071]

Evidence-Based Complementary and Alternative Medicine 33

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

171

34-bis[[(2E

)-3-(34-D

ihydroxyph

enyl)-1-

oxo-2-prop

en-1-

yl]oxo]-15

-dihydroxy-

cyclo

hexanecarboxylicacid

45-Di-O

-caffeoylqu

inic

acid

OHO

OH

HO

O

O

OH

OH

O

OH

HO

OWho

le(Taiwan)

[454970]

172

3-[4-[[-6-O-[3-(4-H

ydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]-phenyl]-2-prop

enoica

cid

120573-D

-p-C

oumaricacid4-O

-(6-O

-p-caoum

aroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HOH

HO

O

O

Leaves

(Japan)

[59]

173

3-[4-[[2-O

-Acetyl-6

-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]ph

enyl]2-propeno

icacid

120573-D

-p-C

oumaricacid4-O

-(2-O

-acetyl-6

-O-p-

caou

maroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HO

HO

O

O

Leaves

(Japan)aerial

(China)

[3459]

174

67-Dihydroxy-2-chrom

enon

eEsculetin

cicho

rigenin

OHO HO

O

Not

foun

d(Egypt)

[39]

34 Evidence-Based Complementary and Alternative Medicine

Table 8 Aromatic compounds isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

175 12-Benzenediol PyrocatechinHO

HOWhole(Japan) [99]

176 4-Ethyl-12-benzenediol Pyrocatechol

OH

OH

Whole(Japan) [99]

177 Dimethoxyphenol26-dimethoxyphenol

O

HO

O

Roots (Japan) [99]

178 4-Ethenyl-2-methoxy-phenol p-Vinylguaiacol

O

OHWhole(Japan) [99]

179 2-Hydroxy-6-methylbenzaldehyde

6-Methyl-salicylaldehyde O

OH

Whole(Japan) [99]

180 Benzene-ethanol 2-Phenyl-ethanolHO

Whole(Japan) [57]

181 4-Hydroxy-3-methoxy-benzaldehyde Vanillin O

O

HOAerial(Japan) [99]

182 3-Hydroxy-4-methoxy-benzaldehyde Vanillin iso

OO

OH

Leaves(Japan) [99]

183 4-Hydroxybenzoic acid p-Hydroxybenzoicacid

O

HOOH Whole

(Japan) [99]

184 2-Hydroxybenzoic acid Salicylic acidO

HO

HO

Stem androots (Japan) [99]

185 34-Dihydroxybenzoicacid Protocatechuic acid

O

HOOH

OH

Whole(Japan) [99]

1864-Hydroxy-

3methoxybenzoicacid

Vanillic acid

OOH

O

OH

Aerial(Uganda)

Roots (Japan)[68 99]

Evidence-Based Complementary and Alternative Medicine 35

Table 8 Continued

S N IUPAC names Common names Structure Plant part(country) References

187 345-Trihydroxybenzoicacid Gallic acid

O

OHHO

HO

HO

Whole(China) [102]

[88] This work supports the use of B pilosa as an anti-inflammatory agent however no compounds responsible forthe anti-inflammatory activity of B pilosa were identified

A further study also reported the anti-inflammatoryactivity as well as the antiallergic activity of B pilosa [75] Inthis study dried powder of the aerial part of B pilosa whichhad been pretreated with the enzyme cellulosine was usedfor further tests The results showed that oral administrationof the cellulosine-treated B pilosa lowered the level of serumIgE in mice 10 days after immunization with DNP (24-dintrophenyl)-Ascaris as an antigen This treatment alsoreduced dye exudation in skin induced by passive cutaneousanaphylaxis and production of inflammatory mediators his-tamine and substance P in rats [75] Phytochemical analysisshowed that cellulosine treatment increased the percentage ofcaffeic acid and flavonoids This study suggests that B pilosaand its phenolics have anti-inflammatory functions

Phenolics and polyynes are major anti-inflammatoryphytochemicals present in B pilosa (Table 11) Unsurpris-6ingly phenolics such as luteolin (103) and ethyl caffeate(161) that are major constituents of B pilosa have alsobeen reported to possess anti-inflammatory activity Lute-olin was reported to exhibit anti-inflammatory activity inmacrophages Xagorari and colleagues showed that luteolininhibited the release of inflammatory cytokines TNF-120572 andinterleukin-6 in RAW 2647 cells following LPS stimulation[87] It inhibited TNF-120572 production with an IC

50value

of 1120583M The underlying anti-inflammatory mechanism ofluteolin was reported to be the inactivation of Akt andNF-120581B activation [87] In addition luteolin was reportedto confer anti-inflammatory activity through inhibition ofLPS-stimulated iNOS expression in BV-2 microglial cellsIt inhibited LPS-activated microglia in a dose-dependentmanner with an IC

50value of 69 120583MMoreover immunoblot

and RT-PCR data proved that luteolin suppressed I120581B-120572degradation and iNOS expression in LPS-activated microglia[85] Kim and colleagues stated that luteolin may havebeneficial effects on inflammatory neural diseases throughinhibition of iNOS expression [85] A related study revealedthat luteolin decreased the transcriptional activity of NF-120581BRelA via partial inhibition of TNF-mediated NF-120581B DNAbinding activity Luteolin also inhibited Akt phosphorylationand induced degradation of a transcription factor interferonregulatory factor (IRF) [86]

Chiang and colleagues showed that ethyl caffeate(161) significantly inhibited NO production in mousemacrophages RAW 2647 cells [84] Based on MTT assays

they concluded that this inhibition was not due to thecytotoxicity of ethyl caffeate The IC

50value of ethyl caffeate

in the inhibition of NO production was 55 120583gmL slightlylower than curcumin (positive control) which has anIC50

value of 65120583gmL They demonstrated that ethylcaffeate exerted anti-inflammatory activity via the reducedtranscription and translation of iNOS (inducible nitric oxidesynthase) in RAW 2467 cells In addition this compoundalso suppressed COX-2 expression in RAW 2467 cellsand MCF-7 cells The in vivo anti-inflammatory effectof ethyl caffeate was verified by testing in TPA-treatedmouse skin Like celecoxib the positive control ethylcaffeate significantly abolished COX-2 expression in adose-dependent manner Ethyl caffeate at 1mg200120583Lsite(24mM) inhibited COX-2 expression at a level comparableto celecoxib at 1mg200120583Lsite (13mM) Remarkably ethylcaffeate at 48mM (2mg200120583Lsite) was more effectivethan celecoxib at 131mM (10mg200120583Lsite) Moreoverthis compound inhibited the activation of nuclear factor-120581B(NF-120581B) by LPS via the prevention of NF-120581B binding toDNA [84] In addition 3 ethyl caffeate analogs (ethyl 34-dihydroxyhydrocinnamate ethyl cinnamate and catechol)also showed different degrees of NF-120581B binding to DNA aslisted in Table 12 Ethyl cinnamate lacks a catechol moietywhich results in ineffective inhibition of NF-120581B binding toDNA [84]

Pereira and colleagues assessed the anti- 7inflammatory and immunomodulatory activities ofB pilosa methanol extract as well as one polyyne2-O-120573-glucosyltrideca-11(E)-en-3 579-tetrayn-12-diol(54) in T lymphocytes and a zymosan-induced arthritismouse model [27] They first examined the in vitro effect of 8the B pilosa extract and compound 54 on cell proliferation ofhuman T cells stimulated with 5120583gmL phytohemagglutinin(PHA) or 100 nM 12-O-tetradecanoyl phorbol-13-acetate(TPA) plus 15 120583M ionomycin and on cell proliferation ofmouse T cells stimulated with 5 120583gmL concanavalin A(Con A) The data demonstrated that both methanol extractand compound 54 suppressed T-cell proliferation in adose-dependent manner The estimated IC

50values of the B

pilosa extract against human T cells stimulated with 5120583gmLPHA and 100 nM TPA plus 15120583M ionomycin were 125 and25 120583gmL respectively In comparison with the methanolextract compound 54 showed 10-fold more inhibition ofhuman T-cell proliferation with an estimated IC

50value of

15 120583gmL Accordingly the B pilosa extract and compound54 dose-dependently suppressed mouse T-cell proliferation

36 Evidence-Based Complementary and Alternative Medicine

Table 9 Porphyrins isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

188

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(15S16S)-10-ethenyl-5-ethyl-1161820-tetrahydro-6111522-tetramethyl-1820-dioxo-15H-912-imino-212-

metheno-471714-dinitrilopyrano[43-

b]azacyclononadecine-16-propanoic

acid

AristophyllC

NH N

HNN

O

O OOOPhytyl

O

Leaves(Taiwan) [100]

189

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -20b-(methoxycarbonyl)-9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-dinitrilo-12-

dioxeto-[310158404101584034]-cyclo-pent[1 2]azacyclononadecine-

19-propanoic acid

Bidenphytin A

NH N

HNN

OPhytylO O O

O O

Leaves(Taiwan) [100]

190

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -2a-

hydroxy-20b-(methoxycarbonyl)-

9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-

dinitrilo-12-dioxeto[310158404101584034]-cyclo-pent[12-b]-

azacyclononadecine-19-propanoic acid

Bidenphytin B

NH N

HNN

OPhytylO O O

O O

OH

Leaves(Taiwan) [100]

191

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO O

HOOO

Leaves(Taiwan) [100]

192

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

Evidence-Based Complementary and Alternative Medicine 37

Table 9 Continued

S N IUPAC names Common names Structure Plant part(country) References

193

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

Leaves(Taiwan) [100]

194

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

195

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3S4S21R)-9-ethenyl-14-ethyl-21-(methoxycarbonyl)-

481318-tetramethyl-20-oxo-3-phorbinepropanoic acid

Pheophytin A

NH N

HNN

PhytylO

O

OO O

Leaves(Taiwan) [100]

with estimated IC50values of 30 and 25 120583gmL respectively

Taken together the data indicate that the B pilosa extractand compound 54 act on human and mouse T cells Totest the in vivo effect of the B pilosa extract and compound54 a zymosan-induced arthritis mouse model was usedThis model was established from B10ASgSnJ mice with aninjection of zymosan (015mg) The zymosan-injected mice9received an intraperitoneal injection of the B pilosa extract(1 5 or 10mg) at one dose a day for 5 days Popliteal lymphnode (PLN) weight was monitored to check the developmentof arthritis The results revealed that 10mg of the methanolextract of B pilosa extract could significantly diminishinflammation as evidenced by PLN weight [27] This worksuggests that B pilosa (and compound 54) can suppressimmune response and inflammation

33 Antidiabetic Activity Anti-diabetic agents are primarilydeveloped from plants and other natural resources [8 24 4951 53] B pilosa is one of 1200 plant species that have beeninvestigated for antidiabetic activity [120 121]B pilosa is usedas an anti-diabetic herb in America Africa and Asia [51 120

122] Many studies have indicated that B pilosa could treattype 1 diabetes (T1D) and type 2 diabetes (T2D) in animals

Etiologically speaking T1D is caused by autoimmune-mediated destruction of pancreatic 120573 cells leading to insulindeficiency hyperglycemia and complications Currentlythere is no cure for T1D Polarization ofTh cell differentiationcontrols the development of T1D Suppression of Th1 celldifferentiation and promotion of Th2 cell differentiationameliorate T1D [123] One study showed that the butanolfraction of B pilosa inhibited T-cell proliferation decreasedTh1 cells and cytokines and increased Th2 cells andcytokines leading to prevention of T1D in nonobesediabetic (NOD) mice [49] Based on a bioactivity-directedisolation strategy 3 polyynes 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne (53) also known ascytopiloyne 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D- Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne (50) were identifiedfrom B pilosa [49 53] The IC

50value of the butanol fraction

was 200120583gmL This inhibition was reported to be partiallyattributed to cytotoxicity because the butanol fraction at

38 Evidence-Based Complementary and Alternative Medicine

Table 10 Other compounds isolated from B pilosa [30]

SN IUPAC names Common names Structure Plant part(country) References

19637-Dihydro-137-

trimethyl-1H-purine-26-dione

CaffeineN

NN

NO

O

Aerial(Uganda) [68]

197

1-((2R4S5R)-Tetrahydro-4-hydroxy-5-(hydroxymethyl)furan-2-yl)-5-

methylpyrimidine-24(1H3H)-dione

ThymidineHO

HO

ON

O

NHO Not found

(China) [37]

198 1-(2-Thienyl)-ethanone 2-Acetyl-thiophene

S

O

Roots(Germany) [41]

199

(2R3S4S5S)-2-(Heptan-2-yloxy)-tetrahydro-6-

((tetrahydro-34-dihydroxy-5-

(hydroxymethyl)furan-2-yloxy)methyl)-

2H-pyran-345-triol

Heptanyl2-O-120573-xylofuranosyl-

(1rarr 6)-120573-glucopyranoside

O

OOH

OH

HO

OHO

H

HO

H

H

HOHH

O

Whole(Taiwan) [70]

200

2-[(3R7R11R)-3-Hydroxy-371115-

tetramethylhexadecyl]-356-trimethyl-

25-cyclohexadiene-14-dione

120572-Tocopherylquinone

OO

HO

Whole(Taiwan) [32]

2017-O-(410158401015840610158401015840-Diacetyl)-

120573-D-glucopyranoside

Not found Leaves(China) [60]

180 120583gmL could cause 50 death of Th1 cells Moreoverthis study suggested that the butanol fraction may preventdiabetes in NOD mice in vivo via downregulation of Th1cells or upregulation Th2 cells that have effects which areantagonistic of those of Th1 cells [49] This was proven byintraperitoneal injection of the butanol fraction at a doseof 3mgkg BW 3 times a week to NOD mice from 4 to 27weeks This dosage resulted in lower incidence of diabetes(33) At a dose of 10mgkg the butanol fraction of Bpilosa totally eliminated (0) the initiation of the diseaseTo further support this result assessment of IgG2a and IgEproduction was performed in the serum of NOD mice Asin vivo results obtained from intracellular cytokine stainingexperiments were not very conclusive levels of IgG2a and IgE

were measured since Th1 cytokine IFN120574 and Th2 cytokineIL-4 favor the production of IgG2a and IgE respectively Asexpected high levels of IgE and some decline in the levels ofIgG2a were observed in the serum Profiling of the butanolextract revealed five compounds 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne(50) 45-Di-O-caffeoyl- quinic acid 35-Di-O-caffeoylquinicacid and 34-Di-O-caffeoylquinic acid Only the first twocompounds showed similar effects on the prevention ofdiabetes in NOD mice as the B pilosa butanol fractionMoreover compound 50 showed greater activity thancompound 69 in terms of enhancement (by 34 comparedto 8) of differentiation of Th0 to Th2 at 15 120583gmL (both

Evidence-Based Complementary and Alternative Medicine 39

Table 11 Chemical constituents of B pilosa and their biological activities

SN Name Classification Molecularformula Biological activities

109 Centaureidin [91] Flavonoid C18H16O8Anti-listerial [69 91]

Cytotoxic [10]

110 Centaurein [91] Flavonoid C24H26O13

Anti-listerial [69 91]Cytotoxic [10]Anti-viral [105]

103 Luteolin [106] Flavonoid C15H10O6

Anti-viral [107 108]Cytotoxic [77]

Anti-inflammatory [109]Anti-allergic [109]

82 Butein [110] Flavonoid C15H12O5Anti-leishmanial [111]

Cytotoxic [78]48 12-Dihydroxytrideca-57911-tetrayne [112] Polyyne C13H12O2 Anti-angiogeneic [112]46 13-Dihyroxy-6(E)-tetradecene-81012-triyne [112] Polyyne C14H16O2 Anti-angiogeneic [112]

45 12-Dihyroxy-5(E)-tridecene-7911-triyne [113] Polyyne C13H14O2Anti-angiogeneic [113]Anti-proliferative [113]

64 1-Phenylhepta-135-triyne [114] Polyyne C13H8

Anti-microbial [115]Anti-malarial [3]Cytotoxic [3]Antifungal [15]

27 Linoleic acid [79] Fatty acid C18H32O2Anti-viral (100)Cytotoxic [17]

161 Ethyl caffeate [84] Phenylpropanoid C11H12O4 Anti-inflammatory [84]

54 2-O-120573-Glucosyltrideca-11(E)-en-3579-tetrayn-12-diol [26] Polyyne C19H20O7Immunosuppressive andAnti-inflammatory [26]

53 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne [50] Polyyne C19H22O7Anti-diabetic [116]

Anti-inflammatory [50]

69 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne [28] Polyyne C20H26O7Anti-diabetic [28]

Anti-inflammatory [94]

50 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne [28] Polyyne C19H24O7

Anti-diabetic [28]Anti-inflammatory [94]

Anti-malarial andantibacterial [47]

129 Quercetin 3-O-120573-D-galactopyranoside [18] Flavonoid C21H20O12 Anti-inflammatory [117]

170 35-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

171 45-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

169 34-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

126 Quercetin 331015840-dimethyl ether7-O-120572-L-rhamnopyranosyl-(1rarr 6)-120573-D-glucopyranoside [118] Flavonoid C21H20O11 Anti-malarial [119]

125 Quercetin 331015840-dimethyl ether-7-O-120573-D-glucopyranoside [118] Flavonoid C21H20O12 Anti-malarial [119]70 1-Phenyl-13-diyn-5-en-7-ol-acetate [27] Polyyne C15H12O2 Anti-malarial [27]199 Heptanyl 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside [70] Miscellaneous C18H44O10 Antioxidant [70]124 3-O-Rabinobioside [70] Saccharide C27H38O15 Antioxidant [70]130 Quercetin 3-O-rutinoside [70] Flavonoid C27H38O15 Antioxidant [70]167 Chlorogenic acid [70] Phenolic C16H26O9 Antioxidant [70]119 Jacein [70] Flavonoid C24H26O13 Antioxidant [70]

49 (R)-12-dihydroxytrideca-357911-pentayne [47] Polyyne C13H8O2Anti-malarial andAntibacterial [47]

40 Evidence-Based Complementary and Alternative Medicine

Table 12 Structure and activity relationship studies of ethyl caffeateusing in vitro NF-120581BDNA binding assays [84]

Compound Concentration(120583M)

NF-120581BDNA binding

Ethyl caffeate 50 100inhibition

Ethyl 34-dihydroxyhydrocinnamate 100 100inhibition

Catechol 400 100inhibition

Ethyl cinnamate 400 Noinhibition

Table 13 Apoptosis in cocultures of T cells and pancreatic 120573 cells[53]

Cellmedium apoptosis andnecrosis

CD4+ T cellscontrol medium lt4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 2

CD4+ T cellswith cytopiloyne-treated 120573 cells 18CD4+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 7

CD8+ T cellscontrol medium 4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 4

CD8+ T cellswith cytopiloyne-treated 120573 cells 4CD8+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 4

Table 14 Radical scavenging activities of B pilosa extracts [70]

Extractscontrol DPPH assayIC50 (120583gmL)

NBThypoxanthinesuperoxide assayIC50 (120583gmL)

Quercetin 198 15Ascorbic acid 634 Not determined120572-tocopherol 897 Not determinedEthyl acetateextract 1383 597

Butanol extract 1669 114Water extract gt100 gt100

compounds) and inhibition (by 40 compared to 10) ofdifferentiation toTh1 at the same concentration [49]

Among the three polyynes found in B pilosa cytopiloyne(53) had the most potent anti-T1D activity [53] To test the invivo effect of cytopiloyne NODmice received intraperitonealor intramuscular injection of cytopiloyne at 25120583gkg BW3 times per week Twelve-week-old NOD mice started todevelop T1D and 70 of NOD mice aged 23 weeks andover developed T1D Remarkably 12- to 30-week-old NODmice treated with cytopiloyne showed normal levels of bloodglucose (lt200mgdL) and insulin (1-2 ngmL) Consistent

Table 15 Radical scavenging activity of secondarymetabolites fromB pilosa [70]

Metabolite (Table 11)control DPPH assay IC50 (120583gmL)199 Not determined124 53130 68167 105169 33171 38119 Not determined110 Not determinedQuercetin 256Caffeic acid 890

Table 16 Antioxidant activity of the essential oils and water extractsfrom B pilosa [92]

Extract IC50 (120583gmL)Leaf essential oils 47Flower essential oils 50Leaf extract 61Flower extract 172

with T1D incidence cytopiloyne delayed and reduced theinvasion of CD4+ T cells into the pancreatic islets [53]

In vitro study showed that cytopiloyne (53) inhibited thedifferentiation of naıveTh (Th0) cells (ie CD4+ T cells) intoTh1 cells and promoted differentiation of Th0 cells into Th2cells [50] The in vitro data are consistent with the in vivoresults indicating that cytopiloyne reducedTh1differentiationand increased Th2 differentiation as shown by intracellularcytokine staining and FACS analysis [53] In line with theskewing of Th differentiation the level of serum IFN-120574 andIgG2c decreased while that of serum IL-4 and serum IgEincreased compared to the negative controls (PBS-treatedmice) Cytopiloyne also enhanced the expression of GATA-3 a master gene for Th2 cell differentiation but not theexpression of T-bet a master gene forTh1 cell differentiationfurther supporting its role in skewingTh differentiation [53]

Also importantly cytopiloyne partially depleted CD4+rather than CD8+ T cells in NOD mice [53] As shown inTable 13 coculture assays showed that the depletion of CD4+T cells was mediated through the induction of Fas ligandexpression on pancreatic islet cells by cytopiloyne leading toapoptosis of infiltrating CD4+ T cells in the pancreas via theFas and Fas ligand pathways However cytopiloyne did notinduce the expression of TNF-120572 in pancreatic islet cells andthus had no effect on CD8+ T cells [53]

In addition Chang and colleagues showed that cytopi-loyne dose-dependently inhibited T-cell proliferation stimu-lated by IL-2 plus Con A or anti-CD3 antibody using [3H]thymidine incorporation assay [53]

Overall the mechanism of action of cytopiloyne andprobably its derivatives in T1D includes inhibition of T-cellproliferation skewing of Th cell differentiation and partialdepletion of Th cells Due to the anti-diabetic mechanisms

Evidence-Based Complementary and Alternative Medicine 41

Table 17 Antibacterial activity of essential oils and flower extracts from B pilosa [92]

Strain Mean zone of inhibition (mm)Leaf essential oil Flower essential oil Leaf extract Flower extract

Micrococcus flavus 127 plusmn 03 87 plusmn 03 102 plusmn 02 108 plusmn 03Bacillus subtilis 173 plusmn 19 117 plusmn 02 109 plusmn 02 103 plusmn 02Bacillus cereus 190 plusmn 14 112 plusmn 03 118 plusmn 04 185 plusmn 10Bacillus pumilus 123 plusmn 07 108 plusmn 02 105 plusmn 04 77 plusmn 02Escherichia coli 137 plusmn 04 203 plusmn 07 102 plusmn 11 140 plusmn 13Pseudomonas ovalis 125 plusmn 08 137 plusmn 15 102 plusmn 06 125 plusmn 06

Table 18 Antibacterial activity of root extracts from B pilosa [93]

Strain MIC50 (mgmL)Methanol extract Acetone extract

Bacillus cereus 10 mdashEscherichia coli 5 5Klebsilla pneumonia 5 10Micrococcus kristinae 10 mdashPseudomonas aeruginosa 10 10Staphylococcus aureus 5 10Sraphylococcus epidermidis 5 5Serratia marcescens 10 mdashShigelea flexneri 10 mdashStreptococcus faecalis 10 mdash

Table 19 Antibacterial activity of B pilosa of compound 29 [47]

Strain MIC50 (120583gmL)Escherichia coli NIHJ 1Escherichia coli ATCC25922 1Klebsiella pneumoniae ATCC700603 128Serratia marcescens ATCC13880 16Pseudomonas aeruginosa ATCC27853 8Staphylococcus aureus FDA209P 05Staphylococcus aureus ATCC29213 025Staphylococcus aureus N315 (MRSA) 05Enterococcus faecalis ATCC29212 2Enterococcus faecalis NCTC12201 (VRE) 1Bacillus subtilis ATCC6633 05Candida albicans ATCC10231 025

of action it was hypothesized that cytopiloyne protectsNOD mice from diabetes by a generalized suppression ofadaptive immunity To evaluate this hypothesis ovalbumin(Ova) was used as a T-cell dependent antigen to primeNOD mice which had already received cytopiloyne orPBS vehicle Ova priming boosted similar anti-Ova titersin cytopiloyne-treated mice and PBS-treated mice but adifference in immunoglobulin isotype was observed in thetwo groups Thus it was concluded that cytopiloyne is animmunomodulatory compound rather than an immunosup-pressive compound [50 53]

T2D is a chronic metabolic disease with serious com-plications resulting from defects in either insulin secretion

insulin action or both [124] A study by Ubillas et alshowed that the aqueous ethanol extract of the aerial partof B pilosa at 1 gkg body weight (BW) lowered bloodglucose in dbdb mice a T2D mouse model [51] Basedon a bioactivity-guided identification compounds 69 and50 were identified Further the mixture of the compounds(69 50) in a 2 3 ratio significantly decreased blood glucoseconcentration and reduced food intake on the second dayof treatment when administered at doses of 250mgkg twicea day to C5BLKs-dbdb mice When tested at 500mgkga more substantial drop in blood glucose level as wellas the stronger anorexic effect (food intake reduced from58 gmouseday to 25 gmouseday) was observed [51] Inthis study it was suggested that the blood glucose lower-ing effect of B pilosa was caused in part by the hungersuppressing effect of its polyynes [51] However the hungersuppressing effect of the ethanol extract of B pilosa wasnot found in the studies described below In another study[24] water extracts of B pilosa (BPWE) were used indiabetic dbdb mice aged 6ndash8 weeks with postprandialblood glucose levels of 350 to 400mgdL Like oral anti-diabetic glimepiride which stimulates insulin release onesingle dose of BPWE reduced blood glucose levels from374 to 144mgdL The antihyperglycemic effect of BPWEwas inversely correlated to an increase in serum insulinlevels suggesting that BPWE acts to lower blood glucose viaincreased insulin production However BPWE had differentinsulin secretion kinetics to glimepiride [24] One flaw incurrent anti-diabetics is their decreasing efficacy over timeThe authors investigated the long term anti-diabetic effectof BPWE in dbdb mice BPWE reduced blood glucoseincreased blood insulin improved glucose tolerance andreduced the percentage of glycosylated hemoglobin (HbA1c)Both long-term and one-time experiments strongly supportthe anti-diabetic action of BPWE [24] In sharp contrast toglimepiride BPWE protected against islet atrophy in mousepancreas The investigators further evaluated anti-diabeticproperties of 3 B pilosa varieties B pilosa L var radiate(BPR) Bpilosa L var pilosa (BPP) and B pilosa L varminor(BPM) in dbdb mice [8] One single oral dose (10 50 and250mgkg body weight) of BPR BPP or BPM crude extractsdecreased postprandial blood glucose levels in dbdb micefor up to four hours and the reduction of glucose levels inthe blood appeared to be dose-dependent Comparing thethree variants BPR extract resulted in a higher reduction inblood glucose levels when administered at the same dose as

42 Evidence-Based Complementary and Alternative Medicine

Table 20 Antifungal activity of B pilosa [92]

Partextract Concentration (ppm) Strain InhibitionCortiicum rolfsii Fusarium solani Fusarium oxysporum

Leaves

Essentialoils

100 857 plusmn 09 682 plusmn 0 745 plusmn 17250 960 plusmn 08 779 plusmn 18 879 plusmn 04

AqueousExtracts

100 446 plusmn 17 605 plusmn 21 716 plusmn 07250 942 plusmn 03 689 plusmn 07 824 plusmn 19

Flowers

Essentialoils

100 604 plusmn 09 892 plusmn 04 869 plusmn 05250 894 plusmn 12 980 plusmn 03 949 plusmn 06

AqueousExtracts

100 331 plusmn 11 714 plusmn 07 573 plusmn 22250 661 plusmn 14 912 plusmn 0 900 plusmn 07

Table 21 Antifungal activity of B pilosa root extracts [93]

Strain LC50 (mgmL)Acetoneextracts

Methanolextracts

Waterextracts

Aspergillus niger 014 006 007Aspergillus flavus 1091 658 0Penicillium notatum 005 005 005

the other two varieties In terms of serum insulin levels a doseof 50mgkg of each extract was used and the BPR extracttogether with the three polyynes significantly increased theserum insulin level in dbdb mice Long-term experiments(28-day treatment) were then conducted using diabetic micewith postprandial glucose levels from 370 to 420mgdL andglimepiride was used as positive controlThe range of dosagesapplied was from 10mgkg BW to 250mgkg BW Resultsshowed that the positive control as well as the crude extractsof the three varieties lowered the blood glucose levels indbdb mice However only BPR extract containing a higherpercentage of cytopiloyne (53) reduced blood glucose levelsand augmented blood insulin levelsmore than BPP andBPMThe percentage of glycosylated hemoglobin A1c (HbA1c) wasalso measured and found to be 79 plusmn 05 in mice aged 10ndash12 weeks and 66 plusmn 02 61 plusmn 03 and 62 plusmn 03 inthe blood of age-matchedmice following treatment with BPRcrude extract (50mgkg) glimepiride (1mgkg) and com-pound 53 (05mgkg) respectively [8] Among the polyynesfound in B pilosa cytopiloyne was the most effective againstT2D Hence cytopiloyne was used for further study on anti-diabetic action and mechanism [125] The data confirmedthat cytopiloyne reduced postprandial blood glucose lev-els increased blood insulin improved glucose tolerancesuppressed HbA1c level and protected pancreatic islets indbdb mice Nevertheless cytopiloyne failed to decreaseblood glucose in streptoztocin (STZ)-treated mice whose bcells were already destroyed Additionally cytopiloyne dose-dependently increased insulin secretion and expression in bcells as well as calcium influx diacylglycerol and activationof protein kinase C120572 Collectively the mechanistic studiessuggest that cytopiloyne treats T2D via regulation of insulin

production involving the calciumDAGPKC120572 cascade in bcells

The studies detailed above point to the conclusion thatcytopiloyne and related polyynes (compounds 69 and 49)are anti-diabetics in animal models The data uncover a newbiological action of polyynes It should be noted that likeall anti-diabetic drugs cytopiloyne failed to prevent or curediabetes completely but reduced diabetic complications [125]Intriguingly 34 polyynes have been found in B pilosa so farIt remains to be seen whether all the polyynes present in thisplant have anti-diabetic activities

34 Antioxidant Activity Free radicals can damage cellularcomponents via a series of chemical reactions [89] leadingto development and progression of cardiovascular diseasecancer neurodegenerative diseases and ageing [70] Freeradicals nitric oxide (NO) and superoxide anions can beproduced in macrophages to kill microbes However anexcessive generation of the free radicals under pathologicalconditions is associated with a wide range of illnessesPlants are known to be rich in antioxidant phytochemicalsChiang and colleagues evaluated the free radical scaveng-ing activity of crude extract fractions and compounds ofB pilosa using 11-diphenyl-2-picrylhydrazyl (DPPH) andhypoxanthinexanthine oxidase assays [70] Using DPPHand hypoxanthinexanthine oxidase assays they found thatthe B pilosa crude extract and the ethyl acetate butanoland water fractions had free radical scavenging activityNine compounds Heptyl- 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside (199) 3-O-Rabinobioside (124) Quercetin3-O-rutinoside (130) Chlorogenic acid (167) 34-Di-O-caffeoylquinic acid (169) 35-Di-O-caffeoylquinic acid (170)45-Di-O-caffeoylquinic acid (171) Jacein (119) and Cen-taurein (110) had DPPH radical scavenging activity [70]The IC

50values of the B pilosa crude extractfractions and

compounds are summarized in Tables 14 and 15 respectivelyMeasurement of free radical scavenging activities is one

way of assessing the antioxidant activities of B pilosa andits fractions and compounds It is interesting that the ethylacetate and butanol fractions are more active than the waterfraction and B pilosa crude extract [70] Of the secondarymetabolites only phenolic compounds 124 130 167 169 and171 showed significant DPPH-radical scavenging activities

Evidence-Based Complementary and Alternative Medicine 43

Further analysis of the structure-activity relationship of thecompounds suggested that substitution of the C3 hydroxylgroup with glycosides increased the activity approximately2-fold (for example in compounds 124 and 130) relative toquercetin (-OH in its C3) [70] Further modification of thestructures of the active compounds needs to be performed totest the effects of various substituents on activity The reasonwhy most of the antioxidant compounds contain phenolmoieties in their structure could be that the reduction-oxidation (redox) properties of phenols allow them to actas reducing agents singlet oxygen quenchers and hydrogendonors

A complementary study by Muchuweti and colleaguesdetermined phenolic content antioxidant activity and thephenolic profile of B pilosa methanol extract [89] Theyestimated that the phenolic content of the methanol extractof B pilosawas 11028plusmn22mgg [89] Vanillin hydroxyben-zaldehyde caffeic acid coumaric acid and ferulic acid werefound in this extractThe B pilosa extract also showed DPPHradical scavenging activity Furthermore the antioxidantactivity of the flavonoids found in B pilosa was correlatedwith its hepatoprotective effects through their inhibitionof NF-120581B activation which may lessen the oxidative stresscaused by the production of free radicals during liver injury[60]This activity might also be due to the anti-inflammatoryeffects of the aqueous extracts of B pilosa aerial parts on theinhibition of COX-2 and PGE

2production [88]

Essential oils from B pilosa flowers and leaves are alsoreported to possess antioxidant activity With the aim ofreplacing chemically synthesized additives Deba and col-leagues [92] worked on the antioxidant antibacterial andantifungal activities of essential oils and water extracts of Bpilosarsquos leaves and flowers Table 16 summarizes the resultsobtained from DPPH free radical scavenging assay

It can be inferred fromTable 16 that essential oils from theleaves possessed the highest activity It is reported elsewherethat monoterpenes present in essential oils such that of Bpilosa have protective effects and antioxidant properties [92]Beta-carotene bleaching method was also performed Leavesessential oils and aqueous extracts of the leaves and flowersshowed higher activity than the flower essential oils This isdue to the volatility of the flower essential oils The activityexhibited by the aqueous extracts was accounted to thepresence of phenolic compounds that are reported to donatea hydrogen atom to free radicals such that the propagation ofthe chain reaction during lipid oxidation is terminated [92]Overall essential oils and phenolics present in B pilosa canbe thought of as major antioxidant compounds

35 Immunomodulatory Activity B pilosa is thought to bean immunomodulatory plant and is reported to be effectivein the treatment of immune disorders such as allergy [75]arthritis [75] and T1D [48 50 53]

As pointed out in the discussion of its anti-diabeticactivities (Section 33) a combination of phytochemicalsmethod and T-cell activation assays was used to studyimmunomodulatory properties of B pilosa IFN-120574 is a keycytokine released by T and NK cells that mediates immune

cells and sustains immunity against pathogens Defects inIFN-120574 expression regulation and activation result in vulner-ability to diseases caused by bacteria and viruses [69] Anelegant study performed byChang and colleagues using IFN-120574 promoter-driven luciferase reporter construct in JurkatT cells showed that hot water crude extracts of B pilosaincreased IFN-120574 promoter activity two-fold [69] Out of thesubfractions of this extract the butanol fraction but not thewater or ethyl acetate fractions increased IFN-120574 promoteractivity six-fold Centaurein (110) and centaureidin (109)were identified from the butanol fraction and were stated tocause a four-fold increase in IFN-120574 promoter activity withEC50

values of 75 120583gmL and 09120583gmL respectively Themechanism of action of centaurein was determined usingtranscription factors such as AP-1 NFAT and NF120581B whichare reported to bind to IFN-120574 promoter and regulate IFN-120574transcription Unlike with the activity of the positive controlPHA centaurein caused a four-fold increase in NFAT a3-fold increase in NF120581B and had little if any effect onAP-1 enhancer activities (23-fold three-fold and ten-foldincreases respectively were seenwith PHA) [69]The authorsconcluded that centaureinmodulates IFN-120574 expression by theNFAT and NF120581B pathways The article only determined themechanism of action of centaurein Its aglycone centaureidinmay act through the samemechanism though this conclusionneeds to be further verified

B pilosa extract and its compounds are reported to inhibitdifferentiation of naıve CD4+ helper T (Th0) cells into Th1cells [49] Using theThcell differentiation assay as a screeningplatform 3 polyynes 2-120573-D-glucopyranosyloxy-1 -hydroxytrideca-57911-tetrayne (53) 3-120573-D-glucopyranosyl-1-hydroxy-6(E)- tetradecene-81012-triyne (69) and 2-120573-D-glucopyranosyloxy-1-hydroxy-5(E)- tridecene-7911-triyne(49) were discovered from B pilosa [49 53] The data showsthat cytopiloyne and other two polyynes suppressed thedifferentiation of type 1 helper T (Th1) cells and productionof Th1 cytokines and promoted that of type 2 helper T (Th2)cells and production of Th2 cytokines thus explaining theimmunomodulatory and anti-inflammatory effects of Bpilosa and its polyynes

Chang and colleagues were the first to report the effect ofthe butanol extract of B pilosa on the autoimmune diabetesand airway inflammation inmice [48] Imbalance in the levelsofTh1 andTh2 and of various cytokines leads to autoimmunediseases T1D and other autoimmune diseases (rheumatoidarthritis Crohnrsquos disease among others) are exacerbated byan increase Th1 levels (specifically CD4+ Th1 cells) whileTh2 cells antagonize this effect [19] Moreover Th2 cellsmediate asthma in ovalbumin-induced hypersensitivity inBALBc mice [48] In their work [48] Chang and colleaguesshowed that 10mgkg butanol extracts with 15 (ww)compound 69 and 11 (ww) compound 49 (Table 11)ameliorated the development of Th1-mediated diabetes inNOD mice through inhibition of 120573 cell death and leukocyteinfiltration At the same dosage the butanol extracts alsoexacerbated ovalbumin-induced pulmonary inflammationin BALBc mice with an increase in the infiltration ofeosinophils and mast cells into the airway of the mice [48]Despite the different outcomes both mouse models proved

44 Evidence-Based Complementary and Alternative Medicine

the concept that control over the Th1Th2 shift is associatedwith autoimmune diseases and that the B pilosa butanolextract can shift the differentiation of Th0 cells to Th2 cells[48 49]

An extended study presented by Chiang and colleaguesshowed that compound 53 modulates T-cell functions [50]Using CD4+ T cells from BALBc mice they demonstratedthat cytopiloyne decreased levels of IFN-120574 producing cells(Th1) by 122 (from 72 to 598) Since Th1 and Th2 celldifferentiation is antagonistic it was expected that compound53 increased the percentage of mouse IL-4 producing cells(Th2) by 72 (from 237 to 309) Subsequent assess-ment of effect of compound 53 on the modulation of thetranscription of IL-4 and IFN-120574 showed that cytopiloyne asexpected decreased the splenocyte levels of IFN-120574 mRNAand increasing that of IL-4 in a dose-dependent manner Inthe range of 01 to 3 120583gmL of compound 53 the effects werenot attributed to its cytotoxicity Consequently using 3120583gmLcytopiloyne for 72 and 96 hours the protein concentration ofIFN-120574 decreased to 186 and 444 respectively Under thesame conditions cytopiloyne increased IL-4 concentrationsto 1985 and 2470 (for 72 and 96 hours resp) Thismodulation of T-cell differentiation exhibited by compound53 was used to explain its anti-diabetic activity [50]

The anti-diabetic role of cytopiloyne was extensivelydiscussed above (Section 33 anti-diabetic activity) Themolecular basis of the regulation of cytokine expression bycytopiloyne has been described Cytopiloyne directly elevatedthe expression level of IL-4 via GATA-3 upregulation inT cells [53] However reduction of IFN-120574 expression in Tcells seemed to come from the indirect opposing effect IL-4cytokine because the expression level of T-bet was unaltered[53] In this way cytopiloyne skewedTh1 polarization into theTh2 state conferring protection against T1D in NOD miceAside frompolarization ofThcell differentiation cytopiloynealso activated the expression of Fas ligand in pancreaticb cells this increase leading to the partial depletion of Tcells and reduction of immune response in local areas suchas the pancreas Of note cytopiloyne also inhibited T-cellproliferation and activation By targeting T cells from threeimmunomodulatory actions cytopiloyne protects againstT1D and probably other Th1-mediated autoimmune diseases[19]

The phytochemical constituents of B pilosa exert theirfunctions on different immune cells to modulate immuneresponse It is possible that some of the compounds mayhave agonistic or antagonistic effects on immune responseImmune function of B pilosa may depend on its compo-sition and amount of compounds which could explain theapparently conflicting report of B pilosa butanol extractaggravating allergy in mice [48] while cellulosine-treatedextract ameliorated allergy [48 75] IFN-120574 promoter reporterassays and T-cell differentiation assays were used to isolate2 flavonoids [69] and 3 polyynes [49 50] as immunomod-ulatory compounds from the butanol fraction of B pilosaInterestingly the flavonoids promote IFN-120574 expression inNK and T cells In marked contrast the polyynes promoteIL-4 expression and indirectly inhibit IFN-120574 expression indifferentiating T cells Sensitivity appears to be the key to

identifying structure- and bioactivity-related phytochemicalsfrom medicinal plants

36 Antimalarial Activity The use of chemical drugs againstpathogens has resulted in drug-resistant mutants Examplesof drug resistance can be found in the species of thePlasmodium that cause malaria It is important to search fornew compounds to combat Plasmodium parasites [47] Astudy of the anti-malarial activity of the leaf extracts of Bpilosa using a combination of phytochemistry and bioassaysshowed that compound 49 (Table 11) showed activity againsta malaria parasite (P falciparum NF54 strain) with an IC

50

value of 60 120583gmL [77] In addition compound 49 isolatedfrom the aerial parts of B pilosa inhibited growth of the Pfalciparum FCR-3 strain with an IC

50value of 035 120583gmL

This compound was tested for its in vivo effect in miceinfected with P berghei NK-65 strain Results showed thatcompound 49 decreased the average parasitemia in the redblood cells by 207 (from 328 of that of the control to121) after an intravenous injection of 08mgkg BWday forfour days [47] Further studies addressing the anti-malarialmechanism underlying both polyynes and clinical studies areneeded

37 Antibacterial Activity Emergence of multiple antibiotic-resistantmicrobes is becoming a global threat to public healthand a challenge to disease treatment For instance penicillinis commonly used to combat a food-borne intracellularbacteriumListeria however penicillin-resistant bacteria havebeen discovered recently [69] Chang and coworkers iso-lated centaurein (110) and centaureidin (109) from B pilosaextract [69 70] Centaurein enhances expression of IFN-120574a key cytokine for macrophage activation and consequentlyenhances bactericidal activity in macrophages [69 70] Inagreement with observed in vitro effects centaurein wasreported to prevent and treat Listeria infection in C57BL6Jmice [91] Mechanistic studies confirmed that centaureinexerted antilisterial action via IFN-120574 expression in wild-typemice but not IFN-120574 knockout mice [91] Further in vitrostudies on centaurein showed that this compound increasedIFN-120574 expression by 13 (from 17 to 20) 20 (from 21to 41) and 11 (from 6 to 17) in CD4+ T cells CD8+Tcells and NK cells respectively That is to say there wasan increase in IFN-120574 producing immune cells As expectedcentaurein also enhanced the expression level of T-bet a keynuclear factor for IFN-120574 expression Consistently centaureinaugmented the serum IFN-120574 levels inC57BL6Jmice and thisaugmentation peaked 24 hours after compound injectionThe quantity of mouse serum IFN-120574 was sufficient to activatemacrophages in vitro and eradicated GFP-producing Listeriainside macrophages However the entry of Listeria intomacrophages was not affected by centaurein-treated mousesera Centaurein treatment at 20120583g per mouse rescued 30of the mice infected with a lethal dose of Listeria (2 times106 CFU) It is noteworthy that in the presence of ampicillin(5 120583gmouse) centaurein (20 120583gmouse) rescued 70 of themice suggesting an additive effect between ampicillin andcentaurein [91] Despite lower abundance centaureidin was

Evidence-Based Complementary and Alternative Medicine 45

30 times more active than centaurein in terms of IFN-120574production [91]

Aside from the indirect antibacterial action mentionedabove extract andor compounds of B pilosa also showeddirect bacteriostatic andor bactericidal action One studyreported that essential oils and leafflower extracts of Bpilosa could suppress the growth of gram positive and gramnegative bacteria as evidenced by zone of inhibition assaysIn this study antibacterial activity of the essential oils fromthe leaves and flowers of B pilosa was determined in anattempt to identify natural products as food preservatives forprevention of microbial multiplication and food oxidationThe essential oils and extracts of B pilosa leaves and flow-ers showed moderate but different extents of antibacterialactivity (Table 17) In general essential oils had higherantibacterial activity than crude extracts One explanation forthis could be that monoterpenes in the essential oils destroycellular integrity and subsequently inhibit the respirationand ion transport processes The presence of antibacterial120573-caryophyllene could be another explanation as reportedelsewhere [92] Another study reported that the methanoland acetone extract of B pilosa roots displayed antibacterialactivities against the bacteria listed in Table 18 [93] andmethanol extracts from the roots seemed to be the mosteffective

Another study indicated that the polyyne (R)-12-dihydroxytrideca-357911-pentayne (49) from this plantalso suppressed bacterial growth as shown by the minimuminhibitory concentration required to inhibit 50 bacterialgrowth (MIC

50) in Table 19 This compound was highly

effective against several Gram positive and Gram negativebacteria including the drug-resistant bacteria Staphylococcusaureus N315 (MRSA) and Enterococcus faecalis NCTC12201(VRE) [47] Strikingly compound 49 had a similar MIC

50

value to antibiotics (ampicillin tetracycline norfloxacin andamphotericin B) in most of the bacteria tested

Antibacterial activity of B pilosa extracts and compo-nents expressed as MIC

50and the mean zone of inhibition

is tabulated in Tables 17 18 and 19The zone of inhibition forAmpicilline (positive control) ranges from 153 plusmn 03mm to443 plusmn 02mm [92]

38 Antifungal Activity B pilosa has traditionally been usedto treat microbial infection Recently different parts of Bpilosa have been tested for antifungal activities Deba andcolleagues first evaluated the antifungal effect of the hotwater extracts of the B pilosa roots stems and leaves againstCorticium rolfsii Fusarium solani and Fusarium oxysporumThey discovered that C rolfsii was most suppressed bytreatment with B pilosa as its growth was reduced at almostall the tested doses followed by F oxysporum and F solani[99] However the fungicidal activities of the stems androots were greater than the leaves [99] Moreover the samegroup assessed the antifungal activity of the essential oilsand aqueous extracts from B pilosa flowers and leaves [92]They showed that the extracts and oils had antifungal activityagainst C rolfsii F solani and F oxysporum Essential oils

appeared to have better fungicidal activity thanwater extractsas summarized in Table 20

Another study by Ashafa and colleagues showed thatacetone methanol and water extracts of the B pilosa rootsshowed antifungal activities against Aspergillus niger Aflavus and Penicillium notatum using the agar dilutionmethod The results are tabulated in Table 21 [93] Negativecontrols showed 0 growth inhibitionThe methanol extractof the B pilosa roots at 10mgmL was also effective againstCandida albicans [93]Of note B pilosa obtained fromPapuaNew Guinea had no activity against A niger and C albicans[126] but the SouthAfrican (EasternCape) ecotype exhibitedmoderate activity against C albicans [93] This discrepancymay depend on extraction solvents extraction procedureassay techniques different plant parts and abundance ofactive compounds

B pilosa produces a variety of secondary metabolitessuch as flavonoids phenylacetylenes alkaloids sterols ter-penoids and tannis [92 93] However none of them havebeen confirmed as active compounds against fungi Furtherinvestigation of active compounds from B pilosa is necessaryto further understand the antifungal efficacy of this plant

39 Hypotensive and Vasodilatory Activities In early studiesDimo and colleagues used three rat models normotensiveWistar rats (NTR) salt-loading hypertensive rats (SLHR)and spontaneous hypertensive rats (SHR) to investigate thehypotensive effect of the methanol crude extract of B pilosaleaves [28 127] The extract lowered systolic blood pressurein hypertensive rats (SLHR and SHR) to a greater degreethan NTR [28 127] In addition a decrease in urinarysodium ions and an increase in urinary potassium ions wereobserved after treatment with the methanol extract of Bpilosa leaves although neither differences were statisticallysignificant [28 127] Taking the data together the studyproposed that B pilosa leaf extract reduced blood pressurevia vasodilation [28 127] The same group continued totest the antihypertensive effect of aqueous and methylenechloride extracts of B pilosa leaves in a hypertensive ratmodel [28 127] To establish a fructose-induced hypertensionmodel male Wistar rats were given 10 fructose solutionto drink ad libitum for three weeks In addition to freeaccess to 10 fructose the rats were treated with the aqueous(150mgkg) or methyl chloride (350mgkg) extracts of Bpilosa for additional three weeks [28 127] Both extracts ofB pilosa leaves had a hypotensive effect on rats Howeverneither extracts reversed the elevation of serum insulin infructose-fed rats Therefore B pilosa lowered blood pressureirrespective of insulin [28 127] To better understand thehypotensive mechanism the authors investigated the effectof a neutral extract of B pilosa (NBP) a mixture of methanolandmethylene chloride (1 1) extract after neutralization withNaOH and HCl on the heart and the blood pressure ofNTR and SHR [110] This study showed that an intravenousinjection of the NBP resulted in a biphasic reduction insystolic blood pressure In addition one intravenous dose ofthe extract at 10 20 and 30mgkg BW decreased systolicblood pressure in normal rats by 183 425 and 30

46 Evidence-Based Complementary and Alternative Medicine

respectively and the same doses reduced the blood pressurein hypertensive rats by 258 389 and 286 respectivelyOnly the highest dose (30mgkg) affected the force of thecontraction of the heart Atropine and propranolol were usedto interfere with the hypotensive action of the NBP Atropinereduced the initial phase of the hypotensive response in NBPand completely abolished the second phase of hypotensiveresponse in NBP In contrast propranolol increased thefirst hypotensive response but partially abolished the secondhypotensive response provoked by the NBP [110]This mech-anistic study suggested that B pilosa invokes the biphasichypotensive responses via targeting cardiac pump efficiencyduring the first phase and vasodilation at the second phase[110]

A further study was performed to investigate the relaxingeffect of a neutral extract of B pilosa (NBP) on rat aorta con-tractedwithKCl (60mM) andnorepinephrine (01mM) [94]Cumulative addition of NBP relaxed the rat aorta previouslycontracted by KCl in a dose-dependent manner The EC

50

value of theNBP for vasorelaxationwas 032mgmLThedataalso showed that the NBP reduced the contraction of aortapreviously contracted by KCl irrespective of the presence ofaortic endothelium [94]

Pretreatment with glibenclamide an ATP-dependent K+channel blocker did not considerably affect the relaxant effectof the NBP on KCl-induced contraction suggesting thatthe vasodilatory effect of B pilosa was not related to theopening of this ATP-dependent K+ channel [94] On theother hand in the presence of indomethacin or pyrilaminemaleate the relaxant response induced by the plant extractwas significantly inhibited at the lower concentrations Theplant extract was able to reduce the aorta resting tone inhibitthe KCl-induced contractions by 90 at 15mgmL and theCaCl2-induced contractions by 95 at 075mgmL These

results demonstrate that B pilosa can act as a vasodilatorprobably via acting as a calcium antagonist [94]

However no specific compound for the above activityhas been identified from B pilosa to date A bioactivity-guided identification approach may be adopted to identifythe active compounds in B pilosa that possess hypotensiveand vasodilatory effects and understand their mechanism ofaction

310 Wound Healing Activity B pilosa has been traditionallyused to treat tissue injury inCameroon Brazil andVenezuela[26] Hassan and colleagues investigated the wound healingpotential of B pilosa in Wistar rats [128] Mirroring thepositive control neomycin sulfate the ethanol extract of Bpilosa had faster wound closure than control rats 3 6 and 9days after topical application Histological examination alsorevealed better collagenation angiogenesis and organizationof wound tissue seven days after application Epithelializationand total healing time in B pilosa-treated rats were compara-ble to those of neomycin sulfate Together these data suggestthat B pilosa may be a viable alternative to neomycin lotionfor the treatment of wounds

In addition to studying the wound healing effect of Bpilosa on external ulcers Tan and colleagues also examined

the effect of methanol cyclohexane and methyl chlorideextracts of B pilosa on gastric ulcers in Wistar rats fedwith 1mL HClethanol gastric necrotizing solution (150mMHCl in 60 ethanol) and macroscopically visible lesionswere scored [26] Among the three extracts methylene chlo-ride extracts exhibited the highest activity showing 464inhibition of lesion formation at a dose of 500mgkg BWand complete inhibition at 750mgkg [26] The efficacy ofthe ethylene chloride extract was followed by that of themethanol extracts which had inhibition ranging from 304to 822 at concentrations of 500mgkg and 1000mgkgBW respectively [26] The cyclohexane extracts showed thelowest activity against gastric ulcers in rats with 13340 and 797 inhibition at 500 750 and 1000mgkg BWrespectively [26] To better understand the mode of actionof the methylene chloride extract of B pilosa rats werepretreated with indomethacin a COX-2 inhibitor involved inprostaglandin synthesis Pretreatment significantly reducedthe protection against HClethanol-induced ulcers to 313inhibition at 750mgkg BW suggesting a link between theantiulcerative activity of B pilosa and prostaglandin syn-thesis Unexpectedly the methylene chloride extract of Bpilosa showed little gastric mucosal protection against gastriclesions induced by 95 ethanol (1mL) [26] Absolute alcoholis known to cause mucosalsubmucosal tissue destruction viacellular necrosis and the release of tissue-derived mediators(histamine and leukotrieneC4)Thus these data imply thatBpilosa did not prevent the generation or the necrotic action ofthese mediators on the gastric microvasculature In additionpylorus ligation can increase gastric acid secretion withoutan alteration of mucosal histamine content The methylenechloride extract of B pilosa did not possess antisecretoryactivity On the contrary it was observed that increases in thedose of the extract led to elevated gastric juice acidity [26]Results with both absolute ethanol and pylorus ligation ratmodels suggested the possibility that the ineffectiveness of Bpilosa against gastric ulcers was due to lack of antihistaminicactivity in the plant In summary overall the data suggest thatB pilosa protects against HCLethanol-mediated ulcers viainhibition of prostaglandin biosynthesis

Previous phytochemical studies showed that a groupof flavonoids acyclichalcones are present in B pilosa[129 130] and the chalcones were proposed to have anti-ulcerative activity [131] Moreover nine hydroxychalcones 10were reported to possess gastric cytoprotective effects with24-dihydroxychalcone being the most active [132] Sincemethylene chloride extracts appear to be the most active Bpilosa extracts next the specific anti-ulcerative phytochemi-cals in the methylene chloride extracts of B pilosa and theirmodes of action needs to be probed

Despite the claims listed inTable 3 relatively few scientificstudies have been conducted in vitro and in vivo to address thetraditional ethnomedical uses of B pilosa Information aboutthe use of B pilosa as a botanical therapy recorded so far isfar from complete Studies conducted thus far only serve asa starting point for further investigation of B pilosa and theultimate efficacious use of the herb in clinical applications

Evidence-Based Complementary and Alternative Medicine 47

4 Toxicology

Despite its use as an ingredient in food for human con-sumption studies on systemic toxicity (eg acute subacutechronic and subchronic toxicities) of B pilosa in humansand animals are still inadequate and insufficient So faracute andor subchronic toxicities have been evaluated inrats and mice Oral acute and 28-day toxicities of waterextract of B pilosa leaves were evaluated in Wistar rats[133] An oral dose of water extract of B pilosa leaves at10 gkg BW showed no obvious mortality or changes inthe appearance in rats [134] The same extract at 08 gkgBWday once a day showed no obvious sub-chronic toxicityin rats over 28 days as measured by survival rate bodyweight and gross examination of organs [134] These dataare consistent with our data indicating that oral deliveryof the water extract of the B pilosa whole plant at 1 gkgBWday once a day is safe in rats over 28 days (unpublisheddata) Taken together these studies suggest that ingestion ofB pilosa aqueous extract at up to at 1 gkg BWday oncea day is highly safe in rats In addition the acute toxicityof aqueous and ethanol extracts of B pilosa in mice havebeen reported [134] Five- to six-week-old mice with weightsbetween 28 and 35 g received a peritoneal injection of bothextracts at the different doses The LD

50 the dose that causes

50 lethality of the aqueous and ethanol extracts in micewas 1230 gkg BW and 615 gkg BW respectively [134] Acomplete toxicological study has not been completed forhumans Furthermore the drug interactions of B pilosa withother drugs are unknown Further safety verification andclinical trials should be performed before B pilosa can beconsidered for medicinal use

5 Conclusions

B pilosa is an erect perennial plant with green leaves whiteor yellow flowers and tiny black seeds As it is distributedworldwide and is widely used as a folk remedy B pilosacan be thought of as an extraordinary source of food andmedicine However a comprehensive up-to-date review ofresearch on B pilosa has hitherto been unavailable In thisarticle scientific studies on B pilosa have been summarizedand critically discussed from the perspectives of botanyethnomedicine phytochemistry pharmacology and toxicol-ogy B pilosa is claimed to treat more than 40 disordersand 201 compounds have been identified from this plantThe medicinal utility of B pilosa and its modes of actionin relation to its known phytochemicals were discussedherein Polyynes flavonoids phenylpropanoids fatty acidsand phenolics are the primary bioactive compounds of Bpilosa and they have been reported to be effective in thetreatment of tumors inflammationimmune modulationdiabetes viruses microbes protozoans gastrointestinal dis-eases hypertension and cardiovascular diseases Cautionshould be exercised in the therapeutic use of B pilosa forhypoglycemia hypotension bleeding and allergy

Acknowledgments

The authors thank their laboratory members for construc-tive suggestions and the authors whose publications theycited for their contributions The authors also thank MsMiranda Loney of Academia Sinica for English editing of thispaper This work was supported by Grants 99-CDA-L11 and101S0010056 from Academia Sinica Taiwan Arlene P Bar-tolome is a recipient of MECO-TECO Sandwich ScholarshipProgram between Taiwan and the Philippines

References

[1] T Shen G H Li X N Wang and H X Lou ldquoThe genusCommiphora a review of its traditional uses phytochemistryand pharmacologyrdquo Journal of Ethnopharmacology vol 142 no2 pp 319ndash330

[2] C Long P Sauleau B David et al ldquoBioactive flavonoids ofTanacetum parthenium revisitedrdquo Phytochemistry vol 64 no2 pp 567ndash569 2003

[3] P O Karis and O Ryding Asteraceae Cladistics and Classifica-tion Bremer K Eds pp 559ndash569 Timber press Portland OreUSA 1994

[4] O N Pozharitskaya A N Shikov M N Makarova et alldquoAnti-inflammatory activity of a HPLC-fingerprinted aqueousinfusion of aerial part of Bidens tripartita LrdquoPhytomedicine vol17 no 6 pp 463ndash468 2010

[5] F Q Oliveira V Andrade-Neto A U Krettli and MG L Brandao ldquoNew evidences of antimalarial activity ofBidens pilosa roots extract correlated with polyacetylene andflavonoidsrdquo Journal of Ethnopharmacology vol 93 no 1 pp 39ndash42 2004

[6] Agriculture USDA ldquoPlants databaserdquo 2013in Natural Resources Conservation Servicehttpwwwnrcsusdagovwpsportalnrcssitenationalhome

[7] Y Zhou and X Z Yan ldquoExperimental study of qi deficiencysyndrome and Codonopsis pillosulae and Astragalus injectionon the immune response in micerdquo Zhong Xi Yi Jie He Za Zhivol 9 no 5 pp 286ndash262 1989

[8] S C Chien P H Young Y J Hsu et al ldquoAnti-diabeticproperties of three common Bidens pilosa variants in TaiwanrdquoPhytochemistry vol 70 no 10 pp 1246ndash1254 2009

[9] M J Alcaraz and M J Jimenez ldquoFlavonoids as anti-inflammatory agentsrdquo Fitoterapia vol 59 no 1 pp 25ndash38 1988

[10] FAO Agriculture Food and Nutrition for AfricamdashA ResourceBook for Teachers of Agriculture Publishing ManagementGroup FAO Information Division Rome Italy 1997

[11] M B Rokaya Z Munzbergova B Timsina and K R BhattaraildquoRheum australe D Don a review of its botany ethnobotanyphytochemistry and pharmacologyrdquo Journal of Ethnopharma-cology vol 141 no 3 pp 761ndash774 2012

[12] P H Young Y J Hsu and W C Yang ldquoBidens pilosa L andits medicinal userdquo in Recent Progress in Medicinal Plants DrugPlant II A S AwaadVK Singh and JNGovil Eds StandiumPress Houston Tex USA 2010

[13] C Ge ldquoCytologic study of Bidens bipinnata LrdquoZhongguo ZhongYao Za Zhi vol 15 no 2 pp 72ndash125 1990

[14] L C Chiang J S Chang C C Chen L T Ng and C CLin ldquoAnti-herpes simplex virus activity of Bidens pilosa and

48 Evidence-Based Complementary and Alternative Medicine

Houttuynia cordatardquoAmerican Journal of ChineseMedicine vol31 no 3 pp 355ndash362 2003

[15] N P Rybalchenko V A Prykhodko S S Nagorna et alldquoIn vitro antifungal activity of phenylheptatriyne from Bidenscernua L against yeastsrdquo Fitoterapia vol 81 no 5 pp 336ndash3382010

[16] K Redl W Breu B Davis and R Bauer ldquoAnti-inflammatoryactive polyacetylenes from Bidens campylothecardquo Planta Med-ica vol 60 no 1 pp 58ndash62 1994

[17] M Ayyanar and S Ignacimuthu ldquoTraditional knowledge ofKani tribals inKouthalai of Tirunelveli hills TamilNadu IndiardquoJournal of Ethnopharmacology vol 102 no 2 pp 246ndash255 2005

[18] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant intraditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[19] AKAbbas KMMurphy andA Sher ldquoFunctional diversity ofhelper T lymphocytesrdquo Nature vol 383 no 6603 pp 787ndash7931996

[20] J H Cano and G Volpato ldquoHerbal mixtures in the traditionalmedicine of Eastern Cubardquo Journal of Ethnopharmacology vol90 no 2-3 pp 293ndash316 2004

[21] J Namukobe J M Kasenene B T Kiremire et al ldquoTraditionalplants used for medicinal purposes by local communitiesaround the Northern sector of Kibale National Park UgandardquoJournal of Ethnopharmacology vol 136 no 1 pp 236ndash245 2011

[22] C Lans ldquoComparison of plants used for skin and stomachproblems in Trinidad and Tobago with Asian ethnomedicinerdquoJournal of Ethnobiology andEthnomedicine vol 3 article 3 2007

[23] S Dharmananda ldquoA Popular Remedy Ecapes Notice ofWesternPractitionersrdquo 2013 httpwwwitmonlineorgartsbidenshtm

[24] Y J Hsu T H Lee C L T Chang Y T Huang andW C YangldquoAnti-hyperglycemic effects and mechanism of Bidens pilosawater extractrdquo Journal of Ethnopharmacology vol 122 no 2 pp379ndash383 2009

[25] E Noumi F Houngue and D Lontsi ldquoTraditional medicinesin primary health care plants used for the treatment ofhypertension in Bafia Cameroonrdquo Fitoterapia vol 70 no 2 pp134ndash139 1999

[26] P V Tan T Dimo and E Dongo ldquoEffects of methanolcyclohexane and methylene chlo ride extracts of Bidens pilosaon various gastric ulcer models in ratsrdquo Journal of Ethnophar-macology vol 73 no 3 pp 415ndash421 2000

[27] R L C Pereira T Ibrahim L Lucchetti A J R Da Silva and VL G De Moraes ldquoImmunosuppressive and anti-inflammatoryeffects of methanolic extract and the polyacetylene isolatedfrom Bidens pilosa Lrdquo Immunopharmacology vol 43 no 1 pp31ndash37 1999

[28] T Dimo J Azay P V Tan et al ldquoEffects of the aqueous andmethylene chloride extracts of Bidens pilosa leaf on fructose-hypertensive ratsrdquo Journal of Ethnopharmacology vol 76 no 3pp 215ndash221 2001

[29] CWiartMedicinal Plants of Southeast Asia Pelanduk SelabgorDarul Ehsan Malaysia 2000

[30] F L Silva D C H Fischer J F Tavares M S Silva P FDe Athayde-Filho and J M Barbosa-Filho ldquoCompilation ofsecondary metabolites from Bidens pilosa LrdquoMolecules vol 16no 2 pp 1070ndash1102 2011

[31] A H Chen S R Lin and C H Hong ldquoPhytochemical studyon Bidens pilosa L varminorrdquo Huaxue pp 38ndash42 1975

[32] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[33] M C Zulueta M Tada and C Y Ragasa ldquoA diterpene fromBidens pilosardquo Phytochemistry vol 38 no 6 pp 1449ndash14501995

[34] J Wang H Yang Z W Lin and H D Sun ldquoFlavonoids fromBidens pilosa var radiatardquo Phytochemistry vol 46 no 7 pp1275ndash1278 1997

[35] A Zhao Q Zhao L Peng et al ldquoA new chalcone glycoside fromBidens pilosardquo Acta Botanica Yunnanica vol 26 no 1 pp 121ndash126

[36] P Gao Y Wu S Cui and Y Zou ldquoSynthesis and absoluteconfiguration of the 7-phenylhepta-4 6-diyne-1 2-diol isolatedfrom Bidens pilosardquo Synthesis no 13 pp 2131ndash2135 2011

[37] S Wang B Yang D Zhu D He and L Wang ldquoActivecomponents of Bidens pilosa Lrdquo Zhongcaoyao vol 36 pp 20ndash21

[38] F Bohlmann T Burkhardt and C Zdero Naturally OccuringAcetylenes Academic Press New York NY USA 1973

[39] T M Sarg A M Ateya N M Farrag and F A AbbasldquoConstituents and biological activity of Bidens pilosa L grownin EgyptrdquoActa Pharmaceutica Hungarica vol 61 no 6 pp 317ndash323 1991

[40] H A L Valdes and H P Rego ldquoBidens pilosa Linnerdquo RevistaCubana de Plantas Medicinales vol 6 pp 28ndash33

[41] F Bohlmann H Bornowski and K M Kleine ldquoNew polyynesfrom the tribe Heliantheaerdquo Chemische Berichte vol 97 pp2135ndash2138

[42] RWang Q XWu and Y P Shi ldquoPolyacetylenes and flavonoidsfrom the aerial parts ofBidens pilosardquo PlantaMedica vol 76 no9 pp 893ndash896 2010

[43] L W Wu Y M Chiang H C Chuang et al ldquoA novelpolyacetylene significantly inhibits angiogenesis and promotesapoptosis in human endothelial cells through activation of theCDK inhibitors and caspase-7rdquo Planta Medica vol 73 no 7 pp655ndash661 2007

[44] L W Wu Y M Chiang H C Chuang et al ldquoPolyacetylenesfunction as anti-angiogenic agentsrdquo Pharmaceutical Researchvol 21 no 11 pp 2112ndash2119 2004

[45] H L Yang S C Chen N W Chang et al ldquoProtection fromoxidative damage using Bidens pilosa extracts in normal humanerythrocytesrdquo Food and Chemical Toxicology vol 44 no 9 pp1513ndash1521 2006

[46] H Q Wang S J Lu H Li and Z H Yao ldquoEDTA-enhancedphytoremediation of lead contaminated soil by Bidens maxi-mowiczianardquo Journal of Environmental Sciences vol 19 no 12pp 1496ndash1499 2007

[47] S TobinagaMK SharmaWG L Aalbersberg et al ldquoIsolationand identification of a potent antimalarial and antibacterialpolyacetylene from Bidens pilosardquo Planta Medica vol 75 no 6pp 624ndash628 2009

[48] C L T Chang H K Kuo S L Chang et al ldquoThe distincteffects of a butanol fraction of Bidens pilosa plant extract onthe development of Th1-mediated diabetes and Th2-mediatedairway inflammation in micerdquo Journal of Biomedical Sciencevol 12 no 1 pp 79ndash89 2005

[49] S L Chang C L T Chang Y M Chiang et al ldquoPolyacetyleniccompounds and butanol fraction from Bidens pilosa can mod-ulate the differentiation of helper T cells and prevent autoim-mune diabetes in non-obese diabetic micerdquo Planta Medica vol70 no 11 pp 1045ndash1051 2004

Evidence-Based Complementary and Alternative Medicine 49

[50] Y M Chiang C L T Chang S L Chang W C Yang and LF Shyur ldquoCytopiloyne a novel polyacetylenic glucoside fromBidens pilosa functions as a T helper cell modulatorrdquo Journal ofEthnopharmacology vol 110 no 3 pp 532ndash538 2007

[51] R PUbillas CDMendez S D Jolad et al ldquoAntihyperglycemicacetylenic glucosides from Bidens pilosardquo Planta Medica vol66 no 1 pp 82ndash83 2000

[52] L Alvarez S Marquina M L Villarreal D Alonso E Arandaand G Delgado ldquoBioactive polyacetylenes from Bidens pilosardquoPlanta Medica vol 62 no 4 pp 355ndash357 1996

[53] C L T Chang S L Chang Y M Lee et al ldquoCytopiloyne apolyacetylenic glucoside prevents type 1 diabetes in nonobesediabetic micerdquo Journal of Immunology vol 178 no 11 pp 6984ndash6993 2007

[54] G Kusano A Kusano and Y Seyama Novel Hypoglycemicand Anti-Inflammatory Polyacetylenic Compounds Their Com-positions Bidens Plant Extract Fractions and CompositionsContaining the Plant or Fraction JPO Tokyo Japan 2004

[55] M G L Brandao A U Krettli L S R Soares C G CNery and H C Marinuzzi ldquoAntimalarial activity of extractsand fractions from Bidens pilosa and other Bidens species(Asteraceae) correlated with the presence of acetylene andflavonoid compoundsrdquo Journal of Ethnopharmacology vol 57no 2 pp 131ndash138 1997

[56] A U Krettli V F Andrade-Neto M D G L Brandao andW M S Ferrari ldquoThe search for new anti-malarial drugs fromplants used to treat fever and malaria or plants ramdomlyselected a reviewrdquoMemorias do Instituto Oswaldo Cruz vol 96no 8 pp 1033ndash1042 2001

[57] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[58] C K Wat R K Biswas E A Graham L Bohm G H NTowers and E R Waygood ldquoUltraviolet-mediated cytotoxicactivity of phenylheptatriyne from Bidens pilosa Lrdquo Journal ofNatural Products vol 42 no 1 pp 103ndash111 1979

[59] Y Sashida K Ogawa M Kitada H Karikome Y Mimakiand H Shimomura ldquoNew aurone glucosides and new phenyl-propanoid glucosides from Bidens pilosardquo Chemical and Phar-maceutical Bulletin vol 39 no 3 pp 709ndash711 1991

[60] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens pilosa L (TFB) on animal liver injury andliver fibrosisrdquo Journal of Ethnopharmacology vol 116 no 3 pp539ndash546 2008

[61] B Hoffmann and J Holzl ldquoAcylated compounds from Bidenspilosardquo Planta Medica vol 55 pp 108ndash109

[62] B Hoffmann and J Holzl ldquoA methylated chalcone glucosidefrom Bidens pilosardquo Phytochemistry vol 27 no 11 pp 3700ndash3701 1988

[63] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[64] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[65] B Hoffmann and J Holzl ldquoWeitere acylierte chalkone ausBidens pilosardquo Planta Medica vol 54 pp 450ndash451 1988

[66] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant in11traditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[67] V V Pham V V T K P V L Hoang and V K Phan ldquoFlavonoidcompounds from the plant Bidens pilosa L (Asteraceae)rdquo TapChi Duoc Hoc vol 50 pp 48ndash53

[68] S D Sarker B Bartholomew R J Nash and N Robinson ldquo5-O-methylhoslundin an unusual flavonoid from Bidens pilosa(Asteraceae)rdquo Biochemical Systematics and Ecology vol 28 no6 pp 591ndash593 2000

[69] S L Chang Y M Chiang C L T Chang et al ldquoFlavonoidscentaurein and centaureidin from Bidens pilosa stimulate IFN-120574 expressionrdquo Journal of Ethnopharmacology vol 112 no 2 pp232ndash236 2007

[70] Y M Chiang D Y Chuang S Y Wang Y H Kuo P WTsai and L F Shyur ldquoMetabolite profiling and chemopreventivebioactivity of plant extracts from Bidens pilosardquo Journal ofEthnopharmacology vol 95 no 2-3 pp 409ndash419 2004

[71] A Kusano Y Seyama E Usami et al ldquoStudies on the antioxi-dant active constituents of the dried powder from Bidens pilosaL var radiata SchrdquoNaturalMedicines vol 57 no 3 pp 100ndash1042003

[72] I Doleckova L Rarova J Gruz et al ldquoAnti-proliferative andanti-angiogenic effects of flavone eupatorin an active con-stituent of chloroform extract ofOrthosiphon stamineus leavesrdquoFitoterapia vol 83 no 6 pp 1000ndash1007

[73] K Bairwa R Kumar R J Sharma and R K Roy ldquoAn updatedreview on Bidens pilosa Lrdquo Der Pharma Chemica vol 2 no 3pp 325ndash337

[74] Q Xia Y Liu and Y Li ldquoDetermination of hyperoside indifferent parts and different species of Herba Bidens by RP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol 50pp 48ndash53

[75] M Horiuchi and Y Seyama ldquoImprovement of the antiinflam-matory and antiallergic activity of Bidens pilosa L var radiataSCHERFF treated with enzyme (Cellulosine)rdquo Journal of HealthScience vol 54 no 3 pp 294ndash301 2008

[76] M G L Brandao C G C Nery M A S Mamao and AU Krettli ldquoTwo methoxylated flavone glycosides from Bidenspilosardquo Phytochemistry vol 48 no 2 pp 397ndash400 1998

[77] P Kumari K Misra B S Sisodia et al ldquoA promising anticancerand antimalarial component from the leaves of Bidens pilosardquoPlanta Medica vol 75 no 1 pp 59ndash61 2009

[78] G Seelinger I Merfort U Wolfle and C M Schempp ldquoAnti-carcinogenic effects of the flavonoid luteolinrdquoMolecules vol 13no 10 pp 2628ndash2651 2008

[79] G Seelinger I Merfort and C M Schempp ldquoAnti-oxidantanti-inflammatory and anti-allergic activities of luteolinrdquoPlanta Medica vol 74 no 14 pp 1667ndash1677 2008

[80] C C Yit and N P Das ldquoCytotoxic effect of butein on humancolon adenocarcinoma cell proliferationrdquo Cancer Letters vol82 no 1 pp 65ndash72 1994

[81] J A Beutler E Hamel A J Vlietinck et al ldquoStructure-activityrequirements for flavone cytotoxicity and binding to tubulinrdquoJournal of Medicinal Chemistry vol 41 no 13 pp 2333ndash23381998

[82] M R Kviecinski K B Felipe T Schoenfelder et al ldquoStudy ofthe antitumor potential of Bidens pilosa (Asteraceae) used inBrazilian folkmedicinerdquo Journal of Ethnopharmacology vol 117no 1 pp 69ndash75 2008

[83] P Sundararajan A Dey A Smith A G Doss M Rajappanand S Natarajan ldquoStudies of anticancer and antipyretic activityof Bidens pilosawhole plantrdquo African Health Sciences vol 6 no1 pp 27ndash30 2006

[84] YMChiang C P Lo Y P Chen et al ldquoEthyl caffeate suppressesNF-120581B activation and its downstream inflammatory mediatorsiNOS COX-2 and PGE2 in vitro or in mouse skinrdquo BritishJournal of Pharmacology vol 146 no 3 pp 352ndash363 2005

50 Evidence-Based Complementary and Alternative Medicine

[85] J S Kim H J Lee M H Lee J Kim C Jin and J HRyu ldquoLuteolin inhibits LPS-stimulated inducible nitric oxidesynthase expression in BV-2 microglial cellsrdquo Planta Medicavol 72 no 1 pp 65ndash68 2006

[86] P A Ruiz and D Haller ldquoFunctional diversity of flavonoidsin the inhibition of the proinflammatory NF-120581B IRF and Aktsignaling pathways in murine intestinal epithelial cellsrdquo Journalof Nutrition vol 136 no 3 pp 664ndash671 2006

[87] A Xagorari A Papapetropoulos A Mauromatis MEconomou T Fotsis and C Roussos ldquoLuteolin inhibitsan endotoxin-stimulated phosphorylation cascade andproinflammatory cytokine production in macrophagesrdquoJournal of Pharmacology and Experimental Therapeutics vol296 no 1 pp 181ndash187 2001

[88] N Yoshida T Kanekura Y Higashi and T Kanzaki ldquoBidenspilosa suppresses interleukin-1120573-induced cyclooxygenase-2expression through the inhibition of mitogen activated proteinkinases phosphorylation in normal human dermal fibroblastsrdquoJournal of Dermatology vol 33 no 10 pp 676ndash683 2006

[89] M Muchuweti C Mupure A Ndhlala T Murenje and M AN Benhura ldquoScreening of antioxidant and radical scavengingactivity of Vigna ungiculata Bidens pilosa and Cleome gynan-drardquo American Journal of Food Technology vol 2 no 3 pp 161ndash168 2007

[90] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens bipinnata L against carbon tetrachloride-induced liver fibrosis in ratsrdquo Journal of Pharmacy and Pharma-cology vol 60 no 10 pp 1393ndash1402 2008

[91] S L Chang H H Yeh Y S Lin Y M Chiang T K Wuand W C Yang ldquoThe effect of centaurein on interferon-120574 expression and Listeria infection in micerdquo Toxicology andApplied Pharmacology vol 219 no 1 pp 54ndash61 2007

[92] F Deba T D Xuan M Yasuda and S Tawata ldquoChemical com-position and antioxidant antibacterial and antifungal activitiesof the essential oils from Bidens pilosa Linn var Radiatardquo FoodControl vol 19 no 4 pp 346ndash352 2008

[93] A O T Ashafa and A J Afolayan ldquoScreening the root extractsfrom Biden pilosa L var radiata (Asteraceae) for antimicrobialpotentialsrdquo Journal of Medicinal Plant Research vol 3 no 8 pp568ndash572 2009

[94] T B Nguelefack T Dimo E P Nguelefack Mbuyo P V TanS V Rakotonirina and A Kamanyi ldquoRelaxant effects of theneutral extract of the leaves of Bidens pilosa linn on isolated ratvascular smooth musclerdquo Phytotherapy Research vol 19 no 3pp 207ndash210 2005

[95] M T Grombone-Guaratini K L Silva-Brandao V N SolferiniJ Semir and J R Trigo ldquoSesquiterpene and polyacetyleneprofile of the Bidens pilosa complex (Asteraceae Heliantheae)from Southeast of Brazilrdquo Biochemical Systematics and Ecologyvol 33 no 5 pp 479ndash486 2005

[96] L L Lin C Y Wu H C Hsiu M T Wang and H ChuangldquoStudies on diabetes mellitus I The hypoglycemic activity ofphytosterin on alloxan diabetic ratsrdquo Taiwan Yi Xue Hui Za Zhivol 66 no 2 pp 58ndash65 1967

[97] M A N Benhura and I C Chitsiku ldquoThe extractable 120573-carotene content of Guku (Bidens pilosa) leaves after cookingdrying and storagerdquo International Journal of Food Science andTechnology vol 32 no 6 pp 495ndash500 1997

[98] J S Chang L C Chiang C C Chen L T Liu K C Wangand C C Lin ldquoAtileukemic activity of Bidens pilosa l varminor (blume) sherff andHouttuynia cordata thunbrdquo AmericanJournal of Chinese Medicine vol 29 no 2 pp 303ndash312 2001

[99] F Deba T D Xuan M Yasuda and S Tawata ldquoHerbicidal andfungicidal activities and identification of potential phytotoxinsfrom Bidens pilosa L var radiata Scherff research paperrdquoWeedBiology and Management vol 7 no 2 pp 77ndash83 2007

[100] J K Kumar and A K Sinha ldquoA new disubstituted acetylacetonefrom the leaves ofBidens pilosa LinnrdquoNatural Product Researchvol 17 no 1 pp 71ndash74 2003

[101] K Ogawa and Y Sashida ldquoCaffeoyl derivatives of a sugarlactone and its hydroxy acid from the leaves of Bidens pilosardquoPhytochemistry vol 31 no 10 pp 3657ndash3658 1992

[102] Q Xia Y Liu and Y Li ldquoDetermination of gallic acid fromdifferent species and different medical parts of Herba Bidens byRP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol24 no 03 pp 308ndash310

[103] W J Lee L F Wu W K Chen C J Wang and T HTseng ldquoInhibitory effect of luteolin on hepatocyte growth fac-torscatter factor-induced HepG2 cell invasion involving bothMAPKERKs and PI3K-Akt pathwaysrdquo Chemico-BiologicalInteractions vol 160 no 2 pp 123ndash133 2006

[104] J A Beutler J H Cardellina C M Lin E Hamel G MCragg andM R Boyd ldquoCentaureidin a cytotoxic flavone fromPolymnia fruticosa inhibits tubulin polymerizationrdquo BioorganicandMedicinal Chemistry Letters vol 3 no 4 pp 581ndash584 1993

[105] A Verma A Su A M Golin B OrsquoMarrah and J K AmorosaldquoThe lateral view a screening method for knee traumardquo Aca-demic Radiology vol 8 no 5 pp 392ndash397 2001

[106] J Corren M Lemay Y Lin L Rozga and R K RandolphldquoClinical and biochemical effects of a combination botanicalproduct (ClearGuard) for allergy a pilot randomized double-blind placebo-controlled trialrdquo Nutrition Journal vol 7 no 1article 20 2008

[107] M S Gachet J S Lecaro M Kaiser et al ldquoAssessment of anti-protozoal activity of plants traditionally used in Ecuador in thetreatment of leishmaniasisrdquo Journal of Ethnopharmacology vol128 no 1 pp 184ndash197 2010

[108] W C Yang M Ghiotto B Barbarat and D Olive ldquoThe role ofTec protein-tyrosine kinase in T cell signalingrdquo The Journal ofBiological Chemistry vol 274 no 2 pp 607ndash617 1999

[109] S Tewtrakul H Miyashiro N Nakamura et al ldquoHIV-1 inte-grase inhibitory substances from Coleus parvifoliusrdquo Phytother-apy Research vol 17 no 3 pp 232ndash239 2003

[110] T Dimo T B Nguelefack P V Tan et al ldquoPossible mechanismsof action of the neutral extract from Bidens pilosa L leaves onthe cardiovascular system of anaesthetized ratsrdquo PhytotherapyResearch vol 17 no 10 pp 1135ndash1139 2003

[111] T S C Li Chinese and Related North American Herbs CRCPress New York NY USA 2002

[112] H Wu H Chen X Hua Z Shi L Zhang and J ChenldquoClinical therapeutic effect of drug-separated moxibustion onchronic diarrhea and its immunologic mechanismsrdquo Journal ofTraditional Chinese Medicine vol 17 no 4 pp 253ndash258 1997

[113] S W Wright R R Harris J S Kerr et al ldquoSynthesis chemicaland biological properties of vinylogous hydroxamic acids dualinhibitors of 5-lipoxygenase and IL-1 biosynthesisrdquo Journal ofMedicinal Chemistry vol 35 no 22 pp 4061ndash4068 1992

[114] W R Almiron and M E Brewer ldquoClassification of immaturestage habitats of Culicidae (Diptera) collected in CordobaArgentinardquo Memorias do Instituto Oswaldo Cruz vol 91 no 1pp 1ndash9 1996

[115] N L Wang J Wang X S Yao and S Kitanaka ldquoTwo newmonoterpene glycosides and a new (+)-jasmololone glucoside

Evidence-Based Complementary and Alternative Medicine 51

from Bidens parvifloraWilldrdquo Journal of Asian Natural ProductsResearch vol 9 no 5 pp 473ndash479 2007

[116] P Champagnat ldquoRole of the terminal bud in the actionexercised by the cotyledon of Bidens pilosus L var radiatus onits axillary budrdquo Comptes Rendus des Seances et Memoires de laSociete de Biologie vol 145 no 17-18 pp 1374ndash1376 1951

[117] S F Nielsen S B Christensen G Cruciani A Kharazmiand T Liljefors ldquoAntileishmaniai chalcones statistical designsynthesis and three-dimensional quantitative structure-activityrelationship analysisrdquo Journal of Medicinal Chemistry vol 41no 24 pp 4819ndash4832 1998

[118] Y C Hwang J J H Chu P L Yang W Chen and M V YatesldquoRapid identification of inhibitors that interfere with poliovirusreplication using a cell-based assayrdquo Antiviral Research vol 77no 3 pp 232ndash236 2008

[119] V F Andrade-Neto M G L Brandao F Q Oliveira et alldquoAntimalarial activity of Bidens pilosa L (Asteraceae) ethanolextracts fromwild plants collected in various localities or plantscultivated in humus soilrdquo Phytotherapy Research vol 18 no 8pp 634ndash639 2004

[120] R J Marles and N R Farnsworth ldquoAnti-diabetic plants andtheir active constituentsrdquo Phytomedicine vol 2 no 2 pp 137ndash189

[121] M Habeck ldquoDiabetes treatments get sweet help from naturerdquoNature Medicine vol 9 no 10 p 1228 2003

[122] H W Lin G Y Han and S X Liao ldquoStudies on the activeconstituents of the Chinese traditional medicine Polygonatumodoratum (Mill) DrucerdquoActa Pharmaceutica Sinica vol 29 no3 pp 215ndash222 1994

[123] C L T Chang Y C Chen H M Chen N S Yang andW C Yang ldquoNatural cures for type 1 diabetes a reviewof phytochemicals biological actions and clinical potentialrdquoCurrent Medicinal Chemistry vol 20 no 7 pp 899ndash907 2013

[124] L Dey A S Attele and C S Yuan ldquoAlternative therapies fortype 2 diabetesrdquo Alternative Medicine Review vol 7 no 1 pp45ndash58 2002

[125] C L T ChangH Y Liu T F Kuo et al ldquoAnti-diabetic effect andmode of action of cytopiloynerdquo Evidence-Based Complementaryand Alternative Medicine vol 2013 Article ID 685642 13 pages2013

[126] M R Khan M Kihara and A D Omoloso ldquoAnti-microbialactivity of Bidens pilosa Bischofia javanica Elmerillia papuanaand Sigesbekia orientalisrdquo Fitoterapia vol 72 no 6 pp 662ndash6652001

[127] T Dimo T B Nguelefack P Kamtchouing E Dongo ARakotonirina and S V Rakotonirina ldquoHypotensive effects ofmethanol extract from Bidens pilosa Linn on hypertensive ratsrdquoComptes Rendus de lrsquoAcademie des Sciences Serie III vol 322no 4 pp 323ndash329 1999

[128] A K Hassan O Deogratius J F Nyafuono O Francis and OP Engeu ldquoWound healing potential of the ethanolic extracts ofBidens pilosa and Ocimum suaverdquo African Journal of Pharmacyand Pharmacology vol 5 no 2 pp 132ndash136 2011

[129] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[130] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[131] C Kandaswami and EMiddleton ldquoFree radical scavenging andantioxidant activity of plant flavonoidsrdquo Advances in Experi-mental Medicine and Biology vol 366 pp 351ndash376 1994

[132] K Yamamoto H Kakegawa H Ueda et al ldquoGastric cytopro-tective anti-ulcerogenic actions of hydroxychalcones in ratsrdquoPlanta Medica vol 58 no 5 pp 389ndash393 1992

[133] J O C Ezeonwumelu A K Julius C N Muhoho et alldquoBiochemical and histological studies of aqueous extract ofBidens pilosa leaves from Ugandan Rift valley in ratsrdquo BritishJournal of Pharmacology and Toxicology vol 2 no 6 pp 302ndash309

[134] L Frida S Rakotonirina A Rakotonirina and J P SavineauldquoIn vivo and in vitro effects of Bidens pilosa L (Asteraceae)leaf aqueous and ethanol extracts on primed-oestrogenized ratuterine musclerdquo African Journal of Traditional Complementaryand Alternative Medicines vol 5 no 1 pp 79ndash91 2008

52 Evidence-Based Complementary and Alternative Medicine

Composition Comments

1 Please provide valid postal code to the address if

possible

2 We made the highlighted change in Table 4 for the sake

of correctness Please check

3 Please check the correctness of the highlighted part in

Table 5

4 We considered the highlighted part in Table 11 as

reference Please check similar cases throughout

5 Please check the format of Table 18

6 We reformatted Table 20 Please check

7 We made the highlighted change in Table 4 for the sake

of consistency Please check similar cases throughout the

paper

8 We made the highlighted change(s) for the sake

of consistency Please check simila highlighted cases

throughout

9 We made the highlighted change in Table 9 for the sake

of clarity Please check similar cases through out

10 We made the highlighted change(s) for the sake of

correctness Please check

11 Comment on ref [66] This reference is a repetition

of [18] Please check similar cases throughout

Author(s) Name(s)

Author 1

Author 2

Author 3

Evidence-Based Complementary and Alternative Medicine 9

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

4857911-Tridecatetrayne-12-diol

12-Dihydroxytrideca-57911-tetrayne

HOOH

Whole (Taiwan) [43 44 46]

49(R)-357911-Tridecapentayne-12-diol

(R)-12-Dihydroxytrideca-357911-pentayne

HO

OH

Aerial (Japan) [47]

50

(4E)-1-(Hydroxymethyl)-4-dodecene-6810-triyn-1-yl-120573-D-glucopyranoside

2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-

tridecene-7911-triyne

O

HO

H

OH

H

H

HOH

O

OH

HO

H Aerial (USA)whole (Taiwan)leaves (Taiwan)

[8 45 48ndash51]

51

(4E)-1-(2-Hydroxyethyl)-4-dodecene-6810-triyn-1-yl-120573-D-glucopyranoside

3-120573-D-Glucopyranosyloxy-1-hydroxy-6(E)-

tridecene-81012-triyne

OHO

H

OH

H

H

HOH

O

OH

OH

HAerial (USA andChina) whole(Taiwan) leaves(Taiwan)

[8 35 4245 48ndash51]

52

3-Hydroxy-6-tetradecene-81012-triynyl-120573-D-glucopyranoside

120573-D-Glucopyranosyloxy-

3-hydroxy-6E-tetradecence-81012-triyne

OH

O

OHO

H

OH

H

H

HOH

OH

HWhole (Mexico) [52]

53

1-(Hydroxymethyl)-46810-dodecatetrayne-1-yl-120573-D-glucopyranoside

Cytopiloyne2-120573-D-

Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne

O

HO

OHO

H

OH

H

H

HOH

OH

HWhole (Taiwan)leaves (Taiwan) [8 50 53]

54

2-O-D-Glucosyltrideca-11E-en-3579-tetrayn-12-diol

OH

O

O

OHH

HO

H

H

HHOH

OH

Leaves (Brazil) [27]

10 Evidence-Based Complementary and Alternative Medicine

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

55

(R)-1-(Hydroxymethyl)-246810-dodecapentayn-1-yl-a-D-glucopyranoside

2- 120573 -D-Glucopyranosyloxy-1-hydroxytrideca-357911-pentayne

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Chinaand Japan) [35 47]

56

1-[[(Carboxyacetyl)oxy]methyl]46810-dodecatetraynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Japan) [54]

57

(4E)-1-[[(Carboxyacetyl)oxy]-methyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHO

H

OH

Aerial (Japan) [54]

58

(4E)-1-[[(Carboxyacetyl)oxy]-ethyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

O

O

O

OHO

O

OHOH

H

HO

H

H

HHO

HOH

Aerial (Japan) [54]

59 (5E)-5-Heptene-13-diyn-1-yl-benzene

1-Phenylhepta-13-diyn-5en Whole (Taiwan) [32]

60 7-Phenyl-2(E)-heptene-46-diyn-1-ol

OHRoots (notstated) Aerial(China)

[38 42]

617-Phenyl-2(E)-heptene-46-diyn-1-ol-acetate

OO

Roots (notstated Brazil) [38 55 56]

62 7-Phenyl-46-heptadiyn-2-ol Pilosol A

OH

Whole (Taiwan)aerial (China) [42 57]

Evidence-Based Complementary and Alternative Medicine 11

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

63 7-Phenylhepta-46-diyn-12-diol HO

OH

Aerial (China) [42]

64 135-Heptatriyn-1-yl-benzene

1-Phenylhepta-135-triyne

Leaves (notstated) leaves oftissue culture(not stated)aerial (TanzaniaChina)Whole (Taiwan)roots (Brazil)

[18 34 3842 56ndash58]

65 7-Phenyl-246-heptatriyn-1-ol HO

Leaves (notstated) aerial(China)

[38 42]

667-Phenyl-246-heptatriyn-1-ol-acetate O

O Leaves (notstated) [38]

675-(2-Phenylethynyl)-2-thiophenemethanol

S OHAerial (China) [42]

685-(2-Phenylethynyl)-2120573-glucosylmethyl-thiophene

O

O

OH

H

OH

H

H

HOH H

O

HO

S

Aerial (China) [42]

69

3- 120573 -D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne

OH

O

OH

HOHO

H

H

HHOH

OH

Leaves(Cameroon) [28]

701-Phenyl-13-diyn-5-en-7-ol-acetate O

O

Leaves (Brazil) [27]

SN denotes serial number

the chloroform fraction was the most toxic with a halfmaximal inhibitory concentration (IC

50) of 97 plusmn 72 and 83 plusmn

52 120583gmL in NRU and MTT respectively [82] Kumari andcolleagues also reported the anti-cancer and anti-malarialactivities of B pilosa leaves [77] Based on a cytotoxicity-directed fractionation strategy they identified phenyl-135-heptatriene with IC

50values of 8plusmn001 049plusmn045 07plusmn001

and 10 plusmn 001 120583gmL against human oral liver colon andbreast cancer cell lines respectively However phenyl-135-heptatriyne showed lower activity against breast cancer celllines than the chloroform leaf extract which had an IC

50

value of 65 plusmn 001 120583gmL Moreover the positive controltaxol showed higher activity than phenyl-135-heptatriyne[77] Furthermore in vitro comet assays were performed toevaluate the toxicity of n-hexane chloroform and methanolextracts of B pilosa and its ethyl acetate acetone and waterfractions onHela andKB cellsThe ethyl acetate fraction fromthe methanol extract exhibited the highest activity with halfmaximal cytotoxic concentrations (CTC

50) of 9652120583gmL

and 5862120583gmL against Hela and KB cells respectivelyDespite the moderate toxicity these findings suggest thatthese B pilosa extractsfractions could be useful for future

12 Evidence-Based Complementary and Alternative Medicine

Table5Flavon

oids

isolated

from

Bpilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

71

2-[(34-Dihydroxyph

enyl)-

methylene]-

6-hydroxy-3(2H

)-benzofuranon

e

Sulfu

retin

O

OHO

OH

OH

Aeria

l(Ch

ina)

[35]

72

2-[(34-Dihydroxyph

enyl)-

methylene]-

67-dihydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-673101584041015840-

tetrahydroxy

maritimetin

O

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

73

2-[(34-Dihydroxyph

enyl)-

methylene]-6-

(120573-D

-glucopyrano

syloxy)-7-

hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-120573-D

-glucop

yranosyl-

673101584041015840-te

trahydroxy

maritimein

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japanand

China)aerial(Ch

ina)

[355960]

74

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

(120573-D

-glucopyrano

syloxy)-

6-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-7-O-120573-D

-glucop

yranosyl-

673101584041015840-

tetrahydroxy

O

OHO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japan)

[59]

75

6-[(6-O-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-acetyl-

120573-D

-glucopyrano

syl)

-673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OHO

H HO

H

H

HOH

HO

O

O

Leaves

(Not

stated)

aeria

l(Ch

ina)

[3559]

Evidence-Based Complementary and Alternative Medicine 13

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

76

6-[(36-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-

2-[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone(Z)-6-O

-(36-di-O

-acetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

bideno

sideA

O

O

HO

OH

OH

OHOH O

H

H

HOH

HO

O

O

O

Leaves

(China)

[60]

77

6-[(46-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(410158401015840610158401015840-

diacetyl-120573-D

-glucropyrano

syl)-

673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

HO

O

OO

Leaves

(not

stated)

aeria

l(Ch

ina)

[343561]

78

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

hydroxy-6-

[(246-tri-O

-acetyl-120573

-D-

glucop

yranosyl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(210158401015840410158401015840610158401015840-

triacetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

O

OOO

O

Leaves

(not

stated)

aeria

l(Ch

ina)

[3461]

79

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-6-

[(346-tri-O

-acetyl-120573

-D-glucopyrano

syl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(310158401015840410158401015840610158401015840-

triacetyl-120573

-D-glucrop

yranosyl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H OH

H

HOH

HO

OOO O

Aeria

l(Ch

ina)

[3435]

80

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-

6-[[6-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propenyl]-120573-

D-glucopyrano

syl]o

xy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-

p-coum

aroyl-

120573-D

-glucropyrano

syl)-

673101584041015840-te

trahydroxy

OH

O

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OLeaves

(Japan)

[59]

14 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

81

1-[2-(120573-D

-Glucopyrano

syloxy)-

4-hydroxyphenyl]-2-

hydroxy-3-(3-

hydroxyphenyl)-2-

prop

en-1-on

e

Chalcone120572

32101584041015840-

tetrahydroxy-

21015840-O

-120573-D

-glucop

yranosyl

HO

O

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[35]

821-(24-Dihydroxyph

enyl)-

3-(34-dihydroxyph

enyl)-

2-prop

en-1-on

eBu

tein

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

833-(34-D

ihydroxyph

enyl)-

1-(234-trihydroxyph

enyl)-

2-prop

en-1-on

eOkanin

O

HO

HO

OH

OH

OH

Leaves

(China)

[60]

84

3-(34-D

ihydroxyph

enyl)-

1-[3-(120573-D

-glucop

yranosyloxy)-

24-dihydroxyphenyl]-2-

prop

en-1-on

e

Okanin

31015840-O

-120573-D

-glucosid

e

O

HO

OH

OH

OH

OO

HO

H

HO

H

H

HOH

H

OH

Leaves

(Germany)

flowers(Germany)

[61ndash63]

85

3-(34-D

ihydroxyph

enyl)-

1-[4-(120573-D

-glucop

yranosyloxy)-

23-dihydroxyphenyl]-

2-prop

en-1-on

e

Okanin

41015840-O

-120573-D

-glucosid

e

OOH

OH

OH

HOO

OHO

H

HO

H

H

HOH

H

OH

Flow

ers(Germany)

leaves

(Japan)

[5963]

Evidence-Based Complementary and Alternative Medicine 15

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

86Okanin41015840-O

-120573-D

-(610158401015840-

O-acetylglucosid

e)

O

O

OH

OH

OH

OHO

H

HO

H

H

HOH

H

HO

O

O

Flow

ers(Germany)

[63]

87

1-[4-[(46-di-O

-Acetyl-

120573-D

-glucopyrano

syl)-oxy]-

23-dihydroxyphenyl]-3-

(34-dihydroxyph

enyl)-2-

prop

en-1-on

e

Okanin

41015840-O

-120573-D

-(410158401015840610158401015840-diacetyl)-

glucop

yranoside

O

O

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Aeria

l(Ch

ina)

[34]

88Okanin41015840-O

-120573-D

-(210158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[64]

89Okanin41015840-O

-120573-D

-(310158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

O

OAe

rial(Ch

ina)

[34]

16 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

90

1-[23-Dihydroxy-4-

[[6-O-[3-(4-hydroxyph

enyl)-

1-oxo-2-prop

enyl]-120573-D

-glucop

yranosyl]oxy]-ph

enyl]-

3-(34-dihydroxyph

enyl)-

2-prop

en-1on

e

Okanin

41015840-O

-120573-D

-(610158401015840-tr

ans-

p-coum

aroyl)-

glucoside

O

O

OH

OH

OH

OHOH

HO

H

H

HOH

H

HO

OO

OH

Leaves

(Germany)

[64]

91Okanin41015840-O

-120573-D

-(410158401015840-acetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Leaves

(Germany)

[65]

92

Okanin

41015840-O

-120573-D

-(210158401015840410158401015840-diacetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[65]

Evidence-Based Complementary and Alternative Medicine 17

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

93

Okanin

41015840-O

-120573-D

-(310158401015840410158401015840-diacetyl-610158401015840-

trans-p-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

OH

O O

Leaves

(Germany)

[65]

94

Okanin

41015840-O

-[120573-D

-glucopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

OOH

OH

OH

HO

OH

HO

HHO

H

HOH

HO

OH

OH

HO

HHO

H

HOH

HO

Flow

er(G

ermany)

[63]

95Okanin

3101584041015840-di-O

-120573-D

-glucosid

e

OOH

OH

OH

O OO

H HO

HHOH

HOH

HHO

OHO

H

HO

H

H

HOH

H

OH

Flow

er(G

ermany)

[63]

18 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

96

1-[3-(120573-D

-Glucopyrano

syloxy)-

24-dihydroxyphenyl]-

3-(3-hydroxy)-

4-metho

xyph

enyl)-

2-prop

en-1-on

e

Okanin

4-methylether-

31015840-O

-120573-D

-glucop

yranoside

OOH

O

OH

HO

O

H HO

HHOH

HOH

H

OHO

Leaves

(Germany)aerial

(China)

[3562]

97

Okanin4-methylether-

3101584041015840-di-O

-120573-(41015840101584061015840101584041015840101584010158406101584010158401015840-

tetraacetyl)-

glucop

yranoside

OOH

OOH

O

HH

OH

HO

HOH

H

OO

O

Ac

Ac

AcAc

OO

H

HO

H

H

HOH

H

O

Aeria

l(Ch

ina)

[42]

98Ch

alcone210158404101584061015840-tr

imetho

xy-

4-O-D

-glucopyrano

syl-

dihydro

Not

foun

dLeaves

(China)

[60]

992-(34-D

ihydroxyph

enyl)-23-

dihydro-78

-dihydroxy-

4H-1-

benzop

yran-4-one

Okanin

isoOO

OH

HO

OH

OH

Leaves

(China)

[60]

100

2-(34-D

ihydroxyph

enyl)-

23-dihydro-8-hydroxy-

7[(246-tr

i-O-acetyl-120573

-D-

glucop

yranosyl)oxy]-4H

-1-b

enzopyran-4-on

eOO

OH

OH

OH

OH

O

HHO

H

HO

HO

OAc

Ac

Ac

Aeria

l(Ch

ina)

[34]

Evidence-Based Complementary and Alternative Medicine 19

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

101

57-Dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

OO

OH

HO

OH

Aeria

l(TanzaniaC

hina)

[4266]

102

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

7-O-glucopyrano

side

OO

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Tanzania)

[66]

103

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-4H

-1-benzop

yran-4-one

Luteolin

OO

HO

OH

OH

OH

Aeria

l(TanzaniaC

hina

Vietnam)

[343542

6667]

104

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5hydroxy-4H-1-

benzop

yran-4-one

Luteolin

7-O-120573-D

-glucop

yranoside

OOOH

OH

OH

OO

HO

H

HO

H

H

HOH

OH H

Aeria

l(Tanzania)

[66]

105

57-Dim

etho

xy-6-

(5-m

etho

xy-6-m

ethyl-

4-oxo-4H

-pyran-3-yl)-

2-ph

enyl-4H-1-

benzop

yran-4-one

5-O-M

ethylhosludin

O OOO

O

O

O

Aeria

l(Ugand

a)[68]

20 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

106

3-(120573-D

-Glucopyrano

syloxy)-

57-dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Astragalin

Kaem

pferol-3-O

-120573-D

-glucopyrano

side

HO

OH

O O

OH

O

O

HO

HHO

H

H

HOH

OH

H

Aeria

l(Ch

ina)

[35]

107

Kaem

pferol3-

(23-di-E

-p-cou

maroyl-

120572-L-rhamno

pyrano

side)

Not

foun

dAe

rial(Vietnam)

[67]

108

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5-hydroxy-36-dimetho

xy-

4H-1-

benzop

yran-4-one

Axillarosid

eO O

OH

OH

OO

OH

OO

HO

H

HO

H

H

HOH

H

OH

Aeria

l(Ch

ina)

[42]

109

57-Dihydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaureidin

O

O

O

O

O

HO

OH

OH

Who

le(Taiwan)

[69]

110

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaurein

O O

OO

O OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan)w

hole

(Taiwan)

[69ndash

71]

Evidence-Based Complementary and Alternative Medicine 21

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

111

5-Hydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-

67-dimetho

xy-4H-

chromen-4-one

Eupatorin

iso

OO O

OH

O

O OH

Not

foun

d(C

hina)

[3772]

112

2-(34-D

imetho

xyph

enyl)-7-

(120573-D

-glucopyrano

syloxy)-35-

dihydroxy-

8-metho

xy-4H-1-

benzop

yran-4-one

O O

OH

O O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

113

7-(120573-D

-Glucopyrano

syloxy)-

5hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-38-

dimehtoxy-4H-1-

benzop

yran-4-one

O O

O

OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

114

Isorhamnetin

3-[O

-120572-

L-rham

nopyrano

syl-

(1-2)-120573-D

-glucop

yranoside]

Not

foun

dAe

rial(Vietnam)

[67]

115

7-[(6-deoxy-120572-L-

Manno

pyrano

syl)o

xy]-

3-120573-D

-glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-4H

-1-benzop

yran-4-one

Luteoside

O O

OH

O

OH

OH

OH

HOH

H

H HO

HO

HO

OH

HO

H

HO

OH

OH

HH

Aeria

l(Ch

ina)

[42]

22 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

116

Luteolin

3-O-120573-D

-Glucopyrano

side

O O

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

HO

Aeria

l(Tanzania)

[66]

117

57-Dihydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Quercetagetin

3631015840-tr

imethylether

OOO

O

O OH

HO

OH

Aeria

l(Ch

ina)

[42]

118

6-((2S3S4S5S)-

Tetrahydro-345-tr

ihydroxy-

6-(hydroxymethyl)-2H

-pyran-2-yloxy)-5-hydroxy-

2-(4-hydroxy-3-

metho

xyph

enyl)-

37-dimetho

xy-4H-

chromen-4-one

Quercetagetin

3731015840-

trim

ethylether-6-O

-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[42]

119

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

JaceinQ

uercetin

3631015840-

trim

ethyl

ether-7-O-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan

China)

who

le(Taiwan)

[425470]

120

2-(34-D

ihydroxyph

enyl)-

357-trihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

OO

HO

OH

OH

OH

OH

Aeria

l(Ch

ina)leaves

(China)who

le(C

hina)

[356074]

Evidence-Based Complementary and Alternative Medicine 23

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

121

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-galactopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-4-one

Quercetin

3-O-120573-D

-galactosidehyperin

hyperosid

e

O O

HO

OH

OH

OH

OOHH

HOH

H

H HO

HO

OH

Aeria

l(TanzaniaJapan)

Leaves

(China)Who

le(C

hina)

[376066

717475]

122

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-glucopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

3-O-120573-

D-glucopyrano

side

O O

HO

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

Aeria

l(TanzaniaC

hina

Japan)L

eaves(Japan)

[355966

75]

123

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-

4-oxo-4H

-1-benzop

yran-3-yl-

120573-D

-glucopyrano

siduron

icacid

Quercetin

3-O-120573-D

-glucuron

opyranoside

O O

HO

OH

OH

OH

OHOH

HOH

H

H HO

HO

OHO

Aeria

l(TanzaniaJapan)

[6671]

124

3-[[6-O-(6-

Deoxy-120572-L-m

anno

pyrano

syl)-

120573-D

-galactopyrano

syl]o

xy]-

2-(34-dihydroxyph

enyl)-57-

dihydroxy-4H

-1-benzop

yran-4-one

Quercetin

3-O-rob

inob

iosid

e

O O

HO

OH

OH

OH

O

HOHH OH

HO

H HH

OO

OHH

HOH

H

H HO

HO

Aeria

l(Japan)W

hole

(Taiwan)

[7071]

24 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

125

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-

3-metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

OH

Roots(Brazil)

[55676]

126

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-

120573-D

-glucopyrano

syl]o

xy]-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120572-

L-rham

nopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

O

HHO HHO

HO H

HH

ORo

ots(Brazil)

[5676]

127

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-120573-D

-glucop

yranosyl]oxy]-5-

hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-3-metho

xy-

4H-1-

benzop

yran-4-one

Quercetin

341015840-

dimethyl

ether-7-O-rutinoside

O OOH

OH

O

O

OH O

HOH

H

HHO

H

O

HHO HHO

HO H

HH

O

HO

Aeria

l(Ch

ina)

[3435]

128

2-(34-D

ihydroxyph

enyl)-3-

(120573-D

-glucofurano

syloxy)-57-

dihydroxy-4H

-1-benzop

yran-

4-on

e

Isoq

uercitrin

O OOH

OH

OHO

H

OHH

OH

HO

H O HOH

OH

Aeria

l(Japan

China)

[3571]

Evidence-Based Complementary and Alternative Medicine 25

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

129

Quercetin

3-O-120573-D

-galactop

yranoside

O

OH

HO

OH OH

O

O

HO

OH

OHOH

OAe

rial(no

tstated)

[18]

130

Quercetin

3-O-rutinoside

O

O

R

OO

OH

OHO

H

O

OH

OH

H3C

H3C

H3C

OH

HO

Who

le(Taiwan)

[70]

26 Evidence-Based Complementary and Alternative Medicine

Table 6 Terpenoids isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(Country) References

131371111-Tetramethylbicyclo[810]undeca-

26-dieneBicyclogermacrene Leaves

(Brazil) [95]

13241111-Trimethyl-8-

methylenebicyclo[720]undec-4-ene

E-Caryophyllene Leaves(Brazil) [95]

1331-Methyl-5-methylene-8-

(1-methylethyl)-16-cyclodecadiene

Germacrene D Leaves(Brazil) [95]

1344-(15-Dimethyl-4-hexen-1-ylidene)-1-methyl-cyclohexene

Z-120574-Bisabolene Leaves(Brazil) [95]

135Decahydro-114-trimethyl-7-methylene-1H-cycloprop[e]-

azulene120573-Gurjunene Leaves

(Brazil) [95]

136 2669-Tetramethyl-148-cycloundecatriene

120572-Humulene120572-caryophyllene

Leaves(Brazil) [95]

13712344a568a-Octahydro-1-isopropyl-7-methyl-4-methylenenaphthalene

120575-Muurolene Leaves(Brazil) [95]

138

12344a568a-Octahydro-4a8-dimethyl-2-(1-methylethylidene)-

naphthalene

Selina-37(11)-diene

Leaves(Brazil) [95]

139(2E7R11R)-371115-

Tetramethyl-2-hexadecen-1-ol

Phytol

HO

Whole(Taiwan) [57]

Evidence-Based Complementary and Alternative Medicine 27

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

140 371115-Tetramethyl-2-hexadecenoic acid Phytanic acid

O

OH

Whole(Taiwan) [57]

141371115-Tetramethyl-2-

hexadecenyl ester-heptanoicacid

Phythylheptanoate O O

Leaves(Not stated) [33]

142

(3S10R13R)-17-((2R5R)-5-Ethyl-6-methylheptan-2-yl)-

2347891011121314151617-tetradecahydro-

1013-dimethyl-1H-cyclopenta[a]phenanthren-

3-ol

Campestrol

HO

Aerial(Tanzania) [18]

143 Not found Phytosterin-B Not foundNot found(TaiwanEgypt)

[39 96]

144 Stigmast-5-en-3-ol 120573-sitosterol

HOH

H

H

Aerial(Tanzania)

whole(Taiwan)

[18 31 57]

145

1314151617-Tetradecahydro-1013-

dimethyl-1H-cyclopenta[a]phenanthren-

3-yloxy)-tetrahydro-6-(hydroxymethyl)-2H-pyran-345-triol

120573-Sitosterolglucoside

H

H

HOH

HO

HHO

H

HOHH O

OH Not found(Egypt) [39]

146 5120572-Stigmasta-7-en-3120573-ol

HO

Whole(Taiwan) [57]

147 5120572-Stigmasta-722t-dien-3120573-ol

HO

Whole(Taiwan) [57]

148 Stigmasta-522-dien-3-ol Stigmasterol

HOH

H

H

Not found(Taiwan)aerial

(Tanzania)leaves (notstated)whole

(Taiwan)

[18 31 3357]

28 Evidence-Based Complementary and Alternative Medicine

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

149 Lup-20(29)-en-3-ol Lupeol

H

HH

HOH

Not found(Egypt) [39]

150 Lup-20(29)-en-3-ol acetate Lupeol acetate

H

HH

O

OH

Not found(Egypt) [39]

151 Olean-12-en-3-ol 120573-amyrin

H

HOH

Not found(Egypt) [39]

152 5913-Trimethyl-242526-trinoroleanan-3-ol Friedelan-3120573-ol

HO

Aerial(Tanzania) [18]

153 5913-Trimethyl-242526-trinoroleanan-3-one

Friedelinfriedelan-3-one

O

Aerial(Tanzania) [18]

1542610151923-

Hexamethyl-2610141822-tetracosahexaene

Squalene

Aerial(Tanzania)Leaves (Notstated)Whole(Taiwan)

[18 33 57]

155 120573120573-Carotene 120573-Carotene Leaves (Notstated) [97]

Evidence-Based Complementary and Alternative Medicine 29

studies [83] Hot water extracts of B pilosa varminor Sheriffwere also assessed for its antileukemic effects on leukemiccell lines L1210 U937 K562 Raji and P3HR1 using XTT-based colorimetric assays The extract inhibited the five celllines with IC

50values ranging from 145120583gmL to 586120583gmL

L1210 K562 Raji and P3HR1 were more sensitive to B pilosaextract with IC

50values below 200120583gmL [98]

Consistent with the antitumor activities of B pilosaextracts and fractions some of its phytochemicals alsoshowed anticancer activity as outlined in Table 11 Amongthem luteolin (103) a well-studied flavonoid with multiplebioactivities was more effective against tumor cell prolifera-tion than its derivatives with IC

50values ranging from 3 120583M

to 50120583M in cells and 5 to 10mgkg in animals Luteolin wasalso found to fight cancer as a food additive at concentrationsof 50 to 200 ppm [78] and prevent skin cancer [78] and cancer3invasion [103] Lee and colleagues reported that luteolinprevents cancer by inhibiting cell adhesion and invasion[103] Significant inhibition concentration was reported to be5 120583M and complete inhibition concentration was reported tobe 40120583M Moreover luteolin was reported to inhibit hepa-tocyte growth factor (HGF)-induced cell scattering as wellas cytoskeleton changes such as filopodia and lamellipodiawhichwas determined using phase-contrast and fluorescencemicroscopy Furthermore luteolin also inhibited the HGF-induced phosphorylation of c-Met ERK 12 and Akt aswell as the MAPKERK and P13K-Akt pathways [78 103]Other mechanisms underlying the anticancer activities ofluteolin are the inhibition of topoisomerase I and II whichinhibits cell replication and DNA repair thus promotingapoptosis regulation of PI-3-KinaseAktMAPKERKJNKactivation of apoptosis in the mitochondrial pathway byactivating caspase 9 and caspase 3 which were found inmalignant cells but not in normal human peripheral bloodmononuclear cells death receptor-induced apoptosis and acell-cycle arrest mechanism inhibition of fatty acid synthasewhich is upregulated in many cancer cells and sensitizationto chemotherapywhereby luteolin increases the susceptibilityof cancer cells to chemotherapy [78] Butein (82) is anotherflavonoid that showed a cytotoxic effect on human colonadenocarcinoma cell proliferation with a reported IC

50value

of 175 120583M Butein at 2 120583M affected the incorporation of[14C]-labeled leucine thymidine and uridine which cancause the inhibition of DNA RNA and protein synthesis ofhuman colon cancer cells Moreover butein also exhibitednoncompetitive inhibition of 1-chloro-24-dinitrobenzene(CDNB) in glutathione S-transferase (GST) activity Tumorresistance was correlated with high levels of GST thus buteininhibited proliferation of cancer cells [80] Another flavonoidpresent in B pilosa centaureidin (109) also showed anti-cancer activity in B lymphoma cells Centaureidin isolatedfrom Polymnia fruticosa inhibited tubulin polymerization invitro and induced mitotic figure formation in CA46 Burkittlymphoma cells Using turbidimetric assay the IC

50value of

centaureidin in inhibition of mitosis was 3120583M [104] Cyto-toxicity of centaureidin was further analyzed using AmericanNational Cancer Institute (NCI) 60 human tumor cell linesThe cytotoxicity potency of centaureidin expressed as GI

50

(50 growth inhibition in the NCI tumor line panel) was024120583M [81] These data mark centaureidin as a promisingantimitotic agent for tumor therapy

In addition to anti-tumor flavones polyynes found in B 4pilosa have also been shown to possess anti-tumor proper-ties Based on a bioactivity-directed isolation approach Wuand colleagues identified two polyyne aglycones from theethyl acetate fraction of B pilosa [44] 12-Dihydroxytrideca-57911-tetrayne (48) and 13-Dihydroxy-6(E)- tetradecene-81012-triyne (46) exhibited significant anticell prolifera-tion activity in primary human umbilical vein endotheliumcells (HUVEC) with IC

50values of 125 120583M and 173 120583M

respectively They also decreased angiogenesis and pro-moted apoptosis in human endothelial cells Their anti-angiogenic and cytotoxic effects correlated with activationof the CDK inhibitors and caspase-7 [44] In addition 12-Dihyroxy-5(E)-tridecene-7911-triyne (45) showed antian-giogenic effects in HUVECs with an IC

50value of 124 120583M as

evidenced by a decrease in the tube formation and migrationof HUVECs [43] The IC

50value of compound 45 in the

inhibition of basic fibroblast growth factor-induced HUVECgrowth was 282120583M However it had higher IC

50values

than those for lung carcinoma cells and keratinocytes Thiscompound could also inhibit cell proliferation of HUVECslung carcinoma A549 cells and HACAT keratinocytes Themechanism by which compound 45 inhibits HUVEC growthand angiogenesis is complicated and includes decreasing theexpression of cell cycle regulators (CDK4 cyclins D1 and Aretinoblastoma (Rb) and vascular endothelial growth factorreceptor 1) caspase-mediated activation of CDK inhibitorsp21 (Cip1) and p27 (Kip) upregulation of Fas ligand expres-sion downregulation of Bcl-2 expression and activation ofcaspase-7 and poly (ADP-ribose) polymerase [43]

32 Anti-Inflammatory Activity B pilosa is commonly usedto treat inflammatory disorders The anti-inflammatory phy-tochemicals present in B pilosa are listed in Table 11 5Cyclooxygenase-2 (COX-2) is a physiologically importantenzyme that converts arachidonic acid to prostaglandin(PGE2) Its expression is induced by a wide variety of external

stimuli indicating its involvement in inflammatory diseasesand it is used as an inflammatory marker [88] Yoshida andcolleagues studied the effects of the aqueous extracts of Bpilosa aerial parts in the production of COX-2 and PGE

2as

well as on the activation of mitogen activated protein kinases(MAPKs) in normal human dermal fibroblasts (HDFs) inresponse to inflammatory cytokine IL-1120573 This work showedthat IL-1120573 activated MAPKs such as ERK12 p38 and JNKto different extents and induced COX-2 expression TheCOX-2 expression in HDFs was regulated mainly by p38following IL-1120573 stimulation Consistently the p38 inhibitorSB203580 blocked this expression Using this cell platformB pilosa extracts were tested for inhibition of inflammationThe extract dose-dependently suppressed the activation ofp38 and JNK and moderately suppressed ERK12 as wellas suppressing COX-2 expression and PGE

2production

30 Evidence-Based Complementary and Alternative Medicine

Table7Ph

enylprop

anoids

isolatedfro

mB

pilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

156

3-(4-H

ydroxyph

enyl)-2-prop

enoica

cid

p-Cou

maricacid

OHO

OH

Who

le(Ja

pan)

[99]

157

2-Metho

xy-4-(2-prop

en-1-yl)-ph

enol

Eugeno

lOOH

Leaves

androots(Japan)

[99]

158

3-(4-H

ydroxy-3-m

etho

xyph

enyl)-

2-prop

enoica

cid

Ferulic

acid

O

OH

O

HO

Who

le(Ja

pan)

[99]

159

3-(34-D

ihydroxyph

enyl)-2-prop

enoic

acid

Caffeicacid

O

HO

OH

OH

Who

leandaeria

l(Japan)

[7599]

160

3-Prop

yl-3[(245-trim

etho

xyph

enyl)-

metho

xy]-24-pentanedione

3-Prop

yl-3-(245-

trim

etho

xy)benzyloxy-

pentan-24-dione

O

O

O

O

O

O

Leaves

(India)

[100]

161

3-(34-D

ihydroxyph

enyl)-2-

prop

enoica

cidethyleste

rCa

ffeateethyl

O

O

OH

OH

Not

foun

d(Taiwan)

who

le(Taiwan)

[434584]

Evidence-Based Complementary and Alternative Medicine 31Ta

ble7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

162

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

d-Erythron

icacid

2-O-caffeoyl-2-C

-methyl

O

HO

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

163

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

2-O-caffeoyl-2-C

-methyl

O

O

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

164

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-24-

dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

3-O-caffeoyl-2-C

-methyl

O

O

HO

HO

O

O

OH

OH

Leaves

(Japan)

[101]

165

4-(A

cetyloxy)-3-[[3-(34-

dihydroxyphenyl)-

1-oxo-2-propen-1-y

l-]oxy]-2-hydroxy-

2-methyl-b

utanoica

cid

O

HO

OH

O OHO

HO

OO

Not

foun

d(Ja

pan)

[73]

166

3-(34-D

ihydroxyph

enyl)-tetrahydro-4-

hydroxy-4-

methyl-5

-oxo-3-fu

ranyl

ester-2-prop

enoica

cid

3-O-C

affeoyl-2-C

-methyl-

D-erythrono

-14-la

cton

eHO

HO

O

OO

OOH

Leaves

(Japan)

[101]

167

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]-oxy]-145-

trihydroxy-cyclohexanecarboxylicacid

Chlorogenica

cid

O

HO

HO

OH

O

OH

O

OH

HO

Aeria

l(Japan)w

hole

(Taiwan)

[707175]

32 Evidence-Based Complementary and Alternative Medicine

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

168

4-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]oxy]-

135-tr

ihydroxy-cyclohexanecarboxylic

acid

4-O-C

affeoylqu

inicacid

OHO

HO

OH

OH

O

O

OH

OH

Aeria

l(Japan)

[71]

169

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-15-

dihydroxy-cyclo

hexanecarboxylicacid

34-Di-O

-caffeoylqu

inic

acid

HO

HO

O

O

O

O

OH

OH

HO

OH O

H

O

Aeria

l(Japan)W

hole

(Taiwan)

[45497071]

170

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-14-dihydroxy-

cyclo

hexanecarboxylicacid

35-Di-O

-caffeoylqu

inic

acid

HO

HO

O O

HO

O

HO

OH OO

OHOH

Aeria

l(Japan)w

hole

(Taiwan)

[45497071]

Evidence-Based Complementary and Alternative Medicine 33

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

171

34-bis[[(2E

)-3-(34-D

ihydroxyph

enyl)-1-

oxo-2-prop

en-1-

yl]oxo]-15

-dihydroxy-

cyclo

hexanecarboxylicacid

45-Di-O

-caffeoylqu

inic

acid

OHO

OH

HO

O

O

OH

OH

O

OH

HO

OWho

le(Taiwan)

[454970]

172

3-[4-[[-6-O-[3-(4-H

ydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]-phenyl]-2-prop

enoica

cid

120573-D

-p-C

oumaricacid4-O

-(6-O

-p-caoum

aroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HOH

HO

O

O

Leaves

(Japan)

[59]

173

3-[4-[[2-O

-Acetyl-6

-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]ph

enyl]2-propeno

icacid

120573-D

-p-C

oumaricacid4-O

-(2-O

-acetyl-6

-O-p-

caou

maroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HO

HO

O

O

Leaves

(Japan)aerial

(China)

[3459]

174

67-Dihydroxy-2-chrom

enon

eEsculetin

cicho

rigenin

OHO HO

O

Not

foun

d(Egypt)

[39]

34 Evidence-Based Complementary and Alternative Medicine

Table 8 Aromatic compounds isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

175 12-Benzenediol PyrocatechinHO

HOWhole(Japan) [99]

176 4-Ethyl-12-benzenediol Pyrocatechol

OH

OH

Whole(Japan) [99]

177 Dimethoxyphenol26-dimethoxyphenol

O

HO

O

Roots (Japan) [99]

178 4-Ethenyl-2-methoxy-phenol p-Vinylguaiacol

O

OHWhole(Japan) [99]

179 2-Hydroxy-6-methylbenzaldehyde

6-Methyl-salicylaldehyde O

OH

Whole(Japan) [99]

180 Benzene-ethanol 2-Phenyl-ethanolHO

Whole(Japan) [57]

181 4-Hydroxy-3-methoxy-benzaldehyde Vanillin O

O

HOAerial(Japan) [99]

182 3-Hydroxy-4-methoxy-benzaldehyde Vanillin iso

OO

OH

Leaves(Japan) [99]

183 4-Hydroxybenzoic acid p-Hydroxybenzoicacid

O

HOOH Whole

(Japan) [99]

184 2-Hydroxybenzoic acid Salicylic acidO

HO

HO

Stem androots (Japan) [99]

185 34-Dihydroxybenzoicacid Protocatechuic acid

O

HOOH

OH

Whole(Japan) [99]

1864-Hydroxy-

3methoxybenzoicacid

Vanillic acid

OOH

O

OH

Aerial(Uganda)

Roots (Japan)[68 99]

Evidence-Based Complementary and Alternative Medicine 35

Table 8 Continued

S N IUPAC names Common names Structure Plant part(country) References

187 345-Trihydroxybenzoicacid Gallic acid

O

OHHO

HO

HO

Whole(China) [102]

[88] This work supports the use of B pilosa as an anti-inflammatory agent however no compounds responsible forthe anti-inflammatory activity of B pilosa were identified

A further study also reported the anti-inflammatoryactivity as well as the antiallergic activity of B pilosa [75] Inthis study dried powder of the aerial part of B pilosa whichhad been pretreated with the enzyme cellulosine was usedfor further tests The results showed that oral administrationof the cellulosine-treated B pilosa lowered the level of serumIgE in mice 10 days after immunization with DNP (24-dintrophenyl)-Ascaris as an antigen This treatment alsoreduced dye exudation in skin induced by passive cutaneousanaphylaxis and production of inflammatory mediators his-tamine and substance P in rats [75] Phytochemical analysisshowed that cellulosine treatment increased the percentage ofcaffeic acid and flavonoids This study suggests that B pilosaand its phenolics have anti-inflammatory functions

Phenolics and polyynes are major anti-inflammatoryphytochemicals present in B pilosa (Table 11) Unsurpris-6ingly phenolics such as luteolin (103) and ethyl caffeate(161) that are major constituents of B pilosa have alsobeen reported to possess anti-inflammatory activity Lute-olin was reported to exhibit anti-inflammatory activity inmacrophages Xagorari and colleagues showed that luteolininhibited the release of inflammatory cytokines TNF-120572 andinterleukin-6 in RAW 2647 cells following LPS stimulation[87] It inhibited TNF-120572 production with an IC

50value

of 1120583M The underlying anti-inflammatory mechanism ofluteolin was reported to be the inactivation of Akt andNF-120581B activation [87] In addition luteolin was reportedto confer anti-inflammatory activity through inhibition ofLPS-stimulated iNOS expression in BV-2 microglial cellsIt inhibited LPS-activated microglia in a dose-dependentmanner with an IC

50value of 69 120583MMoreover immunoblot

and RT-PCR data proved that luteolin suppressed I120581B-120572degradation and iNOS expression in LPS-activated microglia[85] Kim and colleagues stated that luteolin may havebeneficial effects on inflammatory neural diseases throughinhibition of iNOS expression [85] A related study revealedthat luteolin decreased the transcriptional activity of NF-120581BRelA via partial inhibition of TNF-mediated NF-120581B DNAbinding activity Luteolin also inhibited Akt phosphorylationand induced degradation of a transcription factor interferonregulatory factor (IRF) [86]

Chiang and colleagues showed that ethyl caffeate(161) significantly inhibited NO production in mousemacrophages RAW 2647 cells [84] Based on MTT assays

they concluded that this inhibition was not due to thecytotoxicity of ethyl caffeate The IC

50value of ethyl caffeate

in the inhibition of NO production was 55 120583gmL slightlylower than curcumin (positive control) which has anIC50

value of 65120583gmL They demonstrated that ethylcaffeate exerted anti-inflammatory activity via the reducedtranscription and translation of iNOS (inducible nitric oxidesynthase) in RAW 2467 cells In addition this compoundalso suppressed COX-2 expression in RAW 2467 cellsand MCF-7 cells The in vivo anti-inflammatory effectof ethyl caffeate was verified by testing in TPA-treatedmouse skin Like celecoxib the positive control ethylcaffeate significantly abolished COX-2 expression in adose-dependent manner Ethyl caffeate at 1mg200120583Lsite(24mM) inhibited COX-2 expression at a level comparableto celecoxib at 1mg200120583Lsite (13mM) Remarkably ethylcaffeate at 48mM (2mg200120583Lsite) was more effectivethan celecoxib at 131mM (10mg200120583Lsite) Moreoverthis compound inhibited the activation of nuclear factor-120581B(NF-120581B) by LPS via the prevention of NF-120581B binding toDNA [84] In addition 3 ethyl caffeate analogs (ethyl 34-dihydroxyhydrocinnamate ethyl cinnamate and catechol)also showed different degrees of NF-120581B binding to DNA aslisted in Table 12 Ethyl cinnamate lacks a catechol moietywhich results in ineffective inhibition of NF-120581B binding toDNA [84]

Pereira and colleagues assessed the anti- 7inflammatory and immunomodulatory activities ofB pilosa methanol extract as well as one polyyne2-O-120573-glucosyltrideca-11(E)-en-3 579-tetrayn-12-diol(54) in T lymphocytes and a zymosan-induced arthritismouse model [27] They first examined the in vitro effect of 8the B pilosa extract and compound 54 on cell proliferation ofhuman T cells stimulated with 5120583gmL phytohemagglutinin(PHA) or 100 nM 12-O-tetradecanoyl phorbol-13-acetate(TPA) plus 15 120583M ionomycin and on cell proliferation ofmouse T cells stimulated with 5 120583gmL concanavalin A(Con A) The data demonstrated that both methanol extractand compound 54 suppressed T-cell proliferation in adose-dependent manner The estimated IC

50values of the B

pilosa extract against human T cells stimulated with 5120583gmLPHA and 100 nM TPA plus 15120583M ionomycin were 125 and25 120583gmL respectively In comparison with the methanolextract compound 54 showed 10-fold more inhibition ofhuman T-cell proliferation with an estimated IC

50value of

15 120583gmL Accordingly the B pilosa extract and compound54 dose-dependently suppressed mouse T-cell proliferation

36 Evidence-Based Complementary and Alternative Medicine

Table 9 Porphyrins isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

188

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(15S16S)-10-ethenyl-5-ethyl-1161820-tetrahydro-6111522-tetramethyl-1820-dioxo-15H-912-imino-212-

metheno-471714-dinitrilopyrano[43-

b]azacyclononadecine-16-propanoic

acid

AristophyllC

NH N

HNN

O

O OOOPhytyl

O

Leaves(Taiwan) [100]

189

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -20b-(methoxycarbonyl)-9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-dinitrilo-12-

dioxeto-[310158404101584034]-cyclo-pent[1 2]azacyclononadecine-

19-propanoic acid

Bidenphytin A

NH N

HNN

OPhytylO O O

O O

Leaves(Taiwan) [100]

190

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -2a-

hydroxy-20b-(methoxycarbonyl)-

9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-

dinitrilo-12-dioxeto[310158404101584034]-cyclo-pent[12-b]-

azacyclononadecine-19-propanoic acid

Bidenphytin B

NH N

HNN

OPhytylO O O

O O

OH

Leaves(Taiwan) [100]

191

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO O

HOOO

Leaves(Taiwan) [100]

192

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

Evidence-Based Complementary and Alternative Medicine 37

Table 9 Continued

S N IUPAC names Common names Structure Plant part(country) References

193

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

Leaves(Taiwan) [100]

194

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

195

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3S4S21R)-9-ethenyl-14-ethyl-21-(methoxycarbonyl)-

481318-tetramethyl-20-oxo-3-phorbinepropanoic acid

Pheophytin A

NH N

HNN

PhytylO

O

OO O

Leaves(Taiwan) [100]

with estimated IC50values of 30 and 25 120583gmL respectively

Taken together the data indicate that the B pilosa extractand compound 54 act on human and mouse T cells Totest the in vivo effect of the B pilosa extract and compound54 a zymosan-induced arthritis mouse model was usedThis model was established from B10ASgSnJ mice with aninjection of zymosan (015mg) The zymosan-injected mice9received an intraperitoneal injection of the B pilosa extract(1 5 or 10mg) at one dose a day for 5 days Popliteal lymphnode (PLN) weight was monitored to check the developmentof arthritis The results revealed that 10mg of the methanolextract of B pilosa extract could significantly diminishinflammation as evidenced by PLN weight [27] This worksuggests that B pilosa (and compound 54) can suppressimmune response and inflammation

33 Antidiabetic Activity Anti-diabetic agents are primarilydeveloped from plants and other natural resources [8 24 4951 53] B pilosa is one of 1200 plant species that have beeninvestigated for antidiabetic activity [120 121]B pilosa is usedas an anti-diabetic herb in America Africa and Asia [51 120

122] Many studies have indicated that B pilosa could treattype 1 diabetes (T1D) and type 2 diabetes (T2D) in animals

Etiologically speaking T1D is caused by autoimmune-mediated destruction of pancreatic 120573 cells leading to insulindeficiency hyperglycemia and complications Currentlythere is no cure for T1D Polarization ofTh cell differentiationcontrols the development of T1D Suppression of Th1 celldifferentiation and promotion of Th2 cell differentiationameliorate T1D [123] One study showed that the butanolfraction of B pilosa inhibited T-cell proliferation decreasedTh1 cells and cytokines and increased Th2 cells andcytokines leading to prevention of T1D in nonobesediabetic (NOD) mice [49] Based on a bioactivity-directedisolation strategy 3 polyynes 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne (53) also known ascytopiloyne 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D- Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne (50) were identifiedfrom B pilosa [49 53] The IC

50value of the butanol fraction

was 200120583gmL This inhibition was reported to be partiallyattributed to cytotoxicity because the butanol fraction at

38 Evidence-Based Complementary and Alternative Medicine

Table 10 Other compounds isolated from B pilosa [30]

SN IUPAC names Common names Structure Plant part(country) References

19637-Dihydro-137-

trimethyl-1H-purine-26-dione

CaffeineN

NN

NO

O

Aerial(Uganda) [68]

197

1-((2R4S5R)-Tetrahydro-4-hydroxy-5-(hydroxymethyl)furan-2-yl)-5-

methylpyrimidine-24(1H3H)-dione

ThymidineHO

HO

ON

O

NHO Not found

(China) [37]

198 1-(2-Thienyl)-ethanone 2-Acetyl-thiophene

S

O

Roots(Germany) [41]

199

(2R3S4S5S)-2-(Heptan-2-yloxy)-tetrahydro-6-

((tetrahydro-34-dihydroxy-5-

(hydroxymethyl)furan-2-yloxy)methyl)-

2H-pyran-345-triol

Heptanyl2-O-120573-xylofuranosyl-

(1rarr 6)-120573-glucopyranoside

O

OOH

OH

HO

OHO

H

HO

H

H

HOHH

O

Whole(Taiwan) [70]

200

2-[(3R7R11R)-3-Hydroxy-371115-

tetramethylhexadecyl]-356-trimethyl-

25-cyclohexadiene-14-dione

120572-Tocopherylquinone

OO

HO

Whole(Taiwan) [32]

2017-O-(410158401015840610158401015840-Diacetyl)-

120573-D-glucopyranoside

Not found Leaves(China) [60]

180 120583gmL could cause 50 death of Th1 cells Moreoverthis study suggested that the butanol fraction may preventdiabetes in NOD mice in vivo via downregulation of Th1cells or upregulation Th2 cells that have effects which areantagonistic of those of Th1 cells [49] This was proven byintraperitoneal injection of the butanol fraction at a doseof 3mgkg BW 3 times a week to NOD mice from 4 to 27weeks This dosage resulted in lower incidence of diabetes(33) At a dose of 10mgkg the butanol fraction of Bpilosa totally eliminated (0) the initiation of the diseaseTo further support this result assessment of IgG2a and IgEproduction was performed in the serum of NOD mice Asin vivo results obtained from intracellular cytokine stainingexperiments were not very conclusive levels of IgG2a and IgE

were measured since Th1 cytokine IFN120574 and Th2 cytokineIL-4 favor the production of IgG2a and IgE respectively Asexpected high levels of IgE and some decline in the levels ofIgG2a were observed in the serum Profiling of the butanolextract revealed five compounds 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne(50) 45-Di-O-caffeoyl- quinic acid 35-Di-O-caffeoylquinicacid and 34-Di-O-caffeoylquinic acid Only the first twocompounds showed similar effects on the prevention ofdiabetes in NOD mice as the B pilosa butanol fractionMoreover compound 50 showed greater activity thancompound 69 in terms of enhancement (by 34 comparedto 8) of differentiation of Th0 to Th2 at 15 120583gmL (both

Evidence-Based Complementary and Alternative Medicine 39

Table 11 Chemical constituents of B pilosa and their biological activities

SN Name Classification Molecularformula Biological activities

109 Centaureidin [91] Flavonoid C18H16O8Anti-listerial [69 91]

Cytotoxic [10]

110 Centaurein [91] Flavonoid C24H26O13

Anti-listerial [69 91]Cytotoxic [10]Anti-viral [105]

103 Luteolin [106] Flavonoid C15H10O6

Anti-viral [107 108]Cytotoxic [77]

Anti-inflammatory [109]Anti-allergic [109]

82 Butein [110] Flavonoid C15H12O5Anti-leishmanial [111]

Cytotoxic [78]48 12-Dihydroxytrideca-57911-tetrayne [112] Polyyne C13H12O2 Anti-angiogeneic [112]46 13-Dihyroxy-6(E)-tetradecene-81012-triyne [112] Polyyne C14H16O2 Anti-angiogeneic [112]

45 12-Dihyroxy-5(E)-tridecene-7911-triyne [113] Polyyne C13H14O2Anti-angiogeneic [113]Anti-proliferative [113]

64 1-Phenylhepta-135-triyne [114] Polyyne C13H8

Anti-microbial [115]Anti-malarial [3]Cytotoxic [3]Antifungal [15]

27 Linoleic acid [79] Fatty acid C18H32O2Anti-viral (100)Cytotoxic [17]

161 Ethyl caffeate [84] Phenylpropanoid C11H12O4 Anti-inflammatory [84]

54 2-O-120573-Glucosyltrideca-11(E)-en-3579-tetrayn-12-diol [26] Polyyne C19H20O7Immunosuppressive andAnti-inflammatory [26]

53 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne [50] Polyyne C19H22O7Anti-diabetic [116]

Anti-inflammatory [50]

69 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne [28] Polyyne C20H26O7Anti-diabetic [28]

Anti-inflammatory [94]

50 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne [28] Polyyne C19H24O7

Anti-diabetic [28]Anti-inflammatory [94]

Anti-malarial andantibacterial [47]

129 Quercetin 3-O-120573-D-galactopyranoside [18] Flavonoid C21H20O12 Anti-inflammatory [117]

170 35-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

171 45-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

169 34-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

126 Quercetin 331015840-dimethyl ether7-O-120572-L-rhamnopyranosyl-(1rarr 6)-120573-D-glucopyranoside [118] Flavonoid C21H20O11 Anti-malarial [119]

125 Quercetin 331015840-dimethyl ether-7-O-120573-D-glucopyranoside [118] Flavonoid C21H20O12 Anti-malarial [119]70 1-Phenyl-13-diyn-5-en-7-ol-acetate [27] Polyyne C15H12O2 Anti-malarial [27]199 Heptanyl 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside [70] Miscellaneous C18H44O10 Antioxidant [70]124 3-O-Rabinobioside [70] Saccharide C27H38O15 Antioxidant [70]130 Quercetin 3-O-rutinoside [70] Flavonoid C27H38O15 Antioxidant [70]167 Chlorogenic acid [70] Phenolic C16H26O9 Antioxidant [70]119 Jacein [70] Flavonoid C24H26O13 Antioxidant [70]

49 (R)-12-dihydroxytrideca-357911-pentayne [47] Polyyne C13H8O2Anti-malarial andAntibacterial [47]

40 Evidence-Based Complementary and Alternative Medicine

Table 12 Structure and activity relationship studies of ethyl caffeateusing in vitro NF-120581BDNA binding assays [84]

Compound Concentration(120583M)

NF-120581BDNA binding

Ethyl caffeate 50 100inhibition

Ethyl 34-dihydroxyhydrocinnamate 100 100inhibition

Catechol 400 100inhibition

Ethyl cinnamate 400 Noinhibition

Table 13 Apoptosis in cocultures of T cells and pancreatic 120573 cells[53]

Cellmedium apoptosis andnecrosis

CD4+ T cellscontrol medium lt4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 2

CD4+ T cellswith cytopiloyne-treated 120573 cells 18CD4+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 7

CD8+ T cellscontrol medium 4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 4

CD8+ T cellswith cytopiloyne-treated 120573 cells 4CD8+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 4

Table 14 Radical scavenging activities of B pilosa extracts [70]

Extractscontrol DPPH assayIC50 (120583gmL)

NBThypoxanthinesuperoxide assayIC50 (120583gmL)

Quercetin 198 15Ascorbic acid 634 Not determined120572-tocopherol 897 Not determinedEthyl acetateextract 1383 597

Butanol extract 1669 114Water extract gt100 gt100

compounds) and inhibition (by 40 compared to 10) ofdifferentiation toTh1 at the same concentration [49]

Among the three polyynes found in B pilosa cytopiloyne(53) had the most potent anti-T1D activity [53] To test the invivo effect of cytopiloyne NODmice received intraperitonealor intramuscular injection of cytopiloyne at 25120583gkg BW3 times per week Twelve-week-old NOD mice started todevelop T1D and 70 of NOD mice aged 23 weeks andover developed T1D Remarkably 12- to 30-week-old NODmice treated with cytopiloyne showed normal levels of bloodglucose (lt200mgdL) and insulin (1-2 ngmL) Consistent

Table 15 Radical scavenging activity of secondarymetabolites fromB pilosa [70]

Metabolite (Table 11)control DPPH assay IC50 (120583gmL)199 Not determined124 53130 68167 105169 33171 38119 Not determined110 Not determinedQuercetin 256Caffeic acid 890

Table 16 Antioxidant activity of the essential oils and water extractsfrom B pilosa [92]

Extract IC50 (120583gmL)Leaf essential oils 47Flower essential oils 50Leaf extract 61Flower extract 172

with T1D incidence cytopiloyne delayed and reduced theinvasion of CD4+ T cells into the pancreatic islets [53]

In vitro study showed that cytopiloyne (53) inhibited thedifferentiation of naıveTh (Th0) cells (ie CD4+ T cells) intoTh1 cells and promoted differentiation of Th0 cells into Th2cells [50] The in vitro data are consistent with the in vivoresults indicating that cytopiloyne reducedTh1differentiationand increased Th2 differentiation as shown by intracellularcytokine staining and FACS analysis [53] In line with theskewing of Th differentiation the level of serum IFN-120574 andIgG2c decreased while that of serum IL-4 and serum IgEincreased compared to the negative controls (PBS-treatedmice) Cytopiloyne also enhanced the expression of GATA-3 a master gene for Th2 cell differentiation but not theexpression of T-bet a master gene forTh1 cell differentiationfurther supporting its role in skewingTh differentiation [53]

Also importantly cytopiloyne partially depleted CD4+rather than CD8+ T cells in NOD mice [53] As shown inTable 13 coculture assays showed that the depletion of CD4+T cells was mediated through the induction of Fas ligandexpression on pancreatic islet cells by cytopiloyne leading toapoptosis of infiltrating CD4+ T cells in the pancreas via theFas and Fas ligand pathways However cytopiloyne did notinduce the expression of TNF-120572 in pancreatic islet cells andthus had no effect on CD8+ T cells [53]

In addition Chang and colleagues showed that cytopi-loyne dose-dependently inhibited T-cell proliferation stimu-lated by IL-2 plus Con A or anti-CD3 antibody using [3H]thymidine incorporation assay [53]

Overall the mechanism of action of cytopiloyne andprobably its derivatives in T1D includes inhibition of T-cellproliferation skewing of Th cell differentiation and partialdepletion of Th cells Due to the anti-diabetic mechanisms

Evidence-Based Complementary and Alternative Medicine 41

Table 17 Antibacterial activity of essential oils and flower extracts from B pilosa [92]

Strain Mean zone of inhibition (mm)Leaf essential oil Flower essential oil Leaf extract Flower extract

Micrococcus flavus 127 plusmn 03 87 plusmn 03 102 plusmn 02 108 plusmn 03Bacillus subtilis 173 plusmn 19 117 plusmn 02 109 plusmn 02 103 plusmn 02Bacillus cereus 190 plusmn 14 112 plusmn 03 118 plusmn 04 185 plusmn 10Bacillus pumilus 123 plusmn 07 108 plusmn 02 105 plusmn 04 77 plusmn 02Escherichia coli 137 plusmn 04 203 plusmn 07 102 plusmn 11 140 plusmn 13Pseudomonas ovalis 125 plusmn 08 137 plusmn 15 102 plusmn 06 125 plusmn 06

Table 18 Antibacterial activity of root extracts from B pilosa [93]

Strain MIC50 (mgmL)Methanol extract Acetone extract

Bacillus cereus 10 mdashEscherichia coli 5 5Klebsilla pneumonia 5 10Micrococcus kristinae 10 mdashPseudomonas aeruginosa 10 10Staphylococcus aureus 5 10Sraphylococcus epidermidis 5 5Serratia marcescens 10 mdashShigelea flexneri 10 mdashStreptococcus faecalis 10 mdash

Table 19 Antibacterial activity of B pilosa of compound 29 [47]

Strain MIC50 (120583gmL)Escherichia coli NIHJ 1Escherichia coli ATCC25922 1Klebsiella pneumoniae ATCC700603 128Serratia marcescens ATCC13880 16Pseudomonas aeruginosa ATCC27853 8Staphylococcus aureus FDA209P 05Staphylococcus aureus ATCC29213 025Staphylococcus aureus N315 (MRSA) 05Enterococcus faecalis ATCC29212 2Enterococcus faecalis NCTC12201 (VRE) 1Bacillus subtilis ATCC6633 05Candida albicans ATCC10231 025

of action it was hypothesized that cytopiloyne protectsNOD mice from diabetes by a generalized suppression ofadaptive immunity To evaluate this hypothesis ovalbumin(Ova) was used as a T-cell dependent antigen to primeNOD mice which had already received cytopiloyne orPBS vehicle Ova priming boosted similar anti-Ova titersin cytopiloyne-treated mice and PBS-treated mice but adifference in immunoglobulin isotype was observed in thetwo groups Thus it was concluded that cytopiloyne is animmunomodulatory compound rather than an immunosup-pressive compound [50 53]

T2D is a chronic metabolic disease with serious com-plications resulting from defects in either insulin secretion

insulin action or both [124] A study by Ubillas et alshowed that the aqueous ethanol extract of the aerial partof B pilosa at 1 gkg body weight (BW) lowered bloodglucose in dbdb mice a T2D mouse model [51] Basedon a bioactivity-guided identification compounds 69 and50 were identified Further the mixture of the compounds(69 50) in a 2 3 ratio significantly decreased blood glucoseconcentration and reduced food intake on the second dayof treatment when administered at doses of 250mgkg twicea day to C5BLKs-dbdb mice When tested at 500mgkga more substantial drop in blood glucose level as wellas the stronger anorexic effect (food intake reduced from58 gmouseday to 25 gmouseday) was observed [51] Inthis study it was suggested that the blood glucose lower-ing effect of B pilosa was caused in part by the hungersuppressing effect of its polyynes [51] However the hungersuppressing effect of the ethanol extract of B pilosa wasnot found in the studies described below In another study[24] water extracts of B pilosa (BPWE) were used indiabetic dbdb mice aged 6ndash8 weeks with postprandialblood glucose levels of 350 to 400mgdL Like oral anti-diabetic glimepiride which stimulates insulin release onesingle dose of BPWE reduced blood glucose levels from374 to 144mgdL The antihyperglycemic effect of BPWEwas inversely correlated to an increase in serum insulinlevels suggesting that BPWE acts to lower blood glucose viaincreased insulin production However BPWE had differentinsulin secretion kinetics to glimepiride [24] One flaw incurrent anti-diabetics is their decreasing efficacy over timeThe authors investigated the long term anti-diabetic effectof BPWE in dbdb mice BPWE reduced blood glucoseincreased blood insulin improved glucose tolerance andreduced the percentage of glycosylated hemoglobin (HbA1c)Both long-term and one-time experiments strongly supportthe anti-diabetic action of BPWE [24] In sharp contrast toglimepiride BPWE protected against islet atrophy in mousepancreas The investigators further evaluated anti-diabeticproperties of 3 B pilosa varieties B pilosa L var radiate(BPR) Bpilosa L var pilosa (BPP) and B pilosa L varminor(BPM) in dbdb mice [8] One single oral dose (10 50 and250mgkg body weight) of BPR BPP or BPM crude extractsdecreased postprandial blood glucose levels in dbdb micefor up to four hours and the reduction of glucose levels inthe blood appeared to be dose-dependent Comparing thethree variants BPR extract resulted in a higher reduction inblood glucose levels when administered at the same dose as

42 Evidence-Based Complementary and Alternative Medicine

Table 20 Antifungal activity of B pilosa [92]

Partextract Concentration (ppm) Strain InhibitionCortiicum rolfsii Fusarium solani Fusarium oxysporum

Leaves

Essentialoils

100 857 plusmn 09 682 plusmn 0 745 plusmn 17250 960 plusmn 08 779 plusmn 18 879 plusmn 04

AqueousExtracts

100 446 plusmn 17 605 plusmn 21 716 plusmn 07250 942 plusmn 03 689 plusmn 07 824 plusmn 19

Flowers

Essentialoils

100 604 plusmn 09 892 plusmn 04 869 plusmn 05250 894 plusmn 12 980 plusmn 03 949 plusmn 06

AqueousExtracts

100 331 plusmn 11 714 plusmn 07 573 plusmn 22250 661 plusmn 14 912 plusmn 0 900 plusmn 07

Table 21 Antifungal activity of B pilosa root extracts [93]

Strain LC50 (mgmL)Acetoneextracts

Methanolextracts

Waterextracts

Aspergillus niger 014 006 007Aspergillus flavus 1091 658 0Penicillium notatum 005 005 005

the other two varieties In terms of serum insulin levels a doseof 50mgkg of each extract was used and the BPR extracttogether with the three polyynes significantly increased theserum insulin level in dbdb mice Long-term experiments(28-day treatment) were then conducted using diabetic micewith postprandial glucose levels from 370 to 420mgdL andglimepiride was used as positive controlThe range of dosagesapplied was from 10mgkg BW to 250mgkg BW Resultsshowed that the positive control as well as the crude extractsof the three varieties lowered the blood glucose levels indbdb mice However only BPR extract containing a higherpercentage of cytopiloyne (53) reduced blood glucose levelsand augmented blood insulin levelsmore than BPP andBPMThe percentage of glycosylated hemoglobin A1c (HbA1c) wasalso measured and found to be 79 plusmn 05 in mice aged 10ndash12 weeks and 66 plusmn 02 61 plusmn 03 and 62 plusmn 03 inthe blood of age-matchedmice following treatment with BPRcrude extract (50mgkg) glimepiride (1mgkg) and com-pound 53 (05mgkg) respectively [8] Among the polyynesfound in B pilosa cytopiloyne was the most effective againstT2D Hence cytopiloyne was used for further study on anti-diabetic action and mechanism [125] The data confirmedthat cytopiloyne reduced postprandial blood glucose lev-els increased blood insulin improved glucose tolerancesuppressed HbA1c level and protected pancreatic islets indbdb mice Nevertheless cytopiloyne failed to decreaseblood glucose in streptoztocin (STZ)-treated mice whose bcells were already destroyed Additionally cytopiloyne dose-dependently increased insulin secretion and expression in bcells as well as calcium influx diacylglycerol and activationof protein kinase C120572 Collectively the mechanistic studiessuggest that cytopiloyne treats T2D via regulation of insulin

production involving the calciumDAGPKC120572 cascade in bcells

The studies detailed above point to the conclusion thatcytopiloyne and related polyynes (compounds 69 and 49)are anti-diabetics in animal models The data uncover a newbiological action of polyynes It should be noted that likeall anti-diabetic drugs cytopiloyne failed to prevent or curediabetes completely but reduced diabetic complications [125]Intriguingly 34 polyynes have been found in B pilosa so farIt remains to be seen whether all the polyynes present in thisplant have anti-diabetic activities

34 Antioxidant Activity Free radicals can damage cellularcomponents via a series of chemical reactions [89] leadingto development and progression of cardiovascular diseasecancer neurodegenerative diseases and ageing [70] Freeradicals nitric oxide (NO) and superoxide anions can beproduced in macrophages to kill microbes However anexcessive generation of the free radicals under pathologicalconditions is associated with a wide range of illnessesPlants are known to be rich in antioxidant phytochemicalsChiang and colleagues evaluated the free radical scaveng-ing activity of crude extract fractions and compounds ofB pilosa using 11-diphenyl-2-picrylhydrazyl (DPPH) andhypoxanthinexanthine oxidase assays [70] Using DPPHand hypoxanthinexanthine oxidase assays they found thatthe B pilosa crude extract and the ethyl acetate butanoland water fractions had free radical scavenging activityNine compounds Heptyl- 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside (199) 3-O-Rabinobioside (124) Quercetin3-O-rutinoside (130) Chlorogenic acid (167) 34-Di-O-caffeoylquinic acid (169) 35-Di-O-caffeoylquinic acid (170)45-Di-O-caffeoylquinic acid (171) Jacein (119) and Cen-taurein (110) had DPPH radical scavenging activity [70]The IC

50values of the B pilosa crude extractfractions and

compounds are summarized in Tables 14 and 15 respectivelyMeasurement of free radical scavenging activities is one

way of assessing the antioxidant activities of B pilosa andits fractions and compounds It is interesting that the ethylacetate and butanol fractions are more active than the waterfraction and B pilosa crude extract [70] Of the secondarymetabolites only phenolic compounds 124 130 167 169 and171 showed significant DPPH-radical scavenging activities

Evidence-Based Complementary and Alternative Medicine 43

Further analysis of the structure-activity relationship of thecompounds suggested that substitution of the C3 hydroxylgroup with glycosides increased the activity approximately2-fold (for example in compounds 124 and 130) relative toquercetin (-OH in its C3) [70] Further modification of thestructures of the active compounds needs to be performed totest the effects of various substituents on activity The reasonwhy most of the antioxidant compounds contain phenolmoieties in their structure could be that the reduction-oxidation (redox) properties of phenols allow them to actas reducing agents singlet oxygen quenchers and hydrogendonors

A complementary study by Muchuweti and colleaguesdetermined phenolic content antioxidant activity and thephenolic profile of B pilosa methanol extract [89] Theyestimated that the phenolic content of the methanol extractof B pilosawas 11028plusmn22mgg [89] Vanillin hydroxyben-zaldehyde caffeic acid coumaric acid and ferulic acid werefound in this extractThe B pilosa extract also showed DPPHradical scavenging activity Furthermore the antioxidantactivity of the flavonoids found in B pilosa was correlatedwith its hepatoprotective effects through their inhibitionof NF-120581B activation which may lessen the oxidative stresscaused by the production of free radicals during liver injury[60]This activity might also be due to the anti-inflammatoryeffects of the aqueous extracts of B pilosa aerial parts on theinhibition of COX-2 and PGE

2production [88]

Essential oils from B pilosa flowers and leaves are alsoreported to possess antioxidant activity With the aim ofreplacing chemically synthesized additives Deba and col-leagues [92] worked on the antioxidant antibacterial andantifungal activities of essential oils and water extracts of Bpilosarsquos leaves and flowers Table 16 summarizes the resultsobtained from DPPH free radical scavenging assay

It can be inferred fromTable 16 that essential oils from theleaves possessed the highest activity It is reported elsewherethat monoterpenes present in essential oils such that of Bpilosa have protective effects and antioxidant properties [92]Beta-carotene bleaching method was also performed Leavesessential oils and aqueous extracts of the leaves and flowersshowed higher activity than the flower essential oils This isdue to the volatility of the flower essential oils The activityexhibited by the aqueous extracts was accounted to thepresence of phenolic compounds that are reported to donatea hydrogen atom to free radicals such that the propagation ofthe chain reaction during lipid oxidation is terminated [92]Overall essential oils and phenolics present in B pilosa canbe thought of as major antioxidant compounds

35 Immunomodulatory Activity B pilosa is thought to bean immunomodulatory plant and is reported to be effectivein the treatment of immune disorders such as allergy [75]arthritis [75] and T1D [48 50 53]

As pointed out in the discussion of its anti-diabeticactivities (Section 33) a combination of phytochemicalsmethod and T-cell activation assays was used to studyimmunomodulatory properties of B pilosa IFN-120574 is a keycytokine released by T and NK cells that mediates immune

cells and sustains immunity against pathogens Defects inIFN-120574 expression regulation and activation result in vulner-ability to diseases caused by bacteria and viruses [69] Anelegant study performed byChang and colleagues using IFN-120574 promoter-driven luciferase reporter construct in JurkatT cells showed that hot water crude extracts of B pilosaincreased IFN-120574 promoter activity two-fold [69] Out of thesubfractions of this extract the butanol fraction but not thewater or ethyl acetate fractions increased IFN-120574 promoteractivity six-fold Centaurein (110) and centaureidin (109)were identified from the butanol fraction and were stated tocause a four-fold increase in IFN-120574 promoter activity withEC50

values of 75 120583gmL and 09120583gmL respectively Themechanism of action of centaurein was determined usingtranscription factors such as AP-1 NFAT and NF120581B whichare reported to bind to IFN-120574 promoter and regulate IFN-120574transcription Unlike with the activity of the positive controlPHA centaurein caused a four-fold increase in NFAT a3-fold increase in NF120581B and had little if any effect onAP-1 enhancer activities (23-fold three-fold and ten-foldincreases respectively were seenwith PHA) [69]The authorsconcluded that centaureinmodulates IFN-120574 expression by theNFAT and NF120581B pathways The article only determined themechanism of action of centaurein Its aglycone centaureidinmay act through the samemechanism though this conclusionneeds to be further verified

B pilosa extract and its compounds are reported to inhibitdifferentiation of naıve CD4+ helper T (Th0) cells into Th1cells [49] Using theThcell differentiation assay as a screeningplatform 3 polyynes 2-120573-D-glucopyranosyloxy-1 -hydroxytrideca-57911-tetrayne (53) 3-120573-D-glucopyranosyl-1-hydroxy-6(E)- tetradecene-81012-triyne (69) and 2-120573-D-glucopyranosyloxy-1-hydroxy-5(E)- tridecene-7911-triyne(49) were discovered from B pilosa [49 53] The data showsthat cytopiloyne and other two polyynes suppressed thedifferentiation of type 1 helper T (Th1) cells and productionof Th1 cytokines and promoted that of type 2 helper T (Th2)cells and production of Th2 cytokines thus explaining theimmunomodulatory and anti-inflammatory effects of Bpilosa and its polyynes

Chang and colleagues were the first to report the effect ofthe butanol extract of B pilosa on the autoimmune diabetesand airway inflammation inmice [48] Imbalance in the levelsofTh1 andTh2 and of various cytokines leads to autoimmunediseases T1D and other autoimmune diseases (rheumatoidarthritis Crohnrsquos disease among others) are exacerbated byan increase Th1 levels (specifically CD4+ Th1 cells) whileTh2 cells antagonize this effect [19] Moreover Th2 cellsmediate asthma in ovalbumin-induced hypersensitivity inBALBc mice [48] In their work [48] Chang and colleaguesshowed that 10mgkg butanol extracts with 15 (ww)compound 69 and 11 (ww) compound 49 (Table 11)ameliorated the development of Th1-mediated diabetes inNOD mice through inhibition of 120573 cell death and leukocyteinfiltration At the same dosage the butanol extracts alsoexacerbated ovalbumin-induced pulmonary inflammationin BALBc mice with an increase in the infiltration ofeosinophils and mast cells into the airway of the mice [48]Despite the different outcomes both mouse models proved

44 Evidence-Based Complementary and Alternative Medicine

the concept that control over the Th1Th2 shift is associatedwith autoimmune diseases and that the B pilosa butanolextract can shift the differentiation of Th0 cells to Th2 cells[48 49]

An extended study presented by Chiang and colleaguesshowed that compound 53 modulates T-cell functions [50]Using CD4+ T cells from BALBc mice they demonstratedthat cytopiloyne decreased levels of IFN-120574 producing cells(Th1) by 122 (from 72 to 598) Since Th1 and Th2 celldifferentiation is antagonistic it was expected that compound53 increased the percentage of mouse IL-4 producing cells(Th2) by 72 (from 237 to 309) Subsequent assess-ment of effect of compound 53 on the modulation of thetranscription of IL-4 and IFN-120574 showed that cytopiloyne asexpected decreased the splenocyte levels of IFN-120574 mRNAand increasing that of IL-4 in a dose-dependent manner Inthe range of 01 to 3 120583gmL of compound 53 the effects werenot attributed to its cytotoxicity Consequently using 3120583gmLcytopiloyne for 72 and 96 hours the protein concentration ofIFN-120574 decreased to 186 and 444 respectively Under thesame conditions cytopiloyne increased IL-4 concentrationsto 1985 and 2470 (for 72 and 96 hours resp) Thismodulation of T-cell differentiation exhibited by compound53 was used to explain its anti-diabetic activity [50]

The anti-diabetic role of cytopiloyne was extensivelydiscussed above (Section 33 anti-diabetic activity) Themolecular basis of the regulation of cytokine expression bycytopiloyne has been described Cytopiloyne directly elevatedthe expression level of IL-4 via GATA-3 upregulation inT cells [53] However reduction of IFN-120574 expression in Tcells seemed to come from the indirect opposing effect IL-4cytokine because the expression level of T-bet was unaltered[53] In this way cytopiloyne skewedTh1 polarization into theTh2 state conferring protection against T1D in NOD miceAside frompolarization ofThcell differentiation cytopiloynealso activated the expression of Fas ligand in pancreaticb cells this increase leading to the partial depletion of Tcells and reduction of immune response in local areas suchas the pancreas Of note cytopiloyne also inhibited T-cellproliferation and activation By targeting T cells from threeimmunomodulatory actions cytopiloyne protects againstT1D and probably other Th1-mediated autoimmune diseases[19]

The phytochemical constituents of B pilosa exert theirfunctions on different immune cells to modulate immuneresponse It is possible that some of the compounds mayhave agonistic or antagonistic effects on immune responseImmune function of B pilosa may depend on its compo-sition and amount of compounds which could explain theapparently conflicting report of B pilosa butanol extractaggravating allergy in mice [48] while cellulosine-treatedextract ameliorated allergy [48 75] IFN-120574 promoter reporterassays and T-cell differentiation assays were used to isolate2 flavonoids [69] and 3 polyynes [49 50] as immunomod-ulatory compounds from the butanol fraction of B pilosaInterestingly the flavonoids promote IFN-120574 expression inNK and T cells In marked contrast the polyynes promoteIL-4 expression and indirectly inhibit IFN-120574 expression indifferentiating T cells Sensitivity appears to be the key to

identifying structure- and bioactivity-related phytochemicalsfrom medicinal plants

36 Antimalarial Activity The use of chemical drugs againstpathogens has resulted in drug-resistant mutants Examplesof drug resistance can be found in the species of thePlasmodium that cause malaria It is important to search fornew compounds to combat Plasmodium parasites [47] Astudy of the anti-malarial activity of the leaf extracts of Bpilosa using a combination of phytochemistry and bioassaysshowed that compound 49 (Table 11) showed activity againsta malaria parasite (P falciparum NF54 strain) with an IC

50

value of 60 120583gmL [77] In addition compound 49 isolatedfrom the aerial parts of B pilosa inhibited growth of the Pfalciparum FCR-3 strain with an IC

50value of 035 120583gmL

This compound was tested for its in vivo effect in miceinfected with P berghei NK-65 strain Results showed thatcompound 49 decreased the average parasitemia in the redblood cells by 207 (from 328 of that of the control to121) after an intravenous injection of 08mgkg BWday forfour days [47] Further studies addressing the anti-malarialmechanism underlying both polyynes and clinical studies areneeded

37 Antibacterial Activity Emergence of multiple antibiotic-resistantmicrobes is becoming a global threat to public healthand a challenge to disease treatment For instance penicillinis commonly used to combat a food-borne intracellularbacteriumListeria however penicillin-resistant bacteria havebeen discovered recently [69] Chang and coworkers iso-lated centaurein (110) and centaureidin (109) from B pilosaextract [69 70] Centaurein enhances expression of IFN-120574a key cytokine for macrophage activation and consequentlyenhances bactericidal activity in macrophages [69 70] Inagreement with observed in vitro effects centaurein wasreported to prevent and treat Listeria infection in C57BL6Jmice [91] Mechanistic studies confirmed that centaureinexerted antilisterial action via IFN-120574 expression in wild-typemice but not IFN-120574 knockout mice [91] Further in vitrostudies on centaurein showed that this compound increasedIFN-120574 expression by 13 (from 17 to 20) 20 (from 21to 41) and 11 (from 6 to 17) in CD4+ T cells CD8+Tcells and NK cells respectively That is to say there wasan increase in IFN-120574 producing immune cells As expectedcentaurein also enhanced the expression level of T-bet a keynuclear factor for IFN-120574 expression Consistently centaureinaugmented the serum IFN-120574 levels inC57BL6Jmice and thisaugmentation peaked 24 hours after compound injectionThe quantity of mouse serum IFN-120574 was sufficient to activatemacrophages in vitro and eradicated GFP-producing Listeriainside macrophages However the entry of Listeria intomacrophages was not affected by centaurein-treated mousesera Centaurein treatment at 20120583g per mouse rescued 30of the mice infected with a lethal dose of Listeria (2 times106 CFU) It is noteworthy that in the presence of ampicillin(5 120583gmouse) centaurein (20 120583gmouse) rescued 70 of themice suggesting an additive effect between ampicillin andcentaurein [91] Despite lower abundance centaureidin was

Evidence-Based Complementary and Alternative Medicine 45

30 times more active than centaurein in terms of IFN-120574production [91]

Aside from the indirect antibacterial action mentionedabove extract andor compounds of B pilosa also showeddirect bacteriostatic andor bactericidal action One studyreported that essential oils and leafflower extracts of Bpilosa could suppress the growth of gram positive and gramnegative bacteria as evidenced by zone of inhibition assaysIn this study antibacterial activity of the essential oils fromthe leaves and flowers of B pilosa was determined in anattempt to identify natural products as food preservatives forprevention of microbial multiplication and food oxidationThe essential oils and extracts of B pilosa leaves and flow-ers showed moderate but different extents of antibacterialactivity (Table 17) In general essential oils had higherantibacterial activity than crude extracts One explanation forthis could be that monoterpenes in the essential oils destroycellular integrity and subsequently inhibit the respirationand ion transport processes The presence of antibacterial120573-caryophyllene could be another explanation as reportedelsewhere [92] Another study reported that the methanoland acetone extract of B pilosa roots displayed antibacterialactivities against the bacteria listed in Table 18 [93] andmethanol extracts from the roots seemed to be the mosteffective

Another study indicated that the polyyne (R)-12-dihydroxytrideca-357911-pentayne (49) from this plantalso suppressed bacterial growth as shown by the minimuminhibitory concentration required to inhibit 50 bacterialgrowth (MIC

50) in Table 19 This compound was highly

effective against several Gram positive and Gram negativebacteria including the drug-resistant bacteria Staphylococcusaureus N315 (MRSA) and Enterococcus faecalis NCTC12201(VRE) [47] Strikingly compound 49 had a similar MIC

50

value to antibiotics (ampicillin tetracycline norfloxacin andamphotericin B) in most of the bacteria tested

Antibacterial activity of B pilosa extracts and compo-nents expressed as MIC

50and the mean zone of inhibition

is tabulated in Tables 17 18 and 19The zone of inhibition forAmpicilline (positive control) ranges from 153 plusmn 03mm to443 plusmn 02mm [92]

38 Antifungal Activity B pilosa has traditionally been usedto treat microbial infection Recently different parts of Bpilosa have been tested for antifungal activities Deba andcolleagues first evaluated the antifungal effect of the hotwater extracts of the B pilosa roots stems and leaves againstCorticium rolfsii Fusarium solani and Fusarium oxysporumThey discovered that C rolfsii was most suppressed bytreatment with B pilosa as its growth was reduced at almostall the tested doses followed by F oxysporum and F solani[99] However the fungicidal activities of the stems androots were greater than the leaves [99] Moreover the samegroup assessed the antifungal activity of the essential oilsand aqueous extracts from B pilosa flowers and leaves [92]They showed that the extracts and oils had antifungal activityagainst C rolfsii F solani and F oxysporum Essential oils

appeared to have better fungicidal activity thanwater extractsas summarized in Table 20

Another study by Ashafa and colleagues showed thatacetone methanol and water extracts of the B pilosa rootsshowed antifungal activities against Aspergillus niger Aflavus and Penicillium notatum using the agar dilutionmethod The results are tabulated in Table 21 [93] Negativecontrols showed 0 growth inhibitionThe methanol extractof the B pilosa roots at 10mgmL was also effective againstCandida albicans [93]Of note B pilosa obtained fromPapuaNew Guinea had no activity against A niger and C albicans[126] but the SouthAfrican (EasternCape) ecotype exhibitedmoderate activity against C albicans [93] This discrepancymay depend on extraction solvents extraction procedureassay techniques different plant parts and abundance ofactive compounds

B pilosa produces a variety of secondary metabolitessuch as flavonoids phenylacetylenes alkaloids sterols ter-penoids and tannis [92 93] However none of them havebeen confirmed as active compounds against fungi Furtherinvestigation of active compounds from B pilosa is necessaryto further understand the antifungal efficacy of this plant

39 Hypotensive and Vasodilatory Activities In early studiesDimo and colleagues used three rat models normotensiveWistar rats (NTR) salt-loading hypertensive rats (SLHR)and spontaneous hypertensive rats (SHR) to investigate thehypotensive effect of the methanol crude extract of B pilosaleaves [28 127] The extract lowered systolic blood pressurein hypertensive rats (SLHR and SHR) to a greater degreethan NTR [28 127] In addition a decrease in urinarysodium ions and an increase in urinary potassium ions wereobserved after treatment with the methanol extract of Bpilosa leaves although neither differences were statisticallysignificant [28 127] Taking the data together the studyproposed that B pilosa leaf extract reduced blood pressurevia vasodilation [28 127] The same group continued totest the antihypertensive effect of aqueous and methylenechloride extracts of B pilosa leaves in a hypertensive ratmodel [28 127] To establish a fructose-induced hypertensionmodel male Wistar rats were given 10 fructose solutionto drink ad libitum for three weeks In addition to freeaccess to 10 fructose the rats were treated with the aqueous(150mgkg) or methyl chloride (350mgkg) extracts of Bpilosa for additional three weeks [28 127] Both extracts ofB pilosa leaves had a hypotensive effect on rats Howeverneither extracts reversed the elevation of serum insulin infructose-fed rats Therefore B pilosa lowered blood pressureirrespective of insulin [28 127] To better understand thehypotensive mechanism the authors investigated the effectof a neutral extract of B pilosa (NBP) a mixture of methanolandmethylene chloride (1 1) extract after neutralization withNaOH and HCl on the heart and the blood pressure ofNTR and SHR [110] This study showed that an intravenousinjection of the NBP resulted in a biphasic reduction insystolic blood pressure In addition one intravenous dose ofthe extract at 10 20 and 30mgkg BW decreased systolicblood pressure in normal rats by 183 425 and 30

46 Evidence-Based Complementary and Alternative Medicine

respectively and the same doses reduced the blood pressurein hypertensive rats by 258 389 and 286 respectivelyOnly the highest dose (30mgkg) affected the force of thecontraction of the heart Atropine and propranolol were usedto interfere with the hypotensive action of the NBP Atropinereduced the initial phase of the hypotensive response in NBPand completely abolished the second phase of hypotensiveresponse in NBP In contrast propranolol increased thefirst hypotensive response but partially abolished the secondhypotensive response provoked by the NBP [110]This mech-anistic study suggested that B pilosa invokes the biphasichypotensive responses via targeting cardiac pump efficiencyduring the first phase and vasodilation at the second phase[110]

A further study was performed to investigate the relaxingeffect of a neutral extract of B pilosa (NBP) on rat aorta con-tractedwithKCl (60mM) andnorepinephrine (01mM) [94]Cumulative addition of NBP relaxed the rat aorta previouslycontracted by KCl in a dose-dependent manner The EC

50

value of theNBP for vasorelaxationwas 032mgmLThedataalso showed that the NBP reduced the contraction of aortapreviously contracted by KCl irrespective of the presence ofaortic endothelium [94]

Pretreatment with glibenclamide an ATP-dependent K+channel blocker did not considerably affect the relaxant effectof the NBP on KCl-induced contraction suggesting thatthe vasodilatory effect of B pilosa was not related to theopening of this ATP-dependent K+ channel [94] On theother hand in the presence of indomethacin or pyrilaminemaleate the relaxant response induced by the plant extractwas significantly inhibited at the lower concentrations Theplant extract was able to reduce the aorta resting tone inhibitthe KCl-induced contractions by 90 at 15mgmL and theCaCl2-induced contractions by 95 at 075mgmL These

results demonstrate that B pilosa can act as a vasodilatorprobably via acting as a calcium antagonist [94]

However no specific compound for the above activityhas been identified from B pilosa to date A bioactivity-guided identification approach may be adopted to identifythe active compounds in B pilosa that possess hypotensiveand vasodilatory effects and understand their mechanism ofaction

310 Wound Healing Activity B pilosa has been traditionallyused to treat tissue injury inCameroon Brazil andVenezuela[26] Hassan and colleagues investigated the wound healingpotential of B pilosa in Wistar rats [128] Mirroring thepositive control neomycin sulfate the ethanol extract of Bpilosa had faster wound closure than control rats 3 6 and 9days after topical application Histological examination alsorevealed better collagenation angiogenesis and organizationof wound tissue seven days after application Epithelializationand total healing time in B pilosa-treated rats were compara-ble to those of neomycin sulfate Together these data suggestthat B pilosa may be a viable alternative to neomycin lotionfor the treatment of wounds

In addition to studying the wound healing effect of Bpilosa on external ulcers Tan and colleagues also examined

the effect of methanol cyclohexane and methyl chlorideextracts of B pilosa on gastric ulcers in Wistar rats fedwith 1mL HClethanol gastric necrotizing solution (150mMHCl in 60 ethanol) and macroscopically visible lesionswere scored [26] Among the three extracts methylene chlo-ride extracts exhibited the highest activity showing 464inhibition of lesion formation at a dose of 500mgkg BWand complete inhibition at 750mgkg [26] The efficacy ofthe ethylene chloride extract was followed by that of themethanol extracts which had inhibition ranging from 304to 822 at concentrations of 500mgkg and 1000mgkgBW respectively [26] The cyclohexane extracts showed thelowest activity against gastric ulcers in rats with 13340 and 797 inhibition at 500 750 and 1000mgkg BWrespectively [26] To better understand the mode of actionof the methylene chloride extract of B pilosa rats werepretreated with indomethacin a COX-2 inhibitor involved inprostaglandin synthesis Pretreatment significantly reducedthe protection against HClethanol-induced ulcers to 313inhibition at 750mgkg BW suggesting a link between theantiulcerative activity of B pilosa and prostaglandin syn-thesis Unexpectedly the methylene chloride extract of Bpilosa showed little gastric mucosal protection against gastriclesions induced by 95 ethanol (1mL) [26] Absolute alcoholis known to cause mucosalsubmucosal tissue destruction viacellular necrosis and the release of tissue-derived mediators(histamine and leukotrieneC4)Thus these data imply thatBpilosa did not prevent the generation or the necrotic action ofthese mediators on the gastric microvasculature In additionpylorus ligation can increase gastric acid secretion withoutan alteration of mucosal histamine content The methylenechloride extract of B pilosa did not possess antisecretoryactivity On the contrary it was observed that increases in thedose of the extract led to elevated gastric juice acidity [26]Results with both absolute ethanol and pylorus ligation ratmodels suggested the possibility that the ineffectiveness of Bpilosa against gastric ulcers was due to lack of antihistaminicactivity in the plant In summary overall the data suggest thatB pilosa protects against HCLethanol-mediated ulcers viainhibition of prostaglandin biosynthesis

Previous phytochemical studies showed that a groupof flavonoids acyclichalcones are present in B pilosa[129 130] and the chalcones were proposed to have anti-ulcerative activity [131] Moreover nine hydroxychalcones 10were reported to possess gastric cytoprotective effects with24-dihydroxychalcone being the most active [132] Sincemethylene chloride extracts appear to be the most active Bpilosa extracts next the specific anti-ulcerative phytochemi-cals in the methylene chloride extracts of B pilosa and theirmodes of action needs to be probed

Despite the claims listed inTable 3 relatively few scientificstudies have been conducted in vitro and in vivo to address thetraditional ethnomedical uses of B pilosa Information aboutthe use of B pilosa as a botanical therapy recorded so far isfar from complete Studies conducted thus far only serve asa starting point for further investigation of B pilosa and theultimate efficacious use of the herb in clinical applications

Evidence-Based Complementary and Alternative Medicine 47

4 Toxicology

Despite its use as an ingredient in food for human con-sumption studies on systemic toxicity (eg acute subacutechronic and subchronic toxicities) of B pilosa in humansand animals are still inadequate and insufficient So faracute andor subchronic toxicities have been evaluated inrats and mice Oral acute and 28-day toxicities of waterextract of B pilosa leaves were evaluated in Wistar rats[133] An oral dose of water extract of B pilosa leaves at10 gkg BW showed no obvious mortality or changes inthe appearance in rats [134] The same extract at 08 gkgBWday once a day showed no obvious sub-chronic toxicityin rats over 28 days as measured by survival rate bodyweight and gross examination of organs [134] These dataare consistent with our data indicating that oral deliveryof the water extract of the B pilosa whole plant at 1 gkgBWday once a day is safe in rats over 28 days (unpublisheddata) Taken together these studies suggest that ingestion ofB pilosa aqueous extract at up to at 1 gkg BWday oncea day is highly safe in rats In addition the acute toxicityof aqueous and ethanol extracts of B pilosa in mice havebeen reported [134] Five- to six-week-old mice with weightsbetween 28 and 35 g received a peritoneal injection of bothextracts at the different doses The LD

50 the dose that causes

50 lethality of the aqueous and ethanol extracts in micewas 1230 gkg BW and 615 gkg BW respectively [134] Acomplete toxicological study has not been completed forhumans Furthermore the drug interactions of B pilosa withother drugs are unknown Further safety verification andclinical trials should be performed before B pilosa can beconsidered for medicinal use

5 Conclusions

B pilosa is an erect perennial plant with green leaves whiteor yellow flowers and tiny black seeds As it is distributedworldwide and is widely used as a folk remedy B pilosacan be thought of as an extraordinary source of food andmedicine However a comprehensive up-to-date review ofresearch on B pilosa has hitherto been unavailable In thisarticle scientific studies on B pilosa have been summarizedand critically discussed from the perspectives of botanyethnomedicine phytochemistry pharmacology and toxicol-ogy B pilosa is claimed to treat more than 40 disordersand 201 compounds have been identified from this plantThe medicinal utility of B pilosa and its modes of actionin relation to its known phytochemicals were discussedherein Polyynes flavonoids phenylpropanoids fatty acidsand phenolics are the primary bioactive compounds of Bpilosa and they have been reported to be effective in thetreatment of tumors inflammationimmune modulationdiabetes viruses microbes protozoans gastrointestinal dis-eases hypertension and cardiovascular diseases Cautionshould be exercised in the therapeutic use of B pilosa forhypoglycemia hypotension bleeding and allergy

Acknowledgments

The authors thank their laboratory members for construc-tive suggestions and the authors whose publications theycited for their contributions The authors also thank MsMiranda Loney of Academia Sinica for English editing of thispaper This work was supported by Grants 99-CDA-L11 and101S0010056 from Academia Sinica Taiwan Arlene P Bar-tolome is a recipient of MECO-TECO Sandwich ScholarshipProgram between Taiwan and the Philippines

References

[1] T Shen G H Li X N Wang and H X Lou ldquoThe genusCommiphora a review of its traditional uses phytochemistryand pharmacologyrdquo Journal of Ethnopharmacology vol 142 no2 pp 319ndash330

[2] C Long P Sauleau B David et al ldquoBioactive flavonoids ofTanacetum parthenium revisitedrdquo Phytochemistry vol 64 no2 pp 567ndash569 2003

[3] P O Karis and O Ryding Asteraceae Cladistics and Classifica-tion Bremer K Eds pp 559ndash569 Timber press Portland OreUSA 1994

[4] O N Pozharitskaya A N Shikov M N Makarova et alldquoAnti-inflammatory activity of a HPLC-fingerprinted aqueousinfusion of aerial part of Bidens tripartita LrdquoPhytomedicine vol17 no 6 pp 463ndash468 2010

[5] F Q Oliveira V Andrade-Neto A U Krettli and MG L Brandao ldquoNew evidences of antimalarial activity ofBidens pilosa roots extract correlated with polyacetylene andflavonoidsrdquo Journal of Ethnopharmacology vol 93 no 1 pp 39ndash42 2004

[6] Agriculture USDA ldquoPlants databaserdquo 2013in Natural Resources Conservation Servicehttpwwwnrcsusdagovwpsportalnrcssitenationalhome

[7] Y Zhou and X Z Yan ldquoExperimental study of qi deficiencysyndrome and Codonopsis pillosulae and Astragalus injectionon the immune response in micerdquo Zhong Xi Yi Jie He Za Zhivol 9 no 5 pp 286ndash262 1989

[8] S C Chien P H Young Y J Hsu et al ldquoAnti-diabeticproperties of three common Bidens pilosa variants in TaiwanrdquoPhytochemistry vol 70 no 10 pp 1246ndash1254 2009

[9] M J Alcaraz and M J Jimenez ldquoFlavonoids as anti-inflammatory agentsrdquo Fitoterapia vol 59 no 1 pp 25ndash38 1988

[10] FAO Agriculture Food and Nutrition for AfricamdashA ResourceBook for Teachers of Agriculture Publishing ManagementGroup FAO Information Division Rome Italy 1997

[11] M B Rokaya Z Munzbergova B Timsina and K R BhattaraildquoRheum australe D Don a review of its botany ethnobotanyphytochemistry and pharmacologyrdquo Journal of Ethnopharma-cology vol 141 no 3 pp 761ndash774 2012

[12] P H Young Y J Hsu and W C Yang ldquoBidens pilosa L andits medicinal userdquo in Recent Progress in Medicinal Plants DrugPlant II A S AwaadVK Singh and JNGovil Eds StandiumPress Houston Tex USA 2010

[13] C Ge ldquoCytologic study of Bidens bipinnata LrdquoZhongguo ZhongYao Za Zhi vol 15 no 2 pp 72ndash125 1990

[14] L C Chiang J S Chang C C Chen L T Ng and C CLin ldquoAnti-herpes simplex virus activity of Bidens pilosa and

48 Evidence-Based Complementary and Alternative Medicine

Houttuynia cordatardquoAmerican Journal of ChineseMedicine vol31 no 3 pp 355ndash362 2003

[15] N P Rybalchenko V A Prykhodko S S Nagorna et alldquoIn vitro antifungal activity of phenylheptatriyne from Bidenscernua L against yeastsrdquo Fitoterapia vol 81 no 5 pp 336ndash3382010

[16] K Redl W Breu B Davis and R Bauer ldquoAnti-inflammatoryactive polyacetylenes from Bidens campylothecardquo Planta Med-ica vol 60 no 1 pp 58ndash62 1994

[17] M Ayyanar and S Ignacimuthu ldquoTraditional knowledge ofKani tribals inKouthalai of Tirunelveli hills TamilNadu IndiardquoJournal of Ethnopharmacology vol 102 no 2 pp 246ndash255 2005

[18] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant intraditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[19] AKAbbas KMMurphy andA Sher ldquoFunctional diversity ofhelper T lymphocytesrdquo Nature vol 383 no 6603 pp 787ndash7931996

[20] J H Cano and G Volpato ldquoHerbal mixtures in the traditionalmedicine of Eastern Cubardquo Journal of Ethnopharmacology vol90 no 2-3 pp 293ndash316 2004

[21] J Namukobe J M Kasenene B T Kiremire et al ldquoTraditionalplants used for medicinal purposes by local communitiesaround the Northern sector of Kibale National Park UgandardquoJournal of Ethnopharmacology vol 136 no 1 pp 236ndash245 2011

[22] C Lans ldquoComparison of plants used for skin and stomachproblems in Trinidad and Tobago with Asian ethnomedicinerdquoJournal of Ethnobiology andEthnomedicine vol 3 article 3 2007

[23] S Dharmananda ldquoA Popular Remedy Ecapes Notice ofWesternPractitionersrdquo 2013 httpwwwitmonlineorgartsbidenshtm

[24] Y J Hsu T H Lee C L T Chang Y T Huang andW C YangldquoAnti-hyperglycemic effects and mechanism of Bidens pilosawater extractrdquo Journal of Ethnopharmacology vol 122 no 2 pp379ndash383 2009

[25] E Noumi F Houngue and D Lontsi ldquoTraditional medicinesin primary health care plants used for the treatment ofhypertension in Bafia Cameroonrdquo Fitoterapia vol 70 no 2 pp134ndash139 1999

[26] P V Tan T Dimo and E Dongo ldquoEffects of methanolcyclohexane and methylene chlo ride extracts of Bidens pilosaon various gastric ulcer models in ratsrdquo Journal of Ethnophar-macology vol 73 no 3 pp 415ndash421 2000

[27] R L C Pereira T Ibrahim L Lucchetti A J R Da Silva and VL G De Moraes ldquoImmunosuppressive and anti-inflammatoryeffects of methanolic extract and the polyacetylene isolatedfrom Bidens pilosa Lrdquo Immunopharmacology vol 43 no 1 pp31ndash37 1999

[28] T Dimo J Azay P V Tan et al ldquoEffects of the aqueous andmethylene chloride extracts of Bidens pilosa leaf on fructose-hypertensive ratsrdquo Journal of Ethnopharmacology vol 76 no 3pp 215ndash221 2001

[29] CWiartMedicinal Plants of Southeast Asia Pelanduk SelabgorDarul Ehsan Malaysia 2000

[30] F L Silva D C H Fischer J F Tavares M S Silva P FDe Athayde-Filho and J M Barbosa-Filho ldquoCompilation ofsecondary metabolites from Bidens pilosa LrdquoMolecules vol 16no 2 pp 1070ndash1102 2011

[31] A H Chen S R Lin and C H Hong ldquoPhytochemical studyon Bidens pilosa L varminorrdquo Huaxue pp 38ndash42 1975

[32] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[33] M C Zulueta M Tada and C Y Ragasa ldquoA diterpene fromBidens pilosardquo Phytochemistry vol 38 no 6 pp 1449ndash14501995

[34] J Wang H Yang Z W Lin and H D Sun ldquoFlavonoids fromBidens pilosa var radiatardquo Phytochemistry vol 46 no 7 pp1275ndash1278 1997

[35] A Zhao Q Zhao L Peng et al ldquoA new chalcone glycoside fromBidens pilosardquo Acta Botanica Yunnanica vol 26 no 1 pp 121ndash126

[36] P Gao Y Wu S Cui and Y Zou ldquoSynthesis and absoluteconfiguration of the 7-phenylhepta-4 6-diyne-1 2-diol isolatedfrom Bidens pilosardquo Synthesis no 13 pp 2131ndash2135 2011

[37] S Wang B Yang D Zhu D He and L Wang ldquoActivecomponents of Bidens pilosa Lrdquo Zhongcaoyao vol 36 pp 20ndash21

[38] F Bohlmann T Burkhardt and C Zdero Naturally OccuringAcetylenes Academic Press New York NY USA 1973

[39] T M Sarg A M Ateya N M Farrag and F A AbbasldquoConstituents and biological activity of Bidens pilosa L grownin EgyptrdquoActa Pharmaceutica Hungarica vol 61 no 6 pp 317ndash323 1991

[40] H A L Valdes and H P Rego ldquoBidens pilosa Linnerdquo RevistaCubana de Plantas Medicinales vol 6 pp 28ndash33

[41] F Bohlmann H Bornowski and K M Kleine ldquoNew polyynesfrom the tribe Heliantheaerdquo Chemische Berichte vol 97 pp2135ndash2138

[42] RWang Q XWu and Y P Shi ldquoPolyacetylenes and flavonoidsfrom the aerial parts ofBidens pilosardquo PlantaMedica vol 76 no9 pp 893ndash896 2010

[43] L W Wu Y M Chiang H C Chuang et al ldquoA novelpolyacetylene significantly inhibits angiogenesis and promotesapoptosis in human endothelial cells through activation of theCDK inhibitors and caspase-7rdquo Planta Medica vol 73 no 7 pp655ndash661 2007

[44] L W Wu Y M Chiang H C Chuang et al ldquoPolyacetylenesfunction as anti-angiogenic agentsrdquo Pharmaceutical Researchvol 21 no 11 pp 2112ndash2119 2004

[45] H L Yang S C Chen N W Chang et al ldquoProtection fromoxidative damage using Bidens pilosa extracts in normal humanerythrocytesrdquo Food and Chemical Toxicology vol 44 no 9 pp1513ndash1521 2006

[46] H Q Wang S J Lu H Li and Z H Yao ldquoEDTA-enhancedphytoremediation of lead contaminated soil by Bidens maxi-mowiczianardquo Journal of Environmental Sciences vol 19 no 12pp 1496ndash1499 2007

[47] S TobinagaMK SharmaWG L Aalbersberg et al ldquoIsolationand identification of a potent antimalarial and antibacterialpolyacetylene from Bidens pilosardquo Planta Medica vol 75 no 6pp 624ndash628 2009

[48] C L T Chang H K Kuo S L Chang et al ldquoThe distincteffects of a butanol fraction of Bidens pilosa plant extract onthe development of Th1-mediated diabetes and Th2-mediatedairway inflammation in micerdquo Journal of Biomedical Sciencevol 12 no 1 pp 79ndash89 2005

[49] S L Chang C L T Chang Y M Chiang et al ldquoPolyacetyleniccompounds and butanol fraction from Bidens pilosa can mod-ulate the differentiation of helper T cells and prevent autoim-mune diabetes in non-obese diabetic micerdquo Planta Medica vol70 no 11 pp 1045ndash1051 2004

Evidence-Based Complementary and Alternative Medicine 49

[50] Y M Chiang C L T Chang S L Chang W C Yang and LF Shyur ldquoCytopiloyne a novel polyacetylenic glucoside fromBidens pilosa functions as a T helper cell modulatorrdquo Journal ofEthnopharmacology vol 110 no 3 pp 532ndash538 2007

[51] R PUbillas CDMendez S D Jolad et al ldquoAntihyperglycemicacetylenic glucosides from Bidens pilosardquo Planta Medica vol66 no 1 pp 82ndash83 2000

[52] L Alvarez S Marquina M L Villarreal D Alonso E Arandaand G Delgado ldquoBioactive polyacetylenes from Bidens pilosardquoPlanta Medica vol 62 no 4 pp 355ndash357 1996

[53] C L T Chang S L Chang Y M Lee et al ldquoCytopiloyne apolyacetylenic glucoside prevents type 1 diabetes in nonobesediabetic micerdquo Journal of Immunology vol 178 no 11 pp 6984ndash6993 2007

[54] G Kusano A Kusano and Y Seyama Novel Hypoglycemicand Anti-Inflammatory Polyacetylenic Compounds Their Com-positions Bidens Plant Extract Fractions and CompositionsContaining the Plant or Fraction JPO Tokyo Japan 2004

[55] M G L Brandao A U Krettli L S R Soares C G CNery and H C Marinuzzi ldquoAntimalarial activity of extractsand fractions from Bidens pilosa and other Bidens species(Asteraceae) correlated with the presence of acetylene andflavonoid compoundsrdquo Journal of Ethnopharmacology vol 57no 2 pp 131ndash138 1997

[56] A U Krettli V F Andrade-Neto M D G L Brandao andW M S Ferrari ldquoThe search for new anti-malarial drugs fromplants used to treat fever and malaria or plants ramdomlyselected a reviewrdquoMemorias do Instituto Oswaldo Cruz vol 96no 8 pp 1033ndash1042 2001

[57] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[58] C K Wat R K Biswas E A Graham L Bohm G H NTowers and E R Waygood ldquoUltraviolet-mediated cytotoxicactivity of phenylheptatriyne from Bidens pilosa Lrdquo Journal ofNatural Products vol 42 no 1 pp 103ndash111 1979

[59] Y Sashida K Ogawa M Kitada H Karikome Y Mimakiand H Shimomura ldquoNew aurone glucosides and new phenyl-propanoid glucosides from Bidens pilosardquo Chemical and Phar-maceutical Bulletin vol 39 no 3 pp 709ndash711 1991

[60] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens pilosa L (TFB) on animal liver injury andliver fibrosisrdquo Journal of Ethnopharmacology vol 116 no 3 pp539ndash546 2008

[61] B Hoffmann and J Holzl ldquoAcylated compounds from Bidenspilosardquo Planta Medica vol 55 pp 108ndash109

[62] B Hoffmann and J Holzl ldquoA methylated chalcone glucosidefrom Bidens pilosardquo Phytochemistry vol 27 no 11 pp 3700ndash3701 1988

[63] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[64] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[65] B Hoffmann and J Holzl ldquoWeitere acylierte chalkone ausBidens pilosardquo Planta Medica vol 54 pp 450ndash451 1988

[66] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant in11traditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[67] V V Pham V V T K P V L Hoang and V K Phan ldquoFlavonoidcompounds from the plant Bidens pilosa L (Asteraceae)rdquo TapChi Duoc Hoc vol 50 pp 48ndash53

[68] S D Sarker B Bartholomew R J Nash and N Robinson ldquo5-O-methylhoslundin an unusual flavonoid from Bidens pilosa(Asteraceae)rdquo Biochemical Systematics and Ecology vol 28 no6 pp 591ndash593 2000

[69] S L Chang Y M Chiang C L T Chang et al ldquoFlavonoidscentaurein and centaureidin from Bidens pilosa stimulate IFN-120574 expressionrdquo Journal of Ethnopharmacology vol 112 no 2 pp232ndash236 2007

[70] Y M Chiang D Y Chuang S Y Wang Y H Kuo P WTsai and L F Shyur ldquoMetabolite profiling and chemopreventivebioactivity of plant extracts from Bidens pilosardquo Journal ofEthnopharmacology vol 95 no 2-3 pp 409ndash419 2004

[71] A Kusano Y Seyama E Usami et al ldquoStudies on the antioxi-dant active constituents of the dried powder from Bidens pilosaL var radiata SchrdquoNaturalMedicines vol 57 no 3 pp 100ndash1042003

[72] I Doleckova L Rarova J Gruz et al ldquoAnti-proliferative andanti-angiogenic effects of flavone eupatorin an active con-stituent of chloroform extract ofOrthosiphon stamineus leavesrdquoFitoterapia vol 83 no 6 pp 1000ndash1007

[73] K Bairwa R Kumar R J Sharma and R K Roy ldquoAn updatedreview on Bidens pilosa Lrdquo Der Pharma Chemica vol 2 no 3pp 325ndash337

[74] Q Xia Y Liu and Y Li ldquoDetermination of hyperoside indifferent parts and different species of Herba Bidens by RP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol 50pp 48ndash53

[75] M Horiuchi and Y Seyama ldquoImprovement of the antiinflam-matory and antiallergic activity of Bidens pilosa L var radiataSCHERFF treated with enzyme (Cellulosine)rdquo Journal of HealthScience vol 54 no 3 pp 294ndash301 2008

[76] M G L Brandao C G C Nery M A S Mamao and AU Krettli ldquoTwo methoxylated flavone glycosides from Bidenspilosardquo Phytochemistry vol 48 no 2 pp 397ndash400 1998

[77] P Kumari K Misra B S Sisodia et al ldquoA promising anticancerand antimalarial component from the leaves of Bidens pilosardquoPlanta Medica vol 75 no 1 pp 59ndash61 2009

[78] G Seelinger I Merfort U Wolfle and C M Schempp ldquoAnti-carcinogenic effects of the flavonoid luteolinrdquoMolecules vol 13no 10 pp 2628ndash2651 2008

[79] G Seelinger I Merfort and C M Schempp ldquoAnti-oxidantanti-inflammatory and anti-allergic activities of luteolinrdquoPlanta Medica vol 74 no 14 pp 1667ndash1677 2008

[80] C C Yit and N P Das ldquoCytotoxic effect of butein on humancolon adenocarcinoma cell proliferationrdquo Cancer Letters vol82 no 1 pp 65ndash72 1994

[81] J A Beutler E Hamel A J Vlietinck et al ldquoStructure-activityrequirements for flavone cytotoxicity and binding to tubulinrdquoJournal of Medicinal Chemistry vol 41 no 13 pp 2333ndash23381998

[82] M R Kviecinski K B Felipe T Schoenfelder et al ldquoStudy ofthe antitumor potential of Bidens pilosa (Asteraceae) used inBrazilian folkmedicinerdquo Journal of Ethnopharmacology vol 117no 1 pp 69ndash75 2008

[83] P Sundararajan A Dey A Smith A G Doss M Rajappanand S Natarajan ldquoStudies of anticancer and antipyretic activityof Bidens pilosawhole plantrdquo African Health Sciences vol 6 no1 pp 27ndash30 2006

[84] YMChiang C P Lo Y P Chen et al ldquoEthyl caffeate suppressesNF-120581B activation and its downstream inflammatory mediatorsiNOS COX-2 and PGE2 in vitro or in mouse skinrdquo BritishJournal of Pharmacology vol 146 no 3 pp 352ndash363 2005

50 Evidence-Based Complementary and Alternative Medicine

[85] J S Kim H J Lee M H Lee J Kim C Jin and J HRyu ldquoLuteolin inhibits LPS-stimulated inducible nitric oxidesynthase expression in BV-2 microglial cellsrdquo Planta Medicavol 72 no 1 pp 65ndash68 2006

[86] P A Ruiz and D Haller ldquoFunctional diversity of flavonoidsin the inhibition of the proinflammatory NF-120581B IRF and Aktsignaling pathways in murine intestinal epithelial cellsrdquo Journalof Nutrition vol 136 no 3 pp 664ndash671 2006

[87] A Xagorari A Papapetropoulos A Mauromatis MEconomou T Fotsis and C Roussos ldquoLuteolin inhibitsan endotoxin-stimulated phosphorylation cascade andproinflammatory cytokine production in macrophagesrdquoJournal of Pharmacology and Experimental Therapeutics vol296 no 1 pp 181ndash187 2001

[88] N Yoshida T Kanekura Y Higashi and T Kanzaki ldquoBidenspilosa suppresses interleukin-1120573-induced cyclooxygenase-2expression through the inhibition of mitogen activated proteinkinases phosphorylation in normal human dermal fibroblastsrdquoJournal of Dermatology vol 33 no 10 pp 676ndash683 2006

[89] M Muchuweti C Mupure A Ndhlala T Murenje and M AN Benhura ldquoScreening of antioxidant and radical scavengingactivity of Vigna ungiculata Bidens pilosa and Cleome gynan-drardquo American Journal of Food Technology vol 2 no 3 pp 161ndash168 2007

[90] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens bipinnata L against carbon tetrachloride-induced liver fibrosis in ratsrdquo Journal of Pharmacy and Pharma-cology vol 60 no 10 pp 1393ndash1402 2008

[91] S L Chang H H Yeh Y S Lin Y M Chiang T K Wuand W C Yang ldquoThe effect of centaurein on interferon-120574 expression and Listeria infection in micerdquo Toxicology andApplied Pharmacology vol 219 no 1 pp 54ndash61 2007

[92] F Deba T D Xuan M Yasuda and S Tawata ldquoChemical com-position and antioxidant antibacterial and antifungal activitiesof the essential oils from Bidens pilosa Linn var Radiatardquo FoodControl vol 19 no 4 pp 346ndash352 2008

[93] A O T Ashafa and A J Afolayan ldquoScreening the root extractsfrom Biden pilosa L var radiata (Asteraceae) for antimicrobialpotentialsrdquo Journal of Medicinal Plant Research vol 3 no 8 pp568ndash572 2009

[94] T B Nguelefack T Dimo E P Nguelefack Mbuyo P V TanS V Rakotonirina and A Kamanyi ldquoRelaxant effects of theneutral extract of the leaves of Bidens pilosa linn on isolated ratvascular smooth musclerdquo Phytotherapy Research vol 19 no 3pp 207ndash210 2005

[95] M T Grombone-Guaratini K L Silva-Brandao V N SolferiniJ Semir and J R Trigo ldquoSesquiterpene and polyacetyleneprofile of the Bidens pilosa complex (Asteraceae Heliantheae)from Southeast of Brazilrdquo Biochemical Systematics and Ecologyvol 33 no 5 pp 479ndash486 2005

[96] L L Lin C Y Wu H C Hsiu M T Wang and H ChuangldquoStudies on diabetes mellitus I The hypoglycemic activity ofphytosterin on alloxan diabetic ratsrdquo Taiwan Yi Xue Hui Za Zhivol 66 no 2 pp 58ndash65 1967

[97] M A N Benhura and I C Chitsiku ldquoThe extractable 120573-carotene content of Guku (Bidens pilosa) leaves after cookingdrying and storagerdquo International Journal of Food Science andTechnology vol 32 no 6 pp 495ndash500 1997

[98] J S Chang L C Chiang C C Chen L T Liu K C Wangand C C Lin ldquoAtileukemic activity of Bidens pilosa l varminor (blume) sherff andHouttuynia cordata thunbrdquo AmericanJournal of Chinese Medicine vol 29 no 2 pp 303ndash312 2001

[99] F Deba T D Xuan M Yasuda and S Tawata ldquoHerbicidal andfungicidal activities and identification of potential phytotoxinsfrom Bidens pilosa L var radiata Scherff research paperrdquoWeedBiology and Management vol 7 no 2 pp 77ndash83 2007

[100] J K Kumar and A K Sinha ldquoA new disubstituted acetylacetonefrom the leaves ofBidens pilosa LinnrdquoNatural Product Researchvol 17 no 1 pp 71ndash74 2003

[101] K Ogawa and Y Sashida ldquoCaffeoyl derivatives of a sugarlactone and its hydroxy acid from the leaves of Bidens pilosardquoPhytochemistry vol 31 no 10 pp 3657ndash3658 1992

[102] Q Xia Y Liu and Y Li ldquoDetermination of gallic acid fromdifferent species and different medical parts of Herba Bidens byRP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol24 no 03 pp 308ndash310

[103] W J Lee L F Wu W K Chen C J Wang and T HTseng ldquoInhibitory effect of luteolin on hepatocyte growth fac-torscatter factor-induced HepG2 cell invasion involving bothMAPKERKs and PI3K-Akt pathwaysrdquo Chemico-BiologicalInteractions vol 160 no 2 pp 123ndash133 2006

[104] J A Beutler J H Cardellina C M Lin E Hamel G MCragg andM R Boyd ldquoCentaureidin a cytotoxic flavone fromPolymnia fruticosa inhibits tubulin polymerizationrdquo BioorganicandMedicinal Chemistry Letters vol 3 no 4 pp 581ndash584 1993

[105] A Verma A Su A M Golin B OrsquoMarrah and J K AmorosaldquoThe lateral view a screening method for knee traumardquo Aca-demic Radiology vol 8 no 5 pp 392ndash397 2001

[106] J Corren M Lemay Y Lin L Rozga and R K RandolphldquoClinical and biochemical effects of a combination botanicalproduct (ClearGuard) for allergy a pilot randomized double-blind placebo-controlled trialrdquo Nutrition Journal vol 7 no 1article 20 2008

[107] M S Gachet J S Lecaro M Kaiser et al ldquoAssessment of anti-protozoal activity of plants traditionally used in Ecuador in thetreatment of leishmaniasisrdquo Journal of Ethnopharmacology vol128 no 1 pp 184ndash197 2010

[108] W C Yang M Ghiotto B Barbarat and D Olive ldquoThe role ofTec protein-tyrosine kinase in T cell signalingrdquo The Journal ofBiological Chemistry vol 274 no 2 pp 607ndash617 1999

[109] S Tewtrakul H Miyashiro N Nakamura et al ldquoHIV-1 inte-grase inhibitory substances from Coleus parvifoliusrdquo Phytother-apy Research vol 17 no 3 pp 232ndash239 2003

[110] T Dimo T B Nguelefack P V Tan et al ldquoPossible mechanismsof action of the neutral extract from Bidens pilosa L leaves onthe cardiovascular system of anaesthetized ratsrdquo PhytotherapyResearch vol 17 no 10 pp 1135ndash1139 2003

[111] T S C Li Chinese and Related North American Herbs CRCPress New York NY USA 2002

[112] H Wu H Chen X Hua Z Shi L Zhang and J ChenldquoClinical therapeutic effect of drug-separated moxibustion onchronic diarrhea and its immunologic mechanismsrdquo Journal ofTraditional Chinese Medicine vol 17 no 4 pp 253ndash258 1997

[113] S W Wright R R Harris J S Kerr et al ldquoSynthesis chemicaland biological properties of vinylogous hydroxamic acids dualinhibitors of 5-lipoxygenase and IL-1 biosynthesisrdquo Journal ofMedicinal Chemistry vol 35 no 22 pp 4061ndash4068 1992

[114] W R Almiron and M E Brewer ldquoClassification of immaturestage habitats of Culicidae (Diptera) collected in CordobaArgentinardquo Memorias do Instituto Oswaldo Cruz vol 91 no 1pp 1ndash9 1996

[115] N L Wang J Wang X S Yao and S Kitanaka ldquoTwo newmonoterpene glycosides and a new (+)-jasmololone glucoside

Evidence-Based Complementary and Alternative Medicine 51

from Bidens parvifloraWilldrdquo Journal of Asian Natural ProductsResearch vol 9 no 5 pp 473ndash479 2007

[116] P Champagnat ldquoRole of the terminal bud in the actionexercised by the cotyledon of Bidens pilosus L var radiatus onits axillary budrdquo Comptes Rendus des Seances et Memoires de laSociete de Biologie vol 145 no 17-18 pp 1374ndash1376 1951

[117] S F Nielsen S B Christensen G Cruciani A Kharazmiand T Liljefors ldquoAntileishmaniai chalcones statistical designsynthesis and three-dimensional quantitative structure-activityrelationship analysisrdquo Journal of Medicinal Chemistry vol 41no 24 pp 4819ndash4832 1998

[118] Y C Hwang J J H Chu P L Yang W Chen and M V YatesldquoRapid identification of inhibitors that interfere with poliovirusreplication using a cell-based assayrdquo Antiviral Research vol 77no 3 pp 232ndash236 2008

[119] V F Andrade-Neto M G L Brandao F Q Oliveira et alldquoAntimalarial activity of Bidens pilosa L (Asteraceae) ethanolextracts fromwild plants collected in various localities or plantscultivated in humus soilrdquo Phytotherapy Research vol 18 no 8pp 634ndash639 2004

[120] R J Marles and N R Farnsworth ldquoAnti-diabetic plants andtheir active constituentsrdquo Phytomedicine vol 2 no 2 pp 137ndash189

[121] M Habeck ldquoDiabetes treatments get sweet help from naturerdquoNature Medicine vol 9 no 10 p 1228 2003

[122] H W Lin G Y Han and S X Liao ldquoStudies on the activeconstituents of the Chinese traditional medicine Polygonatumodoratum (Mill) DrucerdquoActa Pharmaceutica Sinica vol 29 no3 pp 215ndash222 1994

[123] C L T Chang Y C Chen H M Chen N S Yang andW C Yang ldquoNatural cures for type 1 diabetes a reviewof phytochemicals biological actions and clinical potentialrdquoCurrent Medicinal Chemistry vol 20 no 7 pp 899ndash907 2013

[124] L Dey A S Attele and C S Yuan ldquoAlternative therapies fortype 2 diabetesrdquo Alternative Medicine Review vol 7 no 1 pp45ndash58 2002

[125] C L T ChangH Y Liu T F Kuo et al ldquoAnti-diabetic effect andmode of action of cytopiloynerdquo Evidence-Based Complementaryand Alternative Medicine vol 2013 Article ID 685642 13 pages2013

[126] M R Khan M Kihara and A D Omoloso ldquoAnti-microbialactivity of Bidens pilosa Bischofia javanica Elmerillia papuanaand Sigesbekia orientalisrdquo Fitoterapia vol 72 no 6 pp 662ndash6652001

[127] T Dimo T B Nguelefack P Kamtchouing E Dongo ARakotonirina and S V Rakotonirina ldquoHypotensive effects ofmethanol extract from Bidens pilosa Linn on hypertensive ratsrdquoComptes Rendus de lrsquoAcademie des Sciences Serie III vol 322no 4 pp 323ndash329 1999

[128] A K Hassan O Deogratius J F Nyafuono O Francis and OP Engeu ldquoWound healing potential of the ethanolic extracts ofBidens pilosa and Ocimum suaverdquo African Journal of Pharmacyand Pharmacology vol 5 no 2 pp 132ndash136 2011

[129] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[130] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[131] C Kandaswami and EMiddleton ldquoFree radical scavenging andantioxidant activity of plant flavonoidsrdquo Advances in Experi-mental Medicine and Biology vol 366 pp 351ndash376 1994

[132] K Yamamoto H Kakegawa H Ueda et al ldquoGastric cytopro-tective anti-ulcerogenic actions of hydroxychalcones in ratsrdquoPlanta Medica vol 58 no 5 pp 389ndash393 1992

[133] J O C Ezeonwumelu A K Julius C N Muhoho et alldquoBiochemical and histological studies of aqueous extract ofBidens pilosa leaves from Ugandan Rift valley in ratsrdquo BritishJournal of Pharmacology and Toxicology vol 2 no 6 pp 302ndash309

[134] L Frida S Rakotonirina A Rakotonirina and J P SavineauldquoIn vivo and in vitro effects of Bidens pilosa L (Asteraceae)leaf aqueous and ethanol extracts on primed-oestrogenized ratuterine musclerdquo African Journal of Traditional Complementaryand Alternative Medicines vol 5 no 1 pp 79ndash91 2008

52 Evidence-Based Complementary and Alternative Medicine

Composition Comments

1 Please provide valid postal code to the address if

possible

2 We made the highlighted change in Table 4 for the sake

of correctness Please check

3 Please check the correctness of the highlighted part in

Table 5

4 We considered the highlighted part in Table 11 as

reference Please check similar cases throughout

5 Please check the format of Table 18

6 We reformatted Table 20 Please check

7 We made the highlighted change in Table 4 for the sake

of consistency Please check similar cases throughout the

paper

8 We made the highlighted change(s) for the sake

of consistency Please check simila highlighted cases

throughout

9 We made the highlighted change in Table 9 for the sake

of clarity Please check similar cases through out

10 We made the highlighted change(s) for the sake of

correctness Please check

11 Comment on ref [66] This reference is a repetition

of [18] Please check similar cases throughout

Author(s) Name(s)

Author 1

Author 2

Author 3

10 Evidence-Based Complementary and Alternative Medicine

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

55

(R)-1-(Hydroxymethyl)-246810-dodecapentayn-1-yl-a-D-glucopyranoside

2- 120573 -D-Glucopyranosyloxy-1-hydroxytrideca-357911-pentayne

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Chinaand Japan) [35 47]

56

1-[[(Carboxyacetyl)oxy]methyl]46810-dodecatetraynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHOH

OH

Aerial (Japan) [54]

57

(4E)-1-[[(Carboxyacetyl)oxy]-methyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

OOO

OHO

O

OHH

HO

H

H

HHO

H

OH

Aerial (Japan) [54]

58

(4E)-1-[[(Carboxyacetyl)oxy]-ethyl]-4-dodecene-6810-triynyl-a-D-glucopyranoside

O

O

O

OHO

O

OHOH

H

HO

H

H

HHO

HOH

Aerial (Japan) [54]

59 (5E)-5-Heptene-13-diyn-1-yl-benzene

1-Phenylhepta-13-diyn-5en Whole (Taiwan) [32]

60 7-Phenyl-2(E)-heptene-46-diyn-1-ol

OHRoots (notstated) Aerial(China)

[38 42]

617-Phenyl-2(E)-heptene-46-diyn-1-ol-acetate

OO

Roots (notstated Brazil) [38 55 56]

62 7-Phenyl-46-heptadiyn-2-ol Pilosol A

OH

Whole (Taiwan)aerial (China) [42 57]

Evidence-Based Complementary and Alternative Medicine 11

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

63 7-Phenylhepta-46-diyn-12-diol HO

OH

Aerial (China) [42]

64 135-Heptatriyn-1-yl-benzene

1-Phenylhepta-135-triyne

Leaves (notstated) leaves oftissue culture(not stated)aerial (TanzaniaChina)Whole (Taiwan)roots (Brazil)

[18 34 3842 56ndash58]

65 7-Phenyl-246-heptatriyn-1-ol HO

Leaves (notstated) aerial(China)

[38 42]

667-Phenyl-246-heptatriyn-1-ol-acetate O

O Leaves (notstated) [38]

675-(2-Phenylethynyl)-2-thiophenemethanol

S OHAerial (China) [42]

685-(2-Phenylethynyl)-2120573-glucosylmethyl-thiophene

O

O

OH

H

OH

H

H

HOH H

O

HO

S

Aerial (China) [42]

69

3- 120573 -D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne

OH

O

OH

HOHO

H

H

HHOH

OH

Leaves(Cameroon) [28]

701-Phenyl-13-diyn-5-en-7-ol-acetate O

O

Leaves (Brazil) [27]

SN denotes serial number

the chloroform fraction was the most toxic with a halfmaximal inhibitory concentration (IC

50) of 97 plusmn 72 and 83 plusmn

52 120583gmL in NRU and MTT respectively [82] Kumari andcolleagues also reported the anti-cancer and anti-malarialactivities of B pilosa leaves [77] Based on a cytotoxicity-directed fractionation strategy they identified phenyl-135-heptatriene with IC

50values of 8plusmn001 049plusmn045 07plusmn001

and 10 plusmn 001 120583gmL against human oral liver colon andbreast cancer cell lines respectively However phenyl-135-heptatriyne showed lower activity against breast cancer celllines than the chloroform leaf extract which had an IC

50

value of 65 plusmn 001 120583gmL Moreover the positive controltaxol showed higher activity than phenyl-135-heptatriyne[77] Furthermore in vitro comet assays were performed toevaluate the toxicity of n-hexane chloroform and methanolextracts of B pilosa and its ethyl acetate acetone and waterfractions onHela andKB cellsThe ethyl acetate fraction fromthe methanol extract exhibited the highest activity with halfmaximal cytotoxic concentrations (CTC

50) of 9652120583gmL

and 5862120583gmL against Hela and KB cells respectivelyDespite the moderate toxicity these findings suggest thatthese B pilosa extractsfractions could be useful for future

12 Evidence-Based Complementary and Alternative Medicine

Table5Flavon

oids

isolated

from

Bpilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

71

2-[(34-Dihydroxyph

enyl)-

methylene]-

6-hydroxy-3(2H

)-benzofuranon

e

Sulfu

retin

O

OHO

OH

OH

Aeria

l(Ch

ina)

[35]

72

2-[(34-Dihydroxyph

enyl)-

methylene]-

67-dihydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-673101584041015840-

tetrahydroxy

maritimetin

O

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

73

2-[(34-Dihydroxyph

enyl)-

methylene]-6-

(120573-D

-glucopyrano

syloxy)-7-

hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-120573-D

-glucop

yranosyl-

673101584041015840-te

trahydroxy

maritimein

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japanand

China)aerial(Ch

ina)

[355960]

74

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

(120573-D

-glucopyrano

syloxy)-

6-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-7-O-120573-D

-glucop

yranosyl-

673101584041015840-

tetrahydroxy

O

OHO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japan)

[59]

75

6-[(6-O-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-acetyl-

120573-D

-glucopyrano

syl)

-673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OHO

H HO

H

H

HOH

HO

O

O

Leaves

(Not

stated)

aeria

l(Ch

ina)

[3559]

Evidence-Based Complementary and Alternative Medicine 13

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

76

6-[(36-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-

2-[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone(Z)-6-O

-(36-di-O

-acetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

bideno

sideA

O

O

HO

OH

OH

OHOH O

H

H

HOH

HO

O

O

O

Leaves

(China)

[60]

77

6-[(46-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(410158401015840610158401015840-

diacetyl-120573-D

-glucropyrano

syl)-

673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

HO

O

OO

Leaves

(not

stated)

aeria

l(Ch

ina)

[343561]

78

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

hydroxy-6-

[(246-tri-O

-acetyl-120573

-D-

glucop

yranosyl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(210158401015840410158401015840610158401015840-

triacetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

O

OOO

O

Leaves

(not

stated)

aeria

l(Ch

ina)

[3461]

79

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-6-

[(346-tri-O

-acetyl-120573

-D-glucopyrano

syl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(310158401015840410158401015840610158401015840-

triacetyl-120573

-D-glucrop

yranosyl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H OH

H

HOH

HO

OOO O

Aeria

l(Ch

ina)

[3435]

80

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-

6-[[6-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propenyl]-120573-

D-glucopyrano

syl]o

xy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-

p-coum

aroyl-

120573-D

-glucropyrano

syl)-

673101584041015840-te

trahydroxy

OH

O

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OLeaves

(Japan)

[59]

14 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

81

1-[2-(120573-D

-Glucopyrano

syloxy)-

4-hydroxyphenyl]-2-

hydroxy-3-(3-

hydroxyphenyl)-2-

prop

en-1-on

e

Chalcone120572

32101584041015840-

tetrahydroxy-

21015840-O

-120573-D

-glucop

yranosyl

HO

O

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[35]

821-(24-Dihydroxyph

enyl)-

3-(34-dihydroxyph

enyl)-

2-prop

en-1-on

eBu

tein

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

833-(34-D

ihydroxyph

enyl)-

1-(234-trihydroxyph

enyl)-

2-prop

en-1-on

eOkanin

O

HO

HO

OH

OH

OH

Leaves

(China)

[60]

84

3-(34-D

ihydroxyph

enyl)-

1-[3-(120573-D

-glucop

yranosyloxy)-

24-dihydroxyphenyl]-2-

prop

en-1-on

e

Okanin

31015840-O

-120573-D

-glucosid

e

O

HO

OH

OH

OH

OO

HO

H

HO

H

H

HOH

H

OH

Leaves

(Germany)

flowers(Germany)

[61ndash63]

85

3-(34-D

ihydroxyph

enyl)-

1-[4-(120573-D

-glucop

yranosyloxy)-

23-dihydroxyphenyl]-

2-prop

en-1-on

e

Okanin

41015840-O

-120573-D

-glucosid

e

OOH

OH

OH

HOO

OHO

H

HO

H

H

HOH

H

OH

Flow

ers(Germany)

leaves

(Japan)

[5963]

Evidence-Based Complementary and Alternative Medicine 15

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

86Okanin41015840-O

-120573-D

-(610158401015840-

O-acetylglucosid

e)

O

O

OH

OH

OH

OHO

H

HO

H

H

HOH

H

HO

O

O

Flow

ers(Germany)

[63]

87

1-[4-[(46-di-O

-Acetyl-

120573-D

-glucopyrano

syl)-oxy]-

23-dihydroxyphenyl]-3-

(34-dihydroxyph

enyl)-2-

prop

en-1-on

e

Okanin

41015840-O

-120573-D

-(410158401015840610158401015840-diacetyl)-

glucop

yranoside

O

O

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Aeria

l(Ch

ina)

[34]

88Okanin41015840-O

-120573-D

-(210158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[64]

89Okanin41015840-O

-120573-D

-(310158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

O

OAe

rial(Ch

ina)

[34]

16 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

90

1-[23-Dihydroxy-4-

[[6-O-[3-(4-hydroxyph

enyl)-

1-oxo-2-prop

enyl]-120573-D

-glucop

yranosyl]oxy]-ph

enyl]-

3-(34-dihydroxyph

enyl)-

2-prop

en-1on

e

Okanin

41015840-O

-120573-D

-(610158401015840-tr

ans-

p-coum

aroyl)-

glucoside

O

O

OH

OH

OH

OHOH

HO

H

H

HOH

H

HO

OO

OH

Leaves

(Germany)

[64]

91Okanin41015840-O

-120573-D

-(410158401015840-acetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Leaves

(Germany)

[65]

92

Okanin

41015840-O

-120573-D

-(210158401015840410158401015840-diacetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[65]

Evidence-Based Complementary and Alternative Medicine 17

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

93

Okanin

41015840-O

-120573-D

-(310158401015840410158401015840-diacetyl-610158401015840-

trans-p-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

OH

O O

Leaves

(Germany)

[65]

94

Okanin

41015840-O

-[120573-D

-glucopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

OOH

OH

OH

HO

OH

HO

HHO

H

HOH

HO

OH

OH

HO

HHO

H

HOH

HO

Flow

er(G

ermany)

[63]

95Okanin

3101584041015840-di-O

-120573-D

-glucosid

e

OOH

OH

OH

O OO

H HO

HHOH

HOH

HHO

OHO

H

HO

H

H

HOH

H

OH

Flow

er(G

ermany)

[63]

18 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

96

1-[3-(120573-D

-Glucopyrano

syloxy)-

24-dihydroxyphenyl]-

3-(3-hydroxy)-

4-metho

xyph

enyl)-

2-prop

en-1-on

e

Okanin

4-methylether-

31015840-O

-120573-D

-glucop

yranoside

OOH

O

OH

HO

O

H HO

HHOH

HOH

H

OHO

Leaves

(Germany)aerial

(China)

[3562]

97

Okanin4-methylether-

3101584041015840-di-O

-120573-(41015840101584061015840101584041015840101584010158406101584010158401015840-

tetraacetyl)-

glucop

yranoside

OOH

OOH

O

HH

OH

HO

HOH

H

OO

O

Ac

Ac

AcAc

OO

H

HO

H

H

HOH

H

O

Aeria

l(Ch

ina)

[42]

98Ch

alcone210158404101584061015840-tr

imetho

xy-

4-O-D

-glucopyrano

syl-

dihydro

Not

foun

dLeaves

(China)

[60]

992-(34-D

ihydroxyph

enyl)-23-

dihydro-78

-dihydroxy-

4H-1-

benzop

yran-4-one

Okanin

isoOO

OH

HO

OH

OH

Leaves

(China)

[60]

100

2-(34-D

ihydroxyph

enyl)-

23-dihydro-8-hydroxy-

7[(246-tr

i-O-acetyl-120573

-D-

glucop

yranosyl)oxy]-4H

-1-b

enzopyran-4-on

eOO

OH

OH

OH

OH

O

HHO

H

HO

HO

OAc

Ac

Ac

Aeria

l(Ch

ina)

[34]

Evidence-Based Complementary and Alternative Medicine 19

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

101

57-Dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

OO

OH

HO

OH

Aeria

l(TanzaniaC

hina)

[4266]

102

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

7-O-glucopyrano

side

OO

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Tanzania)

[66]

103

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-4H

-1-benzop

yran-4-one

Luteolin

OO

HO

OH

OH

OH

Aeria

l(TanzaniaC

hina

Vietnam)

[343542

6667]

104

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5hydroxy-4H-1-

benzop

yran-4-one

Luteolin

7-O-120573-D

-glucop

yranoside

OOOH

OH

OH

OO

HO

H

HO

H

H

HOH

OH H

Aeria

l(Tanzania)

[66]

105

57-Dim

etho

xy-6-

(5-m

etho

xy-6-m

ethyl-

4-oxo-4H

-pyran-3-yl)-

2-ph

enyl-4H-1-

benzop

yran-4-one

5-O-M

ethylhosludin

O OOO

O

O

O

Aeria

l(Ugand

a)[68]

20 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

106

3-(120573-D

-Glucopyrano

syloxy)-

57-dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Astragalin

Kaem

pferol-3-O

-120573-D

-glucopyrano

side

HO

OH

O O

OH

O

O

HO

HHO

H

H

HOH

OH

H

Aeria

l(Ch

ina)

[35]

107

Kaem

pferol3-

(23-di-E

-p-cou

maroyl-

120572-L-rhamno

pyrano

side)

Not

foun

dAe

rial(Vietnam)

[67]

108

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5-hydroxy-36-dimetho

xy-

4H-1-

benzop

yran-4-one

Axillarosid

eO O

OH

OH

OO

OH

OO

HO

H

HO

H

H

HOH

H

OH

Aeria

l(Ch

ina)

[42]

109

57-Dihydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaureidin

O

O

O

O

O

HO

OH

OH

Who

le(Taiwan)

[69]

110

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaurein

O O

OO

O OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan)w

hole

(Taiwan)

[69ndash

71]

Evidence-Based Complementary and Alternative Medicine 21

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

111

5-Hydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-

67-dimetho

xy-4H-

chromen-4-one

Eupatorin

iso

OO O

OH

O

O OH

Not

foun

d(C

hina)

[3772]

112

2-(34-D

imetho

xyph

enyl)-7-

(120573-D

-glucopyrano

syloxy)-35-

dihydroxy-

8-metho

xy-4H-1-

benzop

yran-4-one

O O

OH

O O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

113

7-(120573-D

-Glucopyrano

syloxy)-

5hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-38-

dimehtoxy-4H-1-

benzop

yran-4-one

O O

O

OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

114

Isorhamnetin

3-[O

-120572-

L-rham

nopyrano

syl-

(1-2)-120573-D

-glucop

yranoside]

Not

foun

dAe

rial(Vietnam)

[67]

115

7-[(6-deoxy-120572-L-

Manno

pyrano

syl)o

xy]-

3-120573-D

-glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-4H

-1-benzop

yran-4-one

Luteoside

O O

OH

O

OH

OH

OH

HOH

H

H HO

HO

HO

OH

HO

H

HO

OH

OH

HH

Aeria

l(Ch

ina)

[42]

22 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

116

Luteolin

3-O-120573-D

-Glucopyrano

side

O O

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

HO

Aeria

l(Tanzania)

[66]

117

57-Dihydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Quercetagetin

3631015840-tr

imethylether

OOO

O

O OH

HO

OH

Aeria

l(Ch

ina)

[42]

118

6-((2S3S4S5S)-

Tetrahydro-345-tr

ihydroxy-

6-(hydroxymethyl)-2H

-pyran-2-yloxy)-5-hydroxy-

2-(4-hydroxy-3-

metho

xyph

enyl)-

37-dimetho

xy-4H-

chromen-4-one

Quercetagetin

3731015840-

trim

ethylether-6-O

-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[42]

119

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

JaceinQ

uercetin

3631015840-

trim

ethyl

ether-7-O-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan

China)

who

le(Taiwan)

[425470]

120

2-(34-D

ihydroxyph

enyl)-

357-trihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

OO

HO

OH

OH

OH

OH

Aeria

l(Ch

ina)leaves

(China)who

le(C

hina)

[356074]

Evidence-Based Complementary and Alternative Medicine 23

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

121

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-galactopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-4-one

Quercetin

3-O-120573-D

-galactosidehyperin

hyperosid

e

O O

HO

OH

OH

OH

OOHH

HOH

H

H HO

HO

OH

Aeria

l(TanzaniaJapan)

Leaves

(China)Who

le(C

hina)

[376066

717475]

122

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-glucopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

3-O-120573-

D-glucopyrano

side

O O

HO

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

Aeria

l(TanzaniaC

hina

Japan)L

eaves(Japan)

[355966

75]

123

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-

4-oxo-4H

-1-benzop

yran-3-yl-

120573-D

-glucopyrano

siduron

icacid

Quercetin

3-O-120573-D

-glucuron

opyranoside

O O

HO

OH

OH

OH

OHOH

HOH

H

H HO

HO

OHO

Aeria

l(TanzaniaJapan)

[6671]

124

3-[[6-O-(6-

Deoxy-120572-L-m

anno

pyrano

syl)-

120573-D

-galactopyrano

syl]o

xy]-

2-(34-dihydroxyph

enyl)-57-

dihydroxy-4H

-1-benzop

yran-4-one

Quercetin

3-O-rob

inob

iosid

e

O O

HO

OH

OH

OH

O

HOHH OH

HO

H HH

OO

OHH

HOH

H

H HO

HO

Aeria

l(Japan)W

hole

(Taiwan)

[7071]

24 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

125

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-

3-metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

OH

Roots(Brazil)

[55676]

126

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-

120573-D

-glucopyrano

syl]o

xy]-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120572-

L-rham

nopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

O

HHO HHO

HO H

HH

ORo

ots(Brazil)

[5676]

127

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-120573-D

-glucop

yranosyl]oxy]-5-

hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-3-metho

xy-

4H-1-

benzop

yran-4-one

Quercetin

341015840-

dimethyl

ether-7-O-rutinoside

O OOH

OH

O

O

OH O

HOH

H

HHO

H

O

HHO HHO

HO H

HH

O

HO

Aeria

l(Ch

ina)

[3435]

128

2-(34-D

ihydroxyph

enyl)-3-

(120573-D

-glucofurano

syloxy)-57-

dihydroxy-4H

-1-benzop

yran-

4-on

e

Isoq

uercitrin

O OOH

OH

OHO

H

OHH

OH

HO

H O HOH

OH

Aeria

l(Japan

China)

[3571]

Evidence-Based Complementary and Alternative Medicine 25

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

129

Quercetin

3-O-120573-D

-galactop

yranoside

O

OH

HO

OH OH

O

O

HO

OH

OHOH

OAe

rial(no

tstated)

[18]

130

Quercetin

3-O-rutinoside

O

O

R

OO

OH

OHO

H

O

OH

OH

H3C

H3C

H3C

OH

HO

Who

le(Taiwan)

[70]

26 Evidence-Based Complementary and Alternative Medicine

Table 6 Terpenoids isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(Country) References

131371111-Tetramethylbicyclo[810]undeca-

26-dieneBicyclogermacrene Leaves

(Brazil) [95]

13241111-Trimethyl-8-

methylenebicyclo[720]undec-4-ene

E-Caryophyllene Leaves(Brazil) [95]

1331-Methyl-5-methylene-8-

(1-methylethyl)-16-cyclodecadiene

Germacrene D Leaves(Brazil) [95]

1344-(15-Dimethyl-4-hexen-1-ylidene)-1-methyl-cyclohexene

Z-120574-Bisabolene Leaves(Brazil) [95]

135Decahydro-114-trimethyl-7-methylene-1H-cycloprop[e]-

azulene120573-Gurjunene Leaves

(Brazil) [95]

136 2669-Tetramethyl-148-cycloundecatriene

120572-Humulene120572-caryophyllene

Leaves(Brazil) [95]

13712344a568a-Octahydro-1-isopropyl-7-methyl-4-methylenenaphthalene

120575-Muurolene Leaves(Brazil) [95]

138

12344a568a-Octahydro-4a8-dimethyl-2-(1-methylethylidene)-

naphthalene

Selina-37(11)-diene

Leaves(Brazil) [95]

139(2E7R11R)-371115-

Tetramethyl-2-hexadecen-1-ol

Phytol

HO

Whole(Taiwan) [57]

Evidence-Based Complementary and Alternative Medicine 27

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

140 371115-Tetramethyl-2-hexadecenoic acid Phytanic acid

O

OH

Whole(Taiwan) [57]

141371115-Tetramethyl-2-

hexadecenyl ester-heptanoicacid

Phythylheptanoate O O

Leaves(Not stated) [33]

142

(3S10R13R)-17-((2R5R)-5-Ethyl-6-methylheptan-2-yl)-

2347891011121314151617-tetradecahydro-

1013-dimethyl-1H-cyclopenta[a]phenanthren-

3-ol

Campestrol

HO

Aerial(Tanzania) [18]

143 Not found Phytosterin-B Not foundNot found(TaiwanEgypt)

[39 96]

144 Stigmast-5-en-3-ol 120573-sitosterol

HOH

H

H

Aerial(Tanzania)

whole(Taiwan)

[18 31 57]

145

1314151617-Tetradecahydro-1013-

dimethyl-1H-cyclopenta[a]phenanthren-

3-yloxy)-tetrahydro-6-(hydroxymethyl)-2H-pyran-345-triol

120573-Sitosterolglucoside

H

H

HOH

HO

HHO

H

HOHH O

OH Not found(Egypt) [39]

146 5120572-Stigmasta-7-en-3120573-ol

HO

Whole(Taiwan) [57]

147 5120572-Stigmasta-722t-dien-3120573-ol

HO

Whole(Taiwan) [57]

148 Stigmasta-522-dien-3-ol Stigmasterol

HOH

H

H

Not found(Taiwan)aerial

(Tanzania)leaves (notstated)whole

(Taiwan)

[18 31 3357]

28 Evidence-Based Complementary and Alternative Medicine

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

149 Lup-20(29)-en-3-ol Lupeol

H

HH

HOH

Not found(Egypt) [39]

150 Lup-20(29)-en-3-ol acetate Lupeol acetate

H

HH

O

OH

Not found(Egypt) [39]

151 Olean-12-en-3-ol 120573-amyrin

H

HOH

Not found(Egypt) [39]

152 5913-Trimethyl-242526-trinoroleanan-3-ol Friedelan-3120573-ol

HO

Aerial(Tanzania) [18]

153 5913-Trimethyl-242526-trinoroleanan-3-one

Friedelinfriedelan-3-one

O

Aerial(Tanzania) [18]

1542610151923-

Hexamethyl-2610141822-tetracosahexaene

Squalene

Aerial(Tanzania)Leaves (Notstated)Whole(Taiwan)

[18 33 57]

155 120573120573-Carotene 120573-Carotene Leaves (Notstated) [97]

Evidence-Based Complementary and Alternative Medicine 29

studies [83] Hot water extracts of B pilosa varminor Sheriffwere also assessed for its antileukemic effects on leukemiccell lines L1210 U937 K562 Raji and P3HR1 using XTT-based colorimetric assays The extract inhibited the five celllines with IC

50values ranging from 145120583gmL to 586120583gmL

L1210 K562 Raji and P3HR1 were more sensitive to B pilosaextract with IC

50values below 200120583gmL [98]

Consistent with the antitumor activities of B pilosaextracts and fractions some of its phytochemicals alsoshowed anticancer activity as outlined in Table 11 Amongthem luteolin (103) a well-studied flavonoid with multiplebioactivities was more effective against tumor cell prolifera-tion than its derivatives with IC

50values ranging from 3 120583M

to 50120583M in cells and 5 to 10mgkg in animals Luteolin wasalso found to fight cancer as a food additive at concentrationsof 50 to 200 ppm [78] and prevent skin cancer [78] and cancer3invasion [103] Lee and colleagues reported that luteolinprevents cancer by inhibiting cell adhesion and invasion[103] Significant inhibition concentration was reported to be5 120583M and complete inhibition concentration was reported tobe 40120583M Moreover luteolin was reported to inhibit hepa-tocyte growth factor (HGF)-induced cell scattering as wellas cytoskeleton changes such as filopodia and lamellipodiawhichwas determined using phase-contrast and fluorescencemicroscopy Furthermore luteolin also inhibited the HGF-induced phosphorylation of c-Met ERK 12 and Akt aswell as the MAPKERK and P13K-Akt pathways [78 103]Other mechanisms underlying the anticancer activities ofluteolin are the inhibition of topoisomerase I and II whichinhibits cell replication and DNA repair thus promotingapoptosis regulation of PI-3-KinaseAktMAPKERKJNKactivation of apoptosis in the mitochondrial pathway byactivating caspase 9 and caspase 3 which were found inmalignant cells but not in normal human peripheral bloodmononuclear cells death receptor-induced apoptosis and acell-cycle arrest mechanism inhibition of fatty acid synthasewhich is upregulated in many cancer cells and sensitizationto chemotherapywhereby luteolin increases the susceptibilityof cancer cells to chemotherapy [78] Butein (82) is anotherflavonoid that showed a cytotoxic effect on human colonadenocarcinoma cell proliferation with a reported IC

50value

of 175 120583M Butein at 2 120583M affected the incorporation of[14C]-labeled leucine thymidine and uridine which cancause the inhibition of DNA RNA and protein synthesis ofhuman colon cancer cells Moreover butein also exhibitednoncompetitive inhibition of 1-chloro-24-dinitrobenzene(CDNB) in glutathione S-transferase (GST) activity Tumorresistance was correlated with high levels of GST thus buteininhibited proliferation of cancer cells [80] Another flavonoidpresent in B pilosa centaureidin (109) also showed anti-cancer activity in B lymphoma cells Centaureidin isolatedfrom Polymnia fruticosa inhibited tubulin polymerization invitro and induced mitotic figure formation in CA46 Burkittlymphoma cells Using turbidimetric assay the IC

50value of

centaureidin in inhibition of mitosis was 3120583M [104] Cyto-toxicity of centaureidin was further analyzed using AmericanNational Cancer Institute (NCI) 60 human tumor cell linesThe cytotoxicity potency of centaureidin expressed as GI

50

(50 growth inhibition in the NCI tumor line panel) was024120583M [81] These data mark centaureidin as a promisingantimitotic agent for tumor therapy

In addition to anti-tumor flavones polyynes found in B 4pilosa have also been shown to possess anti-tumor proper-ties Based on a bioactivity-directed isolation approach Wuand colleagues identified two polyyne aglycones from theethyl acetate fraction of B pilosa [44] 12-Dihydroxytrideca-57911-tetrayne (48) and 13-Dihydroxy-6(E)- tetradecene-81012-triyne (46) exhibited significant anticell prolifera-tion activity in primary human umbilical vein endotheliumcells (HUVEC) with IC

50values of 125 120583M and 173 120583M

respectively They also decreased angiogenesis and pro-moted apoptosis in human endothelial cells Their anti-angiogenic and cytotoxic effects correlated with activationof the CDK inhibitors and caspase-7 [44] In addition 12-Dihyroxy-5(E)-tridecene-7911-triyne (45) showed antian-giogenic effects in HUVECs with an IC

50value of 124 120583M as

evidenced by a decrease in the tube formation and migrationof HUVECs [43] The IC

50value of compound 45 in the

inhibition of basic fibroblast growth factor-induced HUVECgrowth was 282120583M However it had higher IC

50values

than those for lung carcinoma cells and keratinocytes Thiscompound could also inhibit cell proliferation of HUVECslung carcinoma A549 cells and HACAT keratinocytes Themechanism by which compound 45 inhibits HUVEC growthand angiogenesis is complicated and includes decreasing theexpression of cell cycle regulators (CDK4 cyclins D1 and Aretinoblastoma (Rb) and vascular endothelial growth factorreceptor 1) caspase-mediated activation of CDK inhibitorsp21 (Cip1) and p27 (Kip) upregulation of Fas ligand expres-sion downregulation of Bcl-2 expression and activation ofcaspase-7 and poly (ADP-ribose) polymerase [43]

32 Anti-Inflammatory Activity B pilosa is commonly usedto treat inflammatory disorders The anti-inflammatory phy-tochemicals present in B pilosa are listed in Table 11 5Cyclooxygenase-2 (COX-2) is a physiologically importantenzyme that converts arachidonic acid to prostaglandin(PGE2) Its expression is induced by a wide variety of external

stimuli indicating its involvement in inflammatory diseasesand it is used as an inflammatory marker [88] Yoshida andcolleagues studied the effects of the aqueous extracts of Bpilosa aerial parts in the production of COX-2 and PGE

2as

well as on the activation of mitogen activated protein kinases(MAPKs) in normal human dermal fibroblasts (HDFs) inresponse to inflammatory cytokine IL-1120573 This work showedthat IL-1120573 activated MAPKs such as ERK12 p38 and JNKto different extents and induced COX-2 expression TheCOX-2 expression in HDFs was regulated mainly by p38following IL-1120573 stimulation Consistently the p38 inhibitorSB203580 blocked this expression Using this cell platformB pilosa extracts were tested for inhibition of inflammationThe extract dose-dependently suppressed the activation ofp38 and JNK and moderately suppressed ERK12 as wellas suppressing COX-2 expression and PGE

2production

30 Evidence-Based Complementary and Alternative Medicine

Table7Ph

enylprop

anoids

isolatedfro

mB

pilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

156

3-(4-H

ydroxyph

enyl)-2-prop

enoica

cid

p-Cou

maricacid

OHO

OH

Who

le(Ja

pan)

[99]

157

2-Metho

xy-4-(2-prop

en-1-yl)-ph

enol

Eugeno

lOOH

Leaves

androots(Japan)

[99]

158

3-(4-H

ydroxy-3-m

etho

xyph

enyl)-

2-prop

enoica

cid

Ferulic

acid

O

OH

O

HO

Who

le(Ja

pan)

[99]

159

3-(34-D

ihydroxyph

enyl)-2-prop

enoic

acid

Caffeicacid

O

HO

OH

OH

Who

leandaeria

l(Japan)

[7599]

160

3-Prop

yl-3[(245-trim

etho

xyph

enyl)-

metho

xy]-24-pentanedione

3-Prop

yl-3-(245-

trim

etho

xy)benzyloxy-

pentan-24-dione

O

O

O

O

O

O

Leaves

(India)

[100]

161

3-(34-D

ihydroxyph

enyl)-2-

prop

enoica

cidethyleste

rCa

ffeateethyl

O

O

OH

OH

Not

foun

d(Taiwan)

who

le(Taiwan)

[434584]

Evidence-Based Complementary and Alternative Medicine 31Ta

ble7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

162

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

d-Erythron

icacid

2-O-caffeoyl-2-C

-methyl

O

HO

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

163

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

2-O-caffeoyl-2-C

-methyl

O

O

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

164

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-24-

dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

3-O-caffeoyl-2-C

-methyl

O

O

HO

HO

O

O

OH

OH

Leaves

(Japan)

[101]

165

4-(A

cetyloxy)-3-[[3-(34-

dihydroxyphenyl)-

1-oxo-2-propen-1-y

l-]oxy]-2-hydroxy-

2-methyl-b

utanoica

cid

O

HO

OH

O OHO

HO

OO

Not

foun

d(Ja

pan)

[73]

166

3-(34-D

ihydroxyph

enyl)-tetrahydro-4-

hydroxy-4-

methyl-5

-oxo-3-fu

ranyl

ester-2-prop

enoica

cid

3-O-C

affeoyl-2-C

-methyl-

D-erythrono

-14-la

cton

eHO

HO

O

OO

OOH

Leaves

(Japan)

[101]

167

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]-oxy]-145-

trihydroxy-cyclohexanecarboxylicacid

Chlorogenica

cid

O

HO

HO

OH

O

OH

O

OH

HO

Aeria

l(Japan)w

hole

(Taiwan)

[707175]

32 Evidence-Based Complementary and Alternative Medicine

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

168

4-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]oxy]-

135-tr

ihydroxy-cyclohexanecarboxylic

acid

4-O-C

affeoylqu

inicacid

OHO

HO

OH

OH

O

O

OH

OH

Aeria

l(Japan)

[71]

169

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-15-

dihydroxy-cyclo

hexanecarboxylicacid

34-Di-O

-caffeoylqu

inic

acid

HO

HO

O

O

O

O

OH

OH

HO

OH O

H

O

Aeria

l(Japan)W

hole

(Taiwan)

[45497071]

170

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-14-dihydroxy-

cyclo

hexanecarboxylicacid

35-Di-O

-caffeoylqu

inic

acid

HO

HO

O O

HO

O

HO

OH OO

OHOH

Aeria

l(Japan)w

hole

(Taiwan)

[45497071]

Evidence-Based Complementary and Alternative Medicine 33

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

171

34-bis[[(2E

)-3-(34-D

ihydroxyph

enyl)-1-

oxo-2-prop

en-1-

yl]oxo]-15

-dihydroxy-

cyclo

hexanecarboxylicacid

45-Di-O

-caffeoylqu

inic

acid

OHO

OH

HO

O

O

OH

OH

O

OH

HO

OWho

le(Taiwan)

[454970]

172

3-[4-[[-6-O-[3-(4-H

ydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]-phenyl]-2-prop

enoica

cid

120573-D

-p-C

oumaricacid4-O

-(6-O

-p-caoum

aroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HOH

HO

O

O

Leaves

(Japan)

[59]

173

3-[4-[[2-O

-Acetyl-6

-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]ph

enyl]2-propeno

icacid

120573-D

-p-C

oumaricacid4-O

-(2-O

-acetyl-6

-O-p-

caou

maroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HO

HO

O

O

Leaves

(Japan)aerial

(China)

[3459]

174

67-Dihydroxy-2-chrom

enon

eEsculetin

cicho

rigenin

OHO HO

O

Not

foun

d(Egypt)

[39]

34 Evidence-Based Complementary and Alternative Medicine

Table 8 Aromatic compounds isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

175 12-Benzenediol PyrocatechinHO

HOWhole(Japan) [99]

176 4-Ethyl-12-benzenediol Pyrocatechol

OH

OH

Whole(Japan) [99]

177 Dimethoxyphenol26-dimethoxyphenol

O

HO

O

Roots (Japan) [99]

178 4-Ethenyl-2-methoxy-phenol p-Vinylguaiacol

O

OHWhole(Japan) [99]

179 2-Hydroxy-6-methylbenzaldehyde

6-Methyl-salicylaldehyde O

OH

Whole(Japan) [99]

180 Benzene-ethanol 2-Phenyl-ethanolHO

Whole(Japan) [57]

181 4-Hydroxy-3-methoxy-benzaldehyde Vanillin O

O

HOAerial(Japan) [99]

182 3-Hydroxy-4-methoxy-benzaldehyde Vanillin iso

OO

OH

Leaves(Japan) [99]

183 4-Hydroxybenzoic acid p-Hydroxybenzoicacid

O

HOOH Whole

(Japan) [99]

184 2-Hydroxybenzoic acid Salicylic acidO

HO

HO

Stem androots (Japan) [99]

185 34-Dihydroxybenzoicacid Protocatechuic acid

O

HOOH

OH

Whole(Japan) [99]

1864-Hydroxy-

3methoxybenzoicacid

Vanillic acid

OOH

O

OH

Aerial(Uganda)

Roots (Japan)[68 99]

Evidence-Based Complementary and Alternative Medicine 35

Table 8 Continued

S N IUPAC names Common names Structure Plant part(country) References

187 345-Trihydroxybenzoicacid Gallic acid

O

OHHO

HO

HO

Whole(China) [102]

[88] This work supports the use of B pilosa as an anti-inflammatory agent however no compounds responsible forthe anti-inflammatory activity of B pilosa were identified

A further study also reported the anti-inflammatoryactivity as well as the antiallergic activity of B pilosa [75] Inthis study dried powder of the aerial part of B pilosa whichhad been pretreated with the enzyme cellulosine was usedfor further tests The results showed that oral administrationof the cellulosine-treated B pilosa lowered the level of serumIgE in mice 10 days after immunization with DNP (24-dintrophenyl)-Ascaris as an antigen This treatment alsoreduced dye exudation in skin induced by passive cutaneousanaphylaxis and production of inflammatory mediators his-tamine and substance P in rats [75] Phytochemical analysisshowed that cellulosine treatment increased the percentage ofcaffeic acid and flavonoids This study suggests that B pilosaand its phenolics have anti-inflammatory functions

Phenolics and polyynes are major anti-inflammatoryphytochemicals present in B pilosa (Table 11) Unsurpris-6ingly phenolics such as luteolin (103) and ethyl caffeate(161) that are major constituents of B pilosa have alsobeen reported to possess anti-inflammatory activity Lute-olin was reported to exhibit anti-inflammatory activity inmacrophages Xagorari and colleagues showed that luteolininhibited the release of inflammatory cytokines TNF-120572 andinterleukin-6 in RAW 2647 cells following LPS stimulation[87] It inhibited TNF-120572 production with an IC

50value

of 1120583M The underlying anti-inflammatory mechanism ofluteolin was reported to be the inactivation of Akt andNF-120581B activation [87] In addition luteolin was reportedto confer anti-inflammatory activity through inhibition ofLPS-stimulated iNOS expression in BV-2 microglial cellsIt inhibited LPS-activated microglia in a dose-dependentmanner with an IC

50value of 69 120583MMoreover immunoblot

and RT-PCR data proved that luteolin suppressed I120581B-120572degradation and iNOS expression in LPS-activated microglia[85] Kim and colleagues stated that luteolin may havebeneficial effects on inflammatory neural diseases throughinhibition of iNOS expression [85] A related study revealedthat luteolin decreased the transcriptional activity of NF-120581BRelA via partial inhibition of TNF-mediated NF-120581B DNAbinding activity Luteolin also inhibited Akt phosphorylationand induced degradation of a transcription factor interferonregulatory factor (IRF) [86]

Chiang and colleagues showed that ethyl caffeate(161) significantly inhibited NO production in mousemacrophages RAW 2647 cells [84] Based on MTT assays

they concluded that this inhibition was not due to thecytotoxicity of ethyl caffeate The IC

50value of ethyl caffeate

in the inhibition of NO production was 55 120583gmL slightlylower than curcumin (positive control) which has anIC50

value of 65120583gmL They demonstrated that ethylcaffeate exerted anti-inflammatory activity via the reducedtranscription and translation of iNOS (inducible nitric oxidesynthase) in RAW 2467 cells In addition this compoundalso suppressed COX-2 expression in RAW 2467 cellsand MCF-7 cells The in vivo anti-inflammatory effectof ethyl caffeate was verified by testing in TPA-treatedmouse skin Like celecoxib the positive control ethylcaffeate significantly abolished COX-2 expression in adose-dependent manner Ethyl caffeate at 1mg200120583Lsite(24mM) inhibited COX-2 expression at a level comparableto celecoxib at 1mg200120583Lsite (13mM) Remarkably ethylcaffeate at 48mM (2mg200120583Lsite) was more effectivethan celecoxib at 131mM (10mg200120583Lsite) Moreoverthis compound inhibited the activation of nuclear factor-120581B(NF-120581B) by LPS via the prevention of NF-120581B binding toDNA [84] In addition 3 ethyl caffeate analogs (ethyl 34-dihydroxyhydrocinnamate ethyl cinnamate and catechol)also showed different degrees of NF-120581B binding to DNA aslisted in Table 12 Ethyl cinnamate lacks a catechol moietywhich results in ineffective inhibition of NF-120581B binding toDNA [84]

Pereira and colleagues assessed the anti- 7inflammatory and immunomodulatory activities ofB pilosa methanol extract as well as one polyyne2-O-120573-glucosyltrideca-11(E)-en-3 579-tetrayn-12-diol(54) in T lymphocytes and a zymosan-induced arthritismouse model [27] They first examined the in vitro effect of 8the B pilosa extract and compound 54 on cell proliferation ofhuman T cells stimulated with 5120583gmL phytohemagglutinin(PHA) or 100 nM 12-O-tetradecanoyl phorbol-13-acetate(TPA) plus 15 120583M ionomycin and on cell proliferation ofmouse T cells stimulated with 5 120583gmL concanavalin A(Con A) The data demonstrated that both methanol extractand compound 54 suppressed T-cell proliferation in adose-dependent manner The estimated IC

50values of the B

pilosa extract against human T cells stimulated with 5120583gmLPHA and 100 nM TPA plus 15120583M ionomycin were 125 and25 120583gmL respectively In comparison with the methanolextract compound 54 showed 10-fold more inhibition ofhuman T-cell proliferation with an estimated IC

50value of

15 120583gmL Accordingly the B pilosa extract and compound54 dose-dependently suppressed mouse T-cell proliferation

36 Evidence-Based Complementary and Alternative Medicine

Table 9 Porphyrins isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

188

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(15S16S)-10-ethenyl-5-ethyl-1161820-tetrahydro-6111522-tetramethyl-1820-dioxo-15H-912-imino-212-

metheno-471714-dinitrilopyrano[43-

b]azacyclononadecine-16-propanoic

acid

AristophyllC

NH N

HNN

O

O OOOPhytyl

O

Leaves(Taiwan) [100]

189

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -20b-(methoxycarbonyl)-9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-dinitrilo-12-

dioxeto-[310158404101584034]-cyclo-pent[1 2]azacyclononadecine-

19-propanoic acid

Bidenphytin A

NH N

HNN

OPhytylO O O

O O

Leaves(Taiwan) [100]

190

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -2a-

hydroxy-20b-(methoxycarbonyl)-

9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-

dinitrilo-12-dioxeto[310158404101584034]-cyclo-pent[12-b]-

azacyclononadecine-19-propanoic acid

Bidenphytin B

NH N

HNN

OPhytylO O O

O O

OH

Leaves(Taiwan) [100]

191

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO O

HOOO

Leaves(Taiwan) [100]

192

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

Evidence-Based Complementary and Alternative Medicine 37

Table 9 Continued

S N IUPAC names Common names Structure Plant part(country) References

193

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

Leaves(Taiwan) [100]

194

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

195

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3S4S21R)-9-ethenyl-14-ethyl-21-(methoxycarbonyl)-

481318-tetramethyl-20-oxo-3-phorbinepropanoic acid

Pheophytin A

NH N

HNN

PhytylO

O

OO O

Leaves(Taiwan) [100]

with estimated IC50values of 30 and 25 120583gmL respectively

Taken together the data indicate that the B pilosa extractand compound 54 act on human and mouse T cells Totest the in vivo effect of the B pilosa extract and compound54 a zymosan-induced arthritis mouse model was usedThis model was established from B10ASgSnJ mice with aninjection of zymosan (015mg) The zymosan-injected mice9received an intraperitoneal injection of the B pilosa extract(1 5 or 10mg) at one dose a day for 5 days Popliteal lymphnode (PLN) weight was monitored to check the developmentof arthritis The results revealed that 10mg of the methanolextract of B pilosa extract could significantly diminishinflammation as evidenced by PLN weight [27] This worksuggests that B pilosa (and compound 54) can suppressimmune response and inflammation

33 Antidiabetic Activity Anti-diabetic agents are primarilydeveloped from plants and other natural resources [8 24 4951 53] B pilosa is one of 1200 plant species that have beeninvestigated for antidiabetic activity [120 121]B pilosa is usedas an anti-diabetic herb in America Africa and Asia [51 120

122] Many studies have indicated that B pilosa could treattype 1 diabetes (T1D) and type 2 diabetes (T2D) in animals

Etiologically speaking T1D is caused by autoimmune-mediated destruction of pancreatic 120573 cells leading to insulindeficiency hyperglycemia and complications Currentlythere is no cure for T1D Polarization ofTh cell differentiationcontrols the development of T1D Suppression of Th1 celldifferentiation and promotion of Th2 cell differentiationameliorate T1D [123] One study showed that the butanolfraction of B pilosa inhibited T-cell proliferation decreasedTh1 cells and cytokines and increased Th2 cells andcytokines leading to prevention of T1D in nonobesediabetic (NOD) mice [49] Based on a bioactivity-directedisolation strategy 3 polyynes 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne (53) also known ascytopiloyne 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D- Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne (50) were identifiedfrom B pilosa [49 53] The IC

50value of the butanol fraction

was 200120583gmL This inhibition was reported to be partiallyattributed to cytotoxicity because the butanol fraction at

38 Evidence-Based Complementary and Alternative Medicine

Table 10 Other compounds isolated from B pilosa [30]

SN IUPAC names Common names Structure Plant part(country) References

19637-Dihydro-137-

trimethyl-1H-purine-26-dione

CaffeineN

NN

NO

O

Aerial(Uganda) [68]

197

1-((2R4S5R)-Tetrahydro-4-hydroxy-5-(hydroxymethyl)furan-2-yl)-5-

methylpyrimidine-24(1H3H)-dione

ThymidineHO

HO

ON

O

NHO Not found

(China) [37]

198 1-(2-Thienyl)-ethanone 2-Acetyl-thiophene

S

O

Roots(Germany) [41]

199

(2R3S4S5S)-2-(Heptan-2-yloxy)-tetrahydro-6-

((tetrahydro-34-dihydroxy-5-

(hydroxymethyl)furan-2-yloxy)methyl)-

2H-pyran-345-triol

Heptanyl2-O-120573-xylofuranosyl-

(1rarr 6)-120573-glucopyranoside

O

OOH

OH

HO

OHO

H

HO

H

H

HOHH

O

Whole(Taiwan) [70]

200

2-[(3R7R11R)-3-Hydroxy-371115-

tetramethylhexadecyl]-356-trimethyl-

25-cyclohexadiene-14-dione

120572-Tocopherylquinone

OO

HO

Whole(Taiwan) [32]

2017-O-(410158401015840610158401015840-Diacetyl)-

120573-D-glucopyranoside

Not found Leaves(China) [60]

180 120583gmL could cause 50 death of Th1 cells Moreoverthis study suggested that the butanol fraction may preventdiabetes in NOD mice in vivo via downregulation of Th1cells or upregulation Th2 cells that have effects which areantagonistic of those of Th1 cells [49] This was proven byintraperitoneal injection of the butanol fraction at a doseof 3mgkg BW 3 times a week to NOD mice from 4 to 27weeks This dosage resulted in lower incidence of diabetes(33) At a dose of 10mgkg the butanol fraction of Bpilosa totally eliminated (0) the initiation of the diseaseTo further support this result assessment of IgG2a and IgEproduction was performed in the serum of NOD mice Asin vivo results obtained from intracellular cytokine stainingexperiments were not very conclusive levels of IgG2a and IgE

were measured since Th1 cytokine IFN120574 and Th2 cytokineIL-4 favor the production of IgG2a and IgE respectively Asexpected high levels of IgE and some decline in the levels ofIgG2a were observed in the serum Profiling of the butanolextract revealed five compounds 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne(50) 45-Di-O-caffeoyl- quinic acid 35-Di-O-caffeoylquinicacid and 34-Di-O-caffeoylquinic acid Only the first twocompounds showed similar effects on the prevention ofdiabetes in NOD mice as the B pilosa butanol fractionMoreover compound 50 showed greater activity thancompound 69 in terms of enhancement (by 34 comparedto 8) of differentiation of Th0 to Th2 at 15 120583gmL (both

Evidence-Based Complementary and Alternative Medicine 39

Table 11 Chemical constituents of B pilosa and their biological activities

SN Name Classification Molecularformula Biological activities

109 Centaureidin [91] Flavonoid C18H16O8Anti-listerial [69 91]

Cytotoxic [10]

110 Centaurein [91] Flavonoid C24H26O13

Anti-listerial [69 91]Cytotoxic [10]Anti-viral [105]

103 Luteolin [106] Flavonoid C15H10O6

Anti-viral [107 108]Cytotoxic [77]

Anti-inflammatory [109]Anti-allergic [109]

82 Butein [110] Flavonoid C15H12O5Anti-leishmanial [111]

Cytotoxic [78]48 12-Dihydroxytrideca-57911-tetrayne [112] Polyyne C13H12O2 Anti-angiogeneic [112]46 13-Dihyroxy-6(E)-tetradecene-81012-triyne [112] Polyyne C14H16O2 Anti-angiogeneic [112]

45 12-Dihyroxy-5(E)-tridecene-7911-triyne [113] Polyyne C13H14O2Anti-angiogeneic [113]Anti-proliferative [113]

64 1-Phenylhepta-135-triyne [114] Polyyne C13H8

Anti-microbial [115]Anti-malarial [3]Cytotoxic [3]Antifungal [15]

27 Linoleic acid [79] Fatty acid C18H32O2Anti-viral (100)Cytotoxic [17]

161 Ethyl caffeate [84] Phenylpropanoid C11H12O4 Anti-inflammatory [84]

54 2-O-120573-Glucosyltrideca-11(E)-en-3579-tetrayn-12-diol [26] Polyyne C19H20O7Immunosuppressive andAnti-inflammatory [26]

53 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne [50] Polyyne C19H22O7Anti-diabetic [116]

Anti-inflammatory [50]

69 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne [28] Polyyne C20H26O7Anti-diabetic [28]

Anti-inflammatory [94]

50 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne [28] Polyyne C19H24O7

Anti-diabetic [28]Anti-inflammatory [94]

Anti-malarial andantibacterial [47]

129 Quercetin 3-O-120573-D-galactopyranoside [18] Flavonoid C21H20O12 Anti-inflammatory [117]

170 35-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

171 45-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

169 34-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

126 Quercetin 331015840-dimethyl ether7-O-120572-L-rhamnopyranosyl-(1rarr 6)-120573-D-glucopyranoside [118] Flavonoid C21H20O11 Anti-malarial [119]

125 Quercetin 331015840-dimethyl ether-7-O-120573-D-glucopyranoside [118] Flavonoid C21H20O12 Anti-malarial [119]70 1-Phenyl-13-diyn-5-en-7-ol-acetate [27] Polyyne C15H12O2 Anti-malarial [27]199 Heptanyl 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside [70] Miscellaneous C18H44O10 Antioxidant [70]124 3-O-Rabinobioside [70] Saccharide C27H38O15 Antioxidant [70]130 Quercetin 3-O-rutinoside [70] Flavonoid C27H38O15 Antioxidant [70]167 Chlorogenic acid [70] Phenolic C16H26O9 Antioxidant [70]119 Jacein [70] Flavonoid C24H26O13 Antioxidant [70]

49 (R)-12-dihydroxytrideca-357911-pentayne [47] Polyyne C13H8O2Anti-malarial andAntibacterial [47]

40 Evidence-Based Complementary and Alternative Medicine

Table 12 Structure and activity relationship studies of ethyl caffeateusing in vitro NF-120581BDNA binding assays [84]

Compound Concentration(120583M)

NF-120581BDNA binding

Ethyl caffeate 50 100inhibition

Ethyl 34-dihydroxyhydrocinnamate 100 100inhibition

Catechol 400 100inhibition

Ethyl cinnamate 400 Noinhibition

Table 13 Apoptosis in cocultures of T cells and pancreatic 120573 cells[53]

Cellmedium apoptosis andnecrosis

CD4+ T cellscontrol medium lt4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 2

CD4+ T cellswith cytopiloyne-treated 120573 cells 18CD4+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 7

CD8+ T cellscontrol medium 4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 4

CD8+ T cellswith cytopiloyne-treated 120573 cells 4CD8+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 4

Table 14 Radical scavenging activities of B pilosa extracts [70]

Extractscontrol DPPH assayIC50 (120583gmL)

NBThypoxanthinesuperoxide assayIC50 (120583gmL)

Quercetin 198 15Ascorbic acid 634 Not determined120572-tocopherol 897 Not determinedEthyl acetateextract 1383 597

Butanol extract 1669 114Water extract gt100 gt100

compounds) and inhibition (by 40 compared to 10) ofdifferentiation toTh1 at the same concentration [49]

Among the three polyynes found in B pilosa cytopiloyne(53) had the most potent anti-T1D activity [53] To test the invivo effect of cytopiloyne NODmice received intraperitonealor intramuscular injection of cytopiloyne at 25120583gkg BW3 times per week Twelve-week-old NOD mice started todevelop T1D and 70 of NOD mice aged 23 weeks andover developed T1D Remarkably 12- to 30-week-old NODmice treated with cytopiloyne showed normal levels of bloodglucose (lt200mgdL) and insulin (1-2 ngmL) Consistent

Table 15 Radical scavenging activity of secondarymetabolites fromB pilosa [70]

Metabolite (Table 11)control DPPH assay IC50 (120583gmL)199 Not determined124 53130 68167 105169 33171 38119 Not determined110 Not determinedQuercetin 256Caffeic acid 890

Table 16 Antioxidant activity of the essential oils and water extractsfrom B pilosa [92]

Extract IC50 (120583gmL)Leaf essential oils 47Flower essential oils 50Leaf extract 61Flower extract 172

with T1D incidence cytopiloyne delayed and reduced theinvasion of CD4+ T cells into the pancreatic islets [53]

In vitro study showed that cytopiloyne (53) inhibited thedifferentiation of naıveTh (Th0) cells (ie CD4+ T cells) intoTh1 cells and promoted differentiation of Th0 cells into Th2cells [50] The in vitro data are consistent with the in vivoresults indicating that cytopiloyne reducedTh1differentiationand increased Th2 differentiation as shown by intracellularcytokine staining and FACS analysis [53] In line with theskewing of Th differentiation the level of serum IFN-120574 andIgG2c decreased while that of serum IL-4 and serum IgEincreased compared to the negative controls (PBS-treatedmice) Cytopiloyne also enhanced the expression of GATA-3 a master gene for Th2 cell differentiation but not theexpression of T-bet a master gene forTh1 cell differentiationfurther supporting its role in skewingTh differentiation [53]

Also importantly cytopiloyne partially depleted CD4+rather than CD8+ T cells in NOD mice [53] As shown inTable 13 coculture assays showed that the depletion of CD4+T cells was mediated through the induction of Fas ligandexpression on pancreatic islet cells by cytopiloyne leading toapoptosis of infiltrating CD4+ T cells in the pancreas via theFas and Fas ligand pathways However cytopiloyne did notinduce the expression of TNF-120572 in pancreatic islet cells andthus had no effect on CD8+ T cells [53]

In addition Chang and colleagues showed that cytopi-loyne dose-dependently inhibited T-cell proliferation stimu-lated by IL-2 plus Con A or anti-CD3 antibody using [3H]thymidine incorporation assay [53]

Overall the mechanism of action of cytopiloyne andprobably its derivatives in T1D includes inhibition of T-cellproliferation skewing of Th cell differentiation and partialdepletion of Th cells Due to the anti-diabetic mechanisms

Evidence-Based Complementary and Alternative Medicine 41

Table 17 Antibacterial activity of essential oils and flower extracts from B pilosa [92]

Strain Mean zone of inhibition (mm)Leaf essential oil Flower essential oil Leaf extract Flower extract

Micrococcus flavus 127 plusmn 03 87 plusmn 03 102 plusmn 02 108 plusmn 03Bacillus subtilis 173 plusmn 19 117 plusmn 02 109 plusmn 02 103 plusmn 02Bacillus cereus 190 plusmn 14 112 plusmn 03 118 plusmn 04 185 plusmn 10Bacillus pumilus 123 plusmn 07 108 plusmn 02 105 plusmn 04 77 plusmn 02Escherichia coli 137 plusmn 04 203 plusmn 07 102 plusmn 11 140 plusmn 13Pseudomonas ovalis 125 plusmn 08 137 plusmn 15 102 plusmn 06 125 plusmn 06

Table 18 Antibacterial activity of root extracts from B pilosa [93]

Strain MIC50 (mgmL)Methanol extract Acetone extract

Bacillus cereus 10 mdashEscherichia coli 5 5Klebsilla pneumonia 5 10Micrococcus kristinae 10 mdashPseudomonas aeruginosa 10 10Staphylococcus aureus 5 10Sraphylococcus epidermidis 5 5Serratia marcescens 10 mdashShigelea flexneri 10 mdashStreptococcus faecalis 10 mdash

Table 19 Antibacterial activity of B pilosa of compound 29 [47]

Strain MIC50 (120583gmL)Escherichia coli NIHJ 1Escherichia coli ATCC25922 1Klebsiella pneumoniae ATCC700603 128Serratia marcescens ATCC13880 16Pseudomonas aeruginosa ATCC27853 8Staphylococcus aureus FDA209P 05Staphylococcus aureus ATCC29213 025Staphylococcus aureus N315 (MRSA) 05Enterococcus faecalis ATCC29212 2Enterococcus faecalis NCTC12201 (VRE) 1Bacillus subtilis ATCC6633 05Candida albicans ATCC10231 025

of action it was hypothesized that cytopiloyne protectsNOD mice from diabetes by a generalized suppression ofadaptive immunity To evaluate this hypothesis ovalbumin(Ova) was used as a T-cell dependent antigen to primeNOD mice which had already received cytopiloyne orPBS vehicle Ova priming boosted similar anti-Ova titersin cytopiloyne-treated mice and PBS-treated mice but adifference in immunoglobulin isotype was observed in thetwo groups Thus it was concluded that cytopiloyne is animmunomodulatory compound rather than an immunosup-pressive compound [50 53]

T2D is a chronic metabolic disease with serious com-plications resulting from defects in either insulin secretion

insulin action or both [124] A study by Ubillas et alshowed that the aqueous ethanol extract of the aerial partof B pilosa at 1 gkg body weight (BW) lowered bloodglucose in dbdb mice a T2D mouse model [51] Basedon a bioactivity-guided identification compounds 69 and50 were identified Further the mixture of the compounds(69 50) in a 2 3 ratio significantly decreased blood glucoseconcentration and reduced food intake on the second dayof treatment when administered at doses of 250mgkg twicea day to C5BLKs-dbdb mice When tested at 500mgkga more substantial drop in blood glucose level as wellas the stronger anorexic effect (food intake reduced from58 gmouseday to 25 gmouseday) was observed [51] Inthis study it was suggested that the blood glucose lower-ing effect of B pilosa was caused in part by the hungersuppressing effect of its polyynes [51] However the hungersuppressing effect of the ethanol extract of B pilosa wasnot found in the studies described below In another study[24] water extracts of B pilosa (BPWE) were used indiabetic dbdb mice aged 6ndash8 weeks with postprandialblood glucose levels of 350 to 400mgdL Like oral anti-diabetic glimepiride which stimulates insulin release onesingle dose of BPWE reduced blood glucose levels from374 to 144mgdL The antihyperglycemic effect of BPWEwas inversely correlated to an increase in serum insulinlevels suggesting that BPWE acts to lower blood glucose viaincreased insulin production However BPWE had differentinsulin secretion kinetics to glimepiride [24] One flaw incurrent anti-diabetics is their decreasing efficacy over timeThe authors investigated the long term anti-diabetic effectof BPWE in dbdb mice BPWE reduced blood glucoseincreased blood insulin improved glucose tolerance andreduced the percentage of glycosylated hemoglobin (HbA1c)Both long-term and one-time experiments strongly supportthe anti-diabetic action of BPWE [24] In sharp contrast toglimepiride BPWE protected against islet atrophy in mousepancreas The investigators further evaluated anti-diabeticproperties of 3 B pilosa varieties B pilosa L var radiate(BPR) Bpilosa L var pilosa (BPP) and B pilosa L varminor(BPM) in dbdb mice [8] One single oral dose (10 50 and250mgkg body weight) of BPR BPP or BPM crude extractsdecreased postprandial blood glucose levels in dbdb micefor up to four hours and the reduction of glucose levels inthe blood appeared to be dose-dependent Comparing thethree variants BPR extract resulted in a higher reduction inblood glucose levels when administered at the same dose as

42 Evidence-Based Complementary and Alternative Medicine

Table 20 Antifungal activity of B pilosa [92]

Partextract Concentration (ppm) Strain InhibitionCortiicum rolfsii Fusarium solani Fusarium oxysporum

Leaves

Essentialoils

100 857 plusmn 09 682 plusmn 0 745 plusmn 17250 960 plusmn 08 779 plusmn 18 879 plusmn 04

AqueousExtracts

100 446 plusmn 17 605 plusmn 21 716 plusmn 07250 942 plusmn 03 689 plusmn 07 824 plusmn 19

Flowers

Essentialoils

100 604 plusmn 09 892 plusmn 04 869 plusmn 05250 894 plusmn 12 980 plusmn 03 949 plusmn 06

AqueousExtracts

100 331 plusmn 11 714 plusmn 07 573 plusmn 22250 661 plusmn 14 912 plusmn 0 900 plusmn 07

Table 21 Antifungal activity of B pilosa root extracts [93]

Strain LC50 (mgmL)Acetoneextracts

Methanolextracts

Waterextracts

Aspergillus niger 014 006 007Aspergillus flavus 1091 658 0Penicillium notatum 005 005 005

the other two varieties In terms of serum insulin levels a doseof 50mgkg of each extract was used and the BPR extracttogether with the three polyynes significantly increased theserum insulin level in dbdb mice Long-term experiments(28-day treatment) were then conducted using diabetic micewith postprandial glucose levels from 370 to 420mgdL andglimepiride was used as positive controlThe range of dosagesapplied was from 10mgkg BW to 250mgkg BW Resultsshowed that the positive control as well as the crude extractsof the three varieties lowered the blood glucose levels indbdb mice However only BPR extract containing a higherpercentage of cytopiloyne (53) reduced blood glucose levelsand augmented blood insulin levelsmore than BPP andBPMThe percentage of glycosylated hemoglobin A1c (HbA1c) wasalso measured and found to be 79 plusmn 05 in mice aged 10ndash12 weeks and 66 plusmn 02 61 plusmn 03 and 62 plusmn 03 inthe blood of age-matchedmice following treatment with BPRcrude extract (50mgkg) glimepiride (1mgkg) and com-pound 53 (05mgkg) respectively [8] Among the polyynesfound in B pilosa cytopiloyne was the most effective againstT2D Hence cytopiloyne was used for further study on anti-diabetic action and mechanism [125] The data confirmedthat cytopiloyne reduced postprandial blood glucose lev-els increased blood insulin improved glucose tolerancesuppressed HbA1c level and protected pancreatic islets indbdb mice Nevertheless cytopiloyne failed to decreaseblood glucose in streptoztocin (STZ)-treated mice whose bcells were already destroyed Additionally cytopiloyne dose-dependently increased insulin secretion and expression in bcells as well as calcium influx diacylglycerol and activationof protein kinase C120572 Collectively the mechanistic studiessuggest that cytopiloyne treats T2D via regulation of insulin

production involving the calciumDAGPKC120572 cascade in bcells

The studies detailed above point to the conclusion thatcytopiloyne and related polyynes (compounds 69 and 49)are anti-diabetics in animal models The data uncover a newbiological action of polyynes It should be noted that likeall anti-diabetic drugs cytopiloyne failed to prevent or curediabetes completely but reduced diabetic complications [125]Intriguingly 34 polyynes have been found in B pilosa so farIt remains to be seen whether all the polyynes present in thisplant have anti-diabetic activities

34 Antioxidant Activity Free radicals can damage cellularcomponents via a series of chemical reactions [89] leadingto development and progression of cardiovascular diseasecancer neurodegenerative diseases and ageing [70] Freeradicals nitric oxide (NO) and superoxide anions can beproduced in macrophages to kill microbes However anexcessive generation of the free radicals under pathologicalconditions is associated with a wide range of illnessesPlants are known to be rich in antioxidant phytochemicalsChiang and colleagues evaluated the free radical scaveng-ing activity of crude extract fractions and compounds ofB pilosa using 11-diphenyl-2-picrylhydrazyl (DPPH) andhypoxanthinexanthine oxidase assays [70] Using DPPHand hypoxanthinexanthine oxidase assays they found thatthe B pilosa crude extract and the ethyl acetate butanoland water fractions had free radical scavenging activityNine compounds Heptyl- 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside (199) 3-O-Rabinobioside (124) Quercetin3-O-rutinoside (130) Chlorogenic acid (167) 34-Di-O-caffeoylquinic acid (169) 35-Di-O-caffeoylquinic acid (170)45-Di-O-caffeoylquinic acid (171) Jacein (119) and Cen-taurein (110) had DPPH radical scavenging activity [70]The IC

50values of the B pilosa crude extractfractions and

compounds are summarized in Tables 14 and 15 respectivelyMeasurement of free radical scavenging activities is one

way of assessing the antioxidant activities of B pilosa andits fractions and compounds It is interesting that the ethylacetate and butanol fractions are more active than the waterfraction and B pilosa crude extract [70] Of the secondarymetabolites only phenolic compounds 124 130 167 169 and171 showed significant DPPH-radical scavenging activities

Evidence-Based Complementary and Alternative Medicine 43

Further analysis of the structure-activity relationship of thecompounds suggested that substitution of the C3 hydroxylgroup with glycosides increased the activity approximately2-fold (for example in compounds 124 and 130) relative toquercetin (-OH in its C3) [70] Further modification of thestructures of the active compounds needs to be performed totest the effects of various substituents on activity The reasonwhy most of the antioxidant compounds contain phenolmoieties in their structure could be that the reduction-oxidation (redox) properties of phenols allow them to actas reducing agents singlet oxygen quenchers and hydrogendonors

A complementary study by Muchuweti and colleaguesdetermined phenolic content antioxidant activity and thephenolic profile of B pilosa methanol extract [89] Theyestimated that the phenolic content of the methanol extractof B pilosawas 11028plusmn22mgg [89] Vanillin hydroxyben-zaldehyde caffeic acid coumaric acid and ferulic acid werefound in this extractThe B pilosa extract also showed DPPHradical scavenging activity Furthermore the antioxidantactivity of the flavonoids found in B pilosa was correlatedwith its hepatoprotective effects through their inhibitionof NF-120581B activation which may lessen the oxidative stresscaused by the production of free radicals during liver injury[60]This activity might also be due to the anti-inflammatoryeffects of the aqueous extracts of B pilosa aerial parts on theinhibition of COX-2 and PGE

2production [88]

Essential oils from B pilosa flowers and leaves are alsoreported to possess antioxidant activity With the aim ofreplacing chemically synthesized additives Deba and col-leagues [92] worked on the antioxidant antibacterial andantifungal activities of essential oils and water extracts of Bpilosarsquos leaves and flowers Table 16 summarizes the resultsobtained from DPPH free radical scavenging assay

It can be inferred fromTable 16 that essential oils from theleaves possessed the highest activity It is reported elsewherethat monoterpenes present in essential oils such that of Bpilosa have protective effects and antioxidant properties [92]Beta-carotene bleaching method was also performed Leavesessential oils and aqueous extracts of the leaves and flowersshowed higher activity than the flower essential oils This isdue to the volatility of the flower essential oils The activityexhibited by the aqueous extracts was accounted to thepresence of phenolic compounds that are reported to donatea hydrogen atom to free radicals such that the propagation ofthe chain reaction during lipid oxidation is terminated [92]Overall essential oils and phenolics present in B pilosa canbe thought of as major antioxidant compounds

35 Immunomodulatory Activity B pilosa is thought to bean immunomodulatory plant and is reported to be effectivein the treatment of immune disorders such as allergy [75]arthritis [75] and T1D [48 50 53]

As pointed out in the discussion of its anti-diabeticactivities (Section 33) a combination of phytochemicalsmethod and T-cell activation assays was used to studyimmunomodulatory properties of B pilosa IFN-120574 is a keycytokine released by T and NK cells that mediates immune

cells and sustains immunity against pathogens Defects inIFN-120574 expression regulation and activation result in vulner-ability to diseases caused by bacteria and viruses [69] Anelegant study performed byChang and colleagues using IFN-120574 promoter-driven luciferase reporter construct in JurkatT cells showed that hot water crude extracts of B pilosaincreased IFN-120574 promoter activity two-fold [69] Out of thesubfractions of this extract the butanol fraction but not thewater or ethyl acetate fractions increased IFN-120574 promoteractivity six-fold Centaurein (110) and centaureidin (109)were identified from the butanol fraction and were stated tocause a four-fold increase in IFN-120574 promoter activity withEC50

values of 75 120583gmL and 09120583gmL respectively Themechanism of action of centaurein was determined usingtranscription factors such as AP-1 NFAT and NF120581B whichare reported to bind to IFN-120574 promoter and regulate IFN-120574transcription Unlike with the activity of the positive controlPHA centaurein caused a four-fold increase in NFAT a3-fold increase in NF120581B and had little if any effect onAP-1 enhancer activities (23-fold three-fold and ten-foldincreases respectively were seenwith PHA) [69]The authorsconcluded that centaureinmodulates IFN-120574 expression by theNFAT and NF120581B pathways The article only determined themechanism of action of centaurein Its aglycone centaureidinmay act through the samemechanism though this conclusionneeds to be further verified

B pilosa extract and its compounds are reported to inhibitdifferentiation of naıve CD4+ helper T (Th0) cells into Th1cells [49] Using theThcell differentiation assay as a screeningplatform 3 polyynes 2-120573-D-glucopyranosyloxy-1 -hydroxytrideca-57911-tetrayne (53) 3-120573-D-glucopyranosyl-1-hydroxy-6(E)- tetradecene-81012-triyne (69) and 2-120573-D-glucopyranosyloxy-1-hydroxy-5(E)- tridecene-7911-triyne(49) were discovered from B pilosa [49 53] The data showsthat cytopiloyne and other two polyynes suppressed thedifferentiation of type 1 helper T (Th1) cells and productionof Th1 cytokines and promoted that of type 2 helper T (Th2)cells and production of Th2 cytokines thus explaining theimmunomodulatory and anti-inflammatory effects of Bpilosa and its polyynes

Chang and colleagues were the first to report the effect ofthe butanol extract of B pilosa on the autoimmune diabetesand airway inflammation inmice [48] Imbalance in the levelsofTh1 andTh2 and of various cytokines leads to autoimmunediseases T1D and other autoimmune diseases (rheumatoidarthritis Crohnrsquos disease among others) are exacerbated byan increase Th1 levels (specifically CD4+ Th1 cells) whileTh2 cells antagonize this effect [19] Moreover Th2 cellsmediate asthma in ovalbumin-induced hypersensitivity inBALBc mice [48] In their work [48] Chang and colleaguesshowed that 10mgkg butanol extracts with 15 (ww)compound 69 and 11 (ww) compound 49 (Table 11)ameliorated the development of Th1-mediated diabetes inNOD mice through inhibition of 120573 cell death and leukocyteinfiltration At the same dosage the butanol extracts alsoexacerbated ovalbumin-induced pulmonary inflammationin BALBc mice with an increase in the infiltration ofeosinophils and mast cells into the airway of the mice [48]Despite the different outcomes both mouse models proved

44 Evidence-Based Complementary and Alternative Medicine

the concept that control over the Th1Th2 shift is associatedwith autoimmune diseases and that the B pilosa butanolextract can shift the differentiation of Th0 cells to Th2 cells[48 49]

An extended study presented by Chiang and colleaguesshowed that compound 53 modulates T-cell functions [50]Using CD4+ T cells from BALBc mice they demonstratedthat cytopiloyne decreased levels of IFN-120574 producing cells(Th1) by 122 (from 72 to 598) Since Th1 and Th2 celldifferentiation is antagonistic it was expected that compound53 increased the percentage of mouse IL-4 producing cells(Th2) by 72 (from 237 to 309) Subsequent assess-ment of effect of compound 53 on the modulation of thetranscription of IL-4 and IFN-120574 showed that cytopiloyne asexpected decreased the splenocyte levels of IFN-120574 mRNAand increasing that of IL-4 in a dose-dependent manner Inthe range of 01 to 3 120583gmL of compound 53 the effects werenot attributed to its cytotoxicity Consequently using 3120583gmLcytopiloyne for 72 and 96 hours the protein concentration ofIFN-120574 decreased to 186 and 444 respectively Under thesame conditions cytopiloyne increased IL-4 concentrationsto 1985 and 2470 (for 72 and 96 hours resp) Thismodulation of T-cell differentiation exhibited by compound53 was used to explain its anti-diabetic activity [50]

The anti-diabetic role of cytopiloyne was extensivelydiscussed above (Section 33 anti-diabetic activity) Themolecular basis of the regulation of cytokine expression bycytopiloyne has been described Cytopiloyne directly elevatedthe expression level of IL-4 via GATA-3 upregulation inT cells [53] However reduction of IFN-120574 expression in Tcells seemed to come from the indirect opposing effect IL-4cytokine because the expression level of T-bet was unaltered[53] In this way cytopiloyne skewedTh1 polarization into theTh2 state conferring protection against T1D in NOD miceAside frompolarization ofThcell differentiation cytopiloynealso activated the expression of Fas ligand in pancreaticb cells this increase leading to the partial depletion of Tcells and reduction of immune response in local areas suchas the pancreas Of note cytopiloyne also inhibited T-cellproliferation and activation By targeting T cells from threeimmunomodulatory actions cytopiloyne protects againstT1D and probably other Th1-mediated autoimmune diseases[19]

The phytochemical constituents of B pilosa exert theirfunctions on different immune cells to modulate immuneresponse It is possible that some of the compounds mayhave agonistic or antagonistic effects on immune responseImmune function of B pilosa may depend on its compo-sition and amount of compounds which could explain theapparently conflicting report of B pilosa butanol extractaggravating allergy in mice [48] while cellulosine-treatedextract ameliorated allergy [48 75] IFN-120574 promoter reporterassays and T-cell differentiation assays were used to isolate2 flavonoids [69] and 3 polyynes [49 50] as immunomod-ulatory compounds from the butanol fraction of B pilosaInterestingly the flavonoids promote IFN-120574 expression inNK and T cells In marked contrast the polyynes promoteIL-4 expression and indirectly inhibit IFN-120574 expression indifferentiating T cells Sensitivity appears to be the key to

identifying structure- and bioactivity-related phytochemicalsfrom medicinal plants

36 Antimalarial Activity The use of chemical drugs againstpathogens has resulted in drug-resistant mutants Examplesof drug resistance can be found in the species of thePlasmodium that cause malaria It is important to search fornew compounds to combat Plasmodium parasites [47] Astudy of the anti-malarial activity of the leaf extracts of Bpilosa using a combination of phytochemistry and bioassaysshowed that compound 49 (Table 11) showed activity againsta malaria parasite (P falciparum NF54 strain) with an IC

50

value of 60 120583gmL [77] In addition compound 49 isolatedfrom the aerial parts of B pilosa inhibited growth of the Pfalciparum FCR-3 strain with an IC

50value of 035 120583gmL

This compound was tested for its in vivo effect in miceinfected with P berghei NK-65 strain Results showed thatcompound 49 decreased the average parasitemia in the redblood cells by 207 (from 328 of that of the control to121) after an intravenous injection of 08mgkg BWday forfour days [47] Further studies addressing the anti-malarialmechanism underlying both polyynes and clinical studies areneeded

37 Antibacterial Activity Emergence of multiple antibiotic-resistantmicrobes is becoming a global threat to public healthand a challenge to disease treatment For instance penicillinis commonly used to combat a food-borne intracellularbacteriumListeria however penicillin-resistant bacteria havebeen discovered recently [69] Chang and coworkers iso-lated centaurein (110) and centaureidin (109) from B pilosaextract [69 70] Centaurein enhances expression of IFN-120574a key cytokine for macrophage activation and consequentlyenhances bactericidal activity in macrophages [69 70] Inagreement with observed in vitro effects centaurein wasreported to prevent and treat Listeria infection in C57BL6Jmice [91] Mechanistic studies confirmed that centaureinexerted antilisterial action via IFN-120574 expression in wild-typemice but not IFN-120574 knockout mice [91] Further in vitrostudies on centaurein showed that this compound increasedIFN-120574 expression by 13 (from 17 to 20) 20 (from 21to 41) and 11 (from 6 to 17) in CD4+ T cells CD8+Tcells and NK cells respectively That is to say there wasan increase in IFN-120574 producing immune cells As expectedcentaurein also enhanced the expression level of T-bet a keynuclear factor for IFN-120574 expression Consistently centaureinaugmented the serum IFN-120574 levels inC57BL6Jmice and thisaugmentation peaked 24 hours after compound injectionThe quantity of mouse serum IFN-120574 was sufficient to activatemacrophages in vitro and eradicated GFP-producing Listeriainside macrophages However the entry of Listeria intomacrophages was not affected by centaurein-treated mousesera Centaurein treatment at 20120583g per mouse rescued 30of the mice infected with a lethal dose of Listeria (2 times106 CFU) It is noteworthy that in the presence of ampicillin(5 120583gmouse) centaurein (20 120583gmouse) rescued 70 of themice suggesting an additive effect between ampicillin andcentaurein [91] Despite lower abundance centaureidin was

Evidence-Based Complementary and Alternative Medicine 45

30 times more active than centaurein in terms of IFN-120574production [91]

Aside from the indirect antibacterial action mentionedabove extract andor compounds of B pilosa also showeddirect bacteriostatic andor bactericidal action One studyreported that essential oils and leafflower extracts of Bpilosa could suppress the growth of gram positive and gramnegative bacteria as evidenced by zone of inhibition assaysIn this study antibacterial activity of the essential oils fromthe leaves and flowers of B pilosa was determined in anattempt to identify natural products as food preservatives forprevention of microbial multiplication and food oxidationThe essential oils and extracts of B pilosa leaves and flow-ers showed moderate but different extents of antibacterialactivity (Table 17) In general essential oils had higherantibacterial activity than crude extracts One explanation forthis could be that monoterpenes in the essential oils destroycellular integrity and subsequently inhibit the respirationand ion transport processes The presence of antibacterial120573-caryophyllene could be another explanation as reportedelsewhere [92] Another study reported that the methanoland acetone extract of B pilosa roots displayed antibacterialactivities against the bacteria listed in Table 18 [93] andmethanol extracts from the roots seemed to be the mosteffective

Another study indicated that the polyyne (R)-12-dihydroxytrideca-357911-pentayne (49) from this plantalso suppressed bacterial growth as shown by the minimuminhibitory concentration required to inhibit 50 bacterialgrowth (MIC

50) in Table 19 This compound was highly

effective against several Gram positive and Gram negativebacteria including the drug-resistant bacteria Staphylococcusaureus N315 (MRSA) and Enterococcus faecalis NCTC12201(VRE) [47] Strikingly compound 49 had a similar MIC

50

value to antibiotics (ampicillin tetracycline norfloxacin andamphotericin B) in most of the bacteria tested

Antibacterial activity of B pilosa extracts and compo-nents expressed as MIC

50and the mean zone of inhibition

is tabulated in Tables 17 18 and 19The zone of inhibition forAmpicilline (positive control) ranges from 153 plusmn 03mm to443 plusmn 02mm [92]

38 Antifungal Activity B pilosa has traditionally been usedto treat microbial infection Recently different parts of Bpilosa have been tested for antifungal activities Deba andcolleagues first evaluated the antifungal effect of the hotwater extracts of the B pilosa roots stems and leaves againstCorticium rolfsii Fusarium solani and Fusarium oxysporumThey discovered that C rolfsii was most suppressed bytreatment with B pilosa as its growth was reduced at almostall the tested doses followed by F oxysporum and F solani[99] However the fungicidal activities of the stems androots were greater than the leaves [99] Moreover the samegroup assessed the antifungal activity of the essential oilsand aqueous extracts from B pilosa flowers and leaves [92]They showed that the extracts and oils had antifungal activityagainst C rolfsii F solani and F oxysporum Essential oils

appeared to have better fungicidal activity thanwater extractsas summarized in Table 20

Another study by Ashafa and colleagues showed thatacetone methanol and water extracts of the B pilosa rootsshowed antifungal activities against Aspergillus niger Aflavus and Penicillium notatum using the agar dilutionmethod The results are tabulated in Table 21 [93] Negativecontrols showed 0 growth inhibitionThe methanol extractof the B pilosa roots at 10mgmL was also effective againstCandida albicans [93]Of note B pilosa obtained fromPapuaNew Guinea had no activity against A niger and C albicans[126] but the SouthAfrican (EasternCape) ecotype exhibitedmoderate activity against C albicans [93] This discrepancymay depend on extraction solvents extraction procedureassay techniques different plant parts and abundance ofactive compounds

B pilosa produces a variety of secondary metabolitessuch as flavonoids phenylacetylenes alkaloids sterols ter-penoids and tannis [92 93] However none of them havebeen confirmed as active compounds against fungi Furtherinvestigation of active compounds from B pilosa is necessaryto further understand the antifungal efficacy of this plant

39 Hypotensive and Vasodilatory Activities In early studiesDimo and colleagues used three rat models normotensiveWistar rats (NTR) salt-loading hypertensive rats (SLHR)and spontaneous hypertensive rats (SHR) to investigate thehypotensive effect of the methanol crude extract of B pilosaleaves [28 127] The extract lowered systolic blood pressurein hypertensive rats (SLHR and SHR) to a greater degreethan NTR [28 127] In addition a decrease in urinarysodium ions and an increase in urinary potassium ions wereobserved after treatment with the methanol extract of Bpilosa leaves although neither differences were statisticallysignificant [28 127] Taking the data together the studyproposed that B pilosa leaf extract reduced blood pressurevia vasodilation [28 127] The same group continued totest the antihypertensive effect of aqueous and methylenechloride extracts of B pilosa leaves in a hypertensive ratmodel [28 127] To establish a fructose-induced hypertensionmodel male Wistar rats were given 10 fructose solutionto drink ad libitum for three weeks In addition to freeaccess to 10 fructose the rats were treated with the aqueous(150mgkg) or methyl chloride (350mgkg) extracts of Bpilosa for additional three weeks [28 127] Both extracts ofB pilosa leaves had a hypotensive effect on rats Howeverneither extracts reversed the elevation of serum insulin infructose-fed rats Therefore B pilosa lowered blood pressureirrespective of insulin [28 127] To better understand thehypotensive mechanism the authors investigated the effectof a neutral extract of B pilosa (NBP) a mixture of methanolandmethylene chloride (1 1) extract after neutralization withNaOH and HCl on the heart and the blood pressure ofNTR and SHR [110] This study showed that an intravenousinjection of the NBP resulted in a biphasic reduction insystolic blood pressure In addition one intravenous dose ofthe extract at 10 20 and 30mgkg BW decreased systolicblood pressure in normal rats by 183 425 and 30

46 Evidence-Based Complementary and Alternative Medicine

respectively and the same doses reduced the blood pressurein hypertensive rats by 258 389 and 286 respectivelyOnly the highest dose (30mgkg) affected the force of thecontraction of the heart Atropine and propranolol were usedto interfere with the hypotensive action of the NBP Atropinereduced the initial phase of the hypotensive response in NBPand completely abolished the second phase of hypotensiveresponse in NBP In contrast propranolol increased thefirst hypotensive response but partially abolished the secondhypotensive response provoked by the NBP [110]This mech-anistic study suggested that B pilosa invokes the biphasichypotensive responses via targeting cardiac pump efficiencyduring the first phase and vasodilation at the second phase[110]

A further study was performed to investigate the relaxingeffect of a neutral extract of B pilosa (NBP) on rat aorta con-tractedwithKCl (60mM) andnorepinephrine (01mM) [94]Cumulative addition of NBP relaxed the rat aorta previouslycontracted by KCl in a dose-dependent manner The EC

50

value of theNBP for vasorelaxationwas 032mgmLThedataalso showed that the NBP reduced the contraction of aortapreviously contracted by KCl irrespective of the presence ofaortic endothelium [94]

Pretreatment with glibenclamide an ATP-dependent K+channel blocker did not considerably affect the relaxant effectof the NBP on KCl-induced contraction suggesting thatthe vasodilatory effect of B pilosa was not related to theopening of this ATP-dependent K+ channel [94] On theother hand in the presence of indomethacin or pyrilaminemaleate the relaxant response induced by the plant extractwas significantly inhibited at the lower concentrations Theplant extract was able to reduce the aorta resting tone inhibitthe KCl-induced contractions by 90 at 15mgmL and theCaCl2-induced contractions by 95 at 075mgmL These

results demonstrate that B pilosa can act as a vasodilatorprobably via acting as a calcium antagonist [94]

However no specific compound for the above activityhas been identified from B pilosa to date A bioactivity-guided identification approach may be adopted to identifythe active compounds in B pilosa that possess hypotensiveand vasodilatory effects and understand their mechanism ofaction

310 Wound Healing Activity B pilosa has been traditionallyused to treat tissue injury inCameroon Brazil andVenezuela[26] Hassan and colleagues investigated the wound healingpotential of B pilosa in Wistar rats [128] Mirroring thepositive control neomycin sulfate the ethanol extract of Bpilosa had faster wound closure than control rats 3 6 and 9days after topical application Histological examination alsorevealed better collagenation angiogenesis and organizationof wound tissue seven days after application Epithelializationand total healing time in B pilosa-treated rats were compara-ble to those of neomycin sulfate Together these data suggestthat B pilosa may be a viable alternative to neomycin lotionfor the treatment of wounds

In addition to studying the wound healing effect of Bpilosa on external ulcers Tan and colleagues also examined

the effect of methanol cyclohexane and methyl chlorideextracts of B pilosa on gastric ulcers in Wistar rats fedwith 1mL HClethanol gastric necrotizing solution (150mMHCl in 60 ethanol) and macroscopically visible lesionswere scored [26] Among the three extracts methylene chlo-ride extracts exhibited the highest activity showing 464inhibition of lesion formation at a dose of 500mgkg BWand complete inhibition at 750mgkg [26] The efficacy ofthe ethylene chloride extract was followed by that of themethanol extracts which had inhibition ranging from 304to 822 at concentrations of 500mgkg and 1000mgkgBW respectively [26] The cyclohexane extracts showed thelowest activity against gastric ulcers in rats with 13340 and 797 inhibition at 500 750 and 1000mgkg BWrespectively [26] To better understand the mode of actionof the methylene chloride extract of B pilosa rats werepretreated with indomethacin a COX-2 inhibitor involved inprostaglandin synthesis Pretreatment significantly reducedthe protection against HClethanol-induced ulcers to 313inhibition at 750mgkg BW suggesting a link between theantiulcerative activity of B pilosa and prostaglandin syn-thesis Unexpectedly the methylene chloride extract of Bpilosa showed little gastric mucosal protection against gastriclesions induced by 95 ethanol (1mL) [26] Absolute alcoholis known to cause mucosalsubmucosal tissue destruction viacellular necrosis and the release of tissue-derived mediators(histamine and leukotrieneC4)Thus these data imply thatBpilosa did not prevent the generation or the necrotic action ofthese mediators on the gastric microvasculature In additionpylorus ligation can increase gastric acid secretion withoutan alteration of mucosal histamine content The methylenechloride extract of B pilosa did not possess antisecretoryactivity On the contrary it was observed that increases in thedose of the extract led to elevated gastric juice acidity [26]Results with both absolute ethanol and pylorus ligation ratmodels suggested the possibility that the ineffectiveness of Bpilosa against gastric ulcers was due to lack of antihistaminicactivity in the plant In summary overall the data suggest thatB pilosa protects against HCLethanol-mediated ulcers viainhibition of prostaglandin biosynthesis

Previous phytochemical studies showed that a groupof flavonoids acyclichalcones are present in B pilosa[129 130] and the chalcones were proposed to have anti-ulcerative activity [131] Moreover nine hydroxychalcones 10were reported to possess gastric cytoprotective effects with24-dihydroxychalcone being the most active [132] Sincemethylene chloride extracts appear to be the most active Bpilosa extracts next the specific anti-ulcerative phytochemi-cals in the methylene chloride extracts of B pilosa and theirmodes of action needs to be probed

Despite the claims listed inTable 3 relatively few scientificstudies have been conducted in vitro and in vivo to address thetraditional ethnomedical uses of B pilosa Information aboutthe use of B pilosa as a botanical therapy recorded so far isfar from complete Studies conducted thus far only serve asa starting point for further investigation of B pilosa and theultimate efficacious use of the herb in clinical applications

Evidence-Based Complementary and Alternative Medicine 47

4 Toxicology

Despite its use as an ingredient in food for human con-sumption studies on systemic toxicity (eg acute subacutechronic and subchronic toxicities) of B pilosa in humansand animals are still inadequate and insufficient So faracute andor subchronic toxicities have been evaluated inrats and mice Oral acute and 28-day toxicities of waterextract of B pilosa leaves were evaluated in Wistar rats[133] An oral dose of water extract of B pilosa leaves at10 gkg BW showed no obvious mortality or changes inthe appearance in rats [134] The same extract at 08 gkgBWday once a day showed no obvious sub-chronic toxicityin rats over 28 days as measured by survival rate bodyweight and gross examination of organs [134] These dataare consistent with our data indicating that oral deliveryof the water extract of the B pilosa whole plant at 1 gkgBWday once a day is safe in rats over 28 days (unpublisheddata) Taken together these studies suggest that ingestion ofB pilosa aqueous extract at up to at 1 gkg BWday oncea day is highly safe in rats In addition the acute toxicityof aqueous and ethanol extracts of B pilosa in mice havebeen reported [134] Five- to six-week-old mice with weightsbetween 28 and 35 g received a peritoneal injection of bothextracts at the different doses The LD

50 the dose that causes

50 lethality of the aqueous and ethanol extracts in micewas 1230 gkg BW and 615 gkg BW respectively [134] Acomplete toxicological study has not been completed forhumans Furthermore the drug interactions of B pilosa withother drugs are unknown Further safety verification andclinical trials should be performed before B pilosa can beconsidered for medicinal use

5 Conclusions

B pilosa is an erect perennial plant with green leaves whiteor yellow flowers and tiny black seeds As it is distributedworldwide and is widely used as a folk remedy B pilosacan be thought of as an extraordinary source of food andmedicine However a comprehensive up-to-date review ofresearch on B pilosa has hitherto been unavailable In thisarticle scientific studies on B pilosa have been summarizedand critically discussed from the perspectives of botanyethnomedicine phytochemistry pharmacology and toxicol-ogy B pilosa is claimed to treat more than 40 disordersand 201 compounds have been identified from this plantThe medicinal utility of B pilosa and its modes of actionin relation to its known phytochemicals were discussedherein Polyynes flavonoids phenylpropanoids fatty acidsand phenolics are the primary bioactive compounds of Bpilosa and they have been reported to be effective in thetreatment of tumors inflammationimmune modulationdiabetes viruses microbes protozoans gastrointestinal dis-eases hypertension and cardiovascular diseases Cautionshould be exercised in the therapeutic use of B pilosa forhypoglycemia hypotension bleeding and allergy

Acknowledgments

The authors thank their laboratory members for construc-tive suggestions and the authors whose publications theycited for their contributions The authors also thank MsMiranda Loney of Academia Sinica for English editing of thispaper This work was supported by Grants 99-CDA-L11 and101S0010056 from Academia Sinica Taiwan Arlene P Bar-tolome is a recipient of MECO-TECO Sandwich ScholarshipProgram between Taiwan and the Philippines

References

[1] T Shen G H Li X N Wang and H X Lou ldquoThe genusCommiphora a review of its traditional uses phytochemistryand pharmacologyrdquo Journal of Ethnopharmacology vol 142 no2 pp 319ndash330

[2] C Long P Sauleau B David et al ldquoBioactive flavonoids ofTanacetum parthenium revisitedrdquo Phytochemistry vol 64 no2 pp 567ndash569 2003

[3] P O Karis and O Ryding Asteraceae Cladistics and Classifica-tion Bremer K Eds pp 559ndash569 Timber press Portland OreUSA 1994

[4] O N Pozharitskaya A N Shikov M N Makarova et alldquoAnti-inflammatory activity of a HPLC-fingerprinted aqueousinfusion of aerial part of Bidens tripartita LrdquoPhytomedicine vol17 no 6 pp 463ndash468 2010

[5] F Q Oliveira V Andrade-Neto A U Krettli and MG L Brandao ldquoNew evidences of antimalarial activity ofBidens pilosa roots extract correlated with polyacetylene andflavonoidsrdquo Journal of Ethnopharmacology vol 93 no 1 pp 39ndash42 2004

[6] Agriculture USDA ldquoPlants databaserdquo 2013in Natural Resources Conservation Servicehttpwwwnrcsusdagovwpsportalnrcssitenationalhome

[7] Y Zhou and X Z Yan ldquoExperimental study of qi deficiencysyndrome and Codonopsis pillosulae and Astragalus injectionon the immune response in micerdquo Zhong Xi Yi Jie He Za Zhivol 9 no 5 pp 286ndash262 1989

[8] S C Chien P H Young Y J Hsu et al ldquoAnti-diabeticproperties of three common Bidens pilosa variants in TaiwanrdquoPhytochemistry vol 70 no 10 pp 1246ndash1254 2009

[9] M J Alcaraz and M J Jimenez ldquoFlavonoids as anti-inflammatory agentsrdquo Fitoterapia vol 59 no 1 pp 25ndash38 1988

[10] FAO Agriculture Food and Nutrition for AfricamdashA ResourceBook for Teachers of Agriculture Publishing ManagementGroup FAO Information Division Rome Italy 1997

[11] M B Rokaya Z Munzbergova B Timsina and K R BhattaraildquoRheum australe D Don a review of its botany ethnobotanyphytochemistry and pharmacologyrdquo Journal of Ethnopharma-cology vol 141 no 3 pp 761ndash774 2012

[12] P H Young Y J Hsu and W C Yang ldquoBidens pilosa L andits medicinal userdquo in Recent Progress in Medicinal Plants DrugPlant II A S AwaadVK Singh and JNGovil Eds StandiumPress Houston Tex USA 2010

[13] C Ge ldquoCytologic study of Bidens bipinnata LrdquoZhongguo ZhongYao Za Zhi vol 15 no 2 pp 72ndash125 1990

[14] L C Chiang J S Chang C C Chen L T Ng and C CLin ldquoAnti-herpes simplex virus activity of Bidens pilosa and

48 Evidence-Based Complementary and Alternative Medicine

Houttuynia cordatardquoAmerican Journal of ChineseMedicine vol31 no 3 pp 355ndash362 2003

[15] N P Rybalchenko V A Prykhodko S S Nagorna et alldquoIn vitro antifungal activity of phenylheptatriyne from Bidenscernua L against yeastsrdquo Fitoterapia vol 81 no 5 pp 336ndash3382010

[16] K Redl W Breu B Davis and R Bauer ldquoAnti-inflammatoryactive polyacetylenes from Bidens campylothecardquo Planta Med-ica vol 60 no 1 pp 58ndash62 1994

[17] M Ayyanar and S Ignacimuthu ldquoTraditional knowledge ofKani tribals inKouthalai of Tirunelveli hills TamilNadu IndiardquoJournal of Ethnopharmacology vol 102 no 2 pp 246ndash255 2005

[18] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant intraditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[19] AKAbbas KMMurphy andA Sher ldquoFunctional diversity ofhelper T lymphocytesrdquo Nature vol 383 no 6603 pp 787ndash7931996

[20] J H Cano and G Volpato ldquoHerbal mixtures in the traditionalmedicine of Eastern Cubardquo Journal of Ethnopharmacology vol90 no 2-3 pp 293ndash316 2004

[21] J Namukobe J M Kasenene B T Kiremire et al ldquoTraditionalplants used for medicinal purposes by local communitiesaround the Northern sector of Kibale National Park UgandardquoJournal of Ethnopharmacology vol 136 no 1 pp 236ndash245 2011

[22] C Lans ldquoComparison of plants used for skin and stomachproblems in Trinidad and Tobago with Asian ethnomedicinerdquoJournal of Ethnobiology andEthnomedicine vol 3 article 3 2007

[23] S Dharmananda ldquoA Popular Remedy Ecapes Notice ofWesternPractitionersrdquo 2013 httpwwwitmonlineorgartsbidenshtm

[24] Y J Hsu T H Lee C L T Chang Y T Huang andW C YangldquoAnti-hyperglycemic effects and mechanism of Bidens pilosawater extractrdquo Journal of Ethnopharmacology vol 122 no 2 pp379ndash383 2009

[25] E Noumi F Houngue and D Lontsi ldquoTraditional medicinesin primary health care plants used for the treatment ofhypertension in Bafia Cameroonrdquo Fitoterapia vol 70 no 2 pp134ndash139 1999

[26] P V Tan T Dimo and E Dongo ldquoEffects of methanolcyclohexane and methylene chlo ride extracts of Bidens pilosaon various gastric ulcer models in ratsrdquo Journal of Ethnophar-macology vol 73 no 3 pp 415ndash421 2000

[27] R L C Pereira T Ibrahim L Lucchetti A J R Da Silva and VL G De Moraes ldquoImmunosuppressive and anti-inflammatoryeffects of methanolic extract and the polyacetylene isolatedfrom Bidens pilosa Lrdquo Immunopharmacology vol 43 no 1 pp31ndash37 1999

[28] T Dimo J Azay P V Tan et al ldquoEffects of the aqueous andmethylene chloride extracts of Bidens pilosa leaf on fructose-hypertensive ratsrdquo Journal of Ethnopharmacology vol 76 no 3pp 215ndash221 2001

[29] CWiartMedicinal Plants of Southeast Asia Pelanduk SelabgorDarul Ehsan Malaysia 2000

[30] F L Silva D C H Fischer J F Tavares M S Silva P FDe Athayde-Filho and J M Barbosa-Filho ldquoCompilation ofsecondary metabolites from Bidens pilosa LrdquoMolecules vol 16no 2 pp 1070ndash1102 2011

[31] A H Chen S R Lin and C H Hong ldquoPhytochemical studyon Bidens pilosa L varminorrdquo Huaxue pp 38ndash42 1975

[32] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[33] M C Zulueta M Tada and C Y Ragasa ldquoA diterpene fromBidens pilosardquo Phytochemistry vol 38 no 6 pp 1449ndash14501995

[34] J Wang H Yang Z W Lin and H D Sun ldquoFlavonoids fromBidens pilosa var radiatardquo Phytochemistry vol 46 no 7 pp1275ndash1278 1997

[35] A Zhao Q Zhao L Peng et al ldquoA new chalcone glycoside fromBidens pilosardquo Acta Botanica Yunnanica vol 26 no 1 pp 121ndash126

[36] P Gao Y Wu S Cui and Y Zou ldquoSynthesis and absoluteconfiguration of the 7-phenylhepta-4 6-diyne-1 2-diol isolatedfrom Bidens pilosardquo Synthesis no 13 pp 2131ndash2135 2011

[37] S Wang B Yang D Zhu D He and L Wang ldquoActivecomponents of Bidens pilosa Lrdquo Zhongcaoyao vol 36 pp 20ndash21

[38] F Bohlmann T Burkhardt and C Zdero Naturally OccuringAcetylenes Academic Press New York NY USA 1973

[39] T M Sarg A M Ateya N M Farrag and F A AbbasldquoConstituents and biological activity of Bidens pilosa L grownin EgyptrdquoActa Pharmaceutica Hungarica vol 61 no 6 pp 317ndash323 1991

[40] H A L Valdes and H P Rego ldquoBidens pilosa Linnerdquo RevistaCubana de Plantas Medicinales vol 6 pp 28ndash33

[41] F Bohlmann H Bornowski and K M Kleine ldquoNew polyynesfrom the tribe Heliantheaerdquo Chemische Berichte vol 97 pp2135ndash2138

[42] RWang Q XWu and Y P Shi ldquoPolyacetylenes and flavonoidsfrom the aerial parts ofBidens pilosardquo PlantaMedica vol 76 no9 pp 893ndash896 2010

[43] L W Wu Y M Chiang H C Chuang et al ldquoA novelpolyacetylene significantly inhibits angiogenesis and promotesapoptosis in human endothelial cells through activation of theCDK inhibitors and caspase-7rdquo Planta Medica vol 73 no 7 pp655ndash661 2007

[44] L W Wu Y M Chiang H C Chuang et al ldquoPolyacetylenesfunction as anti-angiogenic agentsrdquo Pharmaceutical Researchvol 21 no 11 pp 2112ndash2119 2004

[45] H L Yang S C Chen N W Chang et al ldquoProtection fromoxidative damage using Bidens pilosa extracts in normal humanerythrocytesrdquo Food and Chemical Toxicology vol 44 no 9 pp1513ndash1521 2006

[46] H Q Wang S J Lu H Li and Z H Yao ldquoEDTA-enhancedphytoremediation of lead contaminated soil by Bidens maxi-mowiczianardquo Journal of Environmental Sciences vol 19 no 12pp 1496ndash1499 2007

[47] S TobinagaMK SharmaWG L Aalbersberg et al ldquoIsolationand identification of a potent antimalarial and antibacterialpolyacetylene from Bidens pilosardquo Planta Medica vol 75 no 6pp 624ndash628 2009

[48] C L T Chang H K Kuo S L Chang et al ldquoThe distincteffects of a butanol fraction of Bidens pilosa plant extract onthe development of Th1-mediated diabetes and Th2-mediatedairway inflammation in micerdquo Journal of Biomedical Sciencevol 12 no 1 pp 79ndash89 2005

[49] S L Chang C L T Chang Y M Chiang et al ldquoPolyacetyleniccompounds and butanol fraction from Bidens pilosa can mod-ulate the differentiation of helper T cells and prevent autoim-mune diabetes in non-obese diabetic micerdquo Planta Medica vol70 no 11 pp 1045ndash1051 2004

Evidence-Based Complementary and Alternative Medicine 49

[50] Y M Chiang C L T Chang S L Chang W C Yang and LF Shyur ldquoCytopiloyne a novel polyacetylenic glucoside fromBidens pilosa functions as a T helper cell modulatorrdquo Journal ofEthnopharmacology vol 110 no 3 pp 532ndash538 2007

[51] R PUbillas CDMendez S D Jolad et al ldquoAntihyperglycemicacetylenic glucosides from Bidens pilosardquo Planta Medica vol66 no 1 pp 82ndash83 2000

[52] L Alvarez S Marquina M L Villarreal D Alonso E Arandaand G Delgado ldquoBioactive polyacetylenes from Bidens pilosardquoPlanta Medica vol 62 no 4 pp 355ndash357 1996

[53] C L T Chang S L Chang Y M Lee et al ldquoCytopiloyne apolyacetylenic glucoside prevents type 1 diabetes in nonobesediabetic micerdquo Journal of Immunology vol 178 no 11 pp 6984ndash6993 2007

[54] G Kusano A Kusano and Y Seyama Novel Hypoglycemicand Anti-Inflammatory Polyacetylenic Compounds Their Com-positions Bidens Plant Extract Fractions and CompositionsContaining the Plant or Fraction JPO Tokyo Japan 2004

[55] M G L Brandao A U Krettli L S R Soares C G CNery and H C Marinuzzi ldquoAntimalarial activity of extractsand fractions from Bidens pilosa and other Bidens species(Asteraceae) correlated with the presence of acetylene andflavonoid compoundsrdquo Journal of Ethnopharmacology vol 57no 2 pp 131ndash138 1997

[56] A U Krettli V F Andrade-Neto M D G L Brandao andW M S Ferrari ldquoThe search for new anti-malarial drugs fromplants used to treat fever and malaria or plants ramdomlyselected a reviewrdquoMemorias do Instituto Oswaldo Cruz vol 96no 8 pp 1033ndash1042 2001

[57] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[58] C K Wat R K Biswas E A Graham L Bohm G H NTowers and E R Waygood ldquoUltraviolet-mediated cytotoxicactivity of phenylheptatriyne from Bidens pilosa Lrdquo Journal ofNatural Products vol 42 no 1 pp 103ndash111 1979

[59] Y Sashida K Ogawa M Kitada H Karikome Y Mimakiand H Shimomura ldquoNew aurone glucosides and new phenyl-propanoid glucosides from Bidens pilosardquo Chemical and Phar-maceutical Bulletin vol 39 no 3 pp 709ndash711 1991

[60] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens pilosa L (TFB) on animal liver injury andliver fibrosisrdquo Journal of Ethnopharmacology vol 116 no 3 pp539ndash546 2008

[61] B Hoffmann and J Holzl ldquoAcylated compounds from Bidenspilosardquo Planta Medica vol 55 pp 108ndash109

[62] B Hoffmann and J Holzl ldquoA methylated chalcone glucosidefrom Bidens pilosardquo Phytochemistry vol 27 no 11 pp 3700ndash3701 1988

[63] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[64] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[65] B Hoffmann and J Holzl ldquoWeitere acylierte chalkone ausBidens pilosardquo Planta Medica vol 54 pp 450ndash451 1988

[66] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant in11traditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[67] V V Pham V V T K P V L Hoang and V K Phan ldquoFlavonoidcompounds from the plant Bidens pilosa L (Asteraceae)rdquo TapChi Duoc Hoc vol 50 pp 48ndash53

[68] S D Sarker B Bartholomew R J Nash and N Robinson ldquo5-O-methylhoslundin an unusual flavonoid from Bidens pilosa(Asteraceae)rdquo Biochemical Systematics and Ecology vol 28 no6 pp 591ndash593 2000

[69] S L Chang Y M Chiang C L T Chang et al ldquoFlavonoidscentaurein and centaureidin from Bidens pilosa stimulate IFN-120574 expressionrdquo Journal of Ethnopharmacology vol 112 no 2 pp232ndash236 2007

[70] Y M Chiang D Y Chuang S Y Wang Y H Kuo P WTsai and L F Shyur ldquoMetabolite profiling and chemopreventivebioactivity of plant extracts from Bidens pilosardquo Journal ofEthnopharmacology vol 95 no 2-3 pp 409ndash419 2004

[71] A Kusano Y Seyama E Usami et al ldquoStudies on the antioxi-dant active constituents of the dried powder from Bidens pilosaL var radiata SchrdquoNaturalMedicines vol 57 no 3 pp 100ndash1042003

[72] I Doleckova L Rarova J Gruz et al ldquoAnti-proliferative andanti-angiogenic effects of flavone eupatorin an active con-stituent of chloroform extract ofOrthosiphon stamineus leavesrdquoFitoterapia vol 83 no 6 pp 1000ndash1007

[73] K Bairwa R Kumar R J Sharma and R K Roy ldquoAn updatedreview on Bidens pilosa Lrdquo Der Pharma Chemica vol 2 no 3pp 325ndash337

[74] Q Xia Y Liu and Y Li ldquoDetermination of hyperoside indifferent parts and different species of Herba Bidens by RP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol 50pp 48ndash53

[75] M Horiuchi and Y Seyama ldquoImprovement of the antiinflam-matory and antiallergic activity of Bidens pilosa L var radiataSCHERFF treated with enzyme (Cellulosine)rdquo Journal of HealthScience vol 54 no 3 pp 294ndash301 2008

[76] M G L Brandao C G C Nery M A S Mamao and AU Krettli ldquoTwo methoxylated flavone glycosides from Bidenspilosardquo Phytochemistry vol 48 no 2 pp 397ndash400 1998

[77] P Kumari K Misra B S Sisodia et al ldquoA promising anticancerand antimalarial component from the leaves of Bidens pilosardquoPlanta Medica vol 75 no 1 pp 59ndash61 2009

[78] G Seelinger I Merfort U Wolfle and C M Schempp ldquoAnti-carcinogenic effects of the flavonoid luteolinrdquoMolecules vol 13no 10 pp 2628ndash2651 2008

[79] G Seelinger I Merfort and C M Schempp ldquoAnti-oxidantanti-inflammatory and anti-allergic activities of luteolinrdquoPlanta Medica vol 74 no 14 pp 1667ndash1677 2008

[80] C C Yit and N P Das ldquoCytotoxic effect of butein on humancolon adenocarcinoma cell proliferationrdquo Cancer Letters vol82 no 1 pp 65ndash72 1994

[81] J A Beutler E Hamel A J Vlietinck et al ldquoStructure-activityrequirements for flavone cytotoxicity and binding to tubulinrdquoJournal of Medicinal Chemistry vol 41 no 13 pp 2333ndash23381998

[82] M R Kviecinski K B Felipe T Schoenfelder et al ldquoStudy ofthe antitumor potential of Bidens pilosa (Asteraceae) used inBrazilian folkmedicinerdquo Journal of Ethnopharmacology vol 117no 1 pp 69ndash75 2008

[83] P Sundararajan A Dey A Smith A G Doss M Rajappanand S Natarajan ldquoStudies of anticancer and antipyretic activityof Bidens pilosawhole plantrdquo African Health Sciences vol 6 no1 pp 27ndash30 2006

[84] YMChiang C P Lo Y P Chen et al ldquoEthyl caffeate suppressesNF-120581B activation and its downstream inflammatory mediatorsiNOS COX-2 and PGE2 in vitro or in mouse skinrdquo BritishJournal of Pharmacology vol 146 no 3 pp 352ndash363 2005

50 Evidence-Based Complementary and Alternative Medicine

[85] J S Kim H J Lee M H Lee J Kim C Jin and J HRyu ldquoLuteolin inhibits LPS-stimulated inducible nitric oxidesynthase expression in BV-2 microglial cellsrdquo Planta Medicavol 72 no 1 pp 65ndash68 2006

[86] P A Ruiz and D Haller ldquoFunctional diversity of flavonoidsin the inhibition of the proinflammatory NF-120581B IRF and Aktsignaling pathways in murine intestinal epithelial cellsrdquo Journalof Nutrition vol 136 no 3 pp 664ndash671 2006

[87] A Xagorari A Papapetropoulos A Mauromatis MEconomou T Fotsis and C Roussos ldquoLuteolin inhibitsan endotoxin-stimulated phosphorylation cascade andproinflammatory cytokine production in macrophagesrdquoJournal of Pharmacology and Experimental Therapeutics vol296 no 1 pp 181ndash187 2001

[88] N Yoshida T Kanekura Y Higashi and T Kanzaki ldquoBidenspilosa suppresses interleukin-1120573-induced cyclooxygenase-2expression through the inhibition of mitogen activated proteinkinases phosphorylation in normal human dermal fibroblastsrdquoJournal of Dermatology vol 33 no 10 pp 676ndash683 2006

[89] M Muchuweti C Mupure A Ndhlala T Murenje and M AN Benhura ldquoScreening of antioxidant and radical scavengingactivity of Vigna ungiculata Bidens pilosa and Cleome gynan-drardquo American Journal of Food Technology vol 2 no 3 pp 161ndash168 2007

[90] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens bipinnata L against carbon tetrachloride-induced liver fibrosis in ratsrdquo Journal of Pharmacy and Pharma-cology vol 60 no 10 pp 1393ndash1402 2008

[91] S L Chang H H Yeh Y S Lin Y M Chiang T K Wuand W C Yang ldquoThe effect of centaurein on interferon-120574 expression and Listeria infection in micerdquo Toxicology andApplied Pharmacology vol 219 no 1 pp 54ndash61 2007

[92] F Deba T D Xuan M Yasuda and S Tawata ldquoChemical com-position and antioxidant antibacterial and antifungal activitiesof the essential oils from Bidens pilosa Linn var Radiatardquo FoodControl vol 19 no 4 pp 346ndash352 2008

[93] A O T Ashafa and A J Afolayan ldquoScreening the root extractsfrom Biden pilosa L var radiata (Asteraceae) for antimicrobialpotentialsrdquo Journal of Medicinal Plant Research vol 3 no 8 pp568ndash572 2009

[94] T B Nguelefack T Dimo E P Nguelefack Mbuyo P V TanS V Rakotonirina and A Kamanyi ldquoRelaxant effects of theneutral extract of the leaves of Bidens pilosa linn on isolated ratvascular smooth musclerdquo Phytotherapy Research vol 19 no 3pp 207ndash210 2005

[95] M T Grombone-Guaratini K L Silva-Brandao V N SolferiniJ Semir and J R Trigo ldquoSesquiterpene and polyacetyleneprofile of the Bidens pilosa complex (Asteraceae Heliantheae)from Southeast of Brazilrdquo Biochemical Systematics and Ecologyvol 33 no 5 pp 479ndash486 2005

[96] L L Lin C Y Wu H C Hsiu M T Wang and H ChuangldquoStudies on diabetes mellitus I The hypoglycemic activity ofphytosterin on alloxan diabetic ratsrdquo Taiwan Yi Xue Hui Za Zhivol 66 no 2 pp 58ndash65 1967

[97] M A N Benhura and I C Chitsiku ldquoThe extractable 120573-carotene content of Guku (Bidens pilosa) leaves after cookingdrying and storagerdquo International Journal of Food Science andTechnology vol 32 no 6 pp 495ndash500 1997

[98] J S Chang L C Chiang C C Chen L T Liu K C Wangand C C Lin ldquoAtileukemic activity of Bidens pilosa l varminor (blume) sherff andHouttuynia cordata thunbrdquo AmericanJournal of Chinese Medicine vol 29 no 2 pp 303ndash312 2001

[99] F Deba T D Xuan M Yasuda and S Tawata ldquoHerbicidal andfungicidal activities and identification of potential phytotoxinsfrom Bidens pilosa L var radiata Scherff research paperrdquoWeedBiology and Management vol 7 no 2 pp 77ndash83 2007

[100] J K Kumar and A K Sinha ldquoA new disubstituted acetylacetonefrom the leaves ofBidens pilosa LinnrdquoNatural Product Researchvol 17 no 1 pp 71ndash74 2003

[101] K Ogawa and Y Sashida ldquoCaffeoyl derivatives of a sugarlactone and its hydroxy acid from the leaves of Bidens pilosardquoPhytochemistry vol 31 no 10 pp 3657ndash3658 1992

[102] Q Xia Y Liu and Y Li ldquoDetermination of gallic acid fromdifferent species and different medical parts of Herba Bidens byRP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol24 no 03 pp 308ndash310

[103] W J Lee L F Wu W K Chen C J Wang and T HTseng ldquoInhibitory effect of luteolin on hepatocyte growth fac-torscatter factor-induced HepG2 cell invasion involving bothMAPKERKs and PI3K-Akt pathwaysrdquo Chemico-BiologicalInteractions vol 160 no 2 pp 123ndash133 2006

[104] J A Beutler J H Cardellina C M Lin E Hamel G MCragg andM R Boyd ldquoCentaureidin a cytotoxic flavone fromPolymnia fruticosa inhibits tubulin polymerizationrdquo BioorganicandMedicinal Chemistry Letters vol 3 no 4 pp 581ndash584 1993

[105] A Verma A Su A M Golin B OrsquoMarrah and J K AmorosaldquoThe lateral view a screening method for knee traumardquo Aca-demic Radiology vol 8 no 5 pp 392ndash397 2001

[106] J Corren M Lemay Y Lin L Rozga and R K RandolphldquoClinical and biochemical effects of a combination botanicalproduct (ClearGuard) for allergy a pilot randomized double-blind placebo-controlled trialrdquo Nutrition Journal vol 7 no 1article 20 2008

[107] M S Gachet J S Lecaro M Kaiser et al ldquoAssessment of anti-protozoal activity of plants traditionally used in Ecuador in thetreatment of leishmaniasisrdquo Journal of Ethnopharmacology vol128 no 1 pp 184ndash197 2010

[108] W C Yang M Ghiotto B Barbarat and D Olive ldquoThe role ofTec protein-tyrosine kinase in T cell signalingrdquo The Journal ofBiological Chemistry vol 274 no 2 pp 607ndash617 1999

[109] S Tewtrakul H Miyashiro N Nakamura et al ldquoHIV-1 inte-grase inhibitory substances from Coleus parvifoliusrdquo Phytother-apy Research vol 17 no 3 pp 232ndash239 2003

[110] T Dimo T B Nguelefack P V Tan et al ldquoPossible mechanismsof action of the neutral extract from Bidens pilosa L leaves onthe cardiovascular system of anaesthetized ratsrdquo PhytotherapyResearch vol 17 no 10 pp 1135ndash1139 2003

[111] T S C Li Chinese and Related North American Herbs CRCPress New York NY USA 2002

[112] H Wu H Chen X Hua Z Shi L Zhang and J ChenldquoClinical therapeutic effect of drug-separated moxibustion onchronic diarrhea and its immunologic mechanismsrdquo Journal ofTraditional Chinese Medicine vol 17 no 4 pp 253ndash258 1997

[113] S W Wright R R Harris J S Kerr et al ldquoSynthesis chemicaland biological properties of vinylogous hydroxamic acids dualinhibitors of 5-lipoxygenase and IL-1 biosynthesisrdquo Journal ofMedicinal Chemistry vol 35 no 22 pp 4061ndash4068 1992

[114] W R Almiron and M E Brewer ldquoClassification of immaturestage habitats of Culicidae (Diptera) collected in CordobaArgentinardquo Memorias do Instituto Oswaldo Cruz vol 91 no 1pp 1ndash9 1996

[115] N L Wang J Wang X S Yao and S Kitanaka ldquoTwo newmonoterpene glycosides and a new (+)-jasmololone glucoside

Evidence-Based Complementary and Alternative Medicine 51

from Bidens parvifloraWilldrdquo Journal of Asian Natural ProductsResearch vol 9 no 5 pp 473ndash479 2007

[116] P Champagnat ldquoRole of the terminal bud in the actionexercised by the cotyledon of Bidens pilosus L var radiatus onits axillary budrdquo Comptes Rendus des Seances et Memoires de laSociete de Biologie vol 145 no 17-18 pp 1374ndash1376 1951

[117] S F Nielsen S B Christensen G Cruciani A Kharazmiand T Liljefors ldquoAntileishmaniai chalcones statistical designsynthesis and three-dimensional quantitative structure-activityrelationship analysisrdquo Journal of Medicinal Chemistry vol 41no 24 pp 4819ndash4832 1998

[118] Y C Hwang J J H Chu P L Yang W Chen and M V YatesldquoRapid identification of inhibitors that interfere with poliovirusreplication using a cell-based assayrdquo Antiviral Research vol 77no 3 pp 232ndash236 2008

[119] V F Andrade-Neto M G L Brandao F Q Oliveira et alldquoAntimalarial activity of Bidens pilosa L (Asteraceae) ethanolextracts fromwild plants collected in various localities or plantscultivated in humus soilrdquo Phytotherapy Research vol 18 no 8pp 634ndash639 2004

[120] R J Marles and N R Farnsworth ldquoAnti-diabetic plants andtheir active constituentsrdquo Phytomedicine vol 2 no 2 pp 137ndash189

[121] M Habeck ldquoDiabetes treatments get sweet help from naturerdquoNature Medicine vol 9 no 10 p 1228 2003

[122] H W Lin G Y Han and S X Liao ldquoStudies on the activeconstituents of the Chinese traditional medicine Polygonatumodoratum (Mill) DrucerdquoActa Pharmaceutica Sinica vol 29 no3 pp 215ndash222 1994

[123] C L T Chang Y C Chen H M Chen N S Yang andW C Yang ldquoNatural cures for type 1 diabetes a reviewof phytochemicals biological actions and clinical potentialrdquoCurrent Medicinal Chemistry vol 20 no 7 pp 899ndash907 2013

[124] L Dey A S Attele and C S Yuan ldquoAlternative therapies fortype 2 diabetesrdquo Alternative Medicine Review vol 7 no 1 pp45ndash58 2002

[125] C L T ChangH Y Liu T F Kuo et al ldquoAnti-diabetic effect andmode of action of cytopiloynerdquo Evidence-Based Complementaryand Alternative Medicine vol 2013 Article ID 685642 13 pages2013

[126] M R Khan M Kihara and A D Omoloso ldquoAnti-microbialactivity of Bidens pilosa Bischofia javanica Elmerillia papuanaand Sigesbekia orientalisrdquo Fitoterapia vol 72 no 6 pp 662ndash6652001

[127] T Dimo T B Nguelefack P Kamtchouing E Dongo ARakotonirina and S V Rakotonirina ldquoHypotensive effects ofmethanol extract from Bidens pilosa Linn on hypertensive ratsrdquoComptes Rendus de lrsquoAcademie des Sciences Serie III vol 322no 4 pp 323ndash329 1999

[128] A K Hassan O Deogratius J F Nyafuono O Francis and OP Engeu ldquoWound healing potential of the ethanolic extracts ofBidens pilosa and Ocimum suaverdquo African Journal of Pharmacyand Pharmacology vol 5 no 2 pp 132ndash136 2011

[129] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[130] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[131] C Kandaswami and EMiddleton ldquoFree radical scavenging andantioxidant activity of plant flavonoidsrdquo Advances in Experi-mental Medicine and Biology vol 366 pp 351ndash376 1994

[132] K Yamamoto H Kakegawa H Ueda et al ldquoGastric cytopro-tective anti-ulcerogenic actions of hydroxychalcones in ratsrdquoPlanta Medica vol 58 no 5 pp 389ndash393 1992

[133] J O C Ezeonwumelu A K Julius C N Muhoho et alldquoBiochemical and histological studies of aqueous extract ofBidens pilosa leaves from Ugandan Rift valley in ratsrdquo BritishJournal of Pharmacology and Toxicology vol 2 no 6 pp 302ndash309

[134] L Frida S Rakotonirina A Rakotonirina and J P SavineauldquoIn vivo and in vitro effects of Bidens pilosa L (Asteraceae)leaf aqueous and ethanol extracts on primed-oestrogenized ratuterine musclerdquo African Journal of Traditional Complementaryand Alternative Medicines vol 5 no 1 pp 79ndash91 2008

52 Evidence-Based Complementary and Alternative Medicine

Composition Comments

1 Please provide valid postal code to the address if

possible

2 We made the highlighted change in Table 4 for the sake

of correctness Please check

3 Please check the correctness of the highlighted part in

Table 5

4 We considered the highlighted part in Table 11 as

reference Please check similar cases throughout

5 Please check the format of Table 18

6 We reformatted Table 20 Please check

7 We made the highlighted change in Table 4 for the sake

of consistency Please check similar cases throughout the

paper

8 We made the highlighted change(s) for the sake

of consistency Please check simila highlighted cases

throughout

9 We made the highlighted change in Table 9 for the sake

of clarity Please check similar cases through out

10 We made the highlighted change(s) for the sake of

correctness Please check

11 Comment on ref [66] This reference is a repetition

of [18] Please check similar cases throughout

Author(s) Name(s)

Author 1

Author 2

Author 3

Evidence-Based Complementary and Alternative Medicine 11

Table 4 Continued

S N IUPAC names Common names Structure Plant part(country) References

63 7-Phenylhepta-46-diyn-12-diol HO

OH

Aerial (China) [42]

64 135-Heptatriyn-1-yl-benzene

1-Phenylhepta-135-triyne

Leaves (notstated) leaves oftissue culture(not stated)aerial (TanzaniaChina)Whole (Taiwan)roots (Brazil)

[18 34 3842 56ndash58]

65 7-Phenyl-246-heptatriyn-1-ol HO

Leaves (notstated) aerial(China)

[38 42]

667-Phenyl-246-heptatriyn-1-ol-acetate O

O Leaves (notstated) [38]

675-(2-Phenylethynyl)-2-thiophenemethanol

S OHAerial (China) [42]

685-(2-Phenylethynyl)-2120573-glucosylmethyl-thiophene

O

O

OH

H

OH

H

H

HOH H

O

HO

S

Aerial (China) [42]

69

3- 120573 -D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne

OH

O

OH

HOHO

H

H

HHOH

OH

Leaves(Cameroon) [28]

701-Phenyl-13-diyn-5-en-7-ol-acetate O

O

Leaves (Brazil) [27]

SN denotes serial number

the chloroform fraction was the most toxic with a halfmaximal inhibitory concentration (IC

50) of 97 plusmn 72 and 83 plusmn

52 120583gmL in NRU and MTT respectively [82] Kumari andcolleagues also reported the anti-cancer and anti-malarialactivities of B pilosa leaves [77] Based on a cytotoxicity-directed fractionation strategy they identified phenyl-135-heptatriene with IC

50values of 8plusmn001 049plusmn045 07plusmn001

and 10 plusmn 001 120583gmL against human oral liver colon andbreast cancer cell lines respectively However phenyl-135-heptatriyne showed lower activity against breast cancer celllines than the chloroform leaf extract which had an IC

50

value of 65 plusmn 001 120583gmL Moreover the positive controltaxol showed higher activity than phenyl-135-heptatriyne[77] Furthermore in vitro comet assays were performed toevaluate the toxicity of n-hexane chloroform and methanolextracts of B pilosa and its ethyl acetate acetone and waterfractions onHela andKB cellsThe ethyl acetate fraction fromthe methanol extract exhibited the highest activity with halfmaximal cytotoxic concentrations (CTC

50) of 9652120583gmL

and 5862120583gmL against Hela and KB cells respectivelyDespite the moderate toxicity these findings suggest thatthese B pilosa extractsfractions could be useful for future

12 Evidence-Based Complementary and Alternative Medicine

Table5Flavon

oids

isolated

from

Bpilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

71

2-[(34-Dihydroxyph

enyl)-

methylene]-

6-hydroxy-3(2H

)-benzofuranon

e

Sulfu

retin

O

OHO

OH

OH

Aeria

l(Ch

ina)

[35]

72

2-[(34-Dihydroxyph

enyl)-

methylene]-

67-dihydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-673101584041015840-

tetrahydroxy

maritimetin

O

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

73

2-[(34-Dihydroxyph

enyl)-

methylene]-6-

(120573-D

-glucopyrano

syloxy)-7-

hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-120573-D

-glucop

yranosyl-

673101584041015840-te

trahydroxy

maritimein

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japanand

China)aerial(Ch

ina)

[355960]

74

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

(120573-D

-glucopyrano

syloxy)-

6-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-7-O-120573-D

-glucop

yranosyl-

673101584041015840-

tetrahydroxy

O

OHO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japan)

[59]

75

6-[(6-O-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-acetyl-

120573-D

-glucopyrano

syl)

-673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OHO

H HO

H

H

HOH

HO

O

O

Leaves

(Not

stated)

aeria

l(Ch

ina)

[3559]

Evidence-Based Complementary and Alternative Medicine 13

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

76

6-[(36-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-

2-[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone(Z)-6-O

-(36-di-O

-acetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

bideno

sideA

O

O

HO

OH

OH

OHOH O

H

H

HOH

HO

O

O

O

Leaves

(China)

[60]

77

6-[(46-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(410158401015840610158401015840-

diacetyl-120573-D

-glucropyrano

syl)-

673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

HO

O

OO

Leaves

(not

stated)

aeria

l(Ch

ina)

[343561]

78

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

hydroxy-6-

[(246-tri-O

-acetyl-120573

-D-

glucop

yranosyl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(210158401015840410158401015840610158401015840-

triacetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

O

OOO

O

Leaves

(not

stated)

aeria

l(Ch

ina)

[3461]

79

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-6-

[(346-tri-O

-acetyl-120573

-D-glucopyrano

syl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(310158401015840410158401015840610158401015840-

triacetyl-120573

-D-glucrop

yranosyl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H OH

H

HOH

HO

OOO O

Aeria

l(Ch

ina)

[3435]

80

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-

6-[[6-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propenyl]-120573-

D-glucopyrano

syl]o

xy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-

p-coum

aroyl-

120573-D

-glucropyrano

syl)-

673101584041015840-te

trahydroxy

OH

O

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OLeaves

(Japan)

[59]

14 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

81

1-[2-(120573-D

-Glucopyrano

syloxy)-

4-hydroxyphenyl]-2-

hydroxy-3-(3-

hydroxyphenyl)-2-

prop

en-1-on

e

Chalcone120572

32101584041015840-

tetrahydroxy-

21015840-O

-120573-D

-glucop

yranosyl

HO

O

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[35]

821-(24-Dihydroxyph

enyl)-

3-(34-dihydroxyph

enyl)-

2-prop

en-1-on

eBu

tein

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

833-(34-D

ihydroxyph

enyl)-

1-(234-trihydroxyph

enyl)-

2-prop

en-1-on

eOkanin

O

HO

HO

OH

OH

OH

Leaves

(China)

[60]

84

3-(34-D

ihydroxyph

enyl)-

1-[3-(120573-D

-glucop

yranosyloxy)-

24-dihydroxyphenyl]-2-

prop

en-1-on

e

Okanin

31015840-O

-120573-D

-glucosid

e

O

HO

OH

OH

OH

OO

HO

H

HO

H

H

HOH

H

OH

Leaves

(Germany)

flowers(Germany)

[61ndash63]

85

3-(34-D

ihydroxyph

enyl)-

1-[4-(120573-D

-glucop

yranosyloxy)-

23-dihydroxyphenyl]-

2-prop

en-1-on

e

Okanin

41015840-O

-120573-D

-glucosid

e

OOH

OH

OH

HOO

OHO

H

HO

H

H

HOH

H

OH

Flow

ers(Germany)

leaves

(Japan)

[5963]

Evidence-Based Complementary and Alternative Medicine 15

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

86Okanin41015840-O

-120573-D

-(610158401015840-

O-acetylglucosid

e)

O

O

OH

OH

OH

OHO

H

HO

H

H

HOH

H

HO

O

O

Flow

ers(Germany)

[63]

87

1-[4-[(46-di-O

-Acetyl-

120573-D

-glucopyrano

syl)-oxy]-

23-dihydroxyphenyl]-3-

(34-dihydroxyph

enyl)-2-

prop

en-1-on

e

Okanin

41015840-O

-120573-D

-(410158401015840610158401015840-diacetyl)-

glucop

yranoside

O

O

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Aeria

l(Ch

ina)

[34]

88Okanin41015840-O

-120573-D

-(210158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[64]

89Okanin41015840-O

-120573-D

-(310158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

O

OAe

rial(Ch

ina)

[34]

16 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

90

1-[23-Dihydroxy-4-

[[6-O-[3-(4-hydroxyph

enyl)-

1-oxo-2-prop

enyl]-120573-D

-glucop

yranosyl]oxy]-ph

enyl]-

3-(34-dihydroxyph

enyl)-

2-prop

en-1on

e

Okanin

41015840-O

-120573-D

-(610158401015840-tr

ans-

p-coum

aroyl)-

glucoside

O

O

OH

OH

OH

OHOH

HO

H

H

HOH

H

HO

OO

OH

Leaves

(Germany)

[64]

91Okanin41015840-O

-120573-D

-(410158401015840-acetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Leaves

(Germany)

[65]

92

Okanin

41015840-O

-120573-D

-(210158401015840410158401015840-diacetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[65]

Evidence-Based Complementary and Alternative Medicine 17

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

93

Okanin

41015840-O

-120573-D

-(310158401015840410158401015840-diacetyl-610158401015840-

trans-p-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

OH

O O

Leaves

(Germany)

[65]

94

Okanin

41015840-O

-[120573-D

-glucopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

OOH

OH

OH

HO

OH

HO

HHO

H

HOH

HO

OH

OH

HO

HHO

H

HOH

HO

Flow

er(G

ermany)

[63]

95Okanin

3101584041015840-di-O

-120573-D

-glucosid

e

OOH

OH

OH

O OO

H HO

HHOH

HOH

HHO

OHO

H

HO

H

H

HOH

H

OH

Flow

er(G

ermany)

[63]

18 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

96

1-[3-(120573-D

-Glucopyrano

syloxy)-

24-dihydroxyphenyl]-

3-(3-hydroxy)-

4-metho

xyph

enyl)-

2-prop

en-1-on

e

Okanin

4-methylether-

31015840-O

-120573-D

-glucop

yranoside

OOH

O

OH

HO

O

H HO

HHOH

HOH

H

OHO

Leaves

(Germany)aerial

(China)

[3562]

97

Okanin4-methylether-

3101584041015840-di-O

-120573-(41015840101584061015840101584041015840101584010158406101584010158401015840-

tetraacetyl)-

glucop

yranoside

OOH

OOH

O

HH

OH

HO

HOH

H

OO

O

Ac

Ac

AcAc

OO

H

HO

H

H

HOH

H

O

Aeria

l(Ch

ina)

[42]

98Ch

alcone210158404101584061015840-tr

imetho

xy-

4-O-D

-glucopyrano

syl-

dihydro

Not

foun

dLeaves

(China)

[60]

992-(34-D

ihydroxyph

enyl)-23-

dihydro-78

-dihydroxy-

4H-1-

benzop

yran-4-one

Okanin

isoOO

OH

HO

OH

OH

Leaves

(China)

[60]

100

2-(34-D

ihydroxyph

enyl)-

23-dihydro-8-hydroxy-

7[(246-tr

i-O-acetyl-120573

-D-

glucop

yranosyl)oxy]-4H

-1-b

enzopyran-4-on

eOO

OH

OH

OH

OH

O

HHO

H

HO

HO

OAc

Ac

Ac

Aeria

l(Ch

ina)

[34]

Evidence-Based Complementary and Alternative Medicine 19

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

101

57-Dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

OO

OH

HO

OH

Aeria

l(TanzaniaC

hina)

[4266]

102

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

7-O-glucopyrano

side

OO

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Tanzania)

[66]

103

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-4H

-1-benzop

yran-4-one

Luteolin

OO

HO

OH

OH

OH

Aeria

l(TanzaniaC

hina

Vietnam)

[343542

6667]

104

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5hydroxy-4H-1-

benzop

yran-4-one

Luteolin

7-O-120573-D

-glucop

yranoside

OOOH

OH

OH

OO

HO

H

HO

H

H

HOH

OH H

Aeria

l(Tanzania)

[66]

105

57-Dim

etho

xy-6-

(5-m

etho

xy-6-m

ethyl-

4-oxo-4H

-pyran-3-yl)-

2-ph

enyl-4H-1-

benzop

yran-4-one

5-O-M

ethylhosludin

O OOO

O

O

O

Aeria

l(Ugand

a)[68]

20 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

106

3-(120573-D

-Glucopyrano

syloxy)-

57-dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Astragalin

Kaem

pferol-3-O

-120573-D

-glucopyrano

side

HO

OH

O O

OH

O

O

HO

HHO

H

H

HOH

OH

H

Aeria

l(Ch

ina)

[35]

107

Kaem

pferol3-

(23-di-E

-p-cou

maroyl-

120572-L-rhamno

pyrano

side)

Not

foun

dAe

rial(Vietnam)

[67]

108

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5-hydroxy-36-dimetho

xy-

4H-1-

benzop

yran-4-one

Axillarosid

eO O

OH

OH

OO

OH

OO

HO

H

HO

H

H

HOH

H

OH

Aeria

l(Ch

ina)

[42]

109

57-Dihydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaureidin

O

O

O

O

O

HO

OH

OH

Who

le(Taiwan)

[69]

110

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaurein

O O

OO

O OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan)w

hole

(Taiwan)

[69ndash

71]

Evidence-Based Complementary and Alternative Medicine 21

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

111

5-Hydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-

67-dimetho

xy-4H-

chromen-4-one

Eupatorin

iso

OO O

OH

O

O OH

Not

foun

d(C

hina)

[3772]

112

2-(34-D

imetho

xyph

enyl)-7-

(120573-D

-glucopyrano

syloxy)-35-

dihydroxy-

8-metho

xy-4H-1-

benzop

yran-4-one

O O

OH

O O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

113

7-(120573-D

-Glucopyrano

syloxy)-

5hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-38-

dimehtoxy-4H-1-

benzop

yran-4-one

O O

O

OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

114

Isorhamnetin

3-[O

-120572-

L-rham

nopyrano

syl-

(1-2)-120573-D

-glucop

yranoside]

Not

foun

dAe

rial(Vietnam)

[67]

115

7-[(6-deoxy-120572-L-

Manno

pyrano

syl)o

xy]-

3-120573-D

-glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-4H

-1-benzop

yran-4-one

Luteoside

O O

OH

O

OH

OH

OH

HOH

H

H HO

HO

HO

OH

HO

H

HO

OH

OH

HH

Aeria

l(Ch

ina)

[42]

22 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

116

Luteolin

3-O-120573-D

-Glucopyrano

side

O O

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

HO

Aeria

l(Tanzania)

[66]

117

57-Dihydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Quercetagetin

3631015840-tr

imethylether

OOO

O

O OH

HO

OH

Aeria

l(Ch

ina)

[42]

118

6-((2S3S4S5S)-

Tetrahydro-345-tr

ihydroxy-

6-(hydroxymethyl)-2H

-pyran-2-yloxy)-5-hydroxy-

2-(4-hydroxy-3-

metho

xyph

enyl)-

37-dimetho

xy-4H-

chromen-4-one

Quercetagetin

3731015840-

trim

ethylether-6-O

-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[42]

119

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

JaceinQ

uercetin

3631015840-

trim

ethyl

ether-7-O-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan

China)

who

le(Taiwan)

[425470]

120

2-(34-D

ihydroxyph

enyl)-

357-trihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

OO

HO

OH

OH

OH

OH

Aeria

l(Ch

ina)leaves

(China)who

le(C

hina)

[356074]

Evidence-Based Complementary and Alternative Medicine 23

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

121

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-galactopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-4-one

Quercetin

3-O-120573-D

-galactosidehyperin

hyperosid

e

O O

HO

OH

OH

OH

OOHH

HOH

H

H HO

HO

OH

Aeria

l(TanzaniaJapan)

Leaves

(China)Who

le(C

hina)

[376066

717475]

122

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-glucopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

3-O-120573-

D-glucopyrano

side

O O

HO

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

Aeria

l(TanzaniaC

hina

Japan)L

eaves(Japan)

[355966

75]

123

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-

4-oxo-4H

-1-benzop

yran-3-yl-

120573-D

-glucopyrano

siduron

icacid

Quercetin

3-O-120573-D

-glucuron

opyranoside

O O

HO

OH

OH

OH

OHOH

HOH

H

H HO

HO

OHO

Aeria

l(TanzaniaJapan)

[6671]

124

3-[[6-O-(6-

Deoxy-120572-L-m

anno

pyrano

syl)-

120573-D

-galactopyrano

syl]o

xy]-

2-(34-dihydroxyph

enyl)-57-

dihydroxy-4H

-1-benzop

yran-4-one

Quercetin

3-O-rob

inob

iosid

e

O O

HO

OH

OH

OH

O

HOHH OH

HO

H HH

OO

OHH

HOH

H

H HO

HO

Aeria

l(Japan)W

hole

(Taiwan)

[7071]

24 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

125

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-

3-metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

OH

Roots(Brazil)

[55676]

126

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-

120573-D

-glucopyrano

syl]o

xy]-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120572-

L-rham

nopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

O

HHO HHO

HO H

HH

ORo

ots(Brazil)

[5676]

127

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-120573-D

-glucop

yranosyl]oxy]-5-

hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-3-metho

xy-

4H-1-

benzop

yran-4-one

Quercetin

341015840-

dimethyl

ether-7-O-rutinoside

O OOH

OH

O

O

OH O

HOH

H

HHO

H

O

HHO HHO

HO H

HH

O

HO

Aeria

l(Ch

ina)

[3435]

128

2-(34-D

ihydroxyph

enyl)-3-

(120573-D

-glucofurano

syloxy)-57-

dihydroxy-4H

-1-benzop

yran-

4-on

e

Isoq

uercitrin

O OOH

OH

OHO

H

OHH

OH

HO

H O HOH

OH

Aeria

l(Japan

China)

[3571]

Evidence-Based Complementary and Alternative Medicine 25

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

129

Quercetin

3-O-120573-D

-galactop

yranoside

O

OH

HO

OH OH

O

O

HO

OH

OHOH

OAe

rial(no

tstated)

[18]

130

Quercetin

3-O-rutinoside

O

O

R

OO

OH

OHO

H

O

OH

OH

H3C

H3C

H3C

OH

HO

Who

le(Taiwan)

[70]

26 Evidence-Based Complementary and Alternative Medicine

Table 6 Terpenoids isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(Country) References

131371111-Tetramethylbicyclo[810]undeca-

26-dieneBicyclogermacrene Leaves

(Brazil) [95]

13241111-Trimethyl-8-

methylenebicyclo[720]undec-4-ene

E-Caryophyllene Leaves(Brazil) [95]

1331-Methyl-5-methylene-8-

(1-methylethyl)-16-cyclodecadiene

Germacrene D Leaves(Brazil) [95]

1344-(15-Dimethyl-4-hexen-1-ylidene)-1-methyl-cyclohexene

Z-120574-Bisabolene Leaves(Brazil) [95]

135Decahydro-114-trimethyl-7-methylene-1H-cycloprop[e]-

azulene120573-Gurjunene Leaves

(Brazil) [95]

136 2669-Tetramethyl-148-cycloundecatriene

120572-Humulene120572-caryophyllene

Leaves(Brazil) [95]

13712344a568a-Octahydro-1-isopropyl-7-methyl-4-methylenenaphthalene

120575-Muurolene Leaves(Brazil) [95]

138

12344a568a-Octahydro-4a8-dimethyl-2-(1-methylethylidene)-

naphthalene

Selina-37(11)-diene

Leaves(Brazil) [95]

139(2E7R11R)-371115-

Tetramethyl-2-hexadecen-1-ol

Phytol

HO

Whole(Taiwan) [57]

Evidence-Based Complementary and Alternative Medicine 27

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

140 371115-Tetramethyl-2-hexadecenoic acid Phytanic acid

O

OH

Whole(Taiwan) [57]

141371115-Tetramethyl-2-

hexadecenyl ester-heptanoicacid

Phythylheptanoate O O

Leaves(Not stated) [33]

142

(3S10R13R)-17-((2R5R)-5-Ethyl-6-methylheptan-2-yl)-

2347891011121314151617-tetradecahydro-

1013-dimethyl-1H-cyclopenta[a]phenanthren-

3-ol

Campestrol

HO

Aerial(Tanzania) [18]

143 Not found Phytosterin-B Not foundNot found(TaiwanEgypt)

[39 96]

144 Stigmast-5-en-3-ol 120573-sitosterol

HOH

H

H

Aerial(Tanzania)

whole(Taiwan)

[18 31 57]

145

1314151617-Tetradecahydro-1013-

dimethyl-1H-cyclopenta[a]phenanthren-

3-yloxy)-tetrahydro-6-(hydroxymethyl)-2H-pyran-345-triol

120573-Sitosterolglucoside

H

H

HOH

HO

HHO

H

HOHH O

OH Not found(Egypt) [39]

146 5120572-Stigmasta-7-en-3120573-ol

HO

Whole(Taiwan) [57]

147 5120572-Stigmasta-722t-dien-3120573-ol

HO

Whole(Taiwan) [57]

148 Stigmasta-522-dien-3-ol Stigmasterol

HOH

H

H

Not found(Taiwan)aerial

(Tanzania)leaves (notstated)whole

(Taiwan)

[18 31 3357]

28 Evidence-Based Complementary and Alternative Medicine

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

149 Lup-20(29)-en-3-ol Lupeol

H

HH

HOH

Not found(Egypt) [39]

150 Lup-20(29)-en-3-ol acetate Lupeol acetate

H

HH

O

OH

Not found(Egypt) [39]

151 Olean-12-en-3-ol 120573-amyrin

H

HOH

Not found(Egypt) [39]

152 5913-Trimethyl-242526-trinoroleanan-3-ol Friedelan-3120573-ol

HO

Aerial(Tanzania) [18]

153 5913-Trimethyl-242526-trinoroleanan-3-one

Friedelinfriedelan-3-one

O

Aerial(Tanzania) [18]

1542610151923-

Hexamethyl-2610141822-tetracosahexaene

Squalene

Aerial(Tanzania)Leaves (Notstated)Whole(Taiwan)

[18 33 57]

155 120573120573-Carotene 120573-Carotene Leaves (Notstated) [97]

Evidence-Based Complementary and Alternative Medicine 29

studies [83] Hot water extracts of B pilosa varminor Sheriffwere also assessed for its antileukemic effects on leukemiccell lines L1210 U937 K562 Raji and P3HR1 using XTT-based colorimetric assays The extract inhibited the five celllines with IC

50values ranging from 145120583gmL to 586120583gmL

L1210 K562 Raji and P3HR1 were more sensitive to B pilosaextract with IC

50values below 200120583gmL [98]

Consistent with the antitumor activities of B pilosaextracts and fractions some of its phytochemicals alsoshowed anticancer activity as outlined in Table 11 Amongthem luteolin (103) a well-studied flavonoid with multiplebioactivities was more effective against tumor cell prolifera-tion than its derivatives with IC

50values ranging from 3 120583M

to 50120583M in cells and 5 to 10mgkg in animals Luteolin wasalso found to fight cancer as a food additive at concentrationsof 50 to 200 ppm [78] and prevent skin cancer [78] and cancer3invasion [103] Lee and colleagues reported that luteolinprevents cancer by inhibiting cell adhesion and invasion[103] Significant inhibition concentration was reported to be5 120583M and complete inhibition concentration was reported tobe 40120583M Moreover luteolin was reported to inhibit hepa-tocyte growth factor (HGF)-induced cell scattering as wellas cytoskeleton changes such as filopodia and lamellipodiawhichwas determined using phase-contrast and fluorescencemicroscopy Furthermore luteolin also inhibited the HGF-induced phosphorylation of c-Met ERK 12 and Akt aswell as the MAPKERK and P13K-Akt pathways [78 103]Other mechanisms underlying the anticancer activities ofluteolin are the inhibition of topoisomerase I and II whichinhibits cell replication and DNA repair thus promotingapoptosis regulation of PI-3-KinaseAktMAPKERKJNKactivation of apoptosis in the mitochondrial pathway byactivating caspase 9 and caspase 3 which were found inmalignant cells but not in normal human peripheral bloodmononuclear cells death receptor-induced apoptosis and acell-cycle arrest mechanism inhibition of fatty acid synthasewhich is upregulated in many cancer cells and sensitizationto chemotherapywhereby luteolin increases the susceptibilityof cancer cells to chemotherapy [78] Butein (82) is anotherflavonoid that showed a cytotoxic effect on human colonadenocarcinoma cell proliferation with a reported IC

50value

of 175 120583M Butein at 2 120583M affected the incorporation of[14C]-labeled leucine thymidine and uridine which cancause the inhibition of DNA RNA and protein synthesis ofhuman colon cancer cells Moreover butein also exhibitednoncompetitive inhibition of 1-chloro-24-dinitrobenzene(CDNB) in glutathione S-transferase (GST) activity Tumorresistance was correlated with high levels of GST thus buteininhibited proliferation of cancer cells [80] Another flavonoidpresent in B pilosa centaureidin (109) also showed anti-cancer activity in B lymphoma cells Centaureidin isolatedfrom Polymnia fruticosa inhibited tubulin polymerization invitro and induced mitotic figure formation in CA46 Burkittlymphoma cells Using turbidimetric assay the IC

50value of

centaureidin in inhibition of mitosis was 3120583M [104] Cyto-toxicity of centaureidin was further analyzed using AmericanNational Cancer Institute (NCI) 60 human tumor cell linesThe cytotoxicity potency of centaureidin expressed as GI

50

(50 growth inhibition in the NCI tumor line panel) was024120583M [81] These data mark centaureidin as a promisingantimitotic agent for tumor therapy

In addition to anti-tumor flavones polyynes found in B 4pilosa have also been shown to possess anti-tumor proper-ties Based on a bioactivity-directed isolation approach Wuand colleagues identified two polyyne aglycones from theethyl acetate fraction of B pilosa [44] 12-Dihydroxytrideca-57911-tetrayne (48) and 13-Dihydroxy-6(E)- tetradecene-81012-triyne (46) exhibited significant anticell prolifera-tion activity in primary human umbilical vein endotheliumcells (HUVEC) with IC

50values of 125 120583M and 173 120583M

respectively They also decreased angiogenesis and pro-moted apoptosis in human endothelial cells Their anti-angiogenic and cytotoxic effects correlated with activationof the CDK inhibitors and caspase-7 [44] In addition 12-Dihyroxy-5(E)-tridecene-7911-triyne (45) showed antian-giogenic effects in HUVECs with an IC

50value of 124 120583M as

evidenced by a decrease in the tube formation and migrationof HUVECs [43] The IC

50value of compound 45 in the

inhibition of basic fibroblast growth factor-induced HUVECgrowth was 282120583M However it had higher IC

50values

than those for lung carcinoma cells and keratinocytes Thiscompound could also inhibit cell proliferation of HUVECslung carcinoma A549 cells and HACAT keratinocytes Themechanism by which compound 45 inhibits HUVEC growthand angiogenesis is complicated and includes decreasing theexpression of cell cycle regulators (CDK4 cyclins D1 and Aretinoblastoma (Rb) and vascular endothelial growth factorreceptor 1) caspase-mediated activation of CDK inhibitorsp21 (Cip1) and p27 (Kip) upregulation of Fas ligand expres-sion downregulation of Bcl-2 expression and activation ofcaspase-7 and poly (ADP-ribose) polymerase [43]

32 Anti-Inflammatory Activity B pilosa is commonly usedto treat inflammatory disorders The anti-inflammatory phy-tochemicals present in B pilosa are listed in Table 11 5Cyclooxygenase-2 (COX-2) is a physiologically importantenzyme that converts arachidonic acid to prostaglandin(PGE2) Its expression is induced by a wide variety of external

stimuli indicating its involvement in inflammatory diseasesand it is used as an inflammatory marker [88] Yoshida andcolleagues studied the effects of the aqueous extracts of Bpilosa aerial parts in the production of COX-2 and PGE

2as

well as on the activation of mitogen activated protein kinases(MAPKs) in normal human dermal fibroblasts (HDFs) inresponse to inflammatory cytokine IL-1120573 This work showedthat IL-1120573 activated MAPKs such as ERK12 p38 and JNKto different extents and induced COX-2 expression TheCOX-2 expression in HDFs was regulated mainly by p38following IL-1120573 stimulation Consistently the p38 inhibitorSB203580 blocked this expression Using this cell platformB pilosa extracts were tested for inhibition of inflammationThe extract dose-dependently suppressed the activation ofp38 and JNK and moderately suppressed ERK12 as wellas suppressing COX-2 expression and PGE

2production

30 Evidence-Based Complementary and Alternative Medicine

Table7Ph

enylprop

anoids

isolatedfro

mB

pilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

156

3-(4-H

ydroxyph

enyl)-2-prop

enoica

cid

p-Cou

maricacid

OHO

OH

Who

le(Ja

pan)

[99]

157

2-Metho

xy-4-(2-prop

en-1-yl)-ph

enol

Eugeno

lOOH

Leaves

androots(Japan)

[99]

158

3-(4-H

ydroxy-3-m

etho

xyph

enyl)-

2-prop

enoica

cid

Ferulic

acid

O

OH

O

HO

Who

le(Ja

pan)

[99]

159

3-(34-D

ihydroxyph

enyl)-2-prop

enoic

acid

Caffeicacid

O

HO

OH

OH

Who

leandaeria

l(Japan)

[7599]

160

3-Prop

yl-3[(245-trim

etho

xyph

enyl)-

metho

xy]-24-pentanedione

3-Prop

yl-3-(245-

trim

etho

xy)benzyloxy-

pentan-24-dione

O

O

O

O

O

O

Leaves

(India)

[100]

161

3-(34-D

ihydroxyph

enyl)-2-

prop

enoica

cidethyleste

rCa

ffeateethyl

O

O

OH

OH

Not

foun

d(Taiwan)

who

le(Taiwan)

[434584]

Evidence-Based Complementary and Alternative Medicine 31Ta

ble7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

162

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

d-Erythron

icacid

2-O-caffeoyl-2-C

-methyl

O

HO

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

163

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

2-O-caffeoyl-2-C

-methyl

O

O

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

164

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-24-

dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

3-O-caffeoyl-2-C

-methyl

O

O

HO

HO

O

O

OH

OH

Leaves

(Japan)

[101]

165

4-(A

cetyloxy)-3-[[3-(34-

dihydroxyphenyl)-

1-oxo-2-propen-1-y

l-]oxy]-2-hydroxy-

2-methyl-b

utanoica

cid

O

HO

OH

O OHO

HO

OO

Not

foun

d(Ja

pan)

[73]

166

3-(34-D

ihydroxyph

enyl)-tetrahydro-4-

hydroxy-4-

methyl-5

-oxo-3-fu

ranyl

ester-2-prop

enoica

cid

3-O-C

affeoyl-2-C

-methyl-

D-erythrono

-14-la

cton

eHO

HO

O

OO

OOH

Leaves

(Japan)

[101]

167

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]-oxy]-145-

trihydroxy-cyclohexanecarboxylicacid

Chlorogenica

cid

O

HO

HO

OH

O

OH

O

OH

HO

Aeria

l(Japan)w

hole

(Taiwan)

[707175]

32 Evidence-Based Complementary and Alternative Medicine

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

168

4-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]oxy]-

135-tr

ihydroxy-cyclohexanecarboxylic

acid

4-O-C

affeoylqu

inicacid

OHO

HO

OH

OH

O

O

OH

OH

Aeria

l(Japan)

[71]

169

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-15-

dihydroxy-cyclo

hexanecarboxylicacid

34-Di-O

-caffeoylqu

inic

acid

HO

HO

O

O

O

O

OH

OH

HO

OH O

H

O

Aeria

l(Japan)W

hole

(Taiwan)

[45497071]

170

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-14-dihydroxy-

cyclo

hexanecarboxylicacid

35-Di-O

-caffeoylqu

inic

acid

HO

HO

O O

HO

O

HO

OH OO

OHOH

Aeria

l(Japan)w

hole

(Taiwan)

[45497071]

Evidence-Based Complementary and Alternative Medicine 33

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

171

34-bis[[(2E

)-3-(34-D

ihydroxyph

enyl)-1-

oxo-2-prop

en-1-

yl]oxo]-15

-dihydroxy-

cyclo

hexanecarboxylicacid

45-Di-O

-caffeoylqu

inic

acid

OHO

OH

HO

O

O

OH

OH

O

OH

HO

OWho

le(Taiwan)

[454970]

172

3-[4-[[-6-O-[3-(4-H

ydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]-phenyl]-2-prop

enoica

cid

120573-D

-p-C

oumaricacid4-O

-(6-O

-p-caoum

aroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HOH

HO

O

O

Leaves

(Japan)

[59]

173

3-[4-[[2-O

-Acetyl-6

-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]ph

enyl]2-propeno

icacid

120573-D

-p-C

oumaricacid4-O

-(2-O

-acetyl-6

-O-p-

caou

maroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HO

HO

O

O

Leaves

(Japan)aerial

(China)

[3459]

174

67-Dihydroxy-2-chrom

enon

eEsculetin

cicho

rigenin

OHO HO

O

Not

foun

d(Egypt)

[39]

34 Evidence-Based Complementary and Alternative Medicine

Table 8 Aromatic compounds isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

175 12-Benzenediol PyrocatechinHO

HOWhole(Japan) [99]

176 4-Ethyl-12-benzenediol Pyrocatechol

OH

OH

Whole(Japan) [99]

177 Dimethoxyphenol26-dimethoxyphenol

O

HO

O

Roots (Japan) [99]

178 4-Ethenyl-2-methoxy-phenol p-Vinylguaiacol

O

OHWhole(Japan) [99]

179 2-Hydroxy-6-methylbenzaldehyde

6-Methyl-salicylaldehyde O

OH

Whole(Japan) [99]

180 Benzene-ethanol 2-Phenyl-ethanolHO

Whole(Japan) [57]

181 4-Hydroxy-3-methoxy-benzaldehyde Vanillin O

O

HOAerial(Japan) [99]

182 3-Hydroxy-4-methoxy-benzaldehyde Vanillin iso

OO

OH

Leaves(Japan) [99]

183 4-Hydroxybenzoic acid p-Hydroxybenzoicacid

O

HOOH Whole

(Japan) [99]

184 2-Hydroxybenzoic acid Salicylic acidO

HO

HO

Stem androots (Japan) [99]

185 34-Dihydroxybenzoicacid Protocatechuic acid

O

HOOH

OH

Whole(Japan) [99]

1864-Hydroxy-

3methoxybenzoicacid

Vanillic acid

OOH

O

OH

Aerial(Uganda)

Roots (Japan)[68 99]

Evidence-Based Complementary and Alternative Medicine 35

Table 8 Continued

S N IUPAC names Common names Structure Plant part(country) References

187 345-Trihydroxybenzoicacid Gallic acid

O

OHHO

HO

HO

Whole(China) [102]

[88] This work supports the use of B pilosa as an anti-inflammatory agent however no compounds responsible forthe anti-inflammatory activity of B pilosa were identified

A further study also reported the anti-inflammatoryactivity as well as the antiallergic activity of B pilosa [75] Inthis study dried powder of the aerial part of B pilosa whichhad been pretreated with the enzyme cellulosine was usedfor further tests The results showed that oral administrationof the cellulosine-treated B pilosa lowered the level of serumIgE in mice 10 days after immunization with DNP (24-dintrophenyl)-Ascaris as an antigen This treatment alsoreduced dye exudation in skin induced by passive cutaneousanaphylaxis and production of inflammatory mediators his-tamine and substance P in rats [75] Phytochemical analysisshowed that cellulosine treatment increased the percentage ofcaffeic acid and flavonoids This study suggests that B pilosaand its phenolics have anti-inflammatory functions

Phenolics and polyynes are major anti-inflammatoryphytochemicals present in B pilosa (Table 11) Unsurpris-6ingly phenolics such as luteolin (103) and ethyl caffeate(161) that are major constituents of B pilosa have alsobeen reported to possess anti-inflammatory activity Lute-olin was reported to exhibit anti-inflammatory activity inmacrophages Xagorari and colleagues showed that luteolininhibited the release of inflammatory cytokines TNF-120572 andinterleukin-6 in RAW 2647 cells following LPS stimulation[87] It inhibited TNF-120572 production with an IC

50value

of 1120583M The underlying anti-inflammatory mechanism ofluteolin was reported to be the inactivation of Akt andNF-120581B activation [87] In addition luteolin was reportedto confer anti-inflammatory activity through inhibition ofLPS-stimulated iNOS expression in BV-2 microglial cellsIt inhibited LPS-activated microglia in a dose-dependentmanner with an IC

50value of 69 120583MMoreover immunoblot

and RT-PCR data proved that luteolin suppressed I120581B-120572degradation and iNOS expression in LPS-activated microglia[85] Kim and colleagues stated that luteolin may havebeneficial effects on inflammatory neural diseases throughinhibition of iNOS expression [85] A related study revealedthat luteolin decreased the transcriptional activity of NF-120581BRelA via partial inhibition of TNF-mediated NF-120581B DNAbinding activity Luteolin also inhibited Akt phosphorylationand induced degradation of a transcription factor interferonregulatory factor (IRF) [86]

Chiang and colleagues showed that ethyl caffeate(161) significantly inhibited NO production in mousemacrophages RAW 2647 cells [84] Based on MTT assays

they concluded that this inhibition was not due to thecytotoxicity of ethyl caffeate The IC

50value of ethyl caffeate

in the inhibition of NO production was 55 120583gmL slightlylower than curcumin (positive control) which has anIC50

value of 65120583gmL They demonstrated that ethylcaffeate exerted anti-inflammatory activity via the reducedtranscription and translation of iNOS (inducible nitric oxidesynthase) in RAW 2467 cells In addition this compoundalso suppressed COX-2 expression in RAW 2467 cellsand MCF-7 cells The in vivo anti-inflammatory effectof ethyl caffeate was verified by testing in TPA-treatedmouse skin Like celecoxib the positive control ethylcaffeate significantly abolished COX-2 expression in adose-dependent manner Ethyl caffeate at 1mg200120583Lsite(24mM) inhibited COX-2 expression at a level comparableto celecoxib at 1mg200120583Lsite (13mM) Remarkably ethylcaffeate at 48mM (2mg200120583Lsite) was more effectivethan celecoxib at 131mM (10mg200120583Lsite) Moreoverthis compound inhibited the activation of nuclear factor-120581B(NF-120581B) by LPS via the prevention of NF-120581B binding toDNA [84] In addition 3 ethyl caffeate analogs (ethyl 34-dihydroxyhydrocinnamate ethyl cinnamate and catechol)also showed different degrees of NF-120581B binding to DNA aslisted in Table 12 Ethyl cinnamate lacks a catechol moietywhich results in ineffective inhibition of NF-120581B binding toDNA [84]

Pereira and colleagues assessed the anti- 7inflammatory and immunomodulatory activities ofB pilosa methanol extract as well as one polyyne2-O-120573-glucosyltrideca-11(E)-en-3 579-tetrayn-12-diol(54) in T lymphocytes and a zymosan-induced arthritismouse model [27] They first examined the in vitro effect of 8the B pilosa extract and compound 54 on cell proliferation ofhuman T cells stimulated with 5120583gmL phytohemagglutinin(PHA) or 100 nM 12-O-tetradecanoyl phorbol-13-acetate(TPA) plus 15 120583M ionomycin and on cell proliferation ofmouse T cells stimulated with 5 120583gmL concanavalin A(Con A) The data demonstrated that both methanol extractand compound 54 suppressed T-cell proliferation in adose-dependent manner The estimated IC

50values of the B

pilosa extract against human T cells stimulated with 5120583gmLPHA and 100 nM TPA plus 15120583M ionomycin were 125 and25 120583gmL respectively In comparison with the methanolextract compound 54 showed 10-fold more inhibition ofhuman T-cell proliferation with an estimated IC

50value of

15 120583gmL Accordingly the B pilosa extract and compound54 dose-dependently suppressed mouse T-cell proliferation

36 Evidence-Based Complementary and Alternative Medicine

Table 9 Porphyrins isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

188

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(15S16S)-10-ethenyl-5-ethyl-1161820-tetrahydro-6111522-tetramethyl-1820-dioxo-15H-912-imino-212-

metheno-471714-dinitrilopyrano[43-

b]azacyclononadecine-16-propanoic

acid

AristophyllC

NH N

HNN

O

O OOOPhytyl

O

Leaves(Taiwan) [100]

189

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -20b-(methoxycarbonyl)-9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-dinitrilo-12-

dioxeto-[310158404101584034]-cyclo-pent[1 2]azacyclononadecine-

19-propanoic acid

Bidenphytin A

NH N

HNN

OPhytylO O O

O O

Leaves(Taiwan) [100]

190

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -2a-

hydroxy-20b-(methoxycarbonyl)-

9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-

dinitrilo-12-dioxeto[310158404101584034]-cyclo-pent[12-b]-

azacyclononadecine-19-propanoic acid

Bidenphytin B

NH N

HNN

OPhytylO O O

O O

OH

Leaves(Taiwan) [100]

191

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO O

HOOO

Leaves(Taiwan) [100]

192

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

Evidence-Based Complementary and Alternative Medicine 37

Table 9 Continued

S N IUPAC names Common names Structure Plant part(country) References

193

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

Leaves(Taiwan) [100]

194

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

195

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3S4S21R)-9-ethenyl-14-ethyl-21-(methoxycarbonyl)-

481318-tetramethyl-20-oxo-3-phorbinepropanoic acid

Pheophytin A

NH N

HNN

PhytylO

O

OO O

Leaves(Taiwan) [100]

with estimated IC50values of 30 and 25 120583gmL respectively

Taken together the data indicate that the B pilosa extractand compound 54 act on human and mouse T cells Totest the in vivo effect of the B pilosa extract and compound54 a zymosan-induced arthritis mouse model was usedThis model was established from B10ASgSnJ mice with aninjection of zymosan (015mg) The zymosan-injected mice9received an intraperitoneal injection of the B pilosa extract(1 5 or 10mg) at one dose a day for 5 days Popliteal lymphnode (PLN) weight was monitored to check the developmentof arthritis The results revealed that 10mg of the methanolextract of B pilosa extract could significantly diminishinflammation as evidenced by PLN weight [27] This worksuggests that B pilosa (and compound 54) can suppressimmune response and inflammation

33 Antidiabetic Activity Anti-diabetic agents are primarilydeveloped from plants and other natural resources [8 24 4951 53] B pilosa is one of 1200 plant species that have beeninvestigated for antidiabetic activity [120 121]B pilosa is usedas an anti-diabetic herb in America Africa and Asia [51 120

122] Many studies have indicated that B pilosa could treattype 1 diabetes (T1D) and type 2 diabetes (T2D) in animals

Etiologically speaking T1D is caused by autoimmune-mediated destruction of pancreatic 120573 cells leading to insulindeficiency hyperglycemia and complications Currentlythere is no cure for T1D Polarization ofTh cell differentiationcontrols the development of T1D Suppression of Th1 celldifferentiation and promotion of Th2 cell differentiationameliorate T1D [123] One study showed that the butanolfraction of B pilosa inhibited T-cell proliferation decreasedTh1 cells and cytokines and increased Th2 cells andcytokines leading to prevention of T1D in nonobesediabetic (NOD) mice [49] Based on a bioactivity-directedisolation strategy 3 polyynes 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne (53) also known ascytopiloyne 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D- Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne (50) were identifiedfrom B pilosa [49 53] The IC

50value of the butanol fraction

was 200120583gmL This inhibition was reported to be partiallyattributed to cytotoxicity because the butanol fraction at

38 Evidence-Based Complementary and Alternative Medicine

Table 10 Other compounds isolated from B pilosa [30]

SN IUPAC names Common names Structure Plant part(country) References

19637-Dihydro-137-

trimethyl-1H-purine-26-dione

CaffeineN

NN

NO

O

Aerial(Uganda) [68]

197

1-((2R4S5R)-Tetrahydro-4-hydroxy-5-(hydroxymethyl)furan-2-yl)-5-

methylpyrimidine-24(1H3H)-dione

ThymidineHO

HO

ON

O

NHO Not found

(China) [37]

198 1-(2-Thienyl)-ethanone 2-Acetyl-thiophene

S

O

Roots(Germany) [41]

199

(2R3S4S5S)-2-(Heptan-2-yloxy)-tetrahydro-6-

((tetrahydro-34-dihydroxy-5-

(hydroxymethyl)furan-2-yloxy)methyl)-

2H-pyran-345-triol

Heptanyl2-O-120573-xylofuranosyl-

(1rarr 6)-120573-glucopyranoside

O

OOH

OH

HO

OHO

H

HO

H

H

HOHH

O

Whole(Taiwan) [70]

200

2-[(3R7R11R)-3-Hydroxy-371115-

tetramethylhexadecyl]-356-trimethyl-

25-cyclohexadiene-14-dione

120572-Tocopherylquinone

OO

HO

Whole(Taiwan) [32]

2017-O-(410158401015840610158401015840-Diacetyl)-

120573-D-glucopyranoside

Not found Leaves(China) [60]

180 120583gmL could cause 50 death of Th1 cells Moreoverthis study suggested that the butanol fraction may preventdiabetes in NOD mice in vivo via downregulation of Th1cells or upregulation Th2 cells that have effects which areantagonistic of those of Th1 cells [49] This was proven byintraperitoneal injection of the butanol fraction at a doseof 3mgkg BW 3 times a week to NOD mice from 4 to 27weeks This dosage resulted in lower incidence of diabetes(33) At a dose of 10mgkg the butanol fraction of Bpilosa totally eliminated (0) the initiation of the diseaseTo further support this result assessment of IgG2a and IgEproduction was performed in the serum of NOD mice Asin vivo results obtained from intracellular cytokine stainingexperiments were not very conclusive levels of IgG2a and IgE

were measured since Th1 cytokine IFN120574 and Th2 cytokineIL-4 favor the production of IgG2a and IgE respectively Asexpected high levels of IgE and some decline in the levels ofIgG2a were observed in the serum Profiling of the butanolextract revealed five compounds 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne(50) 45-Di-O-caffeoyl- quinic acid 35-Di-O-caffeoylquinicacid and 34-Di-O-caffeoylquinic acid Only the first twocompounds showed similar effects on the prevention ofdiabetes in NOD mice as the B pilosa butanol fractionMoreover compound 50 showed greater activity thancompound 69 in terms of enhancement (by 34 comparedto 8) of differentiation of Th0 to Th2 at 15 120583gmL (both

Evidence-Based Complementary and Alternative Medicine 39

Table 11 Chemical constituents of B pilosa and their biological activities

SN Name Classification Molecularformula Biological activities

109 Centaureidin [91] Flavonoid C18H16O8Anti-listerial [69 91]

Cytotoxic [10]

110 Centaurein [91] Flavonoid C24H26O13

Anti-listerial [69 91]Cytotoxic [10]Anti-viral [105]

103 Luteolin [106] Flavonoid C15H10O6

Anti-viral [107 108]Cytotoxic [77]

Anti-inflammatory [109]Anti-allergic [109]

82 Butein [110] Flavonoid C15H12O5Anti-leishmanial [111]

Cytotoxic [78]48 12-Dihydroxytrideca-57911-tetrayne [112] Polyyne C13H12O2 Anti-angiogeneic [112]46 13-Dihyroxy-6(E)-tetradecene-81012-triyne [112] Polyyne C14H16O2 Anti-angiogeneic [112]

45 12-Dihyroxy-5(E)-tridecene-7911-triyne [113] Polyyne C13H14O2Anti-angiogeneic [113]Anti-proliferative [113]

64 1-Phenylhepta-135-triyne [114] Polyyne C13H8

Anti-microbial [115]Anti-malarial [3]Cytotoxic [3]Antifungal [15]

27 Linoleic acid [79] Fatty acid C18H32O2Anti-viral (100)Cytotoxic [17]

161 Ethyl caffeate [84] Phenylpropanoid C11H12O4 Anti-inflammatory [84]

54 2-O-120573-Glucosyltrideca-11(E)-en-3579-tetrayn-12-diol [26] Polyyne C19H20O7Immunosuppressive andAnti-inflammatory [26]

53 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne [50] Polyyne C19H22O7Anti-diabetic [116]

Anti-inflammatory [50]

69 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne [28] Polyyne C20H26O7Anti-diabetic [28]

Anti-inflammatory [94]

50 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne [28] Polyyne C19H24O7

Anti-diabetic [28]Anti-inflammatory [94]

Anti-malarial andantibacterial [47]

129 Quercetin 3-O-120573-D-galactopyranoside [18] Flavonoid C21H20O12 Anti-inflammatory [117]

170 35-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

171 45-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

169 34-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

126 Quercetin 331015840-dimethyl ether7-O-120572-L-rhamnopyranosyl-(1rarr 6)-120573-D-glucopyranoside [118] Flavonoid C21H20O11 Anti-malarial [119]

125 Quercetin 331015840-dimethyl ether-7-O-120573-D-glucopyranoside [118] Flavonoid C21H20O12 Anti-malarial [119]70 1-Phenyl-13-diyn-5-en-7-ol-acetate [27] Polyyne C15H12O2 Anti-malarial [27]199 Heptanyl 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside [70] Miscellaneous C18H44O10 Antioxidant [70]124 3-O-Rabinobioside [70] Saccharide C27H38O15 Antioxidant [70]130 Quercetin 3-O-rutinoside [70] Flavonoid C27H38O15 Antioxidant [70]167 Chlorogenic acid [70] Phenolic C16H26O9 Antioxidant [70]119 Jacein [70] Flavonoid C24H26O13 Antioxidant [70]

49 (R)-12-dihydroxytrideca-357911-pentayne [47] Polyyne C13H8O2Anti-malarial andAntibacterial [47]

40 Evidence-Based Complementary and Alternative Medicine

Table 12 Structure and activity relationship studies of ethyl caffeateusing in vitro NF-120581BDNA binding assays [84]

Compound Concentration(120583M)

NF-120581BDNA binding

Ethyl caffeate 50 100inhibition

Ethyl 34-dihydroxyhydrocinnamate 100 100inhibition

Catechol 400 100inhibition

Ethyl cinnamate 400 Noinhibition

Table 13 Apoptosis in cocultures of T cells and pancreatic 120573 cells[53]

Cellmedium apoptosis andnecrosis

CD4+ T cellscontrol medium lt4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 2

CD4+ T cellswith cytopiloyne-treated 120573 cells 18CD4+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 7

CD8+ T cellscontrol medium 4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 4

CD8+ T cellswith cytopiloyne-treated 120573 cells 4CD8+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 4

Table 14 Radical scavenging activities of B pilosa extracts [70]

Extractscontrol DPPH assayIC50 (120583gmL)

NBThypoxanthinesuperoxide assayIC50 (120583gmL)

Quercetin 198 15Ascorbic acid 634 Not determined120572-tocopherol 897 Not determinedEthyl acetateextract 1383 597

Butanol extract 1669 114Water extract gt100 gt100

compounds) and inhibition (by 40 compared to 10) ofdifferentiation toTh1 at the same concentration [49]

Among the three polyynes found in B pilosa cytopiloyne(53) had the most potent anti-T1D activity [53] To test the invivo effect of cytopiloyne NODmice received intraperitonealor intramuscular injection of cytopiloyne at 25120583gkg BW3 times per week Twelve-week-old NOD mice started todevelop T1D and 70 of NOD mice aged 23 weeks andover developed T1D Remarkably 12- to 30-week-old NODmice treated with cytopiloyne showed normal levels of bloodglucose (lt200mgdL) and insulin (1-2 ngmL) Consistent

Table 15 Radical scavenging activity of secondarymetabolites fromB pilosa [70]

Metabolite (Table 11)control DPPH assay IC50 (120583gmL)199 Not determined124 53130 68167 105169 33171 38119 Not determined110 Not determinedQuercetin 256Caffeic acid 890

Table 16 Antioxidant activity of the essential oils and water extractsfrom B pilosa [92]

Extract IC50 (120583gmL)Leaf essential oils 47Flower essential oils 50Leaf extract 61Flower extract 172

with T1D incidence cytopiloyne delayed and reduced theinvasion of CD4+ T cells into the pancreatic islets [53]

In vitro study showed that cytopiloyne (53) inhibited thedifferentiation of naıveTh (Th0) cells (ie CD4+ T cells) intoTh1 cells and promoted differentiation of Th0 cells into Th2cells [50] The in vitro data are consistent with the in vivoresults indicating that cytopiloyne reducedTh1differentiationand increased Th2 differentiation as shown by intracellularcytokine staining and FACS analysis [53] In line with theskewing of Th differentiation the level of serum IFN-120574 andIgG2c decreased while that of serum IL-4 and serum IgEincreased compared to the negative controls (PBS-treatedmice) Cytopiloyne also enhanced the expression of GATA-3 a master gene for Th2 cell differentiation but not theexpression of T-bet a master gene forTh1 cell differentiationfurther supporting its role in skewingTh differentiation [53]

Also importantly cytopiloyne partially depleted CD4+rather than CD8+ T cells in NOD mice [53] As shown inTable 13 coculture assays showed that the depletion of CD4+T cells was mediated through the induction of Fas ligandexpression on pancreatic islet cells by cytopiloyne leading toapoptosis of infiltrating CD4+ T cells in the pancreas via theFas and Fas ligand pathways However cytopiloyne did notinduce the expression of TNF-120572 in pancreatic islet cells andthus had no effect on CD8+ T cells [53]

In addition Chang and colleagues showed that cytopi-loyne dose-dependently inhibited T-cell proliferation stimu-lated by IL-2 plus Con A or anti-CD3 antibody using [3H]thymidine incorporation assay [53]

Overall the mechanism of action of cytopiloyne andprobably its derivatives in T1D includes inhibition of T-cellproliferation skewing of Th cell differentiation and partialdepletion of Th cells Due to the anti-diabetic mechanisms

Evidence-Based Complementary and Alternative Medicine 41

Table 17 Antibacterial activity of essential oils and flower extracts from B pilosa [92]

Strain Mean zone of inhibition (mm)Leaf essential oil Flower essential oil Leaf extract Flower extract

Micrococcus flavus 127 plusmn 03 87 plusmn 03 102 plusmn 02 108 plusmn 03Bacillus subtilis 173 plusmn 19 117 plusmn 02 109 plusmn 02 103 plusmn 02Bacillus cereus 190 plusmn 14 112 plusmn 03 118 plusmn 04 185 plusmn 10Bacillus pumilus 123 plusmn 07 108 plusmn 02 105 plusmn 04 77 plusmn 02Escherichia coli 137 plusmn 04 203 plusmn 07 102 plusmn 11 140 plusmn 13Pseudomonas ovalis 125 plusmn 08 137 plusmn 15 102 plusmn 06 125 plusmn 06

Table 18 Antibacterial activity of root extracts from B pilosa [93]

Strain MIC50 (mgmL)Methanol extract Acetone extract

Bacillus cereus 10 mdashEscherichia coli 5 5Klebsilla pneumonia 5 10Micrococcus kristinae 10 mdashPseudomonas aeruginosa 10 10Staphylococcus aureus 5 10Sraphylococcus epidermidis 5 5Serratia marcescens 10 mdashShigelea flexneri 10 mdashStreptococcus faecalis 10 mdash

Table 19 Antibacterial activity of B pilosa of compound 29 [47]

Strain MIC50 (120583gmL)Escherichia coli NIHJ 1Escherichia coli ATCC25922 1Klebsiella pneumoniae ATCC700603 128Serratia marcescens ATCC13880 16Pseudomonas aeruginosa ATCC27853 8Staphylococcus aureus FDA209P 05Staphylococcus aureus ATCC29213 025Staphylococcus aureus N315 (MRSA) 05Enterococcus faecalis ATCC29212 2Enterococcus faecalis NCTC12201 (VRE) 1Bacillus subtilis ATCC6633 05Candida albicans ATCC10231 025

of action it was hypothesized that cytopiloyne protectsNOD mice from diabetes by a generalized suppression ofadaptive immunity To evaluate this hypothesis ovalbumin(Ova) was used as a T-cell dependent antigen to primeNOD mice which had already received cytopiloyne orPBS vehicle Ova priming boosted similar anti-Ova titersin cytopiloyne-treated mice and PBS-treated mice but adifference in immunoglobulin isotype was observed in thetwo groups Thus it was concluded that cytopiloyne is animmunomodulatory compound rather than an immunosup-pressive compound [50 53]

T2D is a chronic metabolic disease with serious com-plications resulting from defects in either insulin secretion

insulin action or both [124] A study by Ubillas et alshowed that the aqueous ethanol extract of the aerial partof B pilosa at 1 gkg body weight (BW) lowered bloodglucose in dbdb mice a T2D mouse model [51] Basedon a bioactivity-guided identification compounds 69 and50 were identified Further the mixture of the compounds(69 50) in a 2 3 ratio significantly decreased blood glucoseconcentration and reduced food intake on the second dayof treatment when administered at doses of 250mgkg twicea day to C5BLKs-dbdb mice When tested at 500mgkga more substantial drop in blood glucose level as wellas the stronger anorexic effect (food intake reduced from58 gmouseday to 25 gmouseday) was observed [51] Inthis study it was suggested that the blood glucose lower-ing effect of B pilosa was caused in part by the hungersuppressing effect of its polyynes [51] However the hungersuppressing effect of the ethanol extract of B pilosa wasnot found in the studies described below In another study[24] water extracts of B pilosa (BPWE) were used indiabetic dbdb mice aged 6ndash8 weeks with postprandialblood glucose levels of 350 to 400mgdL Like oral anti-diabetic glimepiride which stimulates insulin release onesingle dose of BPWE reduced blood glucose levels from374 to 144mgdL The antihyperglycemic effect of BPWEwas inversely correlated to an increase in serum insulinlevels suggesting that BPWE acts to lower blood glucose viaincreased insulin production However BPWE had differentinsulin secretion kinetics to glimepiride [24] One flaw incurrent anti-diabetics is their decreasing efficacy over timeThe authors investigated the long term anti-diabetic effectof BPWE in dbdb mice BPWE reduced blood glucoseincreased blood insulin improved glucose tolerance andreduced the percentage of glycosylated hemoglobin (HbA1c)Both long-term and one-time experiments strongly supportthe anti-diabetic action of BPWE [24] In sharp contrast toglimepiride BPWE protected against islet atrophy in mousepancreas The investigators further evaluated anti-diabeticproperties of 3 B pilosa varieties B pilosa L var radiate(BPR) Bpilosa L var pilosa (BPP) and B pilosa L varminor(BPM) in dbdb mice [8] One single oral dose (10 50 and250mgkg body weight) of BPR BPP or BPM crude extractsdecreased postprandial blood glucose levels in dbdb micefor up to four hours and the reduction of glucose levels inthe blood appeared to be dose-dependent Comparing thethree variants BPR extract resulted in a higher reduction inblood glucose levels when administered at the same dose as

42 Evidence-Based Complementary and Alternative Medicine

Table 20 Antifungal activity of B pilosa [92]

Partextract Concentration (ppm) Strain InhibitionCortiicum rolfsii Fusarium solani Fusarium oxysporum

Leaves

Essentialoils

100 857 plusmn 09 682 plusmn 0 745 plusmn 17250 960 plusmn 08 779 plusmn 18 879 plusmn 04

AqueousExtracts

100 446 plusmn 17 605 plusmn 21 716 plusmn 07250 942 plusmn 03 689 plusmn 07 824 plusmn 19

Flowers

Essentialoils

100 604 plusmn 09 892 plusmn 04 869 plusmn 05250 894 plusmn 12 980 plusmn 03 949 plusmn 06

AqueousExtracts

100 331 plusmn 11 714 plusmn 07 573 plusmn 22250 661 plusmn 14 912 plusmn 0 900 plusmn 07

Table 21 Antifungal activity of B pilosa root extracts [93]

Strain LC50 (mgmL)Acetoneextracts

Methanolextracts

Waterextracts

Aspergillus niger 014 006 007Aspergillus flavus 1091 658 0Penicillium notatum 005 005 005

the other two varieties In terms of serum insulin levels a doseof 50mgkg of each extract was used and the BPR extracttogether with the three polyynes significantly increased theserum insulin level in dbdb mice Long-term experiments(28-day treatment) were then conducted using diabetic micewith postprandial glucose levels from 370 to 420mgdL andglimepiride was used as positive controlThe range of dosagesapplied was from 10mgkg BW to 250mgkg BW Resultsshowed that the positive control as well as the crude extractsof the three varieties lowered the blood glucose levels indbdb mice However only BPR extract containing a higherpercentage of cytopiloyne (53) reduced blood glucose levelsand augmented blood insulin levelsmore than BPP andBPMThe percentage of glycosylated hemoglobin A1c (HbA1c) wasalso measured and found to be 79 plusmn 05 in mice aged 10ndash12 weeks and 66 plusmn 02 61 plusmn 03 and 62 plusmn 03 inthe blood of age-matchedmice following treatment with BPRcrude extract (50mgkg) glimepiride (1mgkg) and com-pound 53 (05mgkg) respectively [8] Among the polyynesfound in B pilosa cytopiloyne was the most effective againstT2D Hence cytopiloyne was used for further study on anti-diabetic action and mechanism [125] The data confirmedthat cytopiloyne reduced postprandial blood glucose lev-els increased blood insulin improved glucose tolerancesuppressed HbA1c level and protected pancreatic islets indbdb mice Nevertheless cytopiloyne failed to decreaseblood glucose in streptoztocin (STZ)-treated mice whose bcells were already destroyed Additionally cytopiloyne dose-dependently increased insulin secretion and expression in bcells as well as calcium influx diacylglycerol and activationof protein kinase C120572 Collectively the mechanistic studiessuggest that cytopiloyne treats T2D via regulation of insulin

production involving the calciumDAGPKC120572 cascade in bcells

The studies detailed above point to the conclusion thatcytopiloyne and related polyynes (compounds 69 and 49)are anti-diabetics in animal models The data uncover a newbiological action of polyynes It should be noted that likeall anti-diabetic drugs cytopiloyne failed to prevent or curediabetes completely but reduced diabetic complications [125]Intriguingly 34 polyynes have been found in B pilosa so farIt remains to be seen whether all the polyynes present in thisplant have anti-diabetic activities

34 Antioxidant Activity Free radicals can damage cellularcomponents via a series of chemical reactions [89] leadingto development and progression of cardiovascular diseasecancer neurodegenerative diseases and ageing [70] Freeradicals nitric oxide (NO) and superoxide anions can beproduced in macrophages to kill microbes However anexcessive generation of the free radicals under pathologicalconditions is associated with a wide range of illnessesPlants are known to be rich in antioxidant phytochemicalsChiang and colleagues evaluated the free radical scaveng-ing activity of crude extract fractions and compounds ofB pilosa using 11-diphenyl-2-picrylhydrazyl (DPPH) andhypoxanthinexanthine oxidase assays [70] Using DPPHand hypoxanthinexanthine oxidase assays they found thatthe B pilosa crude extract and the ethyl acetate butanoland water fractions had free radical scavenging activityNine compounds Heptyl- 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside (199) 3-O-Rabinobioside (124) Quercetin3-O-rutinoside (130) Chlorogenic acid (167) 34-Di-O-caffeoylquinic acid (169) 35-Di-O-caffeoylquinic acid (170)45-Di-O-caffeoylquinic acid (171) Jacein (119) and Cen-taurein (110) had DPPH radical scavenging activity [70]The IC

50values of the B pilosa crude extractfractions and

compounds are summarized in Tables 14 and 15 respectivelyMeasurement of free radical scavenging activities is one

way of assessing the antioxidant activities of B pilosa andits fractions and compounds It is interesting that the ethylacetate and butanol fractions are more active than the waterfraction and B pilosa crude extract [70] Of the secondarymetabolites only phenolic compounds 124 130 167 169 and171 showed significant DPPH-radical scavenging activities

Evidence-Based Complementary and Alternative Medicine 43

Further analysis of the structure-activity relationship of thecompounds suggested that substitution of the C3 hydroxylgroup with glycosides increased the activity approximately2-fold (for example in compounds 124 and 130) relative toquercetin (-OH in its C3) [70] Further modification of thestructures of the active compounds needs to be performed totest the effects of various substituents on activity The reasonwhy most of the antioxidant compounds contain phenolmoieties in their structure could be that the reduction-oxidation (redox) properties of phenols allow them to actas reducing agents singlet oxygen quenchers and hydrogendonors

A complementary study by Muchuweti and colleaguesdetermined phenolic content antioxidant activity and thephenolic profile of B pilosa methanol extract [89] Theyestimated that the phenolic content of the methanol extractof B pilosawas 11028plusmn22mgg [89] Vanillin hydroxyben-zaldehyde caffeic acid coumaric acid and ferulic acid werefound in this extractThe B pilosa extract also showed DPPHradical scavenging activity Furthermore the antioxidantactivity of the flavonoids found in B pilosa was correlatedwith its hepatoprotective effects through their inhibitionof NF-120581B activation which may lessen the oxidative stresscaused by the production of free radicals during liver injury[60]This activity might also be due to the anti-inflammatoryeffects of the aqueous extracts of B pilosa aerial parts on theinhibition of COX-2 and PGE

2production [88]

Essential oils from B pilosa flowers and leaves are alsoreported to possess antioxidant activity With the aim ofreplacing chemically synthesized additives Deba and col-leagues [92] worked on the antioxidant antibacterial andantifungal activities of essential oils and water extracts of Bpilosarsquos leaves and flowers Table 16 summarizes the resultsobtained from DPPH free radical scavenging assay

It can be inferred fromTable 16 that essential oils from theleaves possessed the highest activity It is reported elsewherethat monoterpenes present in essential oils such that of Bpilosa have protective effects and antioxidant properties [92]Beta-carotene bleaching method was also performed Leavesessential oils and aqueous extracts of the leaves and flowersshowed higher activity than the flower essential oils This isdue to the volatility of the flower essential oils The activityexhibited by the aqueous extracts was accounted to thepresence of phenolic compounds that are reported to donatea hydrogen atom to free radicals such that the propagation ofthe chain reaction during lipid oxidation is terminated [92]Overall essential oils and phenolics present in B pilosa canbe thought of as major antioxidant compounds

35 Immunomodulatory Activity B pilosa is thought to bean immunomodulatory plant and is reported to be effectivein the treatment of immune disorders such as allergy [75]arthritis [75] and T1D [48 50 53]

As pointed out in the discussion of its anti-diabeticactivities (Section 33) a combination of phytochemicalsmethod and T-cell activation assays was used to studyimmunomodulatory properties of B pilosa IFN-120574 is a keycytokine released by T and NK cells that mediates immune

cells and sustains immunity against pathogens Defects inIFN-120574 expression regulation and activation result in vulner-ability to diseases caused by bacteria and viruses [69] Anelegant study performed byChang and colleagues using IFN-120574 promoter-driven luciferase reporter construct in JurkatT cells showed that hot water crude extracts of B pilosaincreased IFN-120574 promoter activity two-fold [69] Out of thesubfractions of this extract the butanol fraction but not thewater or ethyl acetate fractions increased IFN-120574 promoteractivity six-fold Centaurein (110) and centaureidin (109)were identified from the butanol fraction and were stated tocause a four-fold increase in IFN-120574 promoter activity withEC50

values of 75 120583gmL and 09120583gmL respectively Themechanism of action of centaurein was determined usingtranscription factors such as AP-1 NFAT and NF120581B whichare reported to bind to IFN-120574 promoter and regulate IFN-120574transcription Unlike with the activity of the positive controlPHA centaurein caused a four-fold increase in NFAT a3-fold increase in NF120581B and had little if any effect onAP-1 enhancer activities (23-fold three-fold and ten-foldincreases respectively were seenwith PHA) [69]The authorsconcluded that centaureinmodulates IFN-120574 expression by theNFAT and NF120581B pathways The article only determined themechanism of action of centaurein Its aglycone centaureidinmay act through the samemechanism though this conclusionneeds to be further verified

B pilosa extract and its compounds are reported to inhibitdifferentiation of naıve CD4+ helper T (Th0) cells into Th1cells [49] Using theThcell differentiation assay as a screeningplatform 3 polyynes 2-120573-D-glucopyranosyloxy-1 -hydroxytrideca-57911-tetrayne (53) 3-120573-D-glucopyranosyl-1-hydroxy-6(E)- tetradecene-81012-triyne (69) and 2-120573-D-glucopyranosyloxy-1-hydroxy-5(E)- tridecene-7911-triyne(49) were discovered from B pilosa [49 53] The data showsthat cytopiloyne and other two polyynes suppressed thedifferentiation of type 1 helper T (Th1) cells and productionof Th1 cytokines and promoted that of type 2 helper T (Th2)cells and production of Th2 cytokines thus explaining theimmunomodulatory and anti-inflammatory effects of Bpilosa and its polyynes

Chang and colleagues were the first to report the effect ofthe butanol extract of B pilosa on the autoimmune diabetesand airway inflammation inmice [48] Imbalance in the levelsofTh1 andTh2 and of various cytokines leads to autoimmunediseases T1D and other autoimmune diseases (rheumatoidarthritis Crohnrsquos disease among others) are exacerbated byan increase Th1 levels (specifically CD4+ Th1 cells) whileTh2 cells antagonize this effect [19] Moreover Th2 cellsmediate asthma in ovalbumin-induced hypersensitivity inBALBc mice [48] In their work [48] Chang and colleaguesshowed that 10mgkg butanol extracts with 15 (ww)compound 69 and 11 (ww) compound 49 (Table 11)ameliorated the development of Th1-mediated diabetes inNOD mice through inhibition of 120573 cell death and leukocyteinfiltration At the same dosage the butanol extracts alsoexacerbated ovalbumin-induced pulmonary inflammationin BALBc mice with an increase in the infiltration ofeosinophils and mast cells into the airway of the mice [48]Despite the different outcomes both mouse models proved

44 Evidence-Based Complementary and Alternative Medicine

the concept that control over the Th1Th2 shift is associatedwith autoimmune diseases and that the B pilosa butanolextract can shift the differentiation of Th0 cells to Th2 cells[48 49]

An extended study presented by Chiang and colleaguesshowed that compound 53 modulates T-cell functions [50]Using CD4+ T cells from BALBc mice they demonstratedthat cytopiloyne decreased levels of IFN-120574 producing cells(Th1) by 122 (from 72 to 598) Since Th1 and Th2 celldifferentiation is antagonistic it was expected that compound53 increased the percentage of mouse IL-4 producing cells(Th2) by 72 (from 237 to 309) Subsequent assess-ment of effect of compound 53 on the modulation of thetranscription of IL-4 and IFN-120574 showed that cytopiloyne asexpected decreased the splenocyte levels of IFN-120574 mRNAand increasing that of IL-4 in a dose-dependent manner Inthe range of 01 to 3 120583gmL of compound 53 the effects werenot attributed to its cytotoxicity Consequently using 3120583gmLcytopiloyne for 72 and 96 hours the protein concentration ofIFN-120574 decreased to 186 and 444 respectively Under thesame conditions cytopiloyne increased IL-4 concentrationsto 1985 and 2470 (for 72 and 96 hours resp) Thismodulation of T-cell differentiation exhibited by compound53 was used to explain its anti-diabetic activity [50]

The anti-diabetic role of cytopiloyne was extensivelydiscussed above (Section 33 anti-diabetic activity) Themolecular basis of the regulation of cytokine expression bycytopiloyne has been described Cytopiloyne directly elevatedthe expression level of IL-4 via GATA-3 upregulation inT cells [53] However reduction of IFN-120574 expression in Tcells seemed to come from the indirect opposing effect IL-4cytokine because the expression level of T-bet was unaltered[53] In this way cytopiloyne skewedTh1 polarization into theTh2 state conferring protection against T1D in NOD miceAside frompolarization ofThcell differentiation cytopiloynealso activated the expression of Fas ligand in pancreaticb cells this increase leading to the partial depletion of Tcells and reduction of immune response in local areas suchas the pancreas Of note cytopiloyne also inhibited T-cellproliferation and activation By targeting T cells from threeimmunomodulatory actions cytopiloyne protects againstT1D and probably other Th1-mediated autoimmune diseases[19]

The phytochemical constituents of B pilosa exert theirfunctions on different immune cells to modulate immuneresponse It is possible that some of the compounds mayhave agonistic or antagonistic effects on immune responseImmune function of B pilosa may depend on its compo-sition and amount of compounds which could explain theapparently conflicting report of B pilosa butanol extractaggravating allergy in mice [48] while cellulosine-treatedextract ameliorated allergy [48 75] IFN-120574 promoter reporterassays and T-cell differentiation assays were used to isolate2 flavonoids [69] and 3 polyynes [49 50] as immunomod-ulatory compounds from the butanol fraction of B pilosaInterestingly the flavonoids promote IFN-120574 expression inNK and T cells In marked contrast the polyynes promoteIL-4 expression and indirectly inhibit IFN-120574 expression indifferentiating T cells Sensitivity appears to be the key to

identifying structure- and bioactivity-related phytochemicalsfrom medicinal plants

36 Antimalarial Activity The use of chemical drugs againstpathogens has resulted in drug-resistant mutants Examplesof drug resistance can be found in the species of thePlasmodium that cause malaria It is important to search fornew compounds to combat Plasmodium parasites [47] Astudy of the anti-malarial activity of the leaf extracts of Bpilosa using a combination of phytochemistry and bioassaysshowed that compound 49 (Table 11) showed activity againsta malaria parasite (P falciparum NF54 strain) with an IC

50

value of 60 120583gmL [77] In addition compound 49 isolatedfrom the aerial parts of B pilosa inhibited growth of the Pfalciparum FCR-3 strain with an IC

50value of 035 120583gmL

This compound was tested for its in vivo effect in miceinfected with P berghei NK-65 strain Results showed thatcompound 49 decreased the average parasitemia in the redblood cells by 207 (from 328 of that of the control to121) after an intravenous injection of 08mgkg BWday forfour days [47] Further studies addressing the anti-malarialmechanism underlying both polyynes and clinical studies areneeded

37 Antibacterial Activity Emergence of multiple antibiotic-resistantmicrobes is becoming a global threat to public healthand a challenge to disease treatment For instance penicillinis commonly used to combat a food-borne intracellularbacteriumListeria however penicillin-resistant bacteria havebeen discovered recently [69] Chang and coworkers iso-lated centaurein (110) and centaureidin (109) from B pilosaextract [69 70] Centaurein enhances expression of IFN-120574a key cytokine for macrophage activation and consequentlyenhances bactericidal activity in macrophages [69 70] Inagreement with observed in vitro effects centaurein wasreported to prevent and treat Listeria infection in C57BL6Jmice [91] Mechanistic studies confirmed that centaureinexerted antilisterial action via IFN-120574 expression in wild-typemice but not IFN-120574 knockout mice [91] Further in vitrostudies on centaurein showed that this compound increasedIFN-120574 expression by 13 (from 17 to 20) 20 (from 21to 41) and 11 (from 6 to 17) in CD4+ T cells CD8+Tcells and NK cells respectively That is to say there wasan increase in IFN-120574 producing immune cells As expectedcentaurein also enhanced the expression level of T-bet a keynuclear factor for IFN-120574 expression Consistently centaureinaugmented the serum IFN-120574 levels inC57BL6Jmice and thisaugmentation peaked 24 hours after compound injectionThe quantity of mouse serum IFN-120574 was sufficient to activatemacrophages in vitro and eradicated GFP-producing Listeriainside macrophages However the entry of Listeria intomacrophages was not affected by centaurein-treated mousesera Centaurein treatment at 20120583g per mouse rescued 30of the mice infected with a lethal dose of Listeria (2 times106 CFU) It is noteworthy that in the presence of ampicillin(5 120583gmouse) centaurein (20 120583gmouse) rescued 70 of themice suggesting an additive effect between ampicillin andcentaurein [91] Despite lower abundance centaureidin was

Evidence-Based Complementary and Alternative Medicine 45

30 times more active than centaurein in terms of IFN-120574production [91]

Aside from the indirect antibacterial action mentionedabove extract andor compounds of B pilosa also showeddirect bacteriostatic andor bactericidal action One studyreported that essential oils and leafflower extracts of Bpilosa could suppress the growth of gram positive and gramnegative bacteria as evidenced by zone of inhibition assaysIn this study antibacterial activity of the essential oils fromthe leaves and flowers of B pilosa was determined in anattempt to identify natural products as food preservatives forprevention of microbial multiplication and food oxidationThe essential oils and extracts of B pilosa leaves and flow-ers showed moderate but different extents of antibacterialactivity (Table 17) In general essential oils had higherantibacterial activity than crude extracts One explanation forthis could be that monoterpenes in the essential oils destroycellular integrity and subsequently inhibit the respirationand ion transport processes The presence of antibacterial120573-caryophyllene could be another explanation as reportedelsewhere [92] Another study reported that the methanoland acetone extract of B pilosa roots displayed antibacterialactivities against the bacteria listed in Table 18 [93] andmethanol extracts from the roots seemed to be the mosteffective

Another study indicated that the polyyne (R)-12-dihydroxytrideca-357911-pentayne (49) from this plantalso suppressed bacterial growth as shown by the minimuminhibitory concentration required to inhibit 50 bacterialgrowth (MIC

50) in Table 19 This compound was highly

effective against several Gram positive and Gram negativebacteria including the drug-resistant bacteria Staphylococcusaureus N315 (MRSA) and Enterococcus faecalis NCTC12201(VRE) [47] Strikingly compound 49 had a similar MIC

50

value to antibiotics (ampicillin tetracycline norfloxacin andamphotericin B) in most of the bacteria tested

Antibacterial activity of B pilosa extracts and compo-nents expressed as MIC

50and the mean zone of inhibition

is tabulated in Tables 17 18 and 19The zone of inhibition forAmpicilline (positive control) ranges from 153 plusmn 03mm to443 plusmn 02mm [92]

38 Antifungal Activity B pilosa has traditionally been usedto treat microbial infection Recently different parts of Bpilosa have been tested for antifungal activities Deba andcolleagues first evaluated the antifungal effect of the hotwater extracts of the B pilosa roots stems and leaves againstCorticium rolfsii Fusarium solani and Fusarium oxysporumThey discovered that C rolfsii was most suppressed bytreatment with B pilosa as its growth was reduced at almostall the tested doses followed by F oxysporum and F solani[99] However the fungicidal activities of the stems androots were greater than the leaves [99] Moreover the samegroup assessed the antifungal activity of the essential oilsand aqueous extracts from B pilosa flowers and leaves [92]They showed that the extracts and oils had antifungal activityagainst C rolfsii F solani and F oxysporum Essential oils

appeared to have better fungicidal activity thanwater extractsas summarized in Table 20

Another study by Ashafa and colleagues showed thatacetone methanol and water extracts of the B pilosa rootsshowed antifungal activities against Aspergillus niger Aflavus and Penicillium notatum using the agar dilutionmethod The results are tabulated in Table 21 [93] Negativecontrols showed 0 growth inhibitionThe methanol extractof the B pilosa roots at 10mgmL was also effective againstCandida albicans [93]Of note B pilosa obtained fromPapuaNew Guinea had no activity against A niger and C albicans[126] but the SouthAfrican (EasternCape) ecotype exhibitedmoderate activity against C albicans [93] This discrepancymay depend on extraction solvents extraction procedureassay techniques different plant parts and abundance ofactive compounds

B pilosa produces a variety of secondary metabolitessuch as flavonoids phenylacetylenes alkaloids sterols ter-penoids and tannis [92 93] However none of them havebeen confirmed as active compounds against fungi Furtherinvestigation of active compounds from B pilosa is necessaryto further understand the antifungal efficacy of this plant

39 Hypotensive and Vasodilatory Activities In early studiesDimo and colleagues used three rat models normotensiveWistar rats (NTR) salt-loading hypertensive rats (SLHR)and spontaneous hypertensive rats (SHR) to investigate thehypotensive effect of the methanol crude extract of B pilosaleaves [28 127] The extract lowered systolic blood pressurein hypertensive rats (SLHR and SHR) to a greater degreethan NTR [28 127] In addition a decrease in urinarysodium ions and an increase in urinary potassium ions wereobserved after treatment with the methanol extract of Bpilosa leaves although neither differences were statisticallysignificant [28 127] Taking the data together the studyproposed that B pilosa leaf extract reduced blood pressurevia vasodilation [28 127] The same group continued totest the antihypertensive effect of aqueous and methylenechloride extracts of B pilosa leaves in a hypertensive ratmodel [28 127] To establish a fructose-induced hypertensionmodel male Wistar rats were given 10 fructose solutionto drink ad libitum for three weeks In addition to freeaccess to 10 fructose the rats were treated with the aqueous(150mgkg) or methyl chloride (350mgkg) extracts of Bpilosa for additional three weeks [28 127] Both extracts ofB pilosa leaves had a hypotensive effect on rats Howeverneither extracts reversed the elevation of serum insulin infructose-fed rats Therefore B pilosa lowered blood pressureirrespective of insulin [28 127] To better understand thehypotensive mechanism the authors investigated the effectof a neutral extract of B pilosa (NBP) a mixture of methanolandmethylene chloride (1 1) extract after neutralization withNaOH and HCl on the heart and the blood pressure ofNTR and SHR [110] This study showed that an intravenousinjection of the NBP resulted in a biphasic reduction insystolic blood pressure In addition one intravenous dose ofthe extract at 10 20 and 30mgkg BW decreased systolicblood pressure in normal rats by 183 425 and 30

46 Evidence-Based Complementary and Alternative Medicine

respectively and the same doses reduced the blood pressurein hypertensive rats by 258 389 and 286 respectivelyOnly the highest dose (30mgkg) affected the force of thecontraction of the heart Atropine and propranolol were usedto interfere with the hypotensive action of the NBP Atropinereduced the initial phase of the hypotensive response in NBPand completely abolished the second phase of hypotensiveresponse in NBP In contrast propranolol increased thefirst hypotensive response but partially abolished the secondhypotensive response provoked by the NBP [110]This mech-anistic study suggested that B pilosa invokes the biphasichypotensive responses via targeting cardiac pump efficiencyduring the first phase and vasodilation at the second phase[110]

A further study was performed to investigate the relaxingeffect of a neutral extract of B pilosa (NBP) on rat aorta con-tractedwithKCl (60mM) andnorepinephrine (01mM) [94]Cumulative addition of NBP relaxed the rat aorta previouslycontracted by KCl in a dose-dependent manner The EC

50

value of theNBP for vasorelaxationwas 032mgmLThedataalso showed that the NBP reduced the contraction of aortapreviously contracted by KCl irrespective of the presence ofaortic endothelium [94]

Pretreatment with glibenclamide an ATP-dependent K+channel blocker did not considerably affect the relaxant effectof the NBP on KCl-induced contraction suggesting thatthe vasodilatory effect of B pilosa was not related to theopening of this ATP-dependent K+ channel [94] On theother hand in the presence of indomethacin or pyrilaminemaleate the relaxant response induced by the plant extractwas significantly inhibited at the lower concentrations Theplant extract was able to reduce the aorta resting tone inhibitthe KCl-induced contractions by 90 at 15mgmL and theCaCl2-induced contractions by 95 at 075mgmL These

results demonstrate that B pilosa can act as a vasodilatorprobably via acting as a calcium antagonist [94]

However no specific compound for the above activityhas been identified from B pilosa to date A bioactivity-guided identification approach may be adopted to identifythe active compounds in B pilosa that possess hypotensiveand vasodilatory effects and understand their mechanism ofaction

310 Wound Healing Activity B pilosa has been traditionallyused to treat tissue injury inCameroon Brazil andVenezuela[26] Hassan and colleagues investigated the wound healingpotential of B pilosa in Wistar rats [128] Mirroring thepositive control neomycin sulfate the ethanol extract of Bpilosa had faster wound closure than control rats 3 6 and 9days after topical application Histological examination alsorevealed better collagenation angiogenesis and organizationof wound tissue seven days after application Epithelializationand total healing time in B pilosa-treated rats were compara-ble to those of neomycin sulfate Together these data suggestthat B pilosa may be a viable alternative to neomycin lotionfor the treatment of wounds

In addition to studying the wound healing effect of Bpilosa on external ulcers Tan and colleagues also examined

the effect of methanol cyclohexane and methyl chlorideextracts of B pilosa on gastric ulcers in Wistar rats fedwith 1mL HClethanol gastric necrotizing solution (150mMHCl in 60 ethanol) and macroscopically visible lesionswere scored [26] Among the three extracts methylene chlo-ride extracts exhibited the highest activity showing 464inhibition of lesion formation at a dose of 500mgkg BWand complete inhibition at 750mgkg [26] The efficacy ofthe ethylene chloride extract was followed by that of themethanol extracts which had inhibition ranging from 304to 822 at concentrations of 500mgkg and 1000mgkgBW respectively [26] The cyclohexane extracts showed thelowest activity against gastric ulcers in rats with 13340 and 797 inhibition at 500 750 and 1000mgkg BWrespectively [26] To better understand the mode of actionof the methylene chloride extract of B pilosa rats werepretreated with indomethacin a COX-2 inhibitor involved inprostaglandin synthesis Pretreatment significantly reducedthe protection against HClethanol-induced ulcers to 313inhibition at 750mgkg BW suggesting a link between theantiulcerative activity of B pilosa and prostaglandin syn-thesis Unexpectedly the methylene chloride extract of Bpilosa showed little gastric mucosal protection against gastriclesions induced by 95 ethanol (1mL) [26] Absolute alcoholis known to cause mucosalsubmucosal tissue destruction viacellular necrosis and the release of tissue-derived mediators(histamine and leukotrieneC4)Thus these data imply thatBpilosa did not prevent the generation or the necrotic action ofthese mediators on the gastric microvasculature In additionpylorus ligation can increase gastric acid secretion withoutan alteration of mucosal histamine content The methylenechloride extract of B pilosa did not possess antisecretoryactivity On the contrary it was observed that increases in thedose of the extract led to elevated gastric juice acidity [26]Results with both absolute ethanol and pylorus ligation ratmodels suggested the possibility that the ineffectiveness of Bpilosa against gastric ulcers was due to lack of antihistaminicactivity in the plant In summary overall the data suggest thatB pilosa protects against HCLethanol-mediated ulcers viainhibition of prostaglandin biosynthesis

Previous phytochemical studies showed that a groupof flavonoids acyclichalcones are present in B pilosa[129 130] and the chalcones were proposed to have anti-ulcerative activity [131] Moreover nine hydroxychalcones 10were reported to possess gastric cytoprotective effects with24-dihydroxychalcone being the most active [132] Sincemethylene chloride extracts appear to be the most active Bpilosa extracts next the specific anti-ulcerative phytochemi-cals in the methylene chloride extracts of B pilosa and theirmodes of action needs to be probed

Despite the claims listed inTable 3 relatively few scientificstudies have been conducted in vitro and in vivo to address thetraditional ethnomedical uses of B pilosa Information aboutthe use of B pilosa as a botanical therapy recorded so far isfar from complete Studies conducted thus far only serve asa starting point for further investigation of B pilosa and theultimate efficacious use of the herb in clinical applications

Evidence-Based Complementary and Alternative Medicine 47

4 Toxicology

Despite its use as an ingredient in food for human con-sumption studies on systemic toxicity (eg acute subacutechronic and subchronic toxicities) of B pilosa in humansand animals are still inadequate and insufficient So faracute andor subchronic toxicities have been evaluated inrats and mice Oral acute and 28-day toxicities of waterextract of B pilosa leaves were evaluated in Wistar rats[133] An oral dose of water extract of B pilosa leaves at10 gkg BW showed no obvious mortality or changes inthe appearance in rats [134] The same extract at 08 gkgBWday once a day showed no obvious sub-chronic toxicityin rats over 28 days as measured by survival rate bodyweight and gross examination of organs [134] These dataare consistent with our data indicating that oral deliveryof the water extract of the B pilosa whole plant at 1 gkgBWday once a day is safe in rats over 28 days (unpublisheddata) Taken together these studies suggest that ingestion ofB pilosa aqueous extract at up to at 1 gkg BWday oncea day is highly safe in rats In addition the acute toxicityof aqueous and ethanol extracts of B pilosa in mice havebeen reported [134] Five- to six-week-old mice with weightsbetween 28 and 35 g received a peritoneal injection of bothextracts at the different doses The LD

50 the dose that causes

50 lethality of the aqueous and ethanol extracts in micewas 1230 gkg BW and 615 gkg BW respectively [134] Acomplete toxicological study has not been completed forhumans Furthermore the drug interactions of B pilosa withother drugs are unknown Further safety verification andclinical trials should be performed before B pilosa can beconsidered for medicinal use

5 Conclusions

B pilosa is an erect perennial plant with green leaves whiteor yellow flowers and tiny black seeds As it is distributedworldwide and is widely used as a folk remedy B pilosacan be thought of as an extraordinary source of food andmedicine However a comprehensive up-to-date review ofresearch on B pilosa has hitherto been unavailable In thisarticle scientific studies on B pilosa have been summarizedand critically discussed from the perspectives of botanyethnomedicine phytochemistry pharmacology and toxicol-ogy B pilosa is claimed to treat more than 40 disordersand 201 compounds have been identified from this plantThe medicinal utility of B pilosa and its modes of actionin relation to its known phytochemicals were discussedherein Polyynes flavonoids phenylpropanoids fatty acidsand phenolics are the primary bioactive compounds of Bpilosa and they have been reported to be effective in thetreatment of tumors inflammationimmune modulationdiabetes viruses microbes protozoans gastrointestinal dis-eases hypertension and cardiovascular diseases Cautionshould be exercised in the therapeutic use of B pilosa forhypoglycemia hypotension bleeding and allergy

Acknowledgments

The authors thank their laboratory members for construc-tive suggestions and the authors whose publications theycited for their contributions The authors also thank MsMiranda Loney of Academia Sinica for English editing of thispaper This work was supported by Grants 99-CDA-L11 and101S0010056 from Academia Sinica Taiwan Arlene P Bar-tolome is a recipient of MECO-TECO Sandwich ScholarshipProgram between Taiwan and the Philippines

References

[1] T Shen G H Li X N Wang and H X Lou ldquoThe genusCommiphora a review of its traditional uses phytochemistryand pharmacologyrdquo Journal of Ethnopharmacology vol 142 no2 pp 319ndash330

[2] C Long P Sauleau B David et al ldquoBioactive flavonoids ofTanacetum parthenium revisitedrdquo Phytochemistry vol 64 no2 pp 567ndash569 2003

[3] P O Karis and O Ryding Asteraceae Cladistics and Classifica-tion Bremer K Eds pp 559ndash569 Timber press Portland OreUSA 1994

[4] O N Pozharitskaya A N Shikov M N Makarova et alldquoAnti-inflammatory activity of a HPLC-fingerprinted aqueousinfusion of aerial part of Bidens tripartita LrdquoPhytomedicine vol17 no 6 pp 463ndash468 2010

[5] F Q Oliveira V Andrade-Neto A U Krettli and MG L Brandao ldquoNew evidences of antimalarial activity ofBidens pilosa roots extract correlated with polyacetylene andflavonoidsrdquo Journal of Ethnopharmacology vol 93 no 1 pp 39ndash42 2004

[6] Agriculture USDA ldquoPlants databaserdquo 2013in Natural Resources Conservation Servicehttpwwwnrcsusdagovwpsportalnrcssitenationalhome

[7] Y Zhou and X Z Yan ldquoExperimental study of qi deficiencysyndrome and Codonopsis pillosulae and Astragalus injectionon the immune response in micerdquo Zhong Xi Yi Jie He Za Zhivol 9 no 5 pp 286ndash262 1989

[8] S C Chien P H Young Y J Hsu et al ldquoAnti-diabeticproperties of three common Bidens pilosa variants in TaiwanrdquoPhytochemistry vol 70 no 10 pp 1246ndash1254 2009

[9] M J Alcaraz and M J Jimenez ldquoFlavonoids as anti-inflammatory agentsrdquo Fitoterapia vol 59 no 1 pp 25ndash38 1988

[10] FAO Agriculture Food and Nutrition for AfricamdashA ResourceBook for Teachers of Agriculture Publishing ManagementGroup FAO Information Division Rome Italy 1997

[11] M B Rokaya Z Munzbergova B Timsina and K R BhattaraildquoRheum australe D Don a review of its botany ethnobotanyphytochemistry and pharmacologyrdquo Journal of Ethnopharma-cology vol 141 no 3 pp 761ndash774 2012

[12] P H Young Y J Hsu and W C Yang ldquoBidens pilosa L andits medicinal userdquo in Recent Progress in Medicinal Plants DrugPlant II A S AwaadVK Singh and JNGovil Eds StandiumPress Houston Tex USA 2010

[13] C Ge ldquoCytologic study of Bidens bipinnata LrdquoZhongguo ZhongYao Za Zhi vol 15 no 2 pp 72ndash125 1990

[14] L C Chiang J S Chang C C Chen L T Ng and C CLin ldquoAnti-herpes simplex virus activity of Bidens pilosa and

48 Evidence-Based Complementary and Alternative Medicine

Houttuynia cordatardquoAmerican Journal of ChineseMedicine vol31 no 3 pp 355ndash362 2003

[15] N P Rybalchenko V A Prykhodko S S Nagorna et alldquoIn vitro antifungal activity of phenylheptatriyne from Bidenscernua L against yeastsrdquo Fitoterapia vol 81 no 5 pp 336ndash3382010

[16] K Redl W Breu B Davis and R Bauer ldquoAnti-inflammatoryactive polyacetylenes from Bidens campylothecardquo Planta Med-ica vol 60 no 1 pp 58ndash62 1994

[17] M Ayyanar and S Ignacimuthu ldquoTraditional knowledge ofKani tribals inKouthalai of Tirunelveli hills TamilNadu IndiardquoJournal of Ethnopharmacology vol 102 no 2 pp 246ndash255 2005

[18] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant intraditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[19] AKAbbas KMMurphy andA Sher ldquoFunctional diversity ofhelper T lymphocytesrdquo Nature vol 383 no 6603 pp 787ndash7931996

[20] J H Cano and G Volpato ldquoHerbal mixtures in the traditionalmedicine of Eastern Cubardquo Journal of Ethnopharmacology vol90 no 2-3 pp 293ndash316 2004

[21] J Namukobe J M Kasenene B T Kiremire et al ldquoTraditionalplants used for medicinal purposes by local communitiesaround the Northern sector of Kibale National Park UgandardquoJournal of Ethnopharmacology vol 136 no 1 pp 236ndash245 2011

[22] C Lans ldquoComparison of plants used for skin and stomachproblems in Trinidad and Tobago with Asian ethnomedicinerdquoJournal of Ethnobiology andEthnomedicine vol 3 article 3 2007

[23] S Dharmananda ldquoA Popular Remedy Ecapes Notice ofWesternPractitionersrdquo 2013 httpwwwitmonlineorgartsbidenshtm

[24] Y J Hsu T H Lee C L T Chang Y T Huang andW C YangldquoAnti-hyperglycemic effects and mechanism of Bidens pilosawater extractrdquo Journal of Ethnopharmacology vol 122 no 2 pp379ndash383 2009

[25] E Noumi F Houngue and D Lontsi ldquoTraditional medicinesin primary health care plants used for the treatment ofhypertension in Bafia Cameroonrdquo Fitoterapia vol 70 no 2 pp134ndash139 1999

[26] P V Tan T Dimo and E Dongo ldquoEffects of methanolcyclohexane and methylene chlo ride extracts of Bidens pilosaon various gastric ulcer models in ratsrdquo Journal of Ethnophar-macology vol 73 no 3 pp 415ndash421 2000

[27] R L C Pereira T Ibrahim L Lucchetti A J R Da Silva and VL G De Moraes ldquoImmunosuppressive and anti-inflammatoryeffects of methanolic extract and the polyacetylene isolatedfrom Bidens pilosa Lrdquo Immunopharmacology vol 43 no 1 pp31ndash37 1999

[28] T Dimo J Azay P V Tan et al ldquoEffects of the aqueous andmethylene chloride extracts of Bidens pilosa leaf on fructose-hypertensive ratsrdquo Journal of Ethnopharmacology vol 76 no 3pp 215ndash221 2001

[29] CWiartMedicinal Plants of Southeast Asia Pelanduk SelabgorDarul Ehsan Malaysia 2000

[30] F L Silva D C H Fischer J F Tavares M S Silva P FDe Athayde-Filho and J M Barbosa-Filho ldquoCompilation ofsecondary metabolites from Bidens pilosa LrdquoMolecules vol 16no 2 pp 1070ndash1102 2011

[31] A H Chen S R Lin and C H Hong ldquoPhytochemical studyon Bidens pilosa L varminorrdquo Huaxue pp 38ndash42 1975

[32] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[33] M C Zulueta M Tada and C Y Ragasa ldquoA diterpene fromBidens pilosardquo Phytochemistry vol 38 no 6 pp 1449ndash14501995

[34] J Wang H Yang Z W Lin and H D Sun ldquoFlavonoids fromBidens pilosa var radiatardquo Phytochemistry vol 46 no 7 pp1275ndash1278 1997

[35] A Zhao Q Zhao L Peng et al ldquoA new chalcone glycoside fromBidens pilosardquo Acta Botanica Yunnanica vol 26 no 1 pp 121ndash126

[36] P Gao Y Wu S Cui and Y Zou ldquoSynthesis and absoluteconfiguration of the 7-phenylhepta-4 6-diyne-1 2-diol isolatedfrom Bidens pilosardquo Synthesis no 13 pp 2131ndash2135 2011

[37] S Wang B Yang D Zhu D He and L Wang ldquoActivecomponents of Bidens pilosa Lrdquo Zhongcaoyao vol 36 pp 20ndash21

[38] F Bohlmann T Burkhardt and C Zdero Naturally OccuringAcetylenes Academic Press New York NY USA 1973

[39] T M Sarg A M Ateya N M Farrag and F A AbbasldquoConstituents and biological activity of Bidens pilosa L grownin EgyptrdquoActa Pharmaceutica Hungarica vol 61 no 6 pp 317ndash323 1991

[40] H A L Valdes and H P Rego ldquoBidens pilosa Linnerdquo RevistaCubana de Plantas Medicinales vol 6 pp 28ndash33

[41] F Bohlmann H Bornowski and K M Kleine ldquoNew polyynesfrom the tribe Heliantheaerdquo Chemische Berichte vol 97 pp2135ndash2138

[42] RWang Q XWu and Y P Shi ldquoPolyacetylenes and flavonoidsfrom the aerial parts ofBidens pilosardquo PlantaMedica vol 76 no9 pp 893ndash896 2010

[43] L W Wu Y M Chiang H C Chuang et al ldquoA novelpolyacetylene significantly inhibits angiogenesis and promotesapoptosis in human endothelial cells through activation of theCDK inhibitors and caspase-7rdquo Planta Medica vol 73 no 7 pp655ndash661 2007

[44] L W Wu Y M Chiang H C Chuang et al ldquoPolyacetylenesfunction as anti-angiogenic agentsrdquo Pharmaceutical Researchvol 21 no 11 pp 2112ndash2119 2004

[45] H L Yang S C Chen N W Chang et al ldquoProtection fromoxidative damage using Bidens pilosa extracts in normal humanerythrocytesrdquo Food and Chemical Toxicology vol 44 no 9 pp1513ndash1521 2006

[46] H Q Wang S J Lu H Li and Z H Yao ldquoEDTA-enhancedphytoremediation of lead contaminated soil by Bidens maxi-mowiczianardquo Journal of Environmental Sciences vol 19 no 12pp 1496ndash1499 2007

[47] S TobinagaMK SharmaWG L Aalbersberg et al ldquoIsolationand identification of a potent antimalarial and antibacterialpolyacetylene from Bidens pilosardquo Planta Medica vol 75 no 6pp 624ndash628 2009

[48] C L T Chang H K Kuo S L Chang et al ldquoThe distincteffects of a butanol fraction of Bidens pilosa plant extract onthe development of Th1-mediated diabetes and Th2-mediatedairway inflammation in micerdquo Journal of Biomedical Sciencevol 12 no 1 pp 79ndash89 2005

[49] S L Chang C L T Chang Y M Chiang et al ldquoPolyacetyleniccompounds and butanol fraction from Bidens pilosa can mod-ulate the differentiation of helper T cells and prevent autoim-mune diabetes in non-obese diabetic micerdquo Planta Medica vol70 no 11 pp 1045ndash1051 2004

Evidence-Based Complementary and Alternative Medicine 49

[50] Y M Chiang C L T Chang S L Chang W C Yang and LF Shyur ldquoCytopiloyne a novel polyacetylenic glucoside fromBidens pilosa functions as a T helper cell modulatorrdquo Journal ofEthnopharmacology vol 110 no 3 pp 532ndash538 2007

[51] R PUbillas CDMendez S D Jolad et al ldquoAntihyperglycemicacetylenic glucosides from Bidens pilosardquo Planta Medica vol66 no 1 pp 82ndash83 2000

[52] L Alvarez S Marquina M L Villarreal D Alonso E Arandaand G Delgado ldquoBioactive polyacetylenes from Bidens pilosardquoPlanta Medica vol 62 no 4 pp 355ndash357 1996

[53] C L T Chang S L Chang Y M Lee et al ldquoCytopiloyne apolyacetylenic glucoside prevents type 1 diabetes in nonobesediabetic micerdquo Journal of Immunology vol 178 no 11 pp 6984ndash6993 2007

[54] G Kusano A Kusano and Y Seyama Novel Hypoglycemicand Anti-Inflammatory Polyacetylenic Compounds Their Com-positions Bidens Plant Extract Fractions and CompositionsContaining the Plant or Fraction JPO Tokyo Japan 2004

[55] M G L Brandao A U Krettli L S R Soares C G CNery and H C Marinuzzi ldquoAntimalarial activity of extractsand fractions from Bidens pilosa and other Bidens species(Asteraceae) correlated with the presence of acetylene andflavonoid compoundsrdquo Journal of Ethnopharmacology vol 57no 2 pp 131ndash138 1997

[56] A U Krettli V F Andrade-Neto M D G L Brandao andW M S Ferrari ldquoThe search for new anti-malarial drugs fromplants used to treat fever and malaria or plants ramdomlyselected a reviewrdquoMemorias do Instituto Oswaldo Cruz vol 96no 8 pp 1033ndash1042 2001

[57] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[58] C K Wat R K Biswas E A Graham L Bohm G H NTowers and E R Waygood ldquoUltraviolet-mediated cytotoxicactivity of phenylheptatriyne from Bidens pilosa Lrdquo Journal ofNatural Products vol 42 no 1 pp 103ndash111 1979

[59] Y Sashida K Ogawa M Kitada H Karikome Y Mimakiand H Shimomura ldquoNew aurone glucosides and new phenyl-propanoid glucosides from Bidens pilosardquo Chemical and Phar-maceutical Bulletin vol 39 no 3 pp 709ndash711 1991

[60] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens pilosa L (TFB) on animal liver injury andliver fibrosisrdquo Journal of Ethnopharmacology vol 116 no 3 pp539ndash546 2008

[61] B Hoffmann and J Holzl ldquoAcylated compounds from Bidenspilosardquo Planta Medica vol 55 pp 108ndash109

[62] B Hoffmann and J Holzl ldquoA methylated chalcone glucosidefrom Bidens pilosardquo Phytochemistry vol 27 no 11 pp 3700ndash3701 1988

[63] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[64] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[65] B Hoffmann and J Holzl ldquoWeitere acylierte chalkone ausBidens pilosardquo Planta Medica vol 54 pp 450ndash451 1988

[66] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant in11traditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[67] V V Pham V V T K P V L Hoang and V K Phan ldquoFlavonoidcompounds from the plant Bidens pilosa L (Asteraceae)rdquo TapChi Duoc Hoc vol 50 pp 48ndash53

[68] S D Sarker B Bartholomew R J Nash and N Robinson ldquo5-O-methylhoslundin an unusual flavonoid from Bidens pilosa(Asteraceae)rdquo Biochemical Systematics and Ecology vol 28 no6 pp 591ndash593 2000

[69] S L Chang Y M Chiang C L T Chang et al ldquoFlavonoidscentaurein and centaureidin from Bidens pilosa stimulate IFN-120574 expressionrdquo Journal of Ethnopharmacology vol 112 no 2 pp232ndash236 2007

[70] Y M Chiang D Y Chuang S Y Wang Y H Kuo P WTsai and L F Shyur ldquoMetabolite profiling and chemopreventivebioactivity of plant extracts from Bidens pilosardquo Journal ofEthnopharmacology vol 95 no 2-3 pp 409ndash419 2004

[71] A Kusano Y Seyama E Usami et al ldquoStudies on the antioxi-dant active constituents of the dried powder from Bidens pilosaL var radiata SchrdquoNaturalMedicines vol 57 no 3 pp 100ndash1042003

[72] I Doleckova L Rarova J Gruz et al ldquoAnti-proliferative andanti-angiogenic effects of flavone eupatorin an active con-stituent of chloroform extract ofOrthosiphon stamineus leavesrdquoFitoterapia vol 83 no 6 pp 1000ndash1007

[73] K Bairwa R Kumar R J Sharma and R K Roy ldquoAn updatedreview on Bidens pilosa Lrdquo Der Pharma Chemica vol 2 no 3pp 325ndash337

[74] Q Xia Y Liu and Y Li ldquoDetermination of hyperoside indifferent parts and different species of Herba Bidens by RP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol 50pp 48ndash53

[75] M Horiuchi and Y Seyama ldquoImprovement of the antiinflam-matory and antiallergic activity of Bidens pilosa L var radiataSCHERFF treated with enzyme (Cellulosine)rdquo Journal of HealthScience vol 54 no 3 pp 294ndash301 2008

[76] M G L Brandao C G C Nery M A S Mamao and AU Krettli ldquoTwo methoxylated flavone glycosides from Bidenspilosardquo Phytochemistry vol 48 no 2 pp 397ndash400 1998

[77] P Kumari K Misra B S Sisodia et al ldquoA promising anticancerand antimalarial component from the leaves of Bidens pilosardquoPlanta Medica vol 75 no 1 pp 59ndash61 2009

[78] G Seelinger I Merfort U Wolfle and C M Schempp ldquoAnti-carcinogenic effects of the flavonoid luteolinrdquoMolecules vol 13no 10 pp 2628ndash2651 2008

[79] G Seelinger I Merfort and C M Schempp ldquoAnti-oxidantanti-inflammatory and anti-allergic activities of luteolinrdquoPlanta Medica vol 74 no 14 pp 1667ndash1677 2008

[80] C C Yit and N P Das ldquoCytotoxic effect of butein on humancolon adenocarcinoma cell proliferationrdquo Cancer Letters vol82 no 1 pp 65ndash72 1994

[81] J A Beutler E Hamel A J Vlietinck et al ldquoStructure-activityrequirements for flavone cytotoxicity and binding to tubulinrdquoJournal of Medicinal Chemistry vol 41 no 13 pp 2333ndash23381998

[82] M R Kviecinski K B Felipe T Schoenfelder et al ldquoStudy ofthe antitumor potential of Bidens pilosa (Asteraceae) used inBrazilian folkmedicinerdquo Journal of Ethnopharmacology vol 117no 1 pp 69ndash75 2008

[83] P Sundararajan A Dey A Smith A G Doss M Rajappanand S Natarajan ldquoStudies of anticancer and antipyretic activityof Bidens pilosawhole plantrdquo African Health Sciences vol 6 no1 pp 27ndash30 2006

[84] YMChiang C P Lo Y P Chen et al ldquoEthyl caffeate suppressesNF-120581B activation and its downstream inflammatory mediatorsiNOS COX-2 and PGE2 in vitro or in mouse skinrdquo BritishJournal of Pharmacology vol 146 no 3 pp 352ndash363 2005

50 Evidence-Based Complementary and Alternative Medicine

[85] J S Kim H J Lee M H Lee J Kim C Jin and J HRyu ldquoLuteolin inhibits LPS-stimulated inducible nitric oxidesynthase expression in BV-2 microglial cellsrdquo Planta Medicavol 72 no 1 pp 65ndash68 2006

[86] P A Ruiz and D Haller ldquoFunctional diversity of flavonoidsin the inhibition of the proinflammatory NF-120581B IRF and Aktsignaling pathways in murine intestinal epithelial cellsrdquo Journalof Nutrition vol 136 no 3 pp 664ndash671 2006

[87] A Xagorari A Papapetropoulos A Mauromatis MEconomou T Fotsis and C Roussos ldquoLuteolin inhibitsan endotoxin-stimulated phosphorylation cascade andproinflammatory cytokine production in macrophagesrdquoJournal of Pharmacology and Experimental Therapeutics vol296 no 1 pp 181ndash187 2001

[88] N Yoshida T Kanekura Y Higashi and T Kanzaki ldquoBidenspilosa suppresses interleukin-1120573-induced cyclooxygenase-2expression through the inhibition of mitogen activated proteinkinases phosphorylation in normal human dermal fibroblastsrdquoJournal of Dermatology vol 33 no 10 pp 676ndash683 2006

[89] M Muchuweti C Mupure A Ndhlala T Murenje and M AN Benhura ldquoScreening of antioxidant and radical scavengingactivity of Vigna ungiculata Bidens pilosa and Cleome gynan-drardquo American Journal of Food Technology vol 2 no 3 pp 161ndash168 2007

[90] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens bipinnata L against carbon tetrachloride-induced liver fibrosis in ratsrdquo Journal of Pharmacy and Pharma-cology vol 60 no 10 pp 1393ndash1402 2008

[91] S L Chang H H Yeh Y S Lin Y M Chiang T K Wuand W C Yang ldquoThe effect of centaurein on interferon-120574 expression and Listeria infection in micerdquo Toxicology andApplied Pharmacology vol 219 no 1 pp 54ndash61 2007

[92] F Deba T D Xuan M Yasuda and S Tawata ldquoChemical com-position and antioxidant antibacterial and antifungal activitiesof the essential oils from Bidens pilosa Linn var Radiatardquo FoodControl vol 19 no 4 pp 346ndash352 2008

[93] A O T Ashafa and A J Afolayan ldquoScreening the root extractsfrom Biden pilosa L var radiata (Asteraceae) for antimicrobialpotentialsrdquo Journal of Medicinal Plant Research vol 3 no 8 pp568ndash572 2009

[94] T B Nguelefack T Dimo E P Nguelefack Mbuyo P V TanS V Rakotonirina and A Kamanyi ldquoRelaxant effects of theneutral extract of the leaves of Bidens pilosa linn on isolated ratvascular smooth musclerdquo Phytotherapy Research vol 19 no 3pp 207ndash210 2005

[95] M T Grombone-Guaratini K L Silva-Brandao V N SolferiniJ Semir and J R Trigo ldquoSesquiterpene and polyacetyleneprofile of the Bidens pilosa complex (Asteraceae Heliantheae)from Southeast of Brazilrdquo Biochemical Systematics and Ecologyvol 33 no 5 pp 479ndash486 2005

[96] L L Lin C Y Wu H C Hsiu M T Wang and H ChuangldquoStudies on diabetes mellitus I The hypoglycemic activity ofphytosterin on alloxan diabetic ratsrdquo Taiwan Yi Xue Hui Za Zhivol 66 no 2 pp 58ndash65 1967

[97] M A N Benhura and I C Chitsiku ldquoThe extractable 120573-carotene content of Guku (Bidens pilosa) leaves after cookingdrying and storagerdquo International Journal of Food Science andTechnology vol 32 no 6 pp 495ndash500 1997

[98] J S Chang L C Chiang C C Chen L T Liu K C Wangand C C Lin ldquoAtileukemic activity of Bidens pilosa l varminor (blume) sherff andHouttuynia cordata thunbrdquo AmericanJournal of Chinese Medicine vol 29 no 2 pp 303ndash312 2001

[99] F Deba T D Xuan M Yasuda and S Tawata ldquoHerbicidal andfungicidal activities and identification of potential phytotoxinsfrom Bidens pilosa L var radiata Scherff research paperrdquoWeedBiology and Management vol 7 no 2 pp 77ndash83 2007

[100] J K Kumar and A K Sinha ldquoA new disubstituted acetylacetonefrom the leaves ofBidens pilosa LinnrdquoNatural Product Researchvol 17 no 1 pp 71ndash74 2003

[101] K Ogawa and Y Sashida ldquoCaffeoyl derivatives of a sugarlactone and its hydroxy acid from the leaves of Bidens pilosardquoPhytochemistry vol 31 no 10 pp 3657ndash3658 1992

[102] Q Xia Y Liu and Y Li ldquoDetermination of gallic acid fromdifferent species and different medical parts of Herba Bidens byRP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol24 no 03 pp 308ndash310

[103] W J Lee L F Wu W K Chen C J Wang and T HTseng ldquoInhibitory effect of luteolin on hepatocyte growth fac-torscatter factor-induced HepG2 cell invasion involving bothMAPKERKs and PI3K-Akt pathwaysrdquo Chemico-BiologicalInteractions vol 160 no 2 pp 123ndash133 2006

[104] J A Beutler J H Cardellina C M Lin E Hamel G MCragg andM R Boyd ldquoCentaureidin a cytotoxic flavone fromPolymnia fruticosa inhibits tubulin polymerizationrdquo BioorganicandMedicinal Chemistry Letters vol 3 no 4 pp 581ndash584 1993

[105] A Verma A Su A M Golin B OrsquoMarrah and J K AmorosaldquoThe lateral view a screening method for knee traumardquo Aca-demic Radiology vol 8 no 5 pp 392ndash397 2001

[106] J Corren M Lemay Y Lin L Rozga and R K RandolphldquoClinical and biochemical effects of a combination botanicalproduct (ClearGuard) for allergy a pilot randomized double-blind placebo-controlled trialrdquo Nutrition Journal vol 7 no 1article 20 2008

[107] M S Gachet J S Lecaro M Kaiser et al ldquoAssessment of anti-protozoal activity of plants traditionally used in Ecuador in thetreatment of leishmaniasisrdquo Journal of Ethnopharmacology vol128 no 1 pp 184ndash197 2010

[108] W C Yang M Ghiotto B Barbarat and D Olive ldquoThe role ofTec protein-tyrosine kinase in T cell signalingrdquo The Journal ofBiological Chemistry vol 274 no 2 pp 607ndash617 1999

[109] S Tewtrakul H Miyashiro N Nakamura et al ldquoHIV-1 inte-grase inhibitory substances from Coleus parvifoliusrdquo Phytother-apy Research vol 17 no 3 pp 232ndash239 2003

[110] T Dimo T B Nguelefack P V Tan et al ldquoPossible mechanismsof action of the neutral extract from Bidens pilosa L leaves onthe cardiovascular system of anaesthetized ratsrdquo PhytotherapyResearch vol 17 no 10 pp 1135ndash1139 2003

[111] T S C Li Chinese and Related North American Herbs CRCPress New York NY USA 2002

[112] H Wu H Chen X Hua Z Shi L Zhang and J ChenldquoClinical therapeutic effect of drug-separated moxibustion onchronic diarrhea and its immunologic mechanismsrdquo Journal ofTraditional Chinese Medicine vol 17 no 4 pp 253ndash258 1997

[113] S W Wright R R Harris J S Kerr et al ldquoSynthesis chemicaland biological properties of vinylogous hydroxamic acids dualinhibitors of 5-lipoxygenase and IL-1 biosynthesisrdquo Journal ofMedicinal Chemistry vol 35 no 22 pp 4061ndash4068 1992

[114] W R Almiron and M E Brewer ldquoClassification of immaturestage habitats of Culicidae (Diptera) collected in CordobaArgentinardquo Memorias do Instituto Oswaldo Cruz vol 91 no 1pp 1ndash9 1996

[115] N L Wang J Wang X S Yao and S Kitanaka ldquoTwo newmonoterpene glycosides and a new (+)-jasmololone glucoside

Evidence-Based Complementary and Alternative Medicine 51

from Bidens parvifloraWilldrdquo Journal of Asian Natural ProductsResearch vol 9 no 5 pp 473ndash479 2007

[116] P Champagnat ldquoRole of the terminal bud in the actionexercised by the cotyledon of Bidens pilosus L var radiatus onits axillary budrdquo Comptes Rendus des Seances et Memoires de laSociete de Biologie vol 145 no 17-18 pp 1374ndash1376 1951

[117] S F Nielsen S B Christensen G Cruciani A Kharazmiand T Liljefors ldquoAntileishmaniai chalcones statistical designsynthesis and three-dimensional quantitative structure-activityrelationship analysisrdquo Journal of Medicinal Chemistry vol 41no 24 pp 4819ndash4832 1998

[118] Y C Hwang J J H Chu P L Yang W Chen and M V YatesldquoRapid identification of inhibitors that interfere with poliovirusreplication using a cell-based assayrdquo Antiviral Research vol 77no 3 pp 232ndash236 2008

[119] V F Andrade-Neto M G L Brandao F Q Oliveira et alldquoAntimalarial activity of Bidens pilosa L (Asteraceae) ethanolextracts fromwild plants collected in various localities or plantscultivated in humus soilrdquo Phytotherapy Research vol 18 no 8pp 634ndash639 2004

[120] R J Marles and N R Farnsworth ldquoAnti-diabetic plants andtheir active constituentsrdquo Phytomedicine vol 2 no 2 pp 137ndash189

[121] M Habeck ldquoDiabetes treatments get sweet help from naturerdquoNature Medicine vol 9 no 10 p 1228 2003

[122] H W Lin G Y Han and S X Liao ldquoStudies on the activeconstituents of the Chinese traditional medicine Polygonatumodoratum (Mill) DrucerdquoActa Pharmaceutica Sinica vol 29 no3 pp 215ndash222 1994

[123] C L T Chang Y C Chen H M Chen N S Yang andW C Yang ldquoNatural cures for type 1 diabetes a reviewof phytochemicals biological actions and clinical potentialrdquoCurrent Medicinal Chemistry vol 20 no 7 pp 899ndash907 2013

[124] L Dey A S Attele and C S Yuan ldquoAlternative therapies fortype 2 diabetesrdquo Alternative Medicine Review vol 7 no 1 pp45ndash58 2002

[125] C L T ChangH Y Liu T F Kuo et al ldquoAnti-diabetic effect andmode of action of cytopiloynerdquo Evidence-Based Complementaryand Alternative Medicine vol 2013 Article ID 685642 13 pages2013

[126] M R Khan M Kihara and A D Omoloso ldquoAnti-microbialactivity of Bidens pilosa Bischofia javanica Elmerillia papuanaand Sigesbekia orientalisrdquo Fitoterapia vol 72 no 6 pp 662ndash6652001

[127] T Dimo T B Nguelefack P Kamtchouing E Dongo ARakotonirina and S V Rakotonirina ldquoHypotensive effects ofmethanol extract from Bidens pilosa Linn on hypertensive ratsrdquoComptes Rendus de lrsquoAcademie des Sciences Serie III vol 322no 4 pp 323ndash329 1999

[128] A K Hassan O Deogratius J F Nyafuono O Francis and OP Engeu ldquoWound healing potential of the ethanolic extracts ofBidens pilosa and Ocimum suaverdquo African Journal of Pharmacyand Pharmacology vol 5 no 2 pp 132ndash136 2011

[129] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[130] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[131] C Kandaswami and EMiddleton ldquoFree radical scavenging andantioxidant activity of plant flavonoidsrdquo Advances in Experi-mental Medicine and Biology vol 366 pp 351ndash376 1994

[132] K Yamamoto H Kakegawa H Ueda et al ldquoGastric cytopro-tective anti-ulcerogenic actions of hydroxychalcones in ratsrdquoPlanta Medica vol 58 no 5 pp 389ndash393 1992

[133] J O C Ezeonwumelu A K Julius C N Muhoho et alldquoBiochemical and histological studies of aqueous extract ofBidens pilosa leaves from Ugandan Rift valley in ratsrdquo BritishJournal of Pharmacology and Toxicology vol 2 no 6 pp 302ndash309

[134] L Frida S Rakotonirina A Rakotonirina and J P SavineauldquoIn vivo and in vitro effects of Bidens pilosa L (Asteraceae)leaf aqueous and ethanol extracts on primed-oestrogenized ratuterine musclerdquo African Journal of Traditional Complementaryand Alternative Medicines vol 5 no 1 pp 79ndash91 2008

52 Evidence-Based Complementary and Alternative Medicine

Composition Comments

1 Please provide valid postal code to the address if

possible

2 We made the highlighted change in Table 4 for the sake

of correctness Please check

3 Please check the correctness of the highlighted part in

Table 5

4 We considered the highlighted part in Table 11 as

reference Please check similar cases throughout

5 Please check the format of Table 18

6 We reformatted Table 20 Please check

7 We made the highlighted change in Table 4 for the sake

of consistency Please check similar cases throughout the

paper

8 We made the highlighted change(s) for the sake

of consistency Please check simila highlighted cases

throughout

9 We made the highlighted change in Table 9 for the sake

of clarity Please check similar cases through out

10 We made the highlighted change(s) for the sake of

correctness Please check

11 Comment on ref [66] This reference is a repetition

of [18] Please check similar cases throughout

Author(s) Name(s)

Author 1

Author 2

Author 3

12 Evidence-Based Complementary and Alternative Medicine

Table5Flavon

oids

isolated

from

Bpilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

71

2-[(34-Dihydroxyph

enyl)-

methylene]-

6-hydroxy-3(2H

)-benzofuranon

e

Sulfu

retin

O

OHO

OH

OH

Aeria

l(Ch

ina)

[35]

72

2-[(34-Dihydroxyph

enyl)-

methylene]-

67-dihydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-673101584041015840-

tetrahydroxy

maritimetin

O

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

73

2-[(34-Dihydroxyph

enyl)-

methylene]-6-

(120573-D

-glucopyrano

syloxy)-7-

hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-120573-D

-glucop

yranosyl-

673101584041015840-te

trahydroxy

maritimein

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japanand

China)aerial(Ch

ina)

[355960]

74

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

(120573-D

-glucopyrano

syloxy)-

6-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-7-O-120573-D

-glucop

yranosyl-

673101584041015840-

tetrahydroxy

O

OHO

OH

OH

OHO

H

HO

H

H

HOH

HO

OH

Leaves

(Japan)

[59]

75

6-[(6-O-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-acetyl-

120573-D

-glucopyrano

syl)

-673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OHO

H HO

H

H

HOH

HO

O

O

Leaves

(Not

stated)

aeria

l(Ch

ina)

[3559]

Evidence-Based Complementary and Alternative Medicine 13

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

76

6-[(36-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-

2-[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone(Z)-6-O

-(36-di-O

-acetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

bideno

sideA

O

O

HO

OH

OH

OHOH O

H

H

HOH

HO

O

O

O

Leaves

(China)

[60]

77

6-[(46-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(410158401015840610158401015840-

diacetyl-120573-D

-glucropyrano

syl)-

673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

HO

O

OO

Leaves

(not

stated)

aeria

l(Ch

ina)

[343561]

78

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

hydroxy-6-

[(246-tri-O

-acetyl-120573

-D-

glucop

yranosyl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(210158401015840410158401015840610158401015840-

triacetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

O

OOO

O

Leaves

(not

stated)

aeria

l(Ch

ina)

[3461]

79

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-6-

[(346-tri-O

-acetyl-120573

-D-glucopyrano

syl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(310158401015840410158401015840610158401015840-

triacetyl-120573

-D-glucrop

yranosyl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H OH

H

HOH

HO

OOO O

Aeria

l(Ch

ina)

[3435]

80

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-

6-[[6-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propenyl]-120573-

D-glucopyrano

syl]o

xy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-

p-coum

aroyl-

120573-D

-glucropyrano

syl)-

673101584041015840-te

trahydroxy

OH

O

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OLeaves

(Japan)

[59]

14 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

81

1-[2-(120573-D

-Glucopyrano

syloxy)-

4-hydroxyphenyl]-2-

hydroxy-3-(3-

hydroxyphenyl)-2-

prop

en-1-on

e

Chalcone120572

32101584041015840-

tetrahydroxy-

21015840-O

-120573-D

-glucop

yranosyl

HO

O

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[35]

821-(24-Dihydroxyph

enyl)-

3-(34-dihydroxyph

enyl)-

2-prop

en-1-on

eBu

tein

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

833-(34-D

ihydroxyph

enyl)-

1-(234-trihydroxyph

enyl)-

2-prop

en-1-on

eOkanin

O

HO

HO

OH

OH

OH

Leaves

(China)

[60]

84

3-(34-D

ihydroxyph

enyl)-

1-[3-(120573-D

-glucop

yranosyloxy)-

24-dihydroxyphenyl]-2-

prop

en-1-on

e

Okanin

31015840-O

-120573-D

-glucosid

e

O

HO

OH

OH

OH

OO

HO

H

HO

H

H

HOH

H

OH

Leaves

(Germany)

flowers(Germany)

[61ndash63]

85

3-(34-D

ihydroxyph

enyl)-

1-[4-(120573-D

-glucop

yranosyloxy)-

23-dihydroxyphenyl]-

2-prop

en-1-on

e

Okanin

41015840-O

-120573-D

-glucosid

e

OOH

OH

OH

HOO

OHO

H

HO

H

H

HOH

H

OH

Flow

ers(Germany)

leaves

(Japan)

[5963]

Evidence-Based Complementary and Alternative Medicine 15

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

86Okanin41015840-O

-120573-D

-(610158401015840-

O-acetylglucosid

e)

O

O

OH

OH

OH

OHO

H

HO

H

H

HOH

H

HO

O

O

Flow

ers(Germany)

[63]

87

1-[4-[(46-di-O

-Acetyl-

120573-D

-glucopyrano

syl)-oxy]-

23-dihydroxyphenyl]-3-

(34-dihydroxyph

enyl)-2-

prop

en-1-on

e

Okanin

41015840-O

-120573-D

-(410158401015840610158401015840-diacetyl)-

glucop

yranoside

O

O

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Aeria

l(Ch

ina)

[34]

88Okanin41015840-O

-120573-D

-(210158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[64]

89Okanin41015840-O

-120573-D

-(310158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

O

OAe

rial(Ch

ina)

[34]

16 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

90

1-[23-Dihydroxy-4-

[[6-O-[3-(4-hydroxyph

enyl)-

1-oxo-2-prop

enyl]-120573-D

-glucop

yranosyl]oxy]-ph

enyl]-

3-(34-dihydroxyph

enyl)-

2-prop

en-1on

e

Okanin

41015840-O

-120573-D

-(610158401015840-tr

ans-

p-coum

aroyl)-

glucoside

O

O

OH

OH

OH

OHOH

HO

H

H

HOH

H

HO

OO

OH

Leaves

(Germany)

[64]

91Okanin41015840-O

-120573-D

-(410158401015840-acetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Leaves

(Germany)

[65]

92

Okanin

41015840-O

-120573-D

-(210158401015840410158401015840-diacetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[65]

Evidence-Based Complementary and Alternative Medicine 17

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

93

Okanin

41015840-O

-120573-D

-(310158401015840410158401015840-diacetyl-610158401015840-

trans-p-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

OH

O O

Leaves

(Germany)

[65]

94

Okanin

41015840-O

-[120573-D

-glucopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

OOH

OH

OH

HO

OH

HO

HHO

H

HOH

HO

OH

OH

HO

HHO

H

HOH

HO

Flow

er(G

ermany)

[63]

95Okanin

3101584041015840-di-O

-120573-D

-glucosid

e

OOH

OH

OH

O OO

H HO

HHOH

HOH

HHO

OHO

H

HO

H

H

HOH

H

OH

Flow

er(G

ermany)

[63]

18 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

96

1-[3-(120573-D

-Glucopyrano

syloxy)-

24-dihydroxyphenyl]-

3-(3-hydroxy)-

4-metho

xyph

enyl)-

2-prop

en-1-on

e

Okanin

4-methylether-

31015840-O

-120573-D

-glucop

yranoside

OOH

O

OH

HO

O

H HO

HHOH

HOH

H

OHO

Leaves

(Germany)aerial

(China)

[3562]

97

Okanin4-methylether-

3101584041015840-di-O

-120573-(41015840101584061015840101584041015840101584010158406101584010158401015840-

tetraacetyl)-

glucop

yranoside

OOH

OOH

O

HH

OH

HO

HOH

H

OO

O

Ac

Ac

AcAc

OO

H

HO

H

H

HOH

H

O

Aeria

l(Ch

ina)

[42]

98Ch

alcone210158404101584061015840-tr

imetho

xy-

4-O-D

-glucopyrano

syl-

dihydro

Not

foun

dLeaves

(China)

[60]

992-(34-D

ihydroxyph

enyl)-23-

dihydro-78

-dihydroxy-

4H-1-

benzop

yran-4-one

Okanin

isoOO

OH

HO

OH

OH

Leaves

(China)

[60]

100

2-(34-D

ihydroxyph

enyl)-

23-dihydro-8-hydroxy-

7[(246-tr

i-O-acetyl-120573

-D-

glucop

yranosyl)oxy]-4H

-1-b

enzopyran-4-on

eOO

OH

OH

OH

OH

O

HHO

H

HO

HO

OAc

Ac

Ac

Aeria

l(Ch

ina)

[34]

Evidence-Based Complementary and Alternative Medicine 19

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

101

57-Dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

OO

OH

HO

OH

Aeria

l(TanzaniaC

hina)

[4266]

102

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

7-O-glucopyrano

side

OO

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Tanzania)

[66]

103

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-4H

-1-benzop

yran-4-one

Luteolin

OO

HO

OH

OH

OH

Aeria

l(TanzaniaC

hina

Vietnam)

[343542

6667]

104

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5hydroxy-4H-1-

benzop

yran-4-one

Luteolin

7-O-120573-D

-glucop

yranoside

OOOH

OH

OH

OO

HO

H

HO

H

H

HOH

OH H

Aeria

l(Tanzania)

[66]

105

57-Dim

etho

xy-6-

(5-m

etho

xy-6-m

ethyl-

4-oxo-4H

-pyran-3-yl)-

2-ph

enyl-4H-1-

benzop

yran-4-one

5-O-M

ethylhosludin

O OOO

O

O

O

Aeria

l(Ugand

a)[68]

20 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

106

3-(120573-D

-Glucopyrano

syloxy)-

57-dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Astragalin

Kaem

pferol-3-O

-120573-D

-glucopyrano

side

HO

OH

O O

OH

O

O

HO

HHO

H

H

HOH

OH

H

Aeria

l(Ch

ina)

[35]

107

Kaem

pferol3-

(23-di-E

-p-cou

maroyl-

120572-L-rhamno

pyrano

side)

Not

foun

dAe

rial(Vietnam)

[67]

108

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5-hydroxy-36-dimetho

xy-

4H-1-

benzop

yran-4-one

Axillarosid

eO O

OH

OH

OO

OH

OO

HO

H

HO

H

H

HOH

H

OH

Aeria

l(Ch

ina)

[42]

109

57-Dihydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaureidin

O

O

O

O

O

HO

OH

OH

Who

le(Taiwan)

[69]

110

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaurein

O O

OO

O OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan)w

hole

(Taiwan)

[69ndash

71]

Evidence-Based Complementary and Alternative Medicine 21

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

111

5-Hydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-

67-dimetho

xy-4H-

chromen-4-one

Eupatorin

iso

OO O

OH

O

O OH

Not

foun

d(C

hina)

[3772]

112

2-(34-D

imetho

xyph

enyl)-7-

(120573-D

-glucopyrano

syloxy)-35-

dihydroxy-

8-metho

xy-4H-1-

benzop

yran-4-one

O O

OH

O O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

113

7-(120573-D

-Glucopyrano

syloxy)-

5hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-38-

dimehtoxy-4H-1-

benzop

yran-4-one

O O

O

OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

114

Isorhamnetin

3-[O

-120572-

L-rham

nopyrano

syl-

(1-2)-120573-D

-glucop

yranoside]

Not

foun

dAe

rial(Vietnam)

[67]

115

7-[(6-deoxy-120572-L-

Manno

pyrano

syl)o

xy]-

3-120573-D

-glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-4H

-1-benzop

yran-4-one

Luteoside

O O

OH

O

OH

OH

OH

HOH

H

H HO

HO

HO

OH

HO

H

HO

OH

OH

HH

Aeria

l(Ch

ina)

[42]

22 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

116

Luteolin

3-O-120573-D

-Glucopyrano

side

O O

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

HO

Aeria

l(Tanzania)

[66]

117

57-Dihydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Quercetagetin

3631015840-tr

imethylether

OOO

O

O OH

HO

OH

Aeria

l(Ch

ina)

[42]

118

6-((2S3S4S5S)-

Tetrahydro-345-tr

ihydroxy-

6-(hydroxymethyl)-2H

-pyran-2-yloxy)-5-hydroxy-

2-(4-hydroxy-3-

metho

xyph

enyl)-

37-dimetho

xy-4H-

chromen-4-one

Quercetagetin

3731015840-

trim

ethylether-6-O

-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[42]

119

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

JaceinQ

uercetin

3631015840-

trim

ethyl

ether-7-O-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan

China)

who

le(Taiwan)

[425470]

120

2-(34-D

ihydroxyph

enyl)-

357-trihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

OO

HO

OH

OH

OH

OH

Aeria

l(Ch

ina)leaves

(China)who

le(C

hina)

[356074]

Evidence-Based Complementary and Alternative Medicine 23

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

121

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-galactopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-4-one

Quercetin

3-O-120573-D

-galactosidehyperin

hyperosid

e

O O

HO

OH

OH

OH

OOHH

HOH

H

H HO

HO

OH

Aeria

l(TanzaniaJapan)

Leaves

(China)Who

le(C

hina)

[376066

717475]

122

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-glucopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

3-O-120573-

D-glucopyrano

side

O O

HO

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

Aeria

l(TanzaniaC

hina

Japan)L

eaves(Japan)

[355966

75]

123

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-

4-oxo-4H

-1-benzop

yran-3-yl-

120573-D

-glucopyrano

siduron

icacid

Quercetin

3-O-120573-D

-glucuron

opyranoside

O O

HO

OH

OH

OH

OHOH

HOH

H

H HO

HO

OHO

Aeria

l(TanzaniaJapan)

[6671]

124

3-[[6-O-(6-

Deoxy-120572-L-m

anno

pyrano

syl)-

120573-D

-galactopyrano

syl]o

xy]-

2-(34-dihydroxyph

enyl)-57-

dihydroxy-4H

-1-benzop

yran-4-one

Quercetin

3-O-rob

inob

iosid

e

O O

HO

OH

OH

OH

O

HOHH OH

HO

H HH

OO

OHH

HOH

H

H HO

HO

Aeria

l(Japan)W

hole

(Taiwan)

[7071]

24 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

125

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-

3-metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

OH

Roots(Brazil)

[55676]

126

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-

120573-D

-glucopyrano

syl]o

xy]-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120572-

L-rham

nopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

O

HHO HHO

HO H

HH

ORo

ots(Brazil)

[5676]

127

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-120573-D

-glucop

yranosyl]oxy]-5-

hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-3-metho

xy-

4H-1-

benzop

yran-4-one

Quercetin

341015840-

dimethyl

ether-7-O-rutinoside

O OOH

OH

O

O

OH O

HOH

H

HHO

H

O

HHO HHO

HO H

HH

O

HO

Aeria

l(Ch

ina)

[3435]

128

2-(34-D

ihydroxyph

enyl)-3-

(120573-D

-glucofurano

syloxy)-57-

dihydroxy-4H

-1-benzop

yran-

4-on

e

Isoq

uercitrin

O OOH

OH

OHO

H

OHH

OH

HO

H O HOH

OH

Aeria

l(Japan

China)

[3571]

Evidence-Based Complementary and Alternative Medicine 25

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

129

Quercetin

3-O-120573-D

-galactop

yranoside

O

OH

HO

OH OH

O

O

HO

OH

OHOH

OAe

rial(no

tstated)

[18]

130

Quercetin

3-O-rutinoside

O

O

R

OO

OH

OHO

H

O

OH

OH

H3C

H3C

H3C

OH

HO

Who

le(Taiwan)

[70]

26 Evidence-Based Complementary and Alternative Medicine

Table 6 Terpenoids isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(Country) References

131371111-Tetramethylbicyclo[810]undeca-

26-dieneBicyclogermacrene Leaves

(Brazil) [95]

13241111-Trimethyl-8-

methylenebicyclo[720]undec-4-ene

E-Caryophyllene Leaves(Brazil) [95]

1331-Methyl-5-methylene-8-

(1-methylethyl)-16-cyclodecadiene

Germacrene D Leaves(Brazil) [95]

1344-(15-Dimethyl-4-hexen-1-ylidene)-1-methyl-cyclohexene

Z-120574-Bisabolene Leaves(Brazil) [95]

135Decahydro-114-trimethyl-7-methylene-1H-cycloprop[e]-

azulene120573-Gurjunene Leaves

(Brazil) [95]

136 2669-Tetramethyl-148-cycloundecatriene

120572-Humulene120572-caryophyllene

Leaves(Brazil) [95]

13712344a568a-Octahydro-1-isopropyl-7-methyl-4-methylenenaphthalene

120575-Muurolene Leaves(Brazil) [95]

138

12344a568a-Octahydro-4a8-dimethyl-2-(1-methylethylidene)-

naphthalene

Selina-37(11)-diene

Leaves(Brazil) [95]

139(2E7R11R)-371115-

Tetramethyl-2-hexadecen-1-ol

Phytol

HO

Whole(Taiwan) [57]

Evidence-Based Complementary and Alternative Medicine 27

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

140 371115-Tetramethyl-2-hexadecenoic acid Phytanic acid

O

OH

Whole(Taiwan) [57]

141371115-Tetramethyl-2-

hexadecenyl ester-heptanoicacid

Phythylheptanoate O O

Leaves(Not stated) [33]

142

(3S10R13R)-17-((2R5R)-5-Ethyl-6-methylheptan-2-yl)-

2347891011121314151617-tetradecahydro-

1013-dimethyl-1H-cyclopenta[a]phenanthren-

3-ol

Campestrol

HO

Aerial(Tanzania) [18]

143 Not found Phytosterin-B Not foundNot found(TaiwanEgypt)

[39 96]

144 Stigmast-5-en-3-ol 120573-sitosterol

HOH

H

H

Aerial(Tanzania)

whole(Taiwan)

[18 31 57]

145

1314151617-Tetradecahydro-1013-

dimethyl-1H-cyclopenta[a]phenanthren-

3-yloxy)-tetrahydro-6-(hydroxymethyl)-2H-pyran-345-triol

120573-Sitosterolglucoside

H

H

HOH

HO

HHO

H

HOHH O

OH Not found(Egypt) [39]

146 5120572-Stigmasta-7-en-3120573-ol

HO

Whole(Taiwan) [57]

147 5120572-Stigmasta-722t-dien-3120573-ol

HO

Whole(Taiwan) [57]

148 Stigmasta-522-dien-3-ol Stigmasterol

HOH

H

H

Not found(Taiwan)aerial

(Tanzania)leaves (notstated)whole

(Taiwan)

[18 31 3357]

28 Evidence-Based Complementary and Alternative Medicine

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

149 Lup-20(29)-en-3-ol Lupeol

H

HH

HOH

Not found(Egypt) [39]

150 Lup-20(29)-en-3-ol acetate Lupeol acetate

H

HH

O

OH

Not found(Egypt) [39]

151 Olean-12-en-3-ol 120573-amyrin

H

HOH

Not found(Egypt) [39]

152 5913-Trimethyl-242526-trinoroleanan-3-ol Friedelan-3120573-ol

HO

Aerial(Tanzania) [18]

153 5913-Trimethyl-242526-trinoroleanan-3-one

Friedelinfriedelan-3-one

O

Aerial(Tanzania) [18]

1542610151923-

Hexamethyl-2610141822-tetracosahexaene

Squalene

Aerial(Tanzania)Leaves (Notstated)Whole(Taiwan)

[18 33 57]

155 120573120573-Carotene 120573-Carotene Leaves (Notstated) [97]

Evidence-Based Complementary and Alternative Medicine 29

studies [83] Hot water extracts of B pilosa varminor Sheriffwere also assessed for its antileukemic effects on leukemiccell lines L1210 U937 K562 Raji and P3HR1 using XTT-based colorimetric assays The extract inhibited the five celllines with IC

50values ranging from 145120583gmL to 586120583gmL

L1210 K562 Raji and P3HR1 were more sensitive to B pilosaextract with IC

50values below 200120583gmL [98]

Consistent with the antitumor activities of B pilosaextracts and fractions some of its phytochemicals alsoshowed anticancer activity as outlined in Table 11 Amongthem luteolin (103) a well-studied flavonoid with multiplebioactivities was more effective against tumor cell prolifera-tion than its derivatives with IC

50values ranging from 3 120583M

to 50120583M in cells and 5 to 10mgkg in animals Luteolin wasalso found to fight cancer as a food additive at concentrationsof 50 to 200 ppm [78] and prevent skin cancer [78] and cancer3invasion [103] Lee and colleagues reported that luteolinprevents cancer by inhibiting cell adhesion and invasion[103] Significant inhibition concentration was reported to be5 120583M and complete inhibition concentration was reported tobe 40120583M Moreover luteolin was reported to inhibit hepa-tocyte growth factor (HGF)-induced cell scattering as wellas cytoskeleton changes such as filopodia and lamellipodiawhichwas determined using phase-contrast and fluorescencemicroscopy Furthermore luteolin also inhibited the HGF-induced phosphorylation of c-Met ERK 12 and Akt aswell as the MAPKERK and P13K-Akt pathways [78 103]Other mechanisms underlying the anticancer activities ofluteolin are the inhibition of topoisomerase I and II whichinhibits cell replication and DNA repair thus promotingapoptosis regulation of PI-3-KinaseAktMAPKERKJNKactivation of apoptosis in the mitochondrial pathway byactivating caspase 9 and caspase 3 which were found inmalignant cells but not in normal human peripheral bloodmononuclear cells death receptor-induced apoptosis and acell-cycle arrest mechanism inhibition of fatty acid synthasewhich is upregulated in many cancer cells and sensitizationto chemotherapywhereby luteolin increases the susceptibilityof cancer cells to chemotherapy [78] Butein (82) is anotherflavonoid that showed a cytotoxic effect on human colonadenocarcinoma cell proliferation with a reported IC

50value

of 175 120583M Butein at 2 120583M affected the incorporation of[14C]-labeled leucine thymidine and uridine which cancause the inhibition of DNA RNA and protein synthesis ofhuman colon cancer cells Moreover butein also exhibitednoncompetitive inhibition of 1-chloro-24-dinitrobenzene(CDNB) in glutathione S-transferase (GST) activity Tumorresistance was correlated with high levels of GST thus buteininhibited proliferation of cancer cells [80] Another flavonoidpresent in B pilosa centaureidin (109) also showed anti-cancer activity in B lymphoma cells Centaureidin isolatedfrom Polymnia fruticosa inhibited tubulin polymerization invitro and induced mitotic figure formation in CA46 Burkittlymphoma cells Using turbidimetric assay the IC

50value of

centaureidin in inhibition of mitosis was 3120583M [104] Cyto-toxicity of centaureidin was further analyzed using AmericanNational Cancer Institute (NCI) 60 human tumor cell linesThe cytotoxicity potency of centaureidin expressed as GI

50

(50 growth inhibition in the NCI tumor line panel) was024120583M [81] These data mark centaureidin as a promisingantimitotic agent for tumor therapy

In addition to anti-tumor flavones polyynes found in B 4pilosa have also been shown to possess anti-tumor proper-ties Based on a bioactivity-directed isolation approach Wuand colleagues identified two polyyne aglycones from theethyl acetate fraction of B pilosa [44] 12-Dihydroxytrideca-57911-tetrayne (48) and 13-Dihydroxy-6(E)- tetradecene-81012-triyne (46) exhibited significant anticell prolifera-tion activity in primary human umbilical vein endotheliumcells (HUVEC) with IC

50values of 125 120583M and 173 120583M

respectively They also decreased angiogenesis and pro-moted apoptosis in human endothelial cells Their anti-angiogenic and cytotoxic effects correlated with activationof the CDK inhibitors and caspase-7 [44] In addition 12-Dihyroxy-5(E)-tridecene-7911-triyne (45) showed antian-giogenic effects in HUVECs with an IC

50value of 124 120583M as

evidenced by a decrease in the tube formation and migrationof HUVECs [43] The IC

50value of compound 45 in the

inhibition of basic fibroblast growth factor-induced HUVECgrowth was 282120583M However it had higher IC

50values

than those for lung carcinoma cells and keratinocytes Thiscompound could also inhibit cell proliferation of HUVECslung carcinoma A549 cells and HACAT keratinocytes Themechanism by which compound 45 inhibits HUVEC growthand angiogenesis is complicated and includes decreasing theexpression of cell cycle regulators (CDK4 cyclins D1 and Aretinoblastoma (Rb) and vascular endothelial growth factorreceptor 1) caspase-mediated activation of CDK inhibitorsp21 (Cip1) and p27 (Kip) upregulation of Fas ligand expres-sion downregulation of Bcl-2 expression and activation ofcaspase-7 and poly (ADP-ribose) polymerase [43]

32 Anti-Inflammatory Activity B pilosa is commonly usedto treat inflammatory disorders The anti-inflammatory phy-tochemicals present in B pilosa are listed in Table 11 5Cyclooxygenase-2 (COX-2) is a physiologically importantenzyme that converts arachidonic acid to prostaglandin(PGE2) Its expression is induced by a wide variety of external

stimuli indicating its involvement in inflammatory diseasesand it is used as an inflammatory marker [88] Yoshida andcolleagues studied the effects of the aqueous extracts of Bpilosa aerial parts in the production of COX-2 and PGE

2as

well as on the activation of mitogen activated protein kinases(MAPKs) in normal human dermal fibroblasts (HDFs) inresponse to inflammatory cytokine IL-1120573 This work showedthat IL-1120573 activated MAPKs such as ERK12 p38 and JNKto different extents and induced COX-2 expression TheCOX-2 expression in HDFs was regulated mainly by p38following IL-1120573 stimulation Consistently the p38 inhibitorSB203580 blocked this expression Using this cell platformB pilosa extracts were tested for inhibition of inflammationThe extract dose-dependently suppressed the activation ofp38 and JNK and moderately suppressed ERK12 as wellas suppressing COX-2 expression and PGE

2production

30 Evidence-Based Complementary and Alternative Medicine

Table7Ph

enylprop

anoids

isolatedfro

mB

pilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

156

3-(4-H

ydroxyph

enyl)-2-prop

enoica

cid

p-Cou

maricacid

OHO

OH

Who

le(Ja

pan)

[99]

157

2-Metho

xy-4-(2-prop

en-1-yl)-ph

enol

Eugeno

lOOH

Leaves

androots(Japan)

[99]

158

3-(4-H

ydroxy-3-m

etho

xyph

enyl)-

2-prop

enoica

cid

Ferulic

acid

O

OH

O

HO

Who

le(Ja

pan)

[99]

159

3-(34-D

ihydroxyph

enyl)-2-prop

enoic

acid

Caffeicacid

O

HO

OH

OH

Who

leandaeria

l(Japan)

[7599]

160

3-Prop

yl-3[(245-trim

etho

xyph

enyl)-

metho

xy]-24-pentanedione

3-Prop

yl-3-(245-

trim

etho

xy)benzyloxy-

pentan-24-dione

O

O

O

O

O

O

Leaves

(India)

[100]

161

3-(34-D

ihydroxyph

enyl)-2-

prop

enoica

cidethyleste

rCa

ffeateethyl

O

O

OH

OH

Not

foun

d(Taiwan)

who

le(Taiwan)

[434584]

Evidence-Based Complementary and Alternative Medicine 31Ta

ble7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

162

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

d-Erythron

icacid

2-O-caffeoyl-2-C

-methyl

O

HO

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

163

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

2-O-caffeoyl-2-C

-methyl

O

O

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

164

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-24-

dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

3-O-caffeoyl-2-C

-methyl

O

O

HO

HO

O

O

OH

OH

Leaves

(Japan)

[101]

165

4-(A

cetyloxy)-3-[[3-(34-

dihydroxyphenyl)-

1-oxo-2-propen-1-y

l-]oxy]-2-hydroxy-

2-methyl-b

utanoica

cid

O

HO

OH

O OHO

HO

OO

Not

foun

d(Ja

pan)

[73]

166

3-(34-D

ihydroxyph

enyl)-tetrahydro-4-

hydroxy-4-

methyl-5

-oxo-3-fu

ranyl

ester-2-prop

enoica

cid

3-O-C

affeoyl-2-C

-methyl-

D-erythrono

-14-la

cton

eHO

HO

O

OO

OOH

Leaves

(Japan)

[101]

167

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]-oxy]-145-

trihydroxy-cyclohexanecarboxylicacid

Chlorogenica

cid

O

HO

HO

OH

O

OH

O

OH

HO

Aeria

l(Japan)w

hole

(Taiwan)

[707175]

32 Evidence-Based Complementary and Alternative Medicine

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

168

4-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]oxy]-

135-tr

ihydroxy-cyclohexanecarboxylic

acid

4-O-C

affeoylqu

inicacid

OHO

HO

OH

OH

O

O

OH

OH

Aeria

l(Japan)

[71]

169

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-15-

dihydroxy-cyclo

hexanecarboxylicacid

34-Di-O

-caffeoylqu

inic

acid

HO

HO

O

O

O

O

OH

OH

HO

OH O

H

O

Aeria

l(Japan)W

hole

(Taiwan)

[45497071]

170

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-14-dihydroxy-

cyclo

hexanecarboxylicacid

35-Di-O

-caffeoylqu

inic

acid

HO

HO

O O

HO

O

HO

OH OO

OHOH

Aeria

l(Japan)w

hole

(Taiwan)

[45497071]

Evidence-Based Complementary and Alternative Medicine 33

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

171

34-bis[[(2E

)-3-(34-D

ihydroxyph

enyl)-1-

oxo-2-prop

en-1-

yl]oxo]-15

-dihydroxy-

cyclo

hexanecarboxylicacid

45-Di-O

-caffeoylqu

inic

acid

OHO

OH

HO

O

O

OH

OH

O

OH

HO

OWho

le(Taiwan)

[454970]

172

3-[4-[[-6-O-[3-(4-H

ydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]-phenyl]-2-prop

enoica

cid

120573-D

-p-C

oumaricacid4-O

-(6-O

-p-caoum

aroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HOH

HO

O

O

Leaves

(Japan)

[59]

173

3-[4-[[2-O

-Acetyl-6

-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]ph

enyl]2-propeno

icacid

120573-D

-p-C

oumaricacid4-O

-(2-O

-acetyl-6

-O-p-

caou

maroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HO

HO

O

O

Leaves

(Japan)aerial

(China)

[3459]

174

67-Dihydroxy-2-chrom

enon

eEsculetin

cicho

rigenin

OHO HO

O

Not

foun

d(Egypt)

[39]

34 Evidence-Based Complementary and Alternative Medicine

Table 8 Aromatic compounds isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

175 12-Benzenediol PyrocatechinHO

HOWhole(Japan) [99]

176 4-Ethyl-12-benzenediol Pyrocatechol

OH

OH

Whole(Japan) [99]

177 Dimethoxyphenol26-dimethoxyphenol

O

HO

O

Roots (Japan) [99]

178 4-Ethenyl-2-methoxy-phenol p-Vinylguaiacol

O

OHWhole(Japan) [99]

179 2-Hydroxy-6-methylbenzaldehyde

6-Methyl-salicylaldehyde O

OH

Whole(Japan) [99]

180 Benzene-ethanol 2-Phenyl-ethanolHO

Whole(Japan) [57]

181 4-Hydroxy-3-methoxy-benzaldehyde Vanillin O

O

HOAerial(Japan) [99]

182 3-Hydroxy-4-methoxy-benzaldehyde Vanillin iso

OO

OH

Leaves(Japan) [99]

183 4-Hydroxybenzoic acid p-Hydroxybenzoicacid

O

HOOH Whole

(Japan) [99]

184 2-Hydroxybenzoic acid Salicylic acidO

HO

HO

Stem androots (Japan) [99]

185 34-Dihydroxybenzoicacid Protocatechuic acid

O

HOOH

OH

Whole(Japan) [99]

1864-Hydroxy-

3methoxybenzoicacid

Vanillic acid

OOH

O

OH

Aerial(Uganda)

Roots (Japan)[68 99]

Evidence-Based Complementary and Alternative Medicine 35

Table 8 Continued

S N IUPAC names Common names Structure Plant part(country) References

187 345-Trihydroxybenzoicacid Gallic acid

O

OHHO

HO

HO

Whole(China) [102]

[88] This work supports the use of B pilosa as an anti-inflammatory agent however no compounds responsible forthe anti-inflammatory activity of B pilosa were identified

A further study also reported the anti-inflammatoryactivity as well as the antiallergic activity of B pilosa [75] Inthis study dried powder of the aerial part of B pilosa whichhad been pretreated with the enzyme cellulosine was usedfor further tests The results showed that oral administrationof the cellulosine-treated B pilosa lowered the level of serumIgE in mice 10 days after immunization with DNP (24-dintrophenyl)-Ascaris as an antigen This treatment alsoreduced dye exudation in skin induced by passive cutaneousanaphylaxis and production of inflammatory mediators his-tamine and substance P in rats [75] Phytochemical analysisshowed that cellulosine treatment increased the percentage ofcaffeic acid and flavonoids This study suggests that B pilosaand its phenolics have anti-inflammatory functions

Phenolics and polyynes are major anti-inflammatoryphytochemicals present in B pilosa (Table 11) Unsurpris-6ingly phenolics such as luteolin (103) and ethyl caffeate(161) that are major constituents of B pilosa have alsobeen reported to possess anti-inflammatory activity Lute-olin was reported to exhibit anti-inflammatory activity inmacrophages Xagorari and colleagues showed that luteolininhibited the release of inflammatory cytokines TNF-120572 andinterleukin-6 in RAW 2647 cells following LPS stimulation[87] It inhibited TNF-120572 production with an IC

50value

of 1120583M The underlying anti-inflammatory mechanism ofluteolin was reported to be the inactivation of Akt andNF-120581B activation [87] In addition luteolin was reportedto confer anti-inflammatory activity through inhibition ofLPS-stimulated iNOS expression in BV-2 microglial cellsIt inhibited LPS-activated microglia in a dose-dependentmanner with an IC

50value of 69 120583MMoreover immunoblot

and RT-PCR data proved that luteolin suppressed I120581B-120572degradation and iNOS expression in LPS-activated microglia[85] Kim and colleagues stated that luteolin may havebeneficial effects on inflammatory neural diseases throughinhibition of iNOS expression [85] A related study revealedthat luteolin decreased the transcriptional activity of NF-120581BRelA via partial inhibition of TNF-mediated NF-120581B DNAbinding activity Luteolin also inhibited Akt phosphorylationand induced degradation of a transcription factor interferonregulatory factor (IRF) [86]

Chiang and colleagues showed that ethyl caffeate(161) significantly inhibited NO production in mousemacrophages RAW 2647 cells [84] Based on MTT assays

they concluded that this inhibition was not due to thecytotoxicity of ethyl caffeate The IC

50value of ethyl caffeate

in the inhibition of NO production was 55 120583gmL slightlylower than curcumin (positive control) which has anIC50

value of 65120583gmL They demonstrated that ethylcaffeate exerted anti-inflammatory activity via the reducedtranscription and translation of iNOS (inducible nitric oxidesynthase) in RAW 2467 cells In addition this compoundalso suppressed COX-2 expression in RAW 2467 cellsand MCF-7 cells The in vivo anti-inflammatory effectof ethyl caffeate was verified by testing in TPA-treatedmouse skin Like celecoxib the positive control ethylcaffeate significantly abolished COX-2 expression in adose-dependent manner Ethyl caffeate at 1mg200120583Lsite(24mM) inhibited COX-2 expression at a level comparableto celecoxib at 1mg200120583Lsite (13mM) Remarkably ethylcaffeate at 48mM (2mg200120583Lsite) was more effectivethan celecoxib at 131mM (10mg200120583Lsite) Moreoverthis compound inhibited the activation of nuclear factor-120581B(NF-120581B) by LPS via the prevention of NF-120581B binding toDNA [84] In addition 3 ethyl caffeate analogs (ethyl 34-dihydroxyhydrocinnamate ethyl cinnamate and catechol)also showed different degrees of NF-120581B binding to DNA aslisted in Table 12 Ethyl cinnamate lacks a catechol moietywhich results in ineffective inhibition of NF-120581B binding toDNA [84]

Pereira and colleagues assessed the anti- 7inflammatory and immunomodulatory activities ofB pilosa methanol extract as well as one polyyne2-O-120573-glucosyltrideca-11(E)-en-3 579-tetrayn-12-diol(54) in T lymphocytes and a zymosan-induced arthritismouse model [27] They first examined the in vitro effect of 8the B pilosa extract and compound 54 on cell proliferation ofhuman T cells stimulated with 5120583gmL phytohemagglutinin(PHA) or 100 nM 12-O-tetradecanoyl phorbol-13-acetate(TPA) plus 15 120583M ionomycin and on cell proliferation ofmouse T cells stimulated with 5 120583gmL concanavalin A(Con A) The data demonstrated that both methanol extractand compound 54 suppressed T-cell proliferation in adose-dependent manner The estimated IC

50values of the B

pilosa extract against human T cells stimulated with 5120583gmLPHA and 100 nM TPA plus 15120583M ionomycin were 125 and25 120583gmL respectively In comparison with the methanolextract compound 54 showed 10-fold more inhibition ofhuman T-cell proliferation with an estimated IC

50value of

15 120583gmL Accordingly the B pilosa extract and compound54 dose-dependently suppressed mouse T-cell proliferation

36 Evidence-Based Complementary and Alternative Medicine

Table 9 Porphyrins isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

188

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(15S16S)-10-ethenyl-5-ethyl-1161820-tetrahydro-6111522-tetramethyl-1820-dioxo-15H-912-imino-212-

metheno-471714-dinitrilopyrano[43-

b]azacyclononadecine-16-propanoic

acid

AristophyllC

NH N

HNN

O

O OOOPhytyl

O

Leaves(Taiwan) [100]

189

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -20b-(methoxycarbonyl)-9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-dinitrilo-12-

dioxeto-[310158404101584034]-cyclo-pent[1 2]azacyclononadecine-

19-propanoic acid

Bidenphytin A

NH N

HNN

OPhytylO O O

O O

Leaves(Taiwan) [100]

190

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -2a-

hydroxy-20b-(methoxycarbonyl)-

9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-

dinitrilo-12-dioxeto[310158404101584034]-cyclo-pent[12-b]-

azacyclononadecine-19-propanoic acid

Bidenphytin B

NH N

HNN

OPhytylO O O

O O

OH

Leaves(Taiwan) [100]

191

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO O

HOOO

Leaves(Taiwan) [100]

192

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

Evidence-Based Complementary and Alternative Medicine 37

Table 9 Continued

S N IUPAC names Common names Structure Plant part(country) References

193

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

Leaves(Taiwan) [100]

194

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

195

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3S4S21R)-9-ethenyl-14-ethyl-21-(methoxycarbonyl)-

481318-tetramethyl-20-oxo-3-phorbinepropanoic acid

Pheophytin A

NH N

HNN

PhytylO

O

OO O

Leaves(Taiwan) [100]

with estimated IC50values of 30 and 25 120583gmL respectively

Taken together the data indicate that the B pilosa extractand compound 54 act on human and mouse T cells Totest the in vivo effect of the B pilosa extract and compound54 a zymosan-induced arthritis mouse model was usedThis model was established from B10ASgSnJ mice with aninjection of zymosan (015mg) The zymosan-injected mice9received an intraperitoneal injection of the B pilosa extract(1 5 or 10mg) at one dose a day for 5 days Popliteal lymphnode (PLN) weight was monitored to check the developmentof arthritis The results revealed that 10mg of the methanolextract of B pilosa extract could significantly diminishinflammation as evidenced by PLN weight [27] This worksuggests that B pilosa (and compound 54) can suppressimmune response and inflammation

33 Antidiabetic Activity Anti-diabetic agents are primarilydeveloped from plants and other natural resources [8 24 4951 53] B pilosa is one of 1200 plant species that have beeninvestigated for antidiabetic activity [120 121]B pilosa is usedas an anti-diabetic herb in America Africa and Asia [51 120

122] Many studies have indicated that B pilosa could treattype 1 diabetes (T1D) and type 2 diabetes (T2D) in animals

Etiologically speaking T1D is caused by autoimmune-mediated destruction of pancreatic 120573 cells leading to insulindeficiency hyperglycemia and complications Currentlythere is no cure for T1D Polarization ofTh cell differentiationcontrols the development of T1D Suppression of Th1 celldifferentiation and promotion of Th2 cell differentiationameliorate T1D [123] One study showed that the butanolfraction of B pilosa inhibited T-cell proliferation decreasedTh1 cells and cytokines and increased Th2 cells andcytokines leading to prevention of T1D in nonobesediabetic (NOD) mice [49] Based on a bioactivity-directedisolation strategy 3 polyynes 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne (53) also known ascytopiloyne 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D- Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne (50) were identifiedfrom B pilosa [49 53] The IC

50value of the butanol fraction

was 200120583gmL This inhibition was reported to be partiallyattributed to cytotoxicity because the butanol fraction at

38 Evidence-Based Complementary and Alternative Medicine

Table 10 Other compounds isolated from B pilosa [30]

SN IUPAC names Common names Structure Plant part(country) References

19637-Dihydro-137-

trimethyl-1H-purine-26-dione

CaffeineN

NN

NO

O

Aerial(Uganda) [68]

197

1-((2R4S5R)-Tetrahydro-4-hydroxy-5-(hydroxymethyl)furan-2-yl)-5-

methylpyrimidine-24(1H3H)-dione

ThymidineHO

HO

ON

O

NHO Not found

(China) [37]

198 1-(2-Thienyl)-ethanone 2-Acetyl-thiophene

S

O

Roots(Germany) [41]

199

(2R3S4S5S)-2-(Heptan-2-yloxy)-tetrahydro-6-

((tetrahydro-34-dihydroxy-5-

(hydroxymethyl)furan-2-yloxy)methyl)-

2H-pyran-345-triol

Heptanyl2-O-120573-xylofuranosyl-

(1rarr 6)-120573-glucopyranoside

O

OOH

OH

HO

OHO

H

HO

H

H

HOHH

O

Whole(Taiwan) [70]

200

2-[(3R7R11R)-3-Hydroxy-371115-

tetramethylhexadecyl]-356-trimethyl-

25-cyclohexadiene-14-dione

120572-Tocopherylquinone

OO

HO

Whole(Taiwan) [32]

2017-O-(410158401015840610158401015840-Diacetyl)-

120573-D-glucopyranoside

Not found Leaves(China) [60]

180 120583gmL could cause 50 death of Th1 cells Moreoverthis study suggested that the butanol fraction may preventdiabetes in NOD mice in vivo via downregulation of Th1cells or upregulation Th2 cells that have effects which areantagonistic of those of Th1 cells [49] This was proven byintraperitoneal injection of the butanol fraction at a doseof 3mgkg BW 3 times a week to NOD mice from 4 to 27weeks This dosage resulted in lower incidence of diabetes(33) At a dose of 10mgkg the butanol fraction of Bpilosa totally eliminated (0) the initiation of the diseaseTo further support this result assessment of IgG2a and IgEproduction was performed in the serum of NOD mice Asin vivo results obtained from intracellular cytokine stainingexperiments were not very conclusive levels of IgG2a and IgE

were measured since Th1 cytokine IFN120574 and Th2 cytokineIL-4 favor the production of IgG2a and IgE respectively Asexpected high levels of IgE and some decline in the levels ofIgG2a were observed in the serum Profiling of the butanolextract revealed five compounds 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne(50) 45-Di-O-caffeoyl- quinic acid 35-Di-O-caffeoylquinicacid and 34-Di-O-caffeoylquinic acid Only the first twocompounds showed similar effects on the prevention ofdiabetes in NOD mice as the B pilosa butanol fractionMoreover compound 50 showed greater activity thancompound 69 in terms of enhancement (by 34 comparedto 8) of differentiation of Th0 to Th2 at 15 120583gmL (both

Evidence-Based Complementary and Alternative Medicine 39

Table 11 Chemical constituents of B pilosa and their biological activities

SN Name Classification Molecularformula Biological activities

109 Centaureidin [91] Flavonoid C18H16O8Anti-listerial [69 91]

Cytotoxic [10]

110 Centaurein [91] Flavonoid C24H26O13

Anti-listerial [69 91]Cytotoxic [10]Anti-viral [105]

103 Luteolin [106] Flavonoid C15H10O6

Anti-viral [107 108]Cytotoxic [77]

Anti-inflammatory [109]Anti-allergic [109]

82 Butein [110] Flavonoid C15H12O5Anti-leishmanial [111]

Cytotoxic [78]48 12-Dihydroxytrideca-57911-tetrayne [112] Polyyne C13H12O2 Anti-angiogeneic [112]46 13-Dihyroxy-6(E)-tetradecene-81012-triyne [112] Polyyne C14H16O2 Anti-angiogeneic [112]

45 12-Dihyroxy-5(E)-tridecene-7911-triyne [113] Polyyne C13H14O2Anti-angiogeneic [113]Anti-proliferative [113]

64 1-Phenylhepta-135-triyne [114] Polyyne C13H8

Anti-microbial [115]Anti-malarial [3]Cytotoxic [3]Antifungal [15]

27 Linoleic acid [79] Fatty acid C18H32O2Anti-viral (100)Cytotoxic [17]

161 Ethyl caffeate [84] Phenylpropanoid C11H12O4 Anti-inflammatory [84]

54 2-O-120573-Glucosyltrideca-11(E)-en-3579-tetrayn-12-diol [26] Polyyne C19H20O7Immunosuppressive andAnti-inflammatory [26]

53 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne [50] Polyyne C19H22O7Anti-diabetic [116]

Anti-inflammatory [50]

69 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne [28] Polyyne C20H26O7Anti-diabetic [28]

Anti-inflammatory [94]

50 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne [28] Polyyne C19H24O7

Anti-diabetic [28]Anti-inflammatory [94]

Anti-malarial andantibacterial [47]

129 Quercetin 3-O-120573-D-galactopyranoside [18] Flavonoid C21H20O12 Anti-inflammatory [117]

170 35-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

171 45-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

169 34-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

126 Quercetin 331015840-dimethyl ether7-O-120572-L-rhamnopyranosyl-(1rarr 6)-120573-D-glucopyranoside [118] Flavonoid C21H20O11 Anti-malarial [119]

125 Quercetin 331015840-dimethyl ether-7-O-120573-D-glucopyranoside [118] Flavonoid C21H20O12 Anti-malarial [119]70 1-Phenyl-13-diyn-5-en-7-ol-acetate [27] Polyyne C15H12O2 Anti-malarial [27]199 Heptanyl 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside [70] Miscellaneous C18H44O10 Antioxidant [70]124 3-O-Rabinobioside [70] Saccharide C27H38O15 Antioxidant [70]130 Quercetin 3-O-rutinoside [70] Flavonoid C27H38O15 Antioxidant [70]167 Chlorogenic acid [70] Phenolic C16H26O9 Antioxidant [70]119 Jacein [70] Flavonoid C24H26O13 Antioxidant [70]

49 (R)-12-dihydroxytrideca-357911-pentayne [47] Polyyne C13H8O2Anti-malarial andAntibacterial [47]

40 Evidence-Based Complementary and Alternative Medicine

Table 12 Structure and activity relationship studies of ethyl caffeateusing in vitro NF-120581BDNA binding assays [84]

Compound Concentration(120583M)

NF-120581BDNA binding

Ethyl caffeate 50 100inhibition

Ethyl 34-dihydroxyhydrocinnamate 100 100inhibition

Catechol 400 100inhibition

Ethyl cinnamate 400 Noinhibition

Table 13 Apoptosis in cocultures of T cells and pancreatic 120573 cells[53]

Cellmedium apoptosis andnecrosis

CD4+ T cellscontrol medium lt4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 2

CD4+ T cellswith cytopiloyne-treated 120573 cells 18CD4+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 7

CD8+ T cellscontrol medium 4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 4

CD8+ T cellswith cytopiloyne-treated 120573 cells 4CD8+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 4

Table 14 Radical scavenging activities of B pilosa extracts [70]

Extractscontrol DPPH assayIC50 (120583gmL)

NBThypoxanthinesuperoxide assayIC50 (120583gmL)

Quercetin 198 15Ascorbic acid 634 Not determined120572-tocopherol 897 Not determinedEthyl acetateextract 1383 597

Butanol extract 1669 114Water extract gt100 gt100

compounds) and inhibition (by 40 compared to 10) ofdifferentiation toTh1 at the same concentration [49]

Among the three polyynes found in B pilosa cytopiloyne(53) had the most potent anti-T1D activity [53] To test the invivo effect of cytopiloyne NODmice received intraperitonealor intramuscular injection of cytopiloyne at 25120583gkg BW3 times per week Twelve-week-old NOD mice started todevelop T1D and 70 of NOD mice aged 23 weeks andover developed T1D Remarkably 12- to 30-week-old NODmice treated with cytopiloyne showed normal levels of bloodglucose (lt200mgdL) and insulin (1-2 ngmL) Consistent

Table 15 Radical scavenging activity of secondarymetabolites fromB pilosa [70]

Metabolite (Table 11)control DPPH assay IC50 (120583gmL)199 Not determined124 53130 68167 105169 33171 38119 Not determined110 Not determinedQuercetin 256Caffeic acid 890

Table 16 Antioxidant activity of the essential oils and water extractsfrom B pilosa [92]

Extract IC50 (120583gmL)Leaf essential oils 47Flower essential oils 50Leaf extract 61Flower extract 172

with T1D incidence cytopiloyne delayed and reduced theinvasion of CD4+ T cells into the pancreatic islets [53]

In vitro study showed that cytopiloyne (53) inhibited thedifferentiation of naıveTh (Th0) cells (ie CD4+ T cells) intoTh1 cells and promoted differentiation of Th0 cells into Th2cells [50] The in vitro data are consistent with the in vivoresults indicating that cytopiloyne reducedTh1differentiationand increased Th2 differentiation as shown by intracellularcytokine staining and FACS analysis [53] In line with theskewing of Th differentiation the level of serum IFN-120574 andIgG2c decreased while that of serum IL-4 and serum IgEincreased compared to the negative controls (PBS-treatedmice) Cytopiloyne also enhanced the expression of GATA-3 a master gene for Th2 cell differentiation but not theexpression of T-bet a master gene forTh1 cell differentiationfurther supporting its role in skewingTh differentiation [53]

Also importantly cytopiloyne partially depleted CD4+rather than CD8+ T cells in NOD mice [53] As shown inTable 13 coculture assays showed that the depletion of CD4+T cells was mediated through the induction of Fas ligandexpression on pancreatic islet cells by cytopiloyne leading toapoptosis of infiltrating CD4+ T cells in the pancreas via theFas and Fas ligand pathways However cytopiloyne did notinduce the expression of TNF-120572 in pancreatic islet cells andthus had no effect on CD8+ T cells [53]

In addition Chang and colleagues showed that cytopi-loyne dose-dependently inhibited T-cell proliferation stimu-lated by IL-2 plus Con A or anti-CD3 antibody using [3H]thymidine incorporation assay [53]

Overall the mechanism of action of cytopiloyne andprobably its derivatives in T1D includes inhibition of T-cellproliferation skewing of Th cell differentiation and partialdepletion of Th cells Due to the anti-diabetic mechanisms

Evidence-Based Complementary and Alternative Medicine 41

Table 17 Antibacterial activity of essential oils and flower extracts from B pilosa [92]

Strain Mean zone of inhibition (mm)Leaf essential oil Flower essential oil Leaf extract Flower extract

Micrococcus flavus 127 plusmn 03 87 plusmn 03 102 plusmn 02 108 plusmn 03Bacillus subtilis 173 plusmn 19 117 plusmn 02 109 plusmn 02 103 plusmn 02Bacillus cereus 190 plusmn 14 112 plusmn 03 118 plusmn 04 185 plusmn 10Bacillus pumilus 123 plusmn 07 108 plusmn 02 105 plusmn 04 77 plusmn 02Escherichia coli 137 plusmn 04 203 plusmn 07 102 plusmn 11 140 plusmn 13Pseudomonas ovalis 125 plusmn 08 137 plusmn 15 102 plusmn 06 125 plusmn 06

Table 18 Antibacterial activity of root extracts from B pilosa [93]

Strain MIC50 (mgmL)Methanol extract Acetone extract

Bacillus cereus 10 mdashEscherichia coli 5 5Klebsilla pneumonia 5 10Micrococcus kristinae 10 mdashPseudomonas aeruginosa 10 10Staphylococcus aureus 5 10Sraphylococcus epidermidis 5 5Serratia marcescens 10 mdashShigelea flexneri 10 mdashStreptococcus faecalis 10 mdash

Table 19 Antibacterial activity of B pilosa of compound 29 [47]

Strain MIC50 (120583gmL)Escherichia coli NIHJ 1Escherichia coli ATCC25922 1Klebsiella pneumoniae ATCC700603 128Serratia marcescens ATCC13880 16Pseudomonas aeruginosa ATCC27853 8Staphylococcus aureus FDA209P 05Staphylococcus aureus ATCC29213 025Staphylococcus aureus N315 (MRSA) 05Enterococcus faecalis ATCC29212 2Enterococcus faecalis NCTC12201 (VRE) 1Bacillus subtilis ATCC6633 05Candida albicans ATCC10231 025

of action it was hypothesized that cytopiloyne protectsNOD mice from diabetes by a generalized suppression ofadaptive immunity To evaluate this hypothesis ovalbumin(Ova) was used as a T-cell dependent antigen to primeNOD mice which had already received cytopiloyne orPBS vehicle Ova priming boosted similar anti-Ova titersin cytopiloyne-treated mice and PBS-treated mice but adifference in immunoglobulin isotype was observed in thetwo groups Thus it was concluded that cytopiloyne is animmunomodulatory compound rather than an immunosup-pressive compound [50 53]

T2D is a chronic metabolic disease with serious com-plications resulting from defects in either insulin secretion

insulin action or both [124] A study by Ubillas et alshowed that the aqueous ethanol extract of the aerial partof B pilosa at 1 gkg body weight (BW) lowered bloodglucose in dbdb mice a T2D mouse model [51] Basedon a bioactivity-guided identification compounds 69 and50 were identified Further the mixture of the compounds(69 50) in a 2 3 ratio significantly decreased blood glucoseconcentration and reduced food intake on the second dayof treatment when administered at doses of 250mgkg twicea day to C5BLKs-dbdb mice When tested at 500mgkga more substantial drop in blood glucose level as wellas the stronger anorexic effect (food intake reduced from58 gmouseday to 25 gmouseday) was observed [51] Inthis study it was suggested that the blood glucose lower-ing effect of B pilosa was caused in part by the hungersuppressing effect of its polyynes [51] However the hungersuppressing effect of the ethanol extract of B pilosa wasnot found in the studies described below In another study[24] water extracts of B pilosa (BPWE) were used indiabetic dbdb mice aged 6ndash8 weeks with postprandialblood glucose levels of 350 to 400mgdL Like oral anti-diabetic glimepiride which stimulates insulin release onesingle dose of BPWE reduced blood glucose levels from374 to 144mgdL The antihyperglycemic effect of BPWEwas inversely correlated to an increase in serum insulinlevels suggesting that BPWE acts to lower blood glucose viaincreased insulin production However BPWE had differentinsulin secretion kinetics to glimepiride [24] One flaw incurrent anti-diabetics is their decreasing efficacy over timeThe authors investigated the long term anti-diabetic effectof BPWE in dbdb mice BPWE reduced blood glucoseincreased blood insulin improved glucose tolerance andreduced the percentage of glycosylated hemoglobin (HbA1c)Both long-term and one-time experiments strongly supportthe anti-diabetic action of BPWE [24] In sharp contrast toglimepiride BPWE protected against islet atrophy in mousepancreas The investigators further evaluated anti-diabeticproperties of 3 B pilosa varieties B pilosa L var radiate(BPR) Bpilosa L var pilosa (BPP) and B pilosa L varminor(BPM) in dbdb mice [8] One single oral dose (10 50 and250mgkg body weight) of BPR BPP or BPM crude extractsdecreased postprandial blood glucose levels in dbdb micefor up to four hours and the reduction of glucose levels inthe blood appeared to be dose-dependent Comparing thethree variants BPR extract resulted in a higher reduction inblood glucose levels when administered at the same dose as

42 Evidence-Based Complementary and Alternative Medicine

Table 20 Antifungal activity of B pilosa [92]

Partextract Concentration (ppm) Strain InhibitionCortiicum rolfsii Fusarium solani Fusarium oxysporum

Leaves

Essentialoils

100 857 plusmn 09 682 plusmn 0 745 plusmn 17250 960 plusmn 08 779 plusmn 18 879 plusmn 04

AqueousExtracts

100 446 plusmn 17 605 plusmn 21 716 plusmn 07250 942 plusmn 03 689 plusmn 07 824 plusmn 19

Flowers

Essentialoils

100 604 plusmn 09 892 plusmn 04 869 plusmn 05250 894 plusmn 12 980 plusmn 03 949 plusmn 06

AqueousExtracts

100 331 plusmn 11 714 plusmn 07 573 plusmn 22250 661 plusmn 14 912 plusmn 0 900 plusmn 07

Table 21 Antifungal activity of B pilosa root extracts [93]

Strain LC50 (mgmL)Acetoneextracts

Methanolextracts

Waterextracts

Aspergillus niger 014 006 007Aspergillus flavus 1091 658 0Penicillium notatum 005 005 005

the other two varieties In terms of serum insulin levels a doseof 50mgkg of each extract was used and the BPR extracttogether with the three polyynes significantly increased theserum insulin level in dbdb mice Long-term experiments(28-day treatment) were then conducted using diabetic micewith postprandial glucose levels from 370 to 420mgdL andglimepiride was used as positive controlThe range of dosagesapplied was from 10mgkg BW to 250mgkg BW Resultsshowed that the positive control as well as the crude extractsof the three varieties lowered the blood glucose levels indbdb mice However only BPR extract containing a higherpercentage of cytopiloyne (53) reduced blood glucose levelsand augmented blood insulin levelsmore than BPP andBPMThe percentage of glycosylated hemoglobin A1c (HbA1c) wasalso measured and found to be 79 plusmn 05 in mice aged 10ndash12 weeks and 66 plusmn 02 61 plusmn 03 and 62 plusmn 03 inthe blood of age-matchedmice following treatment with BPRcrude extract (50mgkg) glimepiride (1mgkg) and com-pound 53 (05mgkg) respectively [8] Among the polyynesfound in B pilosa cytopiloyne was the most effective againstT2D Hence cytopiloyne was used for further study on anti-diabetic action and mechanism [125] The data confirmedthat cytopiloyne reduced postprandial blood glucose lev-els increased blood insulin improved glucose tolerancesuppressed HbA1c level and protected pancreatic islets indbdb mice Nevertheless cytopiloyne failed to decreaseblood glucose in streptoztocin (STZ)-treated mice whose bcells were already destroyed Additionally cytopiloyne dose-dependently increased insulin secretion and expression in bcells as well as calcium influx diacylglycerol and activationof protein kinase C120572 Collectively the mechanistic studiessuggest that cytopiloyne treats T2D via regulation of insulin

production involving the calciumDAGPKC120572 cascade in bcells

The studies detailed above point to the conclusion thatcytopiloyne and related polyynes (compounds 69 and 49)are anti-diabetics in animal models The data uncover a newbiological action of polyynes It should be noted that likeall anti-diabetic drugs cytopiloyne failed to prevent or curediabetes completely but reduced diabetic complications [125]Intriguingly 34 polyynes have been found in B pilosa so farIt remains to be seen whether all the polyynes present in thisplant have anti-diabetic activities

34 Antioxidant Activity Free radicals can damage cellularcomponents via a series of chemical reactions [89] leadingto development and progression of cardiovascular diseasecancer neurodegenerative diseases and ageing [70] Freeradicals nitric oxide (NO) and superoxide anions can beproduced in macrophages to kill microbes However anexcessive generation of the free radicals under pathologicalconditions is associated with a wide range of illnessesPlants are known to be rich in antioxidant phytochemicalsChiang and colleagues evaluated the free radical scaveng-ing activity of crude extract fractions and compounds ofB pilosa using 11-diphenyl-2-picrylhydrazyl (DPPH) andhypoxanthinexanthine oxidase assays [70] Using DPPHand hypoxanthinexanthine oxidase assays they found thatthe B pilosa crude extract and the ethyl acetate butanoland water fractions had free radical scavenging activityNine compounds Heptyl- 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside (199) 3-O-Rabinobioside (124) Quercetin3-O-rutinoside (130) Chlorogenic acid (167) 34-Di-O-caffeoylquinic acid (169) 35-Di-O-caffeoylquinic acid (170)45-Di-O-caffeoylquinic acid (171) Jacein (119) and Cen-taurein (110) had DPPH radical scavenging activity [70]The IC

50values of the B pilosa crude extractfractions and

compounds are summarized in Tables 14 and 15 respectivelyMeasurement of free radical scavenging activities is one

way of assessing the antioxidant activities of B pilosa andits fractions and compounds It is interesting that the ethylacetate and butanol fractions are more active than the waterfraction and B pilosa crude extract [70] Of the secondarymetabolites only phenolic compounds 124 130 167 169 and171 showed significant DPPH-radical scavenging activities

Evidence-Based Complementary and Alternative Medicine 43

Further analysis of the structure-activity relationship of thecompounds suggested that substitution of the C3 hydroxylgroup with glycosides increased the activity approximately2-fold (for example in compounds 124 and 130) relative toquercetin (-OH in its C3) [70] Further modification of thestructures of the active compounds needs to be performed totest the effects of various substituents on activity The reasonwhy most of the antioxidant compounds contain phenolmoieties in their structure could be that the reduction-oxidation (redox) properties of phenols allow them to actas reducing agents singlet oxygen quenchers and hydrogendonors

A complementary study by Muchuweti and colleaguesdetermined phenolic content antioxidant activity and thephenolic profile of B pilosa methanol extract [89] Theyestimated that the phenolic content of the methanol extractof B pilosawas 11028plusmn22mgg [89] Vanillin hydroxyben-zaldehyde caffeic acid coumaric acid and ferulic acid werefound in this extractThe B pilosa extract also showed DPPHradical scavenging activity Furthermore the antioxidantactivity of the flavonoids found in B pilosa was correlatedwith its hepatoprotective effects through their inhibitionof NF-120581B activation which may lessen the oxidative stresscaused by the production of free radicals during liver injury[60]This activity might also be due to the anti-inflammatoryeffects of the aqueous extracts of B pilosa aerial parts on theinhibition of COX-2 and PGE

2production [88]

Essential oils from B pilosa flowers and leaves are alsoreported to possess antioxidant activity With the aim ofreplacing chemically synthesized additives Deba and col-leagues [92] worked on the antioxidant antibacterial andantifungal activities of essential oils and water extracts of Bpilosarsquos leaves and flowers Table 16 summarizes the resultsobtained from DPPH free radical scavenging assay

It can be inferred fromTable 16 that essential oils from theleaves possessed the highest activity It is reported elsewherethat monoterpenes present in essential oils such that of Bpilosa have protective effects and antioxidant properties [92]Beta-carotene bleaching method was also performed Leavesessential oils and aqueous extracts of the leaves and flowersshowed higher activity than the flower essential oils This isdue to the volatility of the flower essential oils The activityexhibited by the aqueous extracts was accounted to thepresence of phenolic compounds that are reported to donatea hydrogen atom to free radicals such that the propagation ofthe chain reaction during lipid oxidation is terminated [92]Overall essential oils and phenolics present in B pilosa canbe thought of as major antioxidant compounds

35 Immunomodulatory Activity B pilosa is thought to bean immunomodulatory plant and is reported to be effectivein the treatment of immune disorders such as allergy [75]arthritis [75] and T1D [48 50 53]

As pointed out in the discussion of its anti-diabeticactivities (Section 33) a combination of phytochemicalsmethod and T-cell activation assays was used to studyimmunomodulatory properties of B pilosa IFN-120574 is a keycytokine released by T and NK cells that mediates immune

cells and sustains immunity against pathogens Defects inIFN-120574 expression regulation and activation result in vulner-ability to diseases caused by bacteria and viruses [69] Anelegant study performed byChang and colleagues using IFN-120574 promoter-driven luciferase reporter construct in JurkatT cells showed that hot water crude extracts of B pilosaincreased IFN-120574 promoter activity two-fold [69] Out of thesubfractions of this extract the butanol fraction but not thewater or ethyl acetate fractions increased IFN-120574 promoteractivity six-fold Centaurein (110) and centaureidin (109)were identified from the butanol fraction and were stated tocause a four-fold increase in IFN-120574 promoter activity withEC50

values of 75 120583gmL and 09120583gmL respectively Themechanism of action of centaurein was determined usingtranscription factors such as AP-1 NFAT and NF120581B whichare reported to bind to IFN-120574 promoter and regulate IFN-120574transcription Unlike with the activity of the positive controlPHA centaurein caused a four-fold increase in NFAT a3-fold increase in NF120581B and had little if any effect onAP-1 enhancer activities (23-fold three-fold and ten-foldincreases respectively were seenwith PHA) [69]The authorsconcluded that centaureinmodulates IFN-120574 expression by theNFAT and NF120581B pathways The article only determined themechanism of action of centaurein Its aglycone centaureidinmay act through the samemechanism though this conclusionneeds to be further verified

B pilosa extract and its compounds are reported to inhibitdifferentiation of naıve CD4+ helper T (Th0) cells into Th1cells [49] Using theThcell differentiation assay as a screeningplatform 3 polyynes 2-120573-D-glucopyranosyloxy-1 -hydroxytrideca-57911-tetrayne (53) 3-120573-D-glucopyranosyl-1-hydroxy-6(E)- tetradecene-81012-triyne (69) and 2-120573-D-glucopyranosyloxy-1-hydroxy-5(E)- tridecene-7911-triyne(49) were discovered from B pilosa [49 53] The data showsthat cytopiloyne and other two polyynes suppressed thedifferentiation of type 1 helper T (Th1) cells and productionof Th1 cytokines and promoted that of type 2 helper T (Th2)cells and production of Th2 cytokines thus explaining theimmunomodulatory and anti-inflammatory effects of Bpilosa and its polyynes

Chang and colleagues were the first to report the effect ofthe butanol extract of B pilosa on the autoimmune diabetesand airway inflammation inmice [48] Imbalance in the levelsofTh1 andTh2 and of various cytokines leads to autoimmunediseases T1D and other autoimmune diseases (rheumatoidarthritis Crohnrsquos disease among others) are exacerbated byan increase Th1 levels (specifically CD4+ Th1 cells) whileTh2 cells antagonize this effect [19] Moreover Th2 cellsmediate asthma in ovalbumin-induced hypersensitivity inBALBc mice [48] In their work [48] Chang and colleaguesshowed that 10mgkg butanol extracts with 15 (ww)compound 69 and 11 (ww) compound 49 (Table 11)ameliorated the development of Th1-mediated diabetes inNOD mice through inhibition of 120573 cell death and leukocyteinfiltration At the same dosage the butanol extracts alsoexacerbated ovalbumin-induced pulmonary inflammationin BALBc mice with an increase in the infiltration ofeosinophils and mast cells into the airway of the mice [48]Despite the different outcomes both mouse models proved

44 Evidence-Based Complementary and Alternative Medicine

the concept that control over the Th1Th2 shift is associatedwith autoimmune diseases and that the B pilosa butanolextract can shift the differentiation of Th0 cells to Th2 cells[48 49]

An extended study presented by Chiang and colleaguesshowed that compound 53 modulates T-cell functions [50]Using CD4+ T cells from BALBc mice they demonstratedthat cytopiloyne decreased levels of IFN-120574 producing cells(Th1) by 122 (from 72 to 598) Since Th1 and Th2 celldifferentiation is antagonistic it was expected that compound53 increased the percentage of mouse IL-4 producing cells(Th2) by 72 (from 237 to 309) Subsequent assess-ment of effect of compound 53 on the modulation of thetranscription of IL-4 and IFN-120574 showed that cytopiloyne asexpected decreased the splenocyte levels of IFN-120574 mRNAand increasing that of IL-4 in a dose-dependent manner Inthe range of 01 to 3 120583gmL of compound 53 the effects werenot attributed to its cytotoxicity Consequently using 3120583gmLcytopiloyne for 72 and 96 hours the protein concentration ofIFN-120574 decreased to 186 and 444 respectively Under thesame conditions cytopiloyne increased IL-4 concentrationsto 1985 and 2470 (for 72 and 96 hours resp) Thismodulation of T-cell differentiation exhibited by compound53 was used to explain its anti-diabetic activity [50]

The anti-diabetic role of cytopiloyne was extensivelydiscussed above (Section 33 anti-diabetic activity) Themolecular basis of the regulation of cytokine expression bycytopiloyne has been described Cytopiloyne directly elevatedthe expression level of IL-4 via GATA-3 upregulation inT cells [53] However reduction of IFN-120574 expression in Tcells seemed to come from the indirect opposing effect IL-4cytokine because the expression level of T-bet was unaltered[53] In this way cytopiloyne skewedTh1 polarization into theTh2 state conferring protection against T1D in NOD miceAside frompolarization ofThcell differentiation cytopiloynealso activated the expression of Fas ligand in pancreaticb cells this increase leading to the partial depletion of Tcells and reduction of immune response in local areas suchas the pancreas Of note cytopiloyne also inhibited T-cellproliferation and activation By targeting T cells from threeimmunomodulatory actions cytopiloyne protects againstT1D and probably other Th1-mediated autoimmune diseases[19]

The phytochemical constituents of B pilosa exert theirfunctions on different immune cells to modulate immuneresponse It is possible that some of the compounds mayhave agonistic or antagonistic effects on immune responseImmune function of B pilosa may depend on its compo-sition and amount of compounds which could explain theapparently conflicting report of B pilosa butanol extractaggravating allergy in mice [48] while cellulosine-treatedextract ameliorated allergy [48 75] IFN-120574 promoter reporterassays and T-cell differentiation assays were used to isolate2 flavonoids [69] and 3 polyynes [49 50] as immunomod-ulatory compounds from the butanol fraction of B pilosaInterestingly the flavonoids promote IFN-120574 expression inNK and T cells In marked contrast the polyynes promoteIL-4 expression and indirectly inhibit IFN-120574 expression indifferentiating T cells Sensitivity appears to be the key to

identifying structure- and bioactivity-related phytochemicalsfrom medicinal plants

36 Antimalarial Activity The use of chemical drugs againstpathogens has resulted in drug-resistant mutants Examplesof drug resistance can be found in the species of thePlasmodium that cause malaria It is important to search fornew compounds to combat Plasmodium parasites [47] Astudy of the anti-malarial activity of the leaf extracts of Bpilosa using a combination of phytochemistry and bioassaysshowed that compound 49 (Table 11) showed activity againsta malaria parasite (P falciparum NF54 strain) with an IC

50

value of 60 120583gmL [77] In addition compound 49 isolatedfrom the aerial parts of B pilosa inhibited growth of the Pfalciparum FCR-3 strain with an IC

50value of 035 120583gmL

This compound was tested for its in vivo effect in miceinfected with P berghei NK-65 strain Results showed thatcompound 49 decreased the average parasitemia in the redblood cells by 207 (from 328 of that of the control to121) after an intravenous injection of 08mgkg BWday forfour days [47] Further studies addressing the anti-malarialmechanism underlying both polyynes and clinical studies areneeded

37 Antibacterial Activity Emergence of multiple antibiotic-resistantmicrobes is becoming a global threat to public healthand a challenge to disease treatment For instance penicillinis commonly used to combat a food-borne intracellularbacteriumListeria however penicillin-resistant bacteria havebeen discovered recently [69] Chang and coworkers iso-lated centaurein (110) and centaureidin (109) from B pilosaextract [69 70] Centaurein enhances expression of IFN-120574a key cytokine for macrophage activation and consequentlyenhances bactericidal activity in macrophages [69 70] Inagreement with observed in vitro effects centaurein wasreported to prevent and treat Listeria infection in C57BL6Jmice [91] Mechanistic studies confirmed that centaureinexerted antilisterial action via IFN-120574 expression in wild-typemice but not IFN-120574 knockout mice [91] Further in vitrostudies on centaurein showed that this compound increasedIFN-120574 expression by 13 (from 17 to 20) 20 (from 21to 41) and 11 (from 6 to 17) in CD4+ T cells CD8+Tcells and NK cells respectively That is to say there wasan increase in IFN-120574 producing immune cells As expectedcentaurein also enhanced the expression level of T-bet a keynuclear factor for IFN-120574 expression Consistently centaureinaugmented the serum IFN-120574 levels inC57BL6Jmice and thisaugmentation peaked 24 hours after compound injectionThe quantity of mouse serum IFN-120574 was sufficient to activatemacrophages in vitro and eradicated GFP-producing Listeriainside macrophages However the entry of Listeria intomacrophages was not affected by centaurein-treated mousesera Centaurein treatment at 20120583g per mouse rescued 30of the mice infected with a lethal dose of Listeria (2 times106 CFU) It is noteworthy that in the presence of ampicillin(5 120583gmouse) centaurein (20 120583gmouse) rescued 70 of themice suggesting an additive effect between ampicillin andcentaurein [91] Despite lower abundance centaureidin was

Evidence-Based Complementary and Alternative Medicine 45

30 times more active than centaurein in terms of IFN-120574production [91]

Aside from the indirect antibacterial action mentionedabove extract andor compounds of B pilosa also showeddirect bacteriostatic andor bactericidal action One studyreported that essential oils and leafflower extracts of Bpilosa could suppress the growth of gram positive and gramnegative bacteria as evidenced by zone of inhibition assaysIn this study antibacterial activity of the essential oils fromthe leaves and flowers of B pilosa was determined in anattempt to identify natural products as food preservatives forprevention of microbial multiplication and food oxidationThe essential oils and extracts of B pilosa leaves and flow-ers showed moderate but different extents of antibacterialactivity (Table 17) In general essential oils had higherantibacterial activity than crude extracts One explanation forthis could be that monoterpenes in the essential oils destroycellular integrity and subsequently inhibit the respirationand ion transport processes The presence of antibacterial120573-caryophyllene could be another explanation as reportedelsewhere [92] Another study reported that the methanoland acetone extract of B pilosa roots displayed antibacterialactivities against the bacteria listed in Table 18 [93] andmethanol extracts from the roots seemed to be the mosteffective

Another study indicated that the polyyne (R)-12-dihydroxytrideca-357911-pentayne (49) from this plantalso suppressed bacterial growth as shown by the minimuminhibitory concentration required to inhibit 50 bacterialgrowth (MIC

50) in Table 19 This compound was highly

effective against several Gram positive and Gram negativebacteria including the drug-resistant bacteria Staphylococcusaureus N315 (MRSA) and Enterococcus faecalis NCTC12201(VRE) [47] Strikingly compound 49 had a similar MIC

50

value to antibiotics (ampicillin tetracycline norfloxacin andamphotericin B) in most of the bacteria tested

Antibacterial activity of B pilosa extracts and compo-nents expressed as MIC

50and the mean zone of inhibition

is tabulated in Tables 17 18 and 19The zone of inhibition forAmpicilline (positive control) ranges from 153 plusmn 03mm to443 plusmn 02mm [92]

38 Antifungal Activity B pilosa has traditionally been usedto treat microbial infection Recently different parts of Bpilosa have been tested for antifungal activities Deba andcolleagues first evaluated the antifungal effect of the hotwater extracts of the B pilosa roots stems and leaves againstCorticium rolfsii Fusarium solani and Fusarium oxysporumThey discovered that C rolfsii was most suppressed bytreatment with B pilosa as its growth was reduced at almostall the tested doses followed by F oxysporum and F solani[99] However the fungicidal activities of the stems androots were greater than the leaves [99] Moreover the samegroup assessed the antifungal activity of the essential oilsand aqueous extracts from B pilosa flowers and leaves [92]They showed that the extracts and oils had antifungal activityagainst C rolfsii F solani and F oxysporum Essential oils

appeared to have better fungicidal activity thanwater extractsas summarized in Table 20

Another study by Ashafa and colleagues showed thatacetone methanol and water extracts of the B pilosa rootsshowed antifungal activities against Aspergillus niger Aflavus and Penicillium notatum using the agar dilutionmethod The results are tabulated in Table 21 [93] Negativecontrols showed 0 growth inhibitionThe methanol extractof the B pilosa roots at 10mgmL was also effective againstCandida albicans [93]Of note B pilosa obtained fromPapuaNew Guinea had no activity against A niger and C albicans[126] but the SouthAfrican (EasternCape) ecotype exhibitedmoderate activity against C albicans [93] This discrepancymay depend on extraction solvents extraction procedureassay techniques different plant parts and abundance ofactive compounds

B pilosa produces a variety of secondary metabolitessuch as flavonoids phenylacetylenes alkaloids sterols ter-penoids and tannis [92 93] However none of them havebeen confirmed as active compounds against fungi Furtherinvestigation of active compounds from B pilosa is necessaryto further understand the antifungal efficacy of this plant

39 Hypotensive and Vasodilatory Activities In early studiesDimo and colleagues used three rat models normotensiveWistar rats (NTR) salt-loading hypertensive rats (SLHR)and spontaneous hypertensive rats (SHR) to investigate thehypotensive effect of the methanol crude extract of B pilosaleaves [28 127] The extract lowered systolic blood pressurein hypertensive rats (SLHR and SHR) to a greater degreethan NTR [28 127] In addition a decrease in urinarysodium ions and an increase in urinary potassium ions wereobserved after treatment with the methanol extract of Bpilosa leaves although neither differences were statisticallysignificant [28 127] Taking the data together the studyproposed that B pilosa leaf extract reduced blood pressurevia vasodilation [28 127] The same group continued totest the antihypertensive effect of aqueous and methylenechloride extracts of B pilosa leaves in a hypertensive ratmodel [28 127] To establish a fructose-induced hypertensionmodel male Wistar rats were given 10 fructose solutionto drink ad libitum for three weeks In addition to freeaccess to 10 fructose the rats were treated with the aqueous(150mgkg) or methyl chloride (350mgkg) extracts of Bpilosa for additional three weeks [28 127] Both extracts ofB pilosa leaves had a hypotensive effect on rats Howeverneither extracts reversed the elevation of serum insulin infructose-fed rats Therefore B pilosa lowered blood pressureirrespective of insulin [28 127] To better understand thehypotensive mechanism the authors investigated the effectof a neutral extract of B pilosa (NBP) a mixture of methanolandmethylene chloride (1 1) extract after neutralization withNaOH and HCl on the heart and the blood pressure ofNTR and SHR [110] This study showed that an intravenousinjection of the NBP resulted in a biphasic reduction insystolic blood pressure In addition one intravenous dose ofthe extract at 10 20 and 30mgkg BW decreased systolicblood pressure in normal rats by 183 425 and 30

46 Evidence-Based Complementary and Alternative Medicine

respectively and the same doses reduced the blood pressurein hypertensive rats by 258 389 and 286 respectivelyOnly the highest dose (30mgkg) affected the force of thecontraction of the heart Atropine and propranolol were usedto interfere with the hypotensive action of the NBP Atropinereduced the initial phase of the hypotensive response in NBPand completely abolished the second phase of hypotensiveresponse in NBP In contrast propranolol increased thefirst hypotensive response but partially abolished the secondhypotensive response provoked by the NBP [110]This mech-anistic study suggested that B pilosa invokes the biphasichypotensive responses via targeting cardiac pump efficiencyduring the first phase and vasodilation at the second phase[110]

A further study was performed to investigate the relaxingeffect of a neutral extract of B pilosa (NBP) on rat aorta con-tractedwithKCl (60mM) andnorepinephrine (01mM) [94]Cumulative addition of NBP relaxed the rat aorta previouslycontracted by KCl in a dose-dependent manner The EC

50

value of theNBP for vasorelaxationwas 032mgmLThedataalso showed that the NBP reduced the contraction of aortapreviously contracted by KCl irrespective of the presence ofaortic endothelium [94]

Pretreatment with glibenclamide an ATP-dependent K+channel blocker did not considerably affect the relaxant effectof the NBP on KCl-induced contraction suggesting thatthe vasodilatory effect of B pilosa was not related to theopening of this ATP-dependent K+ channel [94] On theother hand in the presence of indomethacin or pyrilaminemaleate the relaxant response induced by the plant extractwas significantly inhibited at the lower concentrations Theplant extract was able to reduce the aorta resting tone inhibitthe KCl-induced contractions by 90 at 15mgmL and theCaCl2-induced contractions by 95 at 075mgmL These

results demonstrate that B pilosa can act as a vasodilatorprobably via acting as a calcium antagonist [94]

However no specific compound for the above activityhas been identified from B pilosa to date A bioactivity-guided identification approach may be adopted to identifythe active compounds in B pilosa that possess hypotensiveand vasodilatory effects and understand their mechanism ofaction

310 Wound Healing Activity B pilosa has been traditionallyused to treat tissue injury inCameroon Brazil andVenezuela[26] Hassan and colleagues investigated the wound healingpotential of B pilosa in Wistar rats [128] Mirroring thepositive control neomycin sulfate the ethanol extract of Bpilosa had faster wound closure than control rats 3 6 and 9days after topical application Histological examination alsorevealed better collagenation angiogenesis and organizationof wound tissue seven days after application Epithelializationand total healing time in B pilosa-treated rats were compara-ble to those of neomycin sulfate Together these data suggestthat B pilosa may be a viable alternative to neomycin lotionfor the treatment of wounds

In addition to studying the wound healing effect of Bpilosa on external ulcers Tan and colleagues also examined

the effect of methanol cyclohexane and methyl chlorideextracts of B pilosa on gastric ulcers in Wistar rats fedwith 1mL HClethanol gastric necrotizing solution (150mMHCl in 60 ethanol) and macroscopically visible lesionswere scored [26] Among the three extracts methylene chlo-ride extracts exhibited the highest activity showing 464inhibition of lesion formation at a dose of 500mgkg BWand complete inhibition at 750mgkg [26] The efficacy ofthe ethylene chloride extract was followed by that of themethanol extracts which had inhibition ranging from 304to 822 at concentrations of 500mgkg and 1000mgkgBW respectively [26] The cyclohexane extracts showed thelowest activity against gastric ulcers in rats with 13340 and 797 inhibition at 500 750 and 1000mgkg BWrespectively [26] To better understand the mode of actionof the methylene chloride extract of B pilosa rats werepretreated with indomethacin a COX-2 inhibitor involved inprostaglandin synthesis Pretreatment significantly reducedthe protection against HClethanol-induced ulcers to 313inhibition at 750mgkg BW suggesting a link between theantiulcerative activity of B pilosa and prostaglandin syn-thesis Unexpectedly the methylene chloride extract of Bpilosa showed little gastric mucosal protection against gastriclesions induced by 95 ethanol (1mL) [26] Absolute alcoholis known to cause mucosalsubmucosal tissue destruction viacellular necrosis and the release of tissue-derived mediators(histamine and leukotrieneC4)Thus these data imply thatBpilosa did not prevent the generation or the necrotic action ofthese mediators on the gastric microvasculature In additionpylorus ligation can increase gastric acid secretion withoutan alteration of mucosal histamine content The methylenechloride extract of B pilosa did not possess antisecretoryactivity On the contrary it was observed that increases in thedose of the extract led to elevated gastric juice acidity [26]Results with both absolute ethanol and pylorus ligation ratmodels suggested the possibility that the ineffectiveness of Bpilosa against gastric ulcers was due to lack of antihistaminicactivity in the plant In summary overall the data suggest thatB pilosa protects against HCLethanol-mediated ulcers viainhibition of prostaglandin biosynthesis

Previous phytochemical studies showed that a groupof flavonoids acyclichalcones are present in B pilosa[129 130] and the chalcones were proposed to have anti-ulcerative activity [131] Moreover nine hydroxychalcones 10were reported to possess gastric cytoprotective effects with24-dihydroxychalcone being the most active [132] Sincemethylene chloride extracts appear to be the most active Bpilosa extracts next the specific anti-ulcerative phytochemi-cals in the methylene chloride extracts of B pilosa and theirmodes of action needs to be probed

Despite the claims listed inTable 3 relatively few scientificstudies have been conducted in vitro and in vivo to address thetraditional ethnomedical uses of B pilosa Information aboutthe use of B pilosa as a botanical therapy recorded so far isfar from complete Studies conducted thus far only serve asa starting point for further investigation of B pilosa and theultimate efficacious use of the herb in clinical applications

Evidence-Based Complementary and Alternative Medicine 47

4 Toxicology

Despite its use as an ingredient in food for human con-sumption studies on systemic toxicity (eg acute subacutechronic and subchronic toxicities) of B pilosa in humansand animals are still inadequate and insufficient So faracute andor subchronic toxicities have been evaluated inrats and mice Oral acute and 28-day toxicities of waterextract of B pilosa leaves were evaluated in Wistar rats[133] An oral dose of water extract of B pilosa leaves at10 gkg BW showed no obvious mortality or changes inthe appearance in rats [134] The same extract at 08 gkgBWday once a day showed no obvious sub-chronic toxicityin rats over 28 days as measured by survival rate bodyweight and gross examination of organs [134] These dataare consistent with our data indicating that oral deliveryof the water extract of the B pilosa whole plant at 1 gkgBWday once a day is safe in rats over 28 days (unpublisheddata) Taken together these studies suggest that ingestion ofB pilosa aqueous extract at up to at 1 gkg BWday oncea day is highly safe in rats In addition the acute toxicityof aqueous and ethanol extracts of B pilosa in mice havebeen reported [134] Five- to six-week-old mice with weightsbetween 28 and 35 g received a peritoneal injection of bothextracts at the different doses The LD

50 the dose that causes

50 lethality of the aqueous and ethanol extracts in micewas 1230 gkg BW and 615 gkg BW respectively [134] Acomplete toxicological study has not been completed forhumans Furthermore the drug interactions of B pilosa withother drugs are unknown Further safety verification andclinical trials should be performed before B pilosa can beconsidered for medicinal use

5 Conclusions

B pilosa is an erect perennial plant with green leaves whiteor yellow flowers and tiny black seeds As it is distributedworldwide and is widely used as a folk remedy B pilosacan be thought of as an extraordinary source of food andmedicine However a comprehensive up-to-date review ofresearch on B pilosa has hitherto been unavailable In thisarticle scientific studies on B pilosa have been summarizedand critically discussed from the perspectives of botanyethnomedicine phytochemistry pharmacology and toxicol-ogy B pilosa is claimed to treat more than 40 disordersand 201 compounds have been identified from this plantThe medicinal utility of B pilosa and its modes of actionin relation to its known phytochemicals were discussedherein Polyynes flavonoids phenylpropanoids fatty acidsand phenolics are the primary bioactive compounds of Bpilosa and they have been reported to be effective in thetreatment of tumors inflammationimmune modulationdiabetes viruses microbes protozoans gastrointestinal dis-eases hypertension and cardiovascular diseases Cautionshould be exercised in the therapeutic use of B pilosa forhypoglycemia hypotension bleeding and allergy

Acknowledgments

The authors thank their laboratory members for construc-tive suggestions and the authors whose publications theycited for their contributions The authors also thank MsMiranda Loney of Academia Sinica for English editing of thispaper This work was supported by Grants 99-CDA-L11 and101S0010056 from Academia Sinica Taiwan Arlene P Bar-tolome is a recipient of MECO-TECO Sandwich ScholarshipProgram between Taiwan and the Philippines

References

[1] T Shen G H Li X N Wang and H X Lou ldquoThe genusCommiphora a review of its traditional uses phytochemistryand pharmacologyrdquo Journal of Ethnopharmacology vol 142 no2 pp 319ndash330

[2] C Long P Sauleau B David et al ldquoBioactive flavonoids ofTanacetum parthenium revisitedrdquo Phytochemistry vol 64 no2 pp 567ndash569 2003

[3] P O Karis and O Ryding Asteraceae Cladistics and Classifica-tion Bremer K Eds pp 559ndash569 Timber press Portland OreUSA 1994

[4] O N Pozharitskaya A N Shikov M N Makarova et alldquoAnti-inflammatory activity of a HPLC-fingerprinted aqueousinfusion of aerial part of Bidens tripartita LrdquoPhytomedicine vol17 no 6 pp 463ndash468 2010

[5] F Q Oliveira V Andrade-Neto A U Krettli and MG L Brandao ldquoNew evidences of antimalarial activity ofBidens pilosa roots extract correlated with polyacetylene andflavonoidsrdquo Journal of Ethnopharmacology vol 93 no 1 pp 39ndash42 2004

[6] Agriculture USDA ldquoPlants databaserdquo 2013in Natural Resources Conservation Servicehttpwwwnrcsusdagovwpsportalnrcssitenationalhome

[7] Y Zhou and X Z Yan ldquoExperimental study of qi deficiencysyndrome and Codonopsis pillosulae and Astragalus injectionon the immune response in micerdquo Zhong Xi Yi Jie He Za Zhivol 9 no 5 pp 286ndash262 1989

[8] S C Chien P H Young Y J Hsu et al ldquoAnti-diabeticproperties of three common Bidens pilosa variants in TaiwanrdquoPhytochemistry vol 70 no 10 pp 1246ndash1254 2009

[9] M J Alcaraz and M J Jimenez ldquoFlavonoids as anti-inflammatory agentsrdquo Fitoterapia vol 59 no 1 pp 25ndash38 1988

[10] FAO Agriculture Food and Nutrition for AfricamdashA ResourceBook for Teachers of Agriculture Publishing ManagementGroup FAO Information Division Rome Italy 1997

[11] M B Rokaya Z Munzbergova B Timsina and K R BhattaraildquoRheum australe D Don a review of its botany ethnobotanyphytochemistry and pharmacologyrdquo Journal of Ethnopharma-cology vol 141 no 3 pp 761ndash774 2012

[12] P H Young Y J Hsu and W C Yang ldquoBidens pilosa L andits medicinal userdquo in Recent Progress in Medicinal Plants DrugPlant II A S AwaadVK Singh and JNGovil Eds StandiumPress Houston Tex USA 2010

[13] C Ge ldquoCytologic study of Bidens bipinnata LrdquoZhongguo ZhongYao Za Zhi vol 15 no 2 pp 72ndash125 1990

[14] L C Chiang J S Chang C C Chen L T Ng and C CLin ldquoAnti-herpes simplex virus activity of Bidens pilosa and

48 Evidence-Based Complementary and Alternative Medicine

Houttuynia cordatardquoAmerican Journal of ChineseMedicine vol31 no 3 pp 355ndash362 2003

[15] N P Rybalchenko V A Prykhodko S S Nagorna et alldquoIn vitro antifungal activity of phenylheptatriyne from Bidenscernua L against yeastsrdquo Fitoterapia vol 81 no 5 pp 336ndash3382010

[16] K Redl W Breu B Davis and R Bauer ldquoAnti-inflammatoryactive polyacetylenes from Bidens campylothecardquo Planta Med-ica vol 60 no 1 pp 58ndash62 1994

[17] M Ayyanar and S Ignacimuthu ldquoTraditional knowledge ofKani tribals inKouthalai of Tirunelveli hills TamilNadu IndiardquoJournal of Ethnopharmacology vol 102 no 2 pp 246ndash255 2005

[18] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant intraditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[19] AKAbbas KMMurphy andA Sher ldquoFunctional diversity ofhelper T lymphocytesrdquo Nature vol 383 no 6603 pp 787ndash7931996

[20] J H Cano and G Volpato ldquoHerbal mixtures in the traditionalmedicine of Eastern Cubardquo Journal of Ethnopharmacology vol90 no 2-3 pp 293ndash316 2004

[21] J Namukobe J M Kasenene B T Kiremire et al ldquoTraditionalplants used for medicinal purposes by local communitiesaround the Northern sector of Kibale National Park UgandardquoJournal of Ethnopharmacology vol 136 no 1 pp 236ndash245 2011

[22] C Lans ldquoComparison of plants used for skin and stomachproblems in Trinidad and Tobago with Asian ethnomedicinerdquoJournal of Ethnobiology andEthnomedicine vol 3 article 3 2007

[23] S Dharmananda ldquoA Popular Remedy Ecapes Notice ofWesternPractitionersrdquo 2013 httpwwwitmonlineorgartsbidenshtm

[24] Y J Hsu T H Lee C L T Chang Y T Huang andW C YangldquoAnti-hyperglycemic effects and mechanism of Bidens pilosawater extractrdquo Journal of Ethnopharmacology vol 122 no 2 pp379ndash383 2009

[25] E Noumi F Houngue and D Lontsi ldquoTraditional medicinesin primary health care plants used for the treatment ofhypertension in Bafia Cameroonrdquo Fitoterapia vol 70 no 2 pp134ndash139 1999

[26] P V Tan T Dimo and E Dongo ldquoEffects of methanolcyclohexane and methylene chlo ride extracts of Bidens pilosaon various gastric ulcer models in ratsrdquo Journal of Ethnophar-macology vol 73 no 3 pp 415ndash421 2000

[27] R L C Pereira T Ibrahim L Lucchetti A J R Da Silva and VL G De Moraes ldquoImmunosuppressive and anti-inflammatoryeffects of methanolic extract and the polyacetylene isolatedfrom Bidens pilosa Lrdquo Immunopharmacology vol 43 no 1 pp31ndash37 1999

[28] T Dimo J Azay P V Tan et al ldquoEffects of the aqueous andmethylene chloride extracts of Bidens pilosa leaf on fructose-hypertensive ratsrdquo Journal of Ethnopharmacology vol 76 no 3pp 215ndash221 2001

[29] CWiartMedicinal Plants of Southeast Asia Pelanduk SelabgorDarul Ehsan Malaysia 2000

[30] F L Silva D C H Fischer J F Tavares M S Silva P FDe Athayde-Filho and J M Barbosa-Filho ldquoCompilation ofsecondary metabolites from Bidens pilosa LrdquoMolecules vol 16no 2 pp 1070ndash1102 2011

[31] A H Chen S R Lin and C H Hong ldquoPhytochemical studyon Bidens pilosa L varminorrdquo Huaxue pp 38ndash42 1975

[32] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[33] M C Zulueta M Tada and C Y Ragasa ldquoA diterpene fromBidens pilosardquo Phytochemistry vol 38 no 6 pp 1449ndash14501995

[34] J Wang H Yang Z W Lin and H D Sun ldquoFlavonoids fromBidens pilosa var radiatardquo Phytochemistry vol 46 no 7 pp1275ndash1278 1997

[35] A Zhao Q Zhao L Peng et al ldquoA new chalcone glycoside fromBidens pilosardquo Acta Botanica Yunnanica vol 26 no 1 pp 121ndash126

[36] P Gao Y Wu S Cui and Y Zou ldquoSynthesis and absoluteconfiguration of the 7-phenylhepta-4 6-diyne-1 2-diol isolatedfrom Bidens pilosardquo Synthesis no 13 pp 2131ndash2135 2011

[37] S Wang B Yang D Zhu D He and L Wang ldquoActivecomponents of Bidens pilosa Lrdquo Zhongcaoyao vol 36 pp 20ndash21

[38] F Bohlmann T Burkhardt and C Zdero Naturally OccuringAcetylenes Academic Press New York NY USA 1973

[39] T M Sarg A M Ateya N M Farrag and F A AbbasldquoConstituents and biological activity of Bidens pilosa L grownin EgyptrdquoActa Pharmaceutica Hungarica vol 61 no 6 pp 317ndash323 1991

[40] H A L Valdes and H P Rego ldquoBidens pilosa Linnerdquo RevistaCubana de Plantas Medicinales vol 6 pp 28ndash33

[41] F Bohlmann H Bornowski and K M Kleine ldquoNew polyynesfrom the tribe Heliantheaerdquo Chemische Berichte vol 97 pp2135ndash2138

[42] RWang Q XWu and Y P Shi ldquoPolyacetylenes and flavonoidsfrom the aerial parts ofBidens pilosardquo PlantaMedica vol 76 no9 pp 893ndash896 2010

[43] L W Wu Y M Chiang H C Chuang et al ldquoA novelpolyacetylene significantly inhibits angiogenesis and promotesapoptosis in human endothelial cells through activation of theCDK inhibitors and caspase-7rdquo Planta Medica vol 73 no 7 pp655ndash661 2007

[44] L W Wu Y M Chiang H C Chuang et al ldquoPolyacetylenesfunction as anti-angiogenic agentsrdquo Pharmaceutical Researchvol 21 no 11 pp 2112ndash2119 2004

[45] H L Yang S C Chen N W Chang et al ldquoProtection fromoxidative damage using Bidens pilosa extracts in normal humanerythrocytesrdquo Food and Chemical Toxicology vol 44 no 9 pp1513ndash1521 2006

[46] H Q Wang S J Lu H Li and Z H Yao ldquoEDTA-enhancedphytoremediation of lead contaminated soil by Bidens maxi-mowiczianardquo Journal of Environmental Sciences vol 19 no 12pp 1496ndash1499 2007

[47] S TobinagaMK SharmaWG L Aalbersberg et al ldquoIsolationand identification of a potent antimalarial and antibacterialpolyacetylene from Bidens pilosardquo Planta Medica vol 75 no 6pp 624ndash628 2009

[48] C L T Chang H K Kuo S L Chang et al ldquoThe distincteffects of a butanol fraction of Bidens pilosa plant extract onthe development of Th1-mediated diabetes and Th2-mediatedairway inflammation in micerdquo Journal of Biomedical Sciencevol 12 no 1 pp 79ndash89 2005

[49] S L Chang C L T Chang Y M Chiang et al ldquoPolyacetyleniccompounds and butanol fraction from Bidens pilosa can mod-ulate the differentiation of helper T cells and prevent autoim-mune diabetes in non-obese diabetic micerdquo Planta Medica vol70 no 11 pp 1045ndash1051 2004

Evidence-Based Complementary and Alternative Medicine 49

[50] Y M Chiang C L T Chang S L Chang W C Yang and LF Shyur ldquoCytopiloyne a novel polyacetylenic glucoside fromBidens pilosa functions as a T helper cell modulatorrdquo Journal ofEthnopharmacology vol 110 no 3 pp 532ndash538 2007

[51] R PUbillas CDMendez S D Jolad et al ldquoAntihyperglycemicacetylenic glucosides from Bidens pilosardquo Planta Medica vol66 no 1 pp 82ndash83 2000

[52] L Alvarez S Marquina M L Villarreal D Alonso E Arandaand G Delgado ldquoBioactive polyacetylenes from Bidens pilosardquoPlanta Medica vol 62 no 4 pp 355ndash357 1996

[53] C L T Chang S L Chang Y M Lee et al ldquoCytopiloyne apolyacetylenic glucoside prevents type 1 diabetes in nonobesediabetic micerdquo Journal of Immunology vol 178 no 11 pp 6984ndash6993 2007

[54] G Kusano A Kusano and Y Seyama Novel Hypoglycemicand Anti-Inflammatory Polyacetylenic Compounds Their Com-positions Bidens Plant Extract Fractions and CompositionsContaining the Plant or Fraction JPO Tokyo Japan 2004

[55] M G L Brandao A U Krettli L S R Soares C G CNery and H C Marinuzzi ldquoAntimalarial activity of extractsand fractions from Bidens pilosa and other Bidens species(Asteraceae) correlated with the presence of acetylene andflavonoid compoundsrdquo Journal of Ethnopharmacology vol 57no 2 pp 131ndash138 1997

[56] A U Krettli V F Andrade-Neto M D G L Brandao andW M S Ferrari ldquoThe search for new anti-malarial drugs fromplants used to treat fever and malaria or plants ramdomlyselected a reviewrdquoMemorias do Instituto Oswaldo Cruz vol 96no 8 pp 1033ndash1042 2001

[57] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[58] C K Wat R K Biswas E A Graham L Bohm G H NTowers and E R Waygood ldquoUltraviolet-mediated cytotoxicactivity of phenylheptatriyne from Bidens pilosa Lrdquo Journal ofNatural Products vol 42 no 1 pp 103ndash111 1979

[59] Y Sashida K Ogawa M Kitada H Karikome Y Mimakiand H Shimomura ldquoNew aurone glucosides and new phenyl-propanoid glucosides from Bidens pilosardquo Chemical and Phar-maceutical Bulletin vol 39 no 3 pp 709ndash711 1991

[60] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens pilosa L (TFB) on animal liver injury andliver fibrosisrdquo Journal of Ethnopharmacology vol 116 no 3 pp539ndash546 2008

[61] B Hoffmann and J Holzl ldquoAcylated compounds from Bidenspilosardquo Planta Medica vol 55 pp 108ndash109

[62] B Hoffmann and J Holzl ldquoA methylated chalcone glucosidefrom Bidens pilosardquo Phytochemistry vol 27 no 11 pp 3700ndash3701 1988

[63] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[64] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[65] B Hoffmann and J Holzl ldquoWeitere acylierte chalkone ausBidens pilosardquo Planta Medica vol 54 pp 450ndash451 1988

[66] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant in11traditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[67] V V Pham V V T K P V L Hoang and V K Phan ldquoFlavonoidcompounds from the plant Bidens pilosa L (Asteraceae)rdquo TapChi Duoc Hoc vol 50 pp 48ndash53

[68] S D Sarker B Bartholomew R J Nash and N Robinson ldquo5-O-methylhoslundin an unusual flavonoid from Bidens pilosa(Asteraceae)rdquo Biochemical Systematics and Ecology vol 28 no6 pp 591ndash593 2000

[69] S L Chang Y M Chiang C L T Chang et al ldquoFlavonoidscentaurein and centaureidin from Bidens pilosa stimulate IFN-120574 expressionrdquo Journal of Ethnopharmacology vol 112 no 2 pp232ndash236 2007

[70] Y M Chiang D Y Chuang S Y Wang Y H Kuo P WTsai and L F Shyur ldquoMetabolite profiling and chemopreventivebioactivity of plant extracts from Bidens pilosardquo Journal ofEthnopharmacology vol 95 no 2-3 pp 409ndash419 2004

[71] A Kusano Y Seyama E Usami et al ldquoStudies on the antioxi-dant active constituents of the dried powder from Bidens pilosaL var radiata SchrdquoNaturalMedicines vol 57 no 3 pp 100ndash1042003

[72] I Doleckova L Rarova J Gruz et al ldquoAnti-proliferative andanti-angiogenic effects of flavone eupatorin an active con-stituent of chloroform extract ofOrthosiphon stamineus leavesrdquoFitoterapia vol 83 no 6 pp 1000ndash1007

[73] K Bairwa R Kumar R J Sharma and R K Roy ldquoAn updatedreview on Bidens pilosa Lrdquo Der Pharma Chemica vol 2 no 3pp 325ndash337

[74] Q Xia Y Liu and Y Li ldquoDetermination of hyperoside indifferent parts and different species of Herba Bidens by RP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol 50pp 48ndash53

[75] M Horiuchi and Y Seyama ldquoImprovement of the antiinflam-matory and antiallergic activity of Bidens pilosa L var radiataSCHERFF treated with enzyme (Cellulosine)rdquo Journal of HealthScience vol 54 no 3 pp 294ndash301 2008

[76] M G L Brandao C G C Nery M A S Mamao and AU Krettli ldquoTwo methoxylated flavone glycosides from Bidenspilosardquo Phytochemistry vol 48 no 2 pp 397ndash400 1998

[77] P Kumari K Misra B S Sisodia et al ldquoA promising anticancerand antimalarial component from the leaves of Bidens pilosardquoPlanta Medica vol 75 no 1 pp 59ndash61 2009

[78] G Seelinger I Merfort U Wolfle and C M Schempp ldquoAnti-carcinogenic effects of the flavonoid luteolinrdquoMolecules vol 13no 10 pp 2628ndash2651 2008

[79] G Seelinger I Merfort and C M Schempp ldquoAnti-oxidantanti-inflammatory and anti-allergic activities of luteolinrdquoPlanta Medica vol 74 no 14 pp 1667ndash1677 2008

[80] C C Yit and N P Das ldquoCytotoxic effect of butein on humancolon adenocarcinoma cell proliferationrdquo Cancer Letters vol82 no 1 pp 65ndash72 1994

[81] J A Beutler E Hamel A J Vlietinck et al ldquoStructure-activityrequirements for flavone cytotoxicity and binding to tubulinrdquoJournal of Medicinal Chemistry vol 41 no 13 pp 2333ndash23381998

[82] M R Kviecinski K B Felipe T Schoenfelder et al ldquoStudy ofthe antitumor potential of Bidens pilosa (Asteraceae) used inBrazilian folkmedicinerdquo Journal of Ethnopharmacology vol 117no 1 pp 69ndash75 2008

[83] P Sundararajan A Dey A Smith A G Doss M Rajappanand S Natarajan ldquoStudies of anticancer and antipyretic activityof Bidens pilosawhole plantrdquo African Health Sciences vol 6 no1 pp 27ndash30 2006

[84] YMChiang C P Lo Y P Chen et al ldquoEthyl caffeate suppressesNF-120581B activation and its downstream inflammatory mediatorsiNOS COX-2 and PGE2 in vitro or in mouse skinrdquo BritishJournal of Pharmacology vol 146 no 3 pp 352ndash363 2005

50 Evidence-Based Complementary and Alternative Medicine

[85] J S Kim H J Lee M H Lee J Kim C Jin and J HRyu ldquoLuteolin inhibits LPS-stimulated inducible nitric oxidesynthase expression in BV-2 microglial cellsrdquo Planta Medicavol 72 no 1 pp 65ndash68 2006

[86] P A Ruiz and D Haller ldquoFunctional diversity of flavonoidsin the inhibition of the proinflammatory NF-120581B IRF and Aktsignaling pathways in murine intestinal epithelial cellsrdquo Journalof Nutrition vol 136 no 3 pp 664ndash671 2006

[87] A Xagorari A Papapetropoulos A Mauromatis MEconomou T Fotsis and C Roussos ldquoLuteolin inhibitsan endotoxin-stimulated phosphorylation cascade andproinflammatory cytokine production in macrophagesrdquoJournal of Pharmacology and Experimental Therapeutics vol296 no 1 pp 181ndash187 2001

[88] N Yoshida T Kanekura Y Higashi and T Kanzaki ldquoBidenspilosa suppresses interleukin-1120573-induced cyclooxygenase-2expression through the inhibition of mitogen activated proteinkinases phosphorylation in normal human dermal fibroblastsrdquoJournal of Dermatology vol 33 no 10 pp 676ndash683 2006

[89] M Muchuweti C Mupure A Ndhlala T Murenje and M AN Benhura ldquoScreening of antioxidant and radical scavengingactivity of Vigna ungiculata Bidens pilosa and Cleome gynan-drardquo American Journal of Food Technology vol 2 no 3 pp 161ndash168 2007

[90] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens bipinnata L against carbon tetrachloride-induced liver fibrosis in ratsrdquo Journal of Pharmacy and Pharma-cology vol 60 no 10 pp 1393ndash1402 2008

[91] S L Chang H H Yeh Y S Lin Y M Chiang T K Wuand W C Yang ldquoThe effect of centaurein on interferon-120574 expression and Listeria infection in micerdquo Toxicology andApplied Pharmacology vol 219 no 1 pp 54ndash61 2007

[92] F Deba T D Xuan M Yasuda and S Tawata ldquoChemical com-position and antioxidant antibacterial and antifungal activitiesof the essential oils from Bidens pilosa Linn var Radiatardquo FoodControl vol 19 no 4 pp 346ndash352 2008

[93] A O T Ashafa and A J Afolayan ldquoScreening the root extractsfrom Biden pilosa L var radiata (Asteraceae) for antimicrobialpotentialsrdquo Journal of Medicinal Plant Research vol 3 no 8 pp568ndash572 2009

[94] T B Nguelefack T Dimo E P Nguelefack Mbuyo P V TanS V Rakotonirina and A Kamanyi ldquoRelaxant effects of theneutral extract of the leaves of Bidens pilosa linn on isolated ratvascular smooth musclerdquo Phytotherapy Research vol 19 no 3pp 207ndash210 2005

[95] M T Grombone-Guaratini K L Silva-Brandao V N SolferiniJ Semir and J R Trigo ldquoSesquiterpene and polyacetyleneprofile of the Bidens pilosa complex (Asteraceae Heliantheae)from Southeast of Brazilrdquo Biochemical Systematics and Ecologyvol 33 no 5 pp 479ndash486 2005

[96] L L Lin C Y Wu H C Hsiu M T Wang and H ChuangldquoStudies on diabetes mellitus I The hypoglycemic activity ofphytosterin on alloxan diabetic ratsrdquo Taiwan Yi Xue Hui Za Zhivol 66 no 2 pp 58ndash65 1967

[97] M A N Benhura and I C Chitsiku ldquoThe extractable 120573-carotene content of Guku (Bidens pilosa) leaves after cookingdrying and storagerdquo International Journal of Food Science andTechnology vol 32 no 6 pp 495ndash500 1997

[98] J S Chang L C Chiang C C Chen L T Liu K C Wangand C C Lin ldquoAtileukemic activity of Bidens pilosa l varminor (blume) sherff andHouttuynia cordata thunbrdquo AmericanJournal of Chinese Medicine vol 29 no 2 pp 303ndash312 2001

[99] F Deba T D Xuan M Yasuda and S Tawata ldquoHerbicidal andfungicidal activities and identification of potential phytotoxinsfrom Bidens pilosa L var radiata Scherff research paperrdquoWeedBiology and Management vol 7 no 2 pp 77ndash83 2007

[100] J K Kumar and A K Sinha ldquoA new disubstituted acetylacetonefrom the leaves ofBidens pilosa LinnrdquoNatural Product Researchvol 17 no 1 pp 71ndash74 2003

[101] K Ogawa and Y Sashida ldquoCaffeoyl derivatives of a sugarlactone and its hydroxy acid from the leaves of Bidens pilosardquoPhytochemistry vol 31 no 10 pp 3657ndash3658 1992

[102] Q Xia Y Liu and Y Li ldquoDetermination of gallic acid fromdifferent species and different medical parts of Herba Bidens byRP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol24 no 03 pp 308ndash310

[103] W J Lee L F Wu W K Chen C J Wang and T HTseng ldquoInhibitory effect of luteolin on hepatocyte growth fac-torscatter factor-induced HepG2 cell invasion involving bothMAPKERKs and PI3K-Akt pathwaysrdquo Chemico-BiologicalInteractions vol 160 no 2 pp 123ndash133 2006

[104] J A Beutler J H Cardellina C M Lin E Hamel G MCragg andM R Boyd ldquoCentaureidin a cytotoxic flavone fromPolymnia fruticosa inhibits tubulin polymerizationrdquo BioorganicandMedicinal Chemistry Letters vol 3 no 4 pp 581ndash584 1993

[105] A Verma A Su A M Golin B OrsquoMarrah and J K AmorosaldquoThe lateral view a screening method for knee traumardquo Aca-demic Radiology vol 8 no 5 pp 392ndash397 2001

[106] J Corren M Lemay Y Lin L Rozga and R K RandolphldquoClinical and biochemical effects of a combination botanicalproduct (ClearGuard) for allergy a pilot randomized double-blind placebo-controlled trialrdquo Nutrition Journal vol 7 no 1article 20 2008

[107] M S Gachet J S Lecaro M Kaiser et al ldquoAssessment of anti-protozoal activity of plants traditionally used in Ecuador in thetreatment of leishmaniasisrdquo Journal of Ethnopharmacology vol128 no 1 pp 184ndash197 2010

[108] W C Yang M Ghiotto B Barbarat and D Olive ldquoThe role ofTec protein-tyrosine kinase in T cell signalingrdquo The Journal ofBiological Chemistry vol 274 no 2 pp 607ndash617 1999

[109] S Tewtrakul H Miyashiro N Nakamura et al ldquoHIV-1 inte-grase inhibitory substances from Coleus parvifoliusrdquo Phytother-apy Research vol 17 no 3 pp 232ndash239 2003

[110] T Dimo T B Nguelefack P V Tan et al ldquoPossible mechanismsof action of the neutral extract from Bidens pilosa L leaves onthe cardiovascular system of anaesthetized ratsrdquo PhytotherapyResearch vol 17 no 10 pp 1135ndash1139 2003

[111] T S C Li Chinese and Related North American Herbs CRCPress New York NY USA 2002

[112] H Wu H Chen X Hua Z Shi L Zhang and J ChenldquoClinical therapeutic effect of drug-separated moxibustion onchronic diarrhea and its immunologic mechanismsrdquo Journal ofTraditional Chinese Medicine vol 17 no 4 pp 253ndash258 1997

[113] S W Wright R R Harris J S Kerr et al ldquoSynthesis chemicaland biological properties of vinylogous hydroxamic acids dualinhibitors of 5-lipoxygenase and IL-1 biosynthesisrdquo Journal ofMedicinal Chemistry vol 35 no 22 pp 4061ndash4068 1992

[114] W R Almiron and M E Brewer ldquoClassification of immaturestage habitats of Culicidae (Diptera) collected in CordobaArgentinardquo Memorias do Instituto Oswaldo Cruz vol 91 no 1pp 1ndash9 1996

[115] N L Wang J Wang X S Yao and S Kitanaka ldquoTwo newmonoterpene glycosides and a new (+)-jasmololone glucoside

Evidence-Based Complementary and Alternative Medicine 51

from Bidens parvifloraWilldrdquo Journal of Asian Natural ProductsResearch vol 9 no 5 pp 473ndash479 2007

[116] P Champagnat ldquoRole of the terminal bud in the actionexercised by the cotyledon of Bidens pilosus L var radiatus onits axillary budrdquo Comptes Rendus des Seances et Memoires de laSociete de Biologie vol 145 no 17-18 pp 1374ndash1376 1951

[117] S F Nielsen S B Christensen G Cruciani A Kharazmiand T Liljefors ldquoAntileishmaniai chalcones statistical designsynthesis and three-dimensional quantitative structure-activityrelationship analysisrdquo Journal of Medicinal Chemistry vol 41no 24 pp 4819ndash4832 1998

[118] Y C Hwang J J H Chu P L Yang W Chen and M V YatesldquoRapid identification of inhibitors that interfere with poliovirusreplication using a cell-based assayrdquo Antiviral Research vol 77no 3 pp 232ndash236 2008

[119] V F Andrade-Neto M G L Brandao F Q Oliveira et alldquoAntimalarial activity of Bidens pilosa L (Asteraceae) ethanolextracts fromwild plants collected in various localities or plantscultivated in humus soilrdquo Phytotherapy Research vol 18 no 8pp 634ndash639 2004

[120] R J Marles and N R Farnsworth ldquoAnti-diabetic plants andtheir active constituentsrdquo Phytomedicine vol 2 no 2 pp 137ndash189

[121] M Habeck ldquoDiabetes treatments get sweet help from naturerdquoNature Medicine vol 9 no 10 p 1228 2003

[122] H W Lin G Y Han and S X Liao ldquoStudies on the activeconstituents of the Chinese traditional medicine Polygonatumodoratum (Mill) DrucerdquoActa Pharmaceutica Sinica vol 29 no3 pp 215ndash222 1994

[123] C L T Chang Y C Chen H M Chen N S Yang andW C Yang ldquoNatural cures for type 1 diabetes a reviewof phytochemicals biological actions and clinical potentialrdquoCurrent Medicinal Chemistry vol 20 no 7 pp 899ndash907 2013

[124] L Dey A S Attele and C S Yuan ldquoAlternative therapies fortype 2 diabetesrdquo Alternative Medicine Review vol 7 no 1 pp45ndash58 2002

[125] C L T ChangH Y Liu T F Kuo et al ldquoAnti-diabetic effect andmode of action of cytopiloynerdquo Evidence-Based Complementaryand Alternative Medicine vol 2013 Article ID 685642 13 pages2013

[126] M R Khan M Kihara and A D Omoloso ldquoAnti-microbialactivity of Bidens pilosa Bischofia javanica Elmerillia papuanaand Sigesbekia orientalisrdquo Fitoterapia vol 72 no 6 pp 662ndash6652001

[127] T Dimo T B Nguelefack P Kamtchouing E Dongo ARakotonirina and S V Rakotonirina ldquoHypotensive effects ofmethanol extract from Bidens pilosa Linn on hypertensive ratsrdquoComptes Rendus de lrsquoAcademie des Sciences Serie III vol 322no 4 pp 323ndash329 1999

[128] A K Hassan O Deogratius J F Nyafuono O Francis and OP Engeu ldquoWound healing potential of the ethanolic extracts ofBidens pilosa and Ocimum suaverdquo African Journal of Pharmacyand Pharmacology vol 5 no 2 pp 132ndash136 2011

[129] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[130] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[131] C Kandaswami and EMiddleton ldquoFree radical scavenging andantioxidant activity of plant flavonoidsrdquo Advances in Experi-mental Medicine and Biology vol 366 pp 351ndash376 1994

[132] K Yamamoto H Kakegawa H Ueda et al ldquoGastric cytopro-tective anti-ulcerogenic actions of hydroxychalcones in ratsrdquoPlanta Medica vol 58 no 5 pp 389ndash393 1992

[133] J O C Ezeonwumelu A K Julius C N Muhoho et alldquoBiochemical and histological studies of aqueous extract ofBidens pilosa leaves from Ugandan Rift valley in ratsrdquo BritishJournal of Pharmacology and Toxicology vol 2 no 6 pp 302ndash309

[134] L Frida S Rakotonirina A Rakotonirina and J P SavineauldquoIn vivo and in vitro effects of Bidens pilosa L (Asteraceae)leaf aqueous and ethanol extracts on primed-oestrogenized ratuterine musclerdquo African Journal of Traditional Complementaryand Alternative Medicines vol 5 no 1 pp 79ndash91 2008

52 Evidence-Based Complementary and Alternative Medicine

Composition Comments

1 Please provide valid postal code to the address if

possible

2 We made the highlighted change in Table 4 for the sake

of correctness Please check

3 Please check the correctness of the highlighted part in

Table 5

4 We considered the highlighted part in Table 11 as

reference Please check similar cases throughout

5 Please check the format of Table 18

6 We reformatted Table 20 Please check

7 We made the highlighted change in Table 4 for the sake

of consistency Please check similar cases throughout the

paper

8 We made the highlighted change(s) for the sake

of consistency Please check simila highlighted cases

throughout

9 We made the highlighted change in Table 9 for the sake

of clarity Please check similar cases through out

10 We made the highlighted change(s) for the sake of

correctness Please check

11 Comment on ref [66] This reference is a repetition

of [18] Please check similar cases throughout

Author(s) Name(s)

Author 1

Author 2

Author 3

Evidence-Based Complementary and Alternative Medicine 13

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

76

6-[(36-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-

2-[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone(Z)-6-O

-(36-di-O

-acetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

bideno

sideA

O

O

HO

OH

OH

OHOH O

H

H

HOH

HO

O

O

O

Leaves

(China)

[60]

77

6-[(46-di-O

-Acetyl-120573

-D-

glucop

yranosyl)oxy]-2-

[(34-dihydroxyphenyl)-

methylene]-7-hydroxy-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(410158401015840610158401015840-

diacetyl-120573-D

-glucropyrano

syl)-

673101584041015840-

tetrahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

HO

O

OO

Leaves

(not

stated)

aeria

l(Ch

ina)

[343561]

78

2-[(34-Dihydroxyph

enyl)-

methylene]-7-

hydroxy-6-

[(246-tri-O

-acetyl-120573

-D-

glucop

yranosyl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(210158401015840410158401015840610158401015840-

triacetyl-120573

-D-

glucropyrano

syl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H

HO

H

H

HOH

O

OOO

O

Leaves

(not

stated)

aeria

l(Ch

ina)

[3461]

79

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-6-

[(346-tri-O

-acetyl-120573

-D-glucopyrano

syl)-oxy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(310158401015840410158401015840610158401015840-

triacetyl-120573

-D-glucrop

yranosyl)-

673101584041015840-te

trahydroxy

O

O

HO

OH

OH

OO

H OH

H

HOH

HO

OOO O

Aeria

l(Ch

ina)

[3435]

80

2-[(34-Dihydroxyph

enyl)-

methylene]-7-hydroxy-

6-[[6-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propenyl]-120573-

D-glucopyrano

syl]o

xy]-

3(2H

)-benzofuranon

e

Aurone

(Z)-6-O-(6-O-

p-coum

aroyl-

120573-D

-glucropyrano

syl)-

673101584041015840-te

trahydroxy

OH

O

O

O

HO

OH

OH

OHO

H

HO

H

H

HOH

HO

OLeaves

(Japan)

[59]

14 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

81

1-[2-(120573-D

-Glucopyrano

syloxy)-

4-hydroxyphenyl]-2-

hydroxy-3-(3-

hydroxyphenyl)-2-

prop

en-1-on

e

Chalcone120572

32101584041015840-

tetrahydroxy-

21015840-O

-120573-D

-glucop

yranosyl

HO

O

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[35]

821-(24-Dihydroxyph

enyl)-

3-(34-dihydroxyph

enyl)-

2-prop

en-1-on

eBu

tein

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

833-(34-D

ihydroxyph

enyl)-

1-(234-trihydroxyph

enyl)-

2-prop

en-1-on

eOkanin

O

HO

HO

OH

OH

OH

Leaves

(China)

[60]

84

3-(34-D

ihydroxyph

enyl)-

1-[3-(120573-D

-glucop

yranosyloxy)-

24-dihydroxyphenyl]-2-

prop

en-1-on

e

Okanin

31015840-O

-120573-D

-glucosid

e

O

HO

OH

OH

OH

OO

HO

H

HO

H

H

HOH

H

OH

Leaves

(Germany)

flowers(Germany)

[61ndash63]

85

3-(34-D

ihydroxyph

enyl)-

1-[4-(120573-D

-glucop

yranosyloxy)-

23-dihydroxyphenyl]-

2-prop

en-1-on

e

Okanin

41015840-O

-120573-D

-glucosid

e

OOH

OH

OH

HOO

OHO

H

HO

H

H

HOH

H

OH

Flow

ers(Germany)

leaves

(Japan)

[5963]

Evidence-Based Complementary and Alternative Medicine 15

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

86Okanin41015840-O

-120573-D

-(610158401015840-

O-acetylglucosid

e)

O

O

OH

OH

OH

OHO

H

HO

H

H

HOH

H

HO

O

O

Flow

ers(Germany)

[63]

87

1-[4-[(46-di-O

-Acetyl-

120573-D

-glucopyrano

syl)-oxy]-

23-dihydroxyphenyl]-3-

(34-dihydroxyph

enyl)-2-

prop

en-1-on

e

Okanin

41015840-O

-120573-D

-(410158401015840610158401015840-diacetyl)-

glucop

yranoside

O

O

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Aeria

l(Ch

ina)

[34]

88Okanin41015840-O

-120573-D

-(210158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[64]

89Okanin41015840-O

-120573-D

-(310158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

O

OAe

rial(Ch

ina)

[34]

16 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

90

1-[23-Dihydroxy-4-

[[6-O-[3-(4-hydroxyph

enyl)-

1-oxo-2-prop

enyl]-120573-D

-glucop

yranosyl]oxy]-ph

enyl]-

3-(34-dihydroxyph

enyl)-

2-prop

en-1on

e

Okanin

41015840-O

-120573-D

-(610158401015840-tr

ans-

p-coum

aroyl)-

glucoside

O

O

OH

OH

OH

OHOH

HO

H

H

HOH

H

HO

OO

OH

Leaves

(Germany)

[64]

91Okanin41015840-O

-120573-D

-(410158401015840-acetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Leaves

(Germany)

[65]

92

Okanin

41015840-O

-120573-D

-(210158401015840410158401015840-diacetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[65]

Evidence-Based Complementary and Alternative Medicine 17

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

93

Okanin

41015840-O

-120573-D

-(310158401015840410158401015840-diacetyl-610158401015840-

trans-p-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

OH

O O

Leaves

(Germany)

[65]

94

Okanin

41015840-O

-[120573-D

-glucopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

OOH

OH

OH

HO

OH

HO

HHO

H

HOH

HO

OH

OH

HO

HHO

H

HOH

HO

Flow

er(G

ermany)

[63]

95Okanin

3101584041015840-di-O

-120573-D

-glucosid

e

OOH

OH

OH

O OO

H HO

HHOH

HOH

HHO

OHO

H

HO

H

H

HOH

H

OH

Flow

er(G

ermany)

[63]

18 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

96

1-[3-(120573-D

-Glucopyrano

syloxy)-

24-dihydroxyphenyl]-

3-(3-hydroxy)-

4-metho

xyph

enyl)-

2-prop

en-1-on

e

Okanin

4-methylether-

31015840-O

-120573-D

-glucop

yranoside

OOH

O

OH

HO

O

H HO

HHOH

HOH

H

OHO

Leaves

(Germany)aerial

(China)

[3562]

97

Okanin4-methylether-

3101584041015840-di-O

-120573-(41015840101584061015840101584041015840101584010158406101584010158401015840-

tetraacetyl)-

glucop

yranoside

OOH

OOH

O

HH

OH

HO

HOH

H

OO

O

Ac

Ac

AcAc

OO

H

HO

H

H

HOH

H

O

Aeria

l(Ch

ina)

[42]

98Ch

alcone210158404101584061015840-tr

imetho

xy-

4-O-D

-glucopyrano

syl-

dihydro

Not

foun

dLeaves

(China)

[60]

992-(34-D

ihydroxyph

enyl)-23-

dihydro-78

-dihydroxy-

4H-1-

benzop

yran-4-one

Okanin

isoOO

OH

HO

OH

OH

Leaves

(China)

[60]

100

2-(34-D

ihydroxyph

enyl)-

23-dihydro-8-hydroxy-

7[(246-tr

i-O-acetyl-120573

-D-

glucop

yranosyl)oxy]-4H

-1-b

enzopyran-4-on

eOO

OH

OH

OH

OH

O

HHO

H

HO

HO

OAc

Ac

Ac

Aeria

l(Ch

ina)

[34]

Evidence-Based Complementary and Alternative Medicine 19

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

101

57-Dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

OO

OH

HO

OH

Aeria

l(TanzaniaC

hina)

[4266]

102

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

7-O-glucopyrano

side

OO

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Tanzania)

[66]

103

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-4H

-1-benzop

yran-4-one

Luteolin

OO

HO

OH

OH

OH

Aeria

l(TanzaniaC

hina

Vietnam)

[343542

6667]

104

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5hydroxy-4H-1-

benzop

yran-4-one

Luteolin

7-O-120573-D

-glucop

yranoside

OOOH

OH

OH

OO

HO

H

HO

H

H

HOH

OH H

Aeria

l(Tanzania)

[66]

105

57-Dim

etho

xy-6-

(5-m

etho

xy-6-m

ethyl-

4-oxo-4H

-pyran-3-yl)-

2-ph

enyl-4H-1-

benzop

yran-4-one

5-O-M

ethylhosludin

O OOO

O

O

O

Aeria

l(Ugand

a)[68]

20 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

106

3-(120573-D

-Glucopyrano

syloxy)-

57-dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Astragalin

Kaem

pferol-3-O

-120573-D

-glucopyrano

side

HO

OH

O O

OH

O

O

HO

HHO

H

H

HOH

OH

H

Aeria

l(Ch

ina)

[35]

107

Kaem

pferol3-

(23-di-E

-p-cou

maroyl-

120572-L-rhamno

pyrano

side)

Not

foun

dAe

rial(Vietnam)

[67]

108

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5-hydroxy-36-dimetho

xy-

4H-1-

benzop

yran-4-one

Axillarosid

eO O

OH

OH

OO

OH

OO

HO

H

HO

H

H

HOH

H

OH

Aeria

l(Ch

ina)

[42]

109

57-Dihydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaureidin

O

O

O

O

O

HO

OH

OH

Who

le(Taiwan)

[69]

110

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaurein

O O

OO

O OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan)w

hole

(Taiwan)

[69ndash

71]

Evidence-Based Complementary and Alternative Medicine 21

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

111

5-Hydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-

67-dimetho

xy-4H-

chromen-4-one

Eupatorin

iso

OO O

OH

O

O OH

Not

foun

d(C

hina)

[3772]

112

2-(34-D

imetho

xyph

enyl)-7-

(120573-D

-glucopyrano

syloxy)-35-

dihydroxy-

8-metho

xy-4H-1-

benzop

yran-4-one

O O

OH

O O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

113

7-(120573-D

-Glucopyrano

syloxy)-

5hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-38-

dimehtoxy-4H-1-

benzop

yran-4-one

O O

O

OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

114

Isorhamnetin

3-[O

-120572-

L-rham

nopyrano

syl-

(1-2)-120573-D

-glucop

yranoside]

Not

foun

dAe

rial(Vietnam)

[67]

115

7-[(6-deoxy-120572-L-

Manno

pyrano

syl)o

xy]-

3-120573-D

-glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-4H

-1-benzop

yran-4-one

Luteoside

O O

OH

O

OH

OH

OH

HOH

H

H HO

HO

HO

OH

HO

H

HO

OH

OH

HH

Aeria

l(Ch

ina)

[42]

22 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

116

Luteolin

3-O-120573-D

-Glucopyrano

side

O O

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

HO

Aeria

l(Tanzania)

[66]

117

57-Dihydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Quercetagetin

3631015840-tr

imethylether

OOO

O

O OH

HO

OH

Aeria

l(Ch

ina)

[42]

118

6-((2S3S4S5S)-

Tetrahydro-345-tr

ihydroxy-

6-(hydroxymethyl)-2H

-pyran-2-yloxy)-5-hydroxy-

2-(4-hydroxy-3-

metho

xyph

enyl)-

37-dimetho

xy-4H-

chromen-4-one

Quercetagetin

3731015840-

trim

ethylether-6-O

-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[42]

119

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

JaceinQ

uercetin

3631015840-

trim

ethyl

ether-7-O-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan

China)

who

le(Taiwan)

[425470]

120

2-(34-D

ihydroxyph

enyl)-

357-trihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

OO

HO

OH

OH

OH

OH

Aeria

l(Ch

ina)leaves

(China)who

le(C

hina)

[356074]

Evidence-Based Complementary and Alternative Medicine 23

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

121

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-galactopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-4-one

Quercetin

3-O-120573-D

-galactosidehyperin

hyperosid

e

O O

HO

OH

OH

OH

OOHH

HOH

H

H HO

HO

OH

Aeria

l(TanzaniaJapan)

Leaves

(China)Who

le(C

hina)

[376066

717475]

122

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-glucopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

3-O-120573-

D-glucopyrano

side

O O

HO

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

Aeria

l(TanzaniaC

hina

Japan)L

eaves(Japan)

[355966

75]

123

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-

4-oxo-4H

-1-benzop

yran-3-yl-

120573-D

-glucopyrano

siduron

icacid

Quercetin

3-O-120573-D

-glucuron

opyranoside

O O

HO

OH

OH

OH

OHOH

HOH

H

H HO

HO

OHO

Aeria

l(TanzaniaJapan)

[6671]

124

3-[[6-O-(6-

Deoxy-120572-L-m

anno

pyrano

syl)-

120573-D

-galactopyrano

syl]o

xy]-

2-(34-dihydroxyph

enyl)-57-

dihydroxy-4H

-1-benzop

yran-4-one

Quercetin

3-O-rob

inob

iosid

e

O O

HO

OH

OH

OH

O

HOHH OH

HO

H HH

OO

OHH

HOH

H

H HO

HO

Aeria

l(Japan)W

hole

(Taiwan)

[7071]

24 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

125

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-

3-metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

OH

Roots(Brazil)

[55676]

126

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-

120573-D

-glucopyrano

syl]o

xy]-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120572-

L-rham

nopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

O

HHO HHO

HO H

HH

ORo

ots(Brazil)

[5676]

127

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-120573-D

-glucop

yranosyl]oxy]-5-

hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-3-metho

xy-

4H-1-

benzop

yran-4-one

Quercetin

341015840-

dimethyl

ether-7-O-rutinoside

O OOH

OH

O

O

OH O

HOH

H

HHO

H

O

HHO HHO

HO H

HH

O

HO

Aeria

l(Ch

ina)

[3435]

128

2-(34-D

ihydroxyph

enyl)-3-

(120573-D

-glucofurano

syloxy)-57-

dihydroxy-4H

-1-benzop

yran-

4-on

e

Isoq

uercitrin

O OOH

OH

OHO

H

OHH

OH

HO

H O HOH

OH

Aeria

l(Japan

China)

[3571]

Evidence-Based Complementary and Alternative Medicine 25

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

129

Quercetin

3-O-120573-D

-galactop

yranoside

O

OH

HO

OH OH

O

O

HO

OH

OHOH

OAe

rial(no

tstated)

[18]

130

Quercetin

3-O-rutinoside

O

O

R

OO

OH

OHO

H

O

OH

OH

H3C

H3C

H3C

OH

HO

Who

le(Taiwan)

[70]

26 Evidence-Based Complementary and Alternative Medicine

Table 6 Terpenoids isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(Country) References

131371111-Tetramethylbicyclo[810]undeca-

26-dieneBicyclogermacrene Leaves

(Brazil) [95]

13241111-Trimethyl-8-

methylenebicyclo[720]undec-4-ene

E-Caryophyllene Leaves(Brazil) [95]

1331-Methyl-5-methylene-8-

(1-methylethyl)-16-cyclodecadiene

Germacrene D Leaves(Brazil) [95]

1344-(15-Dimethyl-4-hexen-1-ylidene)-1-methyl-cyclohexene

Z-120574-Bisabolene Leaves(Brazil) [95]

135Decahydro-114-trimethyl-7-methylene-1H-cycloprop[e]-

azulene120573-Gurjunene Leaves

(Brazil) [95]

136 2669-Tetramethyl-148-cycloundecatriene

120572-Humulene120572-caryophyllene

Leaves(Brazil) [95]

13712344a568a-Octahydro-1-isopropyl-7-methyl-4-methylenenaphthalene

120575-Muurolene Leaves(Brazil) [95]

138

12344a568a-Octahydro-4a8-dimethyl-2-(1-methylethylidene)-

naphthalene

Selina-37(11)-diene

Leaves(Brazil) [95]

139(2E7R11R)-371115-

Tetramethyl-2-hexadecen-1-ol

Phytol

HO

Whole(Taiwan) [57]

Evidence-Based Complementary and Alternative Medicine 27

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

140 371115-Tetramethyl-2-hexadecenoic acid Phytanic acid

O

OH

Whole(Taiwan) [57]

141371115-Tetramethyl-2-

hexadecenyl ester-heptanoicacid

Phythylheptanoate O O

Leaves(Not stated) [33]

142

(3S10R13R)-17-((2R5R)-5-Ethyl-6-methylheptan-2-yl)-

2347891011121314151617-tetradecahydro-

1013-dimethyl-1H-cyclopenta[a]phenanthren-

3-ol

Campestrol

HO

Aerial(Tanzania) [18]

143 Not found Phytosterin-B Not foundNot found(TaiwanEgypt)

[39 96]

144 Stigmast-5-en-3-ol 120573-sitosterol

HOH

H

H

Aerial(Tanzania)

whole(Taiwan)

[18 31 57]

145

1314151617-Tetradecahydro-1013-

dimethyl-1H-cyclopenta[a]phenanthren-

3-yloxy)-tetrahydro-6-(hydroxymethyl)-2H-pyran-345-triol

120573-Sitosterolglucoside

H

H

HOH

HO

HHO

H

HOHH O

OH Not found(Egypt) [39]

146 5120572-Stigmasta-7-en-3120573-ol

HO

Whole(Taiwan) [57]

147 5120572-Stigmasta-722t-dien-3120573-ol

HO

Whole(Taiwan) [57]

148 Stigmasta-522-dien-3-ol Stigmasterol

HOH

H

H

Not found(Taiwan)aerial

(Tanzania)leaves (notstated)whole

(Taiwan)

[18 31 3357]

28 Evidence-Based Complementary and Alternative Medicine

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

149 Lup-20(29)-en-3-ol Lupeol

H

HH

HOH

Not found(Egypt) [39]

150 Lup-20(29)-en-3-ol acetate Lupeol acetate

H

HH

O

OH

Not found(Egypt) [39]

151 Olean-12-en-3-ol 120573-amyrin

H

HOH

Not found(Egypt) [39]

152 5913-Trimethyl-242526-trinoroleanan-3-ol Friedelan-3120573-ol

HO

Aerial(Tanzania) [18]

153 5913-Trimethyl-242526-trinoroleanan-3-one

Friedelinfriedelan-3-one

O

Aerial(Tanzania) [18]

1542610151923-

Hexamethyl-2610141822-tetracosahexaene

Squalene

Aerial(Tanzania)Leaves (Notstated)Whole(Taiwan)

[18 33 57]

155 120573120573-Carotene 120573-Carotene Leaves (Notstated) [97]

Evidence-Based Complementary and Alternative Medicine 29

studies [83] Hot water extracts of B pilosa varminor Sheriffwere also assessed for its antileukemic effects on leukemiccell lines L1210 U937 K562 Raji and P3HR1 using XTT-based colorimetric assays The extract inhibited the five celllines with IC

50values ranging from 145120583gmL to 586120583gmL

L1210 K562 Raji and P3HR1 were more sensitive to B pilosaextract with IC

50values below 200120583gmL [98]

Consistent with the antitumor activities of B pilosaextracts and fractions some of its phytochemicals alsoshowed anticancer activity as outlined in Table 11 Amongthem luteolin (103) a well-studied flavonoid with multiplebioactivities was more effective against tumor cell prolifera-tion than its derivatives with IC

50values ranging from 3 120583M

to 50120583M in cells and 5 to 10mgkg in animals Luteolin wasalso found to fight cancer as a food additive at concentrationsof 50 to 200 ppm [78] and prevent skin cancer [78] and cancer3invasion [103] Lee and colleagues reported that luteolinprevents cancer by inhibiting cell adhesion and invasion[103] Significant inhibition concentration was reported to be5 120583M and complete inhibition concentration was reported tobe 40120583M Moreover luteolin was reported to inhibit hepa-tocyte growth factor (HGF)-induced cell scattering as wellas cytoskeleton changes such as filopodia and lamellipodiawhichwas determined using phase-contrast and fluorescencemicroscopy Furthermore luteolin also inhibited the HGF-induced phosphorylation of c-Met ERK 12 and Akt aswell as the MAPKERK and P13K-Akt pathways [78 103]Other mechanisms underlying the anticancer activities ofluteolin are the inhibition of topoisomerase I and II whichinhibits cell replication and DNA repair thus promotingapoptosis regulation of PI-3-KinaseAktMAPKERKJNKactivation of apoptosis in the mitochondrial pathway byactivating caspase 9 and caspase 3 which were found inmalignant cells but not in normal human peripheral bloodmononuclear cells death receptor-induced apoptosis and acell-cycle arrest mechanism inhibition of fatty acid synthasewhich is upregulated in many cancer cells and sensitizationto chemotherapywhereby luteolin increases the susceptibilityof cancer cells to chemotherapy [78] Butein (82) is anotherflavonoid that showed a cytotoxic effect on human colonadenocarcinoma cell proliferation with a reported IC

50value

of 175 120583M Butein at 2 120583M affected the incorporation of[14C]-labeled leucine thymidine and uridine which cancause the inhibition of DNA RNA and protein synthesis ofhuman colon cancer cells Moreover butein also exhibitednoncompetitive inhibition of 1-chloro-24-dinitrobenzene(CDNB) in glutathione S-transferase (GST) activity Tumorresistance was correlated with high levels of GST thus buteininhibited proliferation of cancer cells [80] Another flavonoidpresent in B pilosa centaureidin (109) also showed anti-cancer activity in B lymphoma cells Centaureidin isolatedfrom Polymnia fruticosa inhibited tubulin polymerization invitro and induced mitotic figure formation in CA46 Burkittlymphoma cells Using turbidimetric assay the IC

50value of

centaureidin in inhibition of mitosis was 3120583M [104] Cyto-toxicity of centaureidin was further analyzed using AmericanNational Cancer Institute (NCI) 60 human tumor cell linesThe cytotoxicity potency of centaureidin expressed as GI

50

(50 growth inhibition in the NCI tumor line panel) was024120583M [81] These data mark centaureidin as a promisingantimitotic agent for tumor therapy

In addition to anti-tumor flavones polyynes found in B 4pilosa have also been shown to possess anti-tumor proper-ties Based on a bioactivity-directed isolation approach Wuand colleagues identified two polyyne aglycones from theethyl acetate fraction of B pilosa [44] 12-Dihydroxytrideca-57911-tetrayne (48) and 13-Dihydroxy-6(E)- tetradecene-81012-triyne (46) exhibited significant anticell prolifera-tion activity in primary human umbilical vein endotheliumcells (HUVEC) with IC

50values of 125 120583M and 173 120583M

respectively They also decreased angiogenesis and pro-moted apoptosis in human endothelial cells Their anti-angiogenic and cytotoxic effects correlated with activationof the CDK inhibitors and caspase-7 [44] In addition 12-Dihyroxy-5(E)-tridecene-7911-triyne (45) showed antian-giogenic effects in HUVECs with an IC

50value of 124 120583M as

evidenced by a decrease in the tube formation and migrationof HUVECs [43] The IC

50value of compound 45 in the

inhibition of basic fibroblast growth factor-induced HUVECgrowth was 282120583M However it had higher IC

50values

than those for lung carcinoma cells and keratinocytes Thiscompound could also inhibit cell proliferation of HUVECslung carcinoma A549 cells and HACAT keratinocytes Themechanism by which compound 45 inhibits HUVEC growthand angiogenesis is complicated and includes decreasing theexpression of cell cycle regulators (CDK4 cyclins D1 and Aretinoblastoma (Rb) and vascular endothelial growth factorreceptor 1) caspase-mediated activation of CDK inhibitorsp21 (Cip1) and p27 (Kip) upregulation of Fas ligand expres-sion downregulation of Bcl-2 expression and activation ofcaspase-7 and poly (ADP-ribose) polymerase [43]

32 Anti-Inflammatory Activity B pilosa is commonly usedto treat inflammatory disorders The anti-inflammatory phy-tochemicals present in B pilosa are listed in Table 11 5Cyclooxygenase-2 (COX-2) is a physiologically importantenzyme that converts arachidonic acid to prostaglandin(PGE2) Its expression is induced by a wide variety of external

stimuli indicating its involvement in inflammatory diseasesand it is used as an inflammatory marker [88] Yoshida andcolleagues studied the effects of the aqueous extracts of Bpilosa aerial parts in the production of COX-2 and PGE

2as

well as on the activation of mitogen activated protein kinases(MAPKs) in normal human dermal fibroblasts (HDFs) inresponse to inflammatory cytokine IL-1120573 This work showedthat IL-1120573 activated MAPKs such as ERK12 p38 and JNKto different extents and induced COX-2 expression TheCOX-2 expression in HDFs was regulated mainly by p38following IL-1120573 stimulation Consistently the p38 inhibitorSB203580 blocked this expression Using this cell platformB pilosa extracts were tested for inhibition of inflammationThe extract dose-dependently suppressed the activation ofp38 and JNK and moderately suppressed ERK12 as wellas suppressing COX-2 expression and PGE

2production

30 Evidence-Based Complementary and Alternative Medicine

Table7Ph

enylprop

anoids

isolatedfro

mB

pilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

156

3-(4-H

ydroxyph

enyl)-2-prop

enoica

cid

p-Cou

maricacid

OHO

OH

Who

le(Ja

pan)

[99]

157

2-Metho

xy-4-(2-prop

en-1-yl)-ph

enol

Eugeno

lOOH

Leaves

androots(Japan)

[99]

158

3-(4-H

ydroxy-3-m

etho

xyph

enyl)-

2-prop

enoica

cid

Ferulic

acid

O

OH

O

HO

Who

le(Ja

pan)

[99]

159

3-(34-D

ihydroxyph

enyl)-2-prop

enoic

acid

Caffeicacid

O

HO

OH

OH

Who

leandaeria

l(Japan)

[7599]

160

3-Prop

yl-3[(245-trim

etho

xyph

enyl)-

metho

xy]-24-pentanedione

3-Prop

yl-3-(245-

trim

etho

xy)benzyloxy-

pentan-24-dione

O

O

O

O

O

O

Leaves

(India)

[100]

161

3-(34-D

ihydroxyph

enyl)-2-

prop

enoica

cidethyleste

rCa

ffeateethyl

O

O

OH

OH

Not

foun

d(Taiwan)

who

le(Taiwan)

[434584]

Evidence-Based Complementary and Alternative Medicine 31Ta

ble7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

162

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

d-Erythron

icacid

2-O-caffeoyl-2-C

-methyl

O

HO

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

163

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

2-O-caffeoyl-2-C

-methyl

O

O

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

164

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-24-

dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

3-O-caffeoyl-2-C

-methyl

O

O

HO

HO

O

O

OH

OH

Leaves

(Japan)

[101]

165

4-(A

cetyloxy)-3-[[3-(34-

dihydroxyphenyl)-

1-oxo-2-propen-1-y

l-]oxy]-2-hydroxy-

2-methyl-b

utanoica

cid

O

HO

OH

O OHO

HO

OO

Not

foun

d(Ja

pan)

[73]

166

3-(34-D

ihydroxyph

enyl)-tetrahydro-4-

hydroxy-4-

methyl-5

-oxo-3-fu

ranyl

ester-2-prop

enoica

cid

3-O-C

affeoyl-2-C

-methyl-

D-erythrono

-14-la

cton

eHO

HO

O

OO

OOH

Leaves

(Japan)

[101]

167

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]-oxy]-145-

trihydroxy-cyclohexanecarboxylicacid

Chlorogenica

cid

O

HO

HO

OH

O

OH

O

OH

HO

Aeria

l(Japan)w

hole

(Taiwan)

[707175]

32 Evidence-Based Complementary and Alternative Medicine

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

168

4-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]oxy]-

135-tr

ihydroxy-cyclohexanecarboxylic

acid

4-O-C

affeoylqu

inicacid

OHO

HO

OH

OH

O

O

OH

OH

Aeria

l(Japan)

[71]

169

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-15-

dihydroxy-cyclo

hexanecarboxylicacid

34-Di-O

-caffeoylqu

inic

acid

HO

HO

O

O

O

O

OH

OH

HO

OH O

H

O

Aeria

l(Japan)W

hole

(Taiwan)

[45497071]

170

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-14-dihydroxy-

cyclo

hexanecarboxylicacid

35-Di-O

-caffeoylqu

inic

acid

HO

HO

O O

HO

O

HO

OH OO

OHOH

Aeria

l(Japan)w

hole

(Taiwan)

[45497071]

Evidence-Based Complementary and Alternative Medicine 33

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

171

34-bis[[(2E

)-3-(34-D

ihydroxyph

enyl)-1-

oxo-2-prop

en-1-

yl]oxo]-15

-dihydroxy-

cyclo

hexanecarboxylicacid

45-Di-O

-caffeoylqu

inic

acid

OHO

OH

HO

O

O

OH

OH

O

OH

HO

OWho

le(Taiwan)

[454970]

172

3-[4-[[-6-O-[3-(4-H

ydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]-phenyl]-2-prop

enoica

cid

120573-D

-p-C

oumaricacid4-O

-(6-O

-p-caoum

aroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HOH

HO

O

O

Leaves

(Japan)

[59]

173

3-[4-[[2-O

-Acetyl-6

-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]ph

enyl]2-propeno

icacid

120573-D

-p-C

oumaricacid4-O

-(2-O

-acetyl-6

-O-p-

caou

maroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HO

HO

O

O

Leaves

(Japan)aerial

(China)

[3459]

174

67-Dihydroxy-2-chrom

enon

eEsculetin

cicho

rigenin

OHO HO

O

Not

foun

d(Egypt)

[39]

34 Evidence-Based Complementary and Alternative Medicine

Table 8 Aromatic compounds isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

175 12-Benzenediol PyrocatechinHO

HOWhole(Japan) [99]

176 4-Ethyl-12-benzenediol Pyrocatechol

OH

OH

Whole(Japan) [99]

177 Dimethoxyphenol26-dimethoxyphenol

O

HO

O

Roots (Japan) [99]

178 4-Ethenyl-2-methoxy-phenol p-Vinylguaiacol

O

OHWhole(Japan) [99]

179 2-Hydroxy-6-methylbenzaldehyde

6-Methyl-salicylaldehyde O

OH

Whole(Japan) [99]

180 Benzene-ethanol 2-Phenyl-ethanolHO

Whole(Japan) [57]

181 4-Hydroxy-3-methoxy-benzaldehyde Vanillin O

O

HOAerial(Japan) [99]

182 3-Hydroxy-4-methoxy-benzaldehyde Vanillin iso

OO

OH

Leaves(Japan) [99]

183 4-Hydroxybenzoic acid p-Hydroxybenzoicacid

O

HOOH Whole

(Japan) [99]

184 2-Hydroxybenzoic acid Salicylic acidO

HO

HO

Stem androots (Japan) [99]

185 34-Dihydroxybenzoicacid Protocatechuic acid

O

HOOH

OH

Whole(Japan) [99]

1864-Hydroxy-

3methoxybenzoicacid

Vanillic acid

OOH

O

OH

Aerial(Uganda)

Roots (Japan)[68 99]

Evidence-Based Complementary and Alternative Medicine 35

Table 8 Continued

S N IUPAC names Common names Structure Plant part(country) References

187 345-Trihydroxybenzoicacid Gallic acid

O

OHHO

HO

HO

Whole(China) [102]

[88] This work supports the use of B pilosa as an anti-inflammatory agent however no compounds responsible forthe anti-inflammatory activity of B pilosa were identified

A further study also reported the anti-inflammatoryactivity as well as the antiallergic activity of B pilosa [75] Inthis study dried powder of the aerial part of B pilosa whichhad been pretreated with the enzyme cellulosine was usedfor further tests The results showed that oral administrationof the cellulosine-treated B pilosa lowered the level of serumIgE in mice 10 days after immunization with DNP (24-dintrophenyl)-Ascaris as an antigen This treatment alsoreduced dye exudation in skin induced by passive cutaneousanaphylaxis and production of inflammatory mediators his-tamine and substance P in rats [75] Phytochemical analysisshowed that cellulosine treatment increased the percentage ofcaffeic acid and flavonoids This study suggests that B pilosaand its phenolics have anti-inflammatory functions

Phenolics and polyynes are major anti-inflammatoryphytochemicals present in B pilosa (Table 11) Unsurpris-6ingly phenolics such as luteolin (103) and ethyl caffeate(161) that are major constituents of B pilosa have alsobeen reported to possess anti-inflammatory activity Lute-olin was reported to exhibit anti-inflammatory activity inmacrophages Xagorari and colleagues showed that luteolininhibited the release of inflammatory cytokines TNF-120572 andinterleukin-6 in RAW 2647 cells following LPS stimulation[87] It inhibited TNF-120572 production with an IC

50value

of 1120583M The underlying anti-inflammatory mechanism ofluteolin was reported to be the inactivation of Akt andNF-120581B activation [87] In addition luteolin was reportedto confer anti-inflammatory activity through inhibition ofLPS-stimulated iNOS expression in BV-2 microglial cellsIt inhibited LPS-activated microglia in a dose-dependentmanner with an IC

50value of 69 120583MMoreover immunoblot

and RT-PCR data proved that luteolin suppressed I120581B-120572degradation and iNOS expression in LPS-activated microglia[85] Kim and colleagues stated that luteolin may havebeneficial effects on inflammatory neural diseases throughinhibition of iNOS expression [85] A related study revealedthat luteolin decreased the transcriptional activity of NF-120581BRelA via partial inhibition of TNF-mediated NF-120581B DNAbinding activity Luteolin also inhibited Akt phosphorylationand induced degradation of a transcription factor interferonregulatory factor (IRF) [86]

Chiang and colleagues showed that ethyl caffeate(161) significantly inhibited NO production in mousemacrophages RAW 2647 cells [84] Based on MTT assays

they concluded that this inhibition was not due to thecytotoxicity of ethyl caffeate The IC

50value of ethyl caffeate

in the inhibition of NO production was 55 120583gmL slightlylower than curcumin (positive control) which has anIC50

value of 65120583gmL They demonstrated that ethylcaffeate exerted anti-inflammatory activity via the reducedtranscription and translation of iNOS (inducible nitric oxidesynthase) in RAW 2467 cells In addition this compoundalso suppressed COX-2 expression in RAW 2467 cellsand MCF-7 cells The in vivo anti-inflammatory effectof ethyl caffeate was verified by testing in TPA-treatedmouse skin Like celecoxib the positive control ethylcaffeate significantly abolished COX-2 expression in adose-dependent manner Ethyl caffeate at 1mg200120583Lsite(24mM) inhibited COX-2 expression at a level comparableto celecoxib at 1mg200120583Lsite (13mM) Remarkably ethylcaffeate at 48mM (2mg200120583Lsite) was more effectivethan celecoxib at 131mM (10mg200120583Lsite) Moreoverthis compound inhibited the activation of nuclear factor-120581B(NF-120581B) by LPS via the prevention of NF-120581B binding toDNA [84] In addition 3 ethyl caffeate analogs (ethyl 34-dihydroxyhydrocinnamate ethyl cinnamate and catechol)also showed different degrees of NF-120581B binding to DNA aslisted in Table 12 Ethyl cinnamate lacks a catechol moietywhich results in ineffective inhibition of NF-120581B binding toDNA [84]

Pereira and colleagues assessed the anti- 7inflammatory and immunomodulatory activities ofB pilosa methanol extract as well as one polyyne2-O-120573-glucosyltrideca-11(E)-en-3 579-tetrayn-12-diol(54) in T lymphocytes and a zymosan-induced arthritismouse model [27] They first examined the in vitro effect of 8the B pilosa extract and compound 54 on cell proliferation ofhuman T cells stimulated with 5120583gmL phytohemagglutinin(PHA) or 100 nM 12-O-tetradecanoyl phorbol-13-acetate(TPA) plus 15 120583M ionomycin and on cell proliferation ofmouse T cells stimulated with 5 120583gmL concanavalin A(Con A) The data demonstrated that both methanol extractand compound 54 suppressed T-cell proliferation in adose-dependent manner The estimated IC

50values of the B

pilosa extract against human T cells stimulated with 5120583gmLPHA and 100 nM TPA plus 15120583M ionomycin were 125 and25 120583gmL respectively In comparison with the methanolextract compound 54 showed 10-fold more inhibition ofhuman T-cell proliferation with an estimated IC

50value of

15 120583gmL Accordingly the B pilosa extract and compound54 dose-dependently suppressed mouse T-cell proliferation

36 Evidence-Based Complementary and Alternative Medicine

Table 9 Porphyrins isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

188

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(15S16S)-10-ethenyl-5-ethyl-1161820-tetrahydro-6111522-tetramethyl-1820-dioxo-15H-912-imino-212-

metheno-471714-dinitrilopyrano[43-

b]azacyclononadecine-16-propanoic

acid

AristophyllC

NH N

HNN

O

O OOOPhytyl

O

Leaves(Taiwan) [100]

189

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -20b-(methoxycarbonyl)-9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-dinitrilo-12-

dioxeto-[310158404101584034]-cyclo-pent[1 2]azacyclononadecine-

19-propanoic acid

Bidenphytin A

NH N

HNN

OPhytylO O O

O O

Leaves(Taiwan) [100]

190

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -2a-

hydroxy-20b-(methoxycarbonyl)-

9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-

dinitrilo-12-dioxeto[310158404101584034]-cyclo-pent[12-b]-

azacyclononadecine-19-propanoic acid

Bidenphytin B

NH N

HNN

OPhytylO O O

O O

OH

Leaves(Taiwan) [100]

191

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO O

HOOO

Leaves(Taiwan) [100]

192

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

Evidence-Based Complementary and Alternative Medicine 37

Table 9 Continued

S N IUPAC names Common names Structure Plant part(country) References

193

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

Leaves(Taiwan) [100]

194

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

195

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3S4S21R)-9-ethenyl-14-ethyl-21-(methoxycarbonyl)-

481318-tetramethyl-20-oxo-3-phorbinepropanoic acid

Pheophytin A

NH N

HNN

PhytylO

O

OO O

Leaves(Taiwan) [100]

with estimated IC50values of 30 and 25 120583gmL respectively

Taken together the data indicate that the B pilosa extractand compound 54 act on human and mouse T cells Totest the in vivo effect of the B pilosa extract and compound54 a zymosan-induced arthritis mouse model was usedThis model was established from B10ASgSnJ mice with aninjection of zymosan (015mg) The zymosan-injected mice9received an intraperitoneal injection of the B pilosa extract(1 5 or 10mg) at one dose a day for 5 days Popliteal lymphnode (PLN) weight was monitored to check the developmentof arthritis The results revealed that 10mg of the methanolextract of B pilosa extract could significantly diminishinflammation as evidenced by PLN weight [27] This worksuggests that B pilosa (and compound 54) can suppressimmune response and inflammation

33 Antidiabetic Activity Anti-diabetic agents are primarilydeveloped from plants and other natural resources [8 24 4951 53] B pilosa is one of 1200 plant species that have beeninvestigated for antidiabetic activity [120 121]B pilosa is usedas an anti-diabetic herb in America Africa and Asia [51 120

122] Many studies have indicated that B pilosa could treattype 1 diabetes (T1D) and type 2 diabetes (T2D) in animals

Etiologically speaking T1D is caused by autoimmune-mediated destruction of pancreatic 120573 cells leading to insulindeficiency hyperglycemia and complications Currentlythere is no cure for T1D Polarization ofTh cell differentiationcontrols the development of T1D Suppression of Th1 celldifferentiation and promotion of Th2 cell differentiationameliorate T1D [123] One study showed that the butanolfraction of B pilosa inhibited T-cell proliferation decreasedTh1 cells and cytokines and increased Th2 cells andcytokines leading to prevention of T1D in nonobesediabetic (NOD) mice [49] Based on a bioactivity-directedisolation strategy 3 polyynes 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne (53) also known ascytopiloyne 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D- Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne (50) were identifiedfrom B pilosa [49 53] The IC

50value of the butanol fraction

was 200120583gmL This inhibition was reported to be partiallyattributed to cytotoxicity because the butanol fraction at

38 Evidence-Based Complementary and Alternative Medicine

Table 10 Other compounds isolated from B pilosa [30]

SN IUPAC names Common names Structure Plant part(country) References

19637-Dihydro-137-

trimethyl-1H-purine-26-dione

CaffeineN

NN

NO

O

Aerial(Uganda) [68]

197

1-((2R4S5R)-Tetrahydro-4-hydroxy-5-(hydroxymethyl)furan-2-yl)-5-

methylpyrimidine-24(1H3H)-dione

ThymidineHO

HO

ON

O

NHO Not found

(China) [37]

198 1-(2-Thienyl)-ethanone 2-Acetyl-thiophene

S

O

Roots(Germany) [41]

199

(2R3S4S5S)-2-(Heptan-2-yloxy)-tetrahydro-6-

((tetrahydro-34-dihydroxy-5-

(hydroxymethyl)furan-2-yloxy)methyl)-

2H-pyran-345-triol

Heptanyl2-O-120573-xylofuranosyl-

(1rarr 6)-120573-glucopyranoside

O

OOH

OH

HO

OHO

H

HO

H

H

HOHH

O

Whole(Taiwan) [70]

200

2-[(3R7R11R)-3-Hydroxy-371115-

tetramethylhexadecyl]-356-trimethyl-

25-cyclohexadiene-14-dione

120572-Tocopherylquinone

OO

HO

Whole(Taiwan) [32]

2017-O-(410158401015840610158401015840-Diacetyl)-

120573-D-glucopyranoside

Not found Leaves(China) [60]

180 120583gmL could cause 50 death of Th1 cells Moreoverthis study suggested that the butanol fraction may preventdiabetes in NOD mice in vivo via downregulation of Th1cells or upregulation Th2 cells that have effects which areantagonistic of those of Th1 cells [49] This was proven byintraperitoneal injection of the butanol fraction at a doseof 3mgkg BW 3 times a week to NOD mice from 4 to 27weeks This dosage resulted in lower incidence of diabetes(33) At a dose of 10mgkg the butanol fraction of Bpilosa totally eliminated (0) the initiation of the diseaseTo further support this result assessment of IgG2a and IgEproduction was performed in the serum of NOD mice Asin vivo results obtained from intracellular cytokine stainingexperiments were not very conclusive levels of IgG2a and IgE

were measured since Th1 cytokine IFN120574 and Th2 cytokineIL-4 favor the production of IgG2a and IgE respectively Asexpected high levels of IgE and some decline in the levels ofIgG2a were observed in the serum Profiling of the butanolextract revealed five compounds 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne(50) 45-Di-O-caffeoyl- quinic acid 35-Di-O-caffeoylquinicacid and 34-Di-O-caffeoylquinic acid Only the first twocompounds showed similar effects on the prevention ofdiabetes in NOD mice as the B pilosa butanol fractionMoreover compound 50 showed greater activity thancompound 69 in terms of enhancement (by 34 comparedto 8) of differentiation of Th0 to Th2 at 15 120583gmL (both

Evidence-Based Complementary and Alternative Medicine 39

Table 11 Chemical constituents of B pilosa and their biological activities

SN Name Classification Molecularformula Biological activities

109 Centaureidin [91] Flavonoid C18H16O8Anti-listerial [69 91]

Cytotoxic [10]

110 Centaurein [91] Flavonoid C24H26O13

Anti-listerial [69 91]Cytotoxic [10]Anti-viral [105]

103 Luteolin [106] Flavonoid C15H10O6

Anti-viral [107 108]Cytotoxic [77]

Anti-inflammatory [109]Anti-allergic [109]

82 Butein [110] Flavonoid C15H12O5Anti-leishmanial [111]

Cytotoxic [78]48 12-Dihydroxytrideca-57911-tetrayne [112] Polyyne C13H12O2 Anti-angiogeneic [112]46 13-Dihyroxy-6(E)-tetradecene-81012-triyne [112] Polyyne C14H16O2 Anti-angiogeneic [112]

45 12-Dihyroxy-5(E)-tridecene-7911-triyne [113] Polyyne C13H14O2Anti-angiogeneic [113]Anti-proliferative [113]

64 1-Phenylhepta-135-triyne [114] Polyyne C13H8

Anti-microbial [115]Anti-malarial [3]Cytotoxic [3]Antifungal [15]

27 Linoleic acid [79] Fatty acid C18H32O2Anti-viral (100)Cytotoxic [17]

161 Ethyl caffeate [84] Phenylpropanoid C11H12O4 Anti-inflammatory [84]

54 2-O-120573-Glucosyltrideca-11(E)-en-3579-tetrayn-12-diol [26] Polyyne C19H20O7Immunosuppressive andAnti-inflammatory [26]

53 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne [50] Polyyne C19H22O7Anti-diabetic [116]

Anti-inflammatory [50]

69 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne [28] Polyyne C20H26O7Anti-diabetic [28]

Anti-inflammatory [94]

50 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne [28] Polyyne C19H24O7

Anti-diabetic [28]Anti-inflammatory [94]

Anti-malarial andantibacterial [47]

129 Quercetin 3-O-120573-D-galactopyranoside [18] Flavonoid C21H20O12 Anti-inflammatory [117]

170 35-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

171 45-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

169 34-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

126 Quercetin 331015840-dimethyl ether7-O-120572-L-rhamnopyranosyl-(1rarr 6)-120573-D-glucopyranoside [118] Flavonoid C21H20O11 Anti-malarial [119]

125 Quercetin 331015840-dimethyl ether-7-O-120573-D-glucopyranoside [118] Flavonoid C21H20O12 Anti-malarial [119]70 1-Phenyl-13-diyn-5-en-7-ol-acetate [27] Polyyne C15H12O2 Anti-malarial [27]199 Heptanyl 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside [70] Miscellaneous C18H44O10 Antioxidant [70]124 3-O-Rabinobioside [70] Saccharide C27H38O15 Antioxidant [70]130 Quercetin 3-O-rutinoside [70] Flavonoid C27H38O15 Antioxidant [70]167 Chlorogenic acid [70] Phenolic C16H26O9 Antioxidant [70]119 Jacein [70] Flavonoid C24H26O13 Antioxidant [70]

49 (R)-12-dihydroxytrideca-357911-pentayne [47] Polyyne C13H8O2Anti-malarial andAntibacterial [47]

40 Evidence-Based Complementary and Alternative Medicine

Table 12 Structure and activity relationship studies of ethyl caffeateusing in vitro NF-120581BDNA binding assays [84]

Compound Concentration(120583M)

NF-120581BDNA binding

Ethyl caffeate 50 100inhibition

Ethyl 34-dihydroxyhydrocinnamate 100 100inhibition

Catechol 400 100inhibition

Ethyl cinnamate 400 Noinhibition

Table 13 Apoptosis in cocultures of T cells and pancreatic 120573 cells[53]

Cellmedium apoptosis andnecrosis

CD4+ T cellscontrol medium lt4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 2

CD4+ T cellswith cytopiloyne-treated 120573 cells 18CD4+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 7

CD8+ T cellscontrol medium 4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 4

CD8+ T cellswith cytopiloyne-treated 120573 cells 4CD8+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 4

Table 14 Radical scavenging activities of B pilosa extracts [70]

Extractscontrol DPPH assayIC50 (120583gmL)

NBThypoxanthinesuperoxide assayIC50 (120583gmL)

Quercetin 198 15Ascorbic acid 634 Not determined120572-tocopherol 897 Not determinedEthyl acetateextract 1383 597

Butanol extract 1669 114Water extract gt100 gt100

compounds) and inhibition (by 40 compared to 10) ofdifferentiation toTh1 at the same concentration [49]

Among the three polyynes found in B pilosa cytopiloyne(53) had the most potent anti-T1D activity [53] To test the invivo effect of cytopiloyne NODmice received intraperitonealor intramuscular injection of cytopiloyne at 25120583gkg BW3 times per week Twelve-week-old NOD mice started todevelop T1D and 70 of NOD mice aged 23 weeks andover developed T1D Remarkably 12- to 30-week-old NODmice treated with cytopiloyne showed normal levels of bloodglucose (lt200mgdL) and insulin (1-2 ngmL) Consistent

Table 15 Radical scavenging activity of secondarymetabolites fromB pilosa [70]

Metabolite (Table 11)control DPPH assay IC50 (120583gmL)199 Not determined124 53130 68167 105169 33171 38119 Not determined110 Not determinedQuercetin 256Caffeic acid 890

Table 16 Antioxidant activity of the essential oils and water extractsfrom B pilosa [92]

Extract IC50 (120583gmL)Leaf essential oils 47Flower essential oils 50Leaf extract 61Flower extract 172

with T1D incidence cytopiloyne delayed and reduced theinvasion of CD4+ T cells into the pancreatic islets [53]

In vitro study showed that cytopiloyne (53) inhibited thedifferentiation of naıveTh (Th0) cells (ie CD4+ T cells) intoTh1 cells and promoted differentiation of Th0 cells into Th2cells [50] The in vitro data are consistent with the in vivoresults indicating that cytopiloyne reducedTh1differentiationand increased Th2 differentiation as shown by intracellularcytokine staining and FACS analysis [53] In line with theskewing of Th differentiation the level of serum IFN-120574 andIgG2c decreased while that of serum IL-4 and serum IgEincreased compared to the negative controls (PBS-treatedmice) Cytopiloyne also enhanced the expression of GATA-3 a master gene for Th2 cell differentiation but not theexpression of T-bet a master gene forTh1 cell differentiationfurther supporting its role in skewingTh differentiation [53]

Also importantly cytopiloyne partially depleted CD4+rather than CD8+ T cells in NOD mice [53] As shown inTable 13 coculture assays showed that the depletion of CD4+T cells was mediated through the induction of Fas ligandexpression on pancreatic islet cells by cytopiloyne leading toapoptosis of infiltrating CD4+ T cells in the pancreas via theFas and Fas ligand pathways However cytopiloyne did notinduce the expression of TNF-120572 in pancreatic islet cells andthus had no effect on CD8+ T cells [53]

In addition Chang and colleagues showed that cytopi-loyne dose-dependently inhibited T-cell proliferation stimu-lated by IL-2 plus Con A or anti-CD3 antibody using [3H]thymidine incorporation assay [53]

Overall the mechanism of action of cytopiloyne andprobably its derivatives in T1D includes inhibition of T-cellproliferation skewing of Th cell differentiation and partialdepletion of Th cells Due to the anti-diabetic mechanisms

Evidence-Based Complementary and Alternative Medicine 41

Table 17 Antibacterial activity of essential oils and flower extracts from B pilosa [92]

Strain Mean zone of inhibition (mm)Leaf essential oil Flower essential oil Leaf extract Flower extract

Micrococcus flavus 127 plusmn 03 87 plusmn 03 102 plusmn 02 108 plusmn 03Bacillus subtilis 173 plusmn 19 117 plusmn 02 109 plusmn 02 103 plusmn 02Bacillus cereus 190 plusmn 14 112 plusmn 03 118 plusmn 04 185 plusmn 10Bacillus pumilus 123 plusmn 07 108 plusmn 02 105 plusmn 04 77 plusmn 02Escherichia coli 137 plusmn 04 203 plusmn 07 102 plusmn 11 140 plusmn 13Pseudomonas ovalis 125 plusmn 08 137 plusmn 15 102 plusmn 06 125 plusmn 06

Table 18 Antibacterial activity of root extracts from B pilosa [93]

Strain MIC50 (mgmL)Methanol extract Acetone extract

Bacillus cereus 10 mdashEscherichia coli 5 5Klebsilla pneumonia 5 10Micrococcus kristinae 10 mdashPseudomonas aeruginosa 10 10Staphylococcus aureus 5 10Sraphylococcus epidermidis 5 5Serratia marcescens 10 mdashShigelea flexneri 10 mdashStreptococcus faecalis 10 mdash

Table 19 Antibacterial activity of B pilosa of compound 29 [47]

Strain MIC50 (120583gmL)Escherichia coli NIHJ 1Escherichia coli ATCC25922 1Klebsiella pneumoniae ATCC700603 128Serratia marcescens ATCC13880 16Pseudomonas aeruginosa ATCC27853 8Staphylococcus aureus FDA209P 05Staphylococcus aureus ATCC29213 025Staphylococcus aureus N315 (MRSA) 05Enterococcus faecalis ATCC29212 2Enterococcus faecalis NCTC12201 (VRE) 1Bacillus subtilis ATCC6633 05Candida albicans ATCC10231 025

of action it was hypothesized that cytopiloyne protectsNOD mice from diabetes by a generalized suppression ofadaptive immunity To evaluate this hypothesis ovalbumin(Ova) was used as a T-cell dependent antigen to primeNOD mice which had already received cytopiloyne orPBS vehicle Ova priming boosted similar anti-Ova titersin cytopiloyne-treated mice and PBS-treated mice but adifference in immunoglobulin isotype was observed in thetwo groups Thus it was concluded that cytopiloyne is animmunomodulatory compound rather than an immunosup-pressive compound [50 53]

T2D is a chronic metabolic disease with serious com-plications resulting from defects in either insulin secretion

insulin action or both [124] A study by Ubillas et alshowed that the aqueous ethanol extract of the aerial partof B pilosa at 1 gkg body weight (BW) lowered bloodglucose in dbdb mice a T2D mouse model [51] Basedon a bioactivity-guided identification compounds 69 and50 were identified Further the mixture of the compounds(69 50) in a 2 3 ratio significantly decreased blood glucoseconcentration and reduced food intake on the second dayof treatment when administered at doses of 250mgkg twicea day to C5BLKs-dbdb mice When tested at 500mgkga more substantial drop in blood glucose level as wellas the stronger anorexic effect (food intake reduced from58 gmouseday to 25 gmouseday) was observed [51] Inthis study it was suggested that the blood glucose lower-ing effect of B pilosa was caused in part by the hungersuppressing effect of its polyynes [51] However the hungersuppressing effect of the ethanol extract of B pilosa wasnot found in the studies described below In another study[24] water extracts of B pilosa (BPWE) were used indiabetic dbdb mice aged 6ndash8 weeks with postprandialblood glucose levels of 350 to 400mgdL Like oral anti-diabetic glimepiride which stimulates insulin release onesingle dose of BPWE reduced blood glucose levels from374 to 144mgdL The antihyperglycemic effect of BPWEwas inversely correlated to an increase in serum insulinlevels suggesting that BPWE acts to lower blood glucose viaincreased insulin production However BPWE had differentinsulin secretion kinetics to glimepiride [24] One flaw incurrent anti-diabetics is their decreasing efficacy over timeThe authors investigated the long term anti-diabetic effectof BPWE in dbdb mice BPWE reduced blood glucoseincreased blood insulin improved glucose tolerance andreduced the percentage of glycosylated hemoglobin (HbA1c)Both long-term and one-time experiments strongly supportthe anti-diabetic action of BPWE [24] In sharp contrast toglimepiride BPWE protected against islet atrophy in mousepancreas The investigators further evaluated anti-diabeticproperties of 3 B pilosa varieties B pilosa L var radiate(BPR) Bpilosa L var pilosa (BPP) and B pilosa L varminor(BPM) in dbdb mice [8] One single oral dose (10 50 and250mgkg body weight) of BPR BPP or BPM crude extractsdecreased postprandial blood glucose levels in dbdb micefor up to four hours and the reduction of glucose levels inthe blood appeared to be dose-dependent Comparing thethree variants BPR extract resulted in a higher reduction inblood glucose levels when administered at the same dose as

42 Evidence-Based Complementary and Alternative Medicine

Table 20 Antifungal activity of B pilosa [92]

Partextract Concentration (ppm) Strain InhibitionCortiicum rolfsii Fusarium solani Fusarium oxysporum

Leaves

Essentialoils

100 857 plusmn 09 682 plusmn 0 745 plusmn 17250 960 plusmn 08 779 plusmn 18 879 plusmn 04

AqueousExtracts

100 446 plusmn 17 605 plusmn 21 716 plusmn 07250 942 plusmn 03 689 plusmn 07 824 plusmn 19

Flowers

Essentialoils

100 604 plusmn 09 892 plusmn 04 869 plusmn 05250 894 plusmn 12 980 plusmn 03 949 plusmn 06

AqueousExtracts

100 331 plusmn 11 714 plusmn 07 573 plusmn 22250 661 plusmn 14 912 plusmn 0 900 plusmn 07

Table 21 Antifungal activity of B pilosa root extracts [93]

Strain LC50 (mgmL)Acetoneextracts

Methanolextracts

Waterextracts

Aspergillus niger 014 006 007Aspergillus flavus 1091 658 0Penicillium notatum 005 005 005

the other two varieties In terms of serum insulin levels a doseof 50mgkg of each extract was used and the BPR extracttogether with the three polyynes significantly increased theserum insulin level in dbdb mice Long-term experiments(28-day treatment) were then conducted using diabetic micewith postprandial glucose levels from 370 to 420mgdL andglimepiride was used as positive controlThe range of dosagesapplied was from 10mgkg BW to 250mgkg BW Resultsshowed that the positive control as well as the crude extractsof the three varieties lowered the blood glucose levels indbdb mice However only BPR extract containing a higherpercentage of cytopiloyne (53) reduced blood glucose levelsand augmented blood insulin levelsmore than BPP andBPMThe percentage of glycosylated hemoglobin A1c (HbA1c) wasalso measured and found to be 79 plusmn 05 in mice aged 10ndash12 weeks and 66 plusmn 02 61 plusmn 03 and 62 plusmn 03 inthe blood of age-matchedmice following treatment with BPRcrude extract (50mgkg) glimepiride (1mgkg) and com-pound 53 (05mgkg) respectively [8] Among the polyynesfound in B pilosa cytopiloyne was the most effective againstT2D Hence cytopiloyne was used for further study on anti-diabetic action and mechanism [125] The data confirmedthat cytopiloyne reduced postprandial blood glucose lev-els increased blood insulin improved glucose tolerancesuppressed HbA1c level and protected pancreatic islets indbdb mice Nevertheless cytopiloyne failed to decreaseblood glucose in streptoztocin (STZ)-treated mice whose bcells were already destroyed Additionally cytopiloyne dose-dependently increased insulin secretion and expression in bcells as well as calcium influx diacylglycerol and activationof protein kinase C120572 Collectively the mechanistic studiessuggest that cytopiloyne treats T2D via regulation of insulin

production involving the calciumDAGPKC120572 cascade in bcells

The studies detailed above point to the conclusion thatcytopiloyne and related polyynes (compounds 69 and 49)are anti-diabetics in animal models The data uncover a newbiological action of polyynes It should be noted that likeall anti-diabetic drugs cytopiloyne failed to prevent or curediabetes completely but reduced diabetic complications [125]Intriguingly 34 polyynes have been found in B pilosa so farIt remains to be seen whether all the polyynes present in thisplant have anti-diabetic activities

34 Antioxidant Activity Free radicals can damage cellularcomponents via a series of chemical reactions [89] leadingto development and progression of cardiovascular diseasecancer neurodegenerative diseases and ageing [70] Freeradicals nitric oxide (NO) and superoxide anions can beproduced in macrophages to kill microbes However anexcessive generation of the free radicals under pathologicalconditions is associated with a wide range of illnessesPlants are known to be rich in antioxidant phytochemicalsChiang and colleagues evaluated the free radical scaveng-ing activity of crude extract fractions and compounds ofB pilosa using 11-diphenyl-2-picrylhydrazyl (DPPH) andhypoxanthinexanthine oxidase assays [70] Using DPPHand hypoxanthinexanthine oxidase assays they found thatthe B pilosa crude extract and the ethyl acetate butanoland water fractions had free radical scavenging activityNine compounds Heptyl- 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside (199) 3-O-Rabinobioside (124) Quercetin3-O-rutinoside (130) Chlorogenic acid (167) 34-Di-O-caffeoylquinic acid (169) 35-Di-O-caffeoylquinic acid (170)45-Di-O-caffeoylquinic acid (171) Jacein (119) and Cen-taurein (110) had DPPH radical scavenging activity [70]The IC

50values of the B pilosa crude extractfractions and

compounds are summarized in Tables 14 and 15 respectivelyMeasurement of free radical scavenging activities is one

way of assessing the antioxidant activities of B pilosa andits fractions and compounds It is interesting that the ethylacetate and butanol fractions are more active than the waterfraction and B pilosa crude extract [70] Of the secondarymetabolites only phenolic compounds 124 130 167 169 and171 showed significant DPPH-radical scavenging activities

Evidence-Based Complementary and Alternative Medicine 43

Further analysis of the structure-activity relationship of thecompounds suggested that substitution of the C3 hydroxylgroup with glycosides increased the activity approximately2-fold (for example in compounds 124 and 130) relative toquercetin (-OH in its C3) [70] Further modification of thestructures of the active compounds needs to be performed totest the effects of various substituents on activity The reasonwhy most of the antioxidant compounds contain phenolmoieties in their structure could be that the reduction-oxidation (redox) properties of phenols allow them to actas reducing agents singlet oxygen quenchers and hydrogendonors

A complementary study by Muchuweti and colleaguesdetermined phenolic content antioxidant activity and thephenolic profile of B pilosa methanol extract [89] Theyestimated that the phenolic content of the methanol extractof B pilosawas 11028plusmn22mgg [89] Vanillin hydroxyben-zaldehyde caffeic acid coumaric acid and ferulic acid werefound in this extractThe B pilosa extract also showed DPPHradical scavenging activity Furthermore the antioxidantactivity of the flavonoids found in B pilosa was correlatedwith its hepatoprotective effects through their inhibitionof NF-120581B activation which may lessen the oxidative stresscaused by the production of free radicals during liver injury[60]This activity might also be due to the anti-inflammatoryeffects of the aqueous extracts of B pilosa aerial parts on theinhibition of COX-2 and PGE

2production [88]

Essential oils from B pilosa flowers and leaves are alsoreported to possess antioxidant activity With the aim ofreplacing chemically synthesized additives Deba and col-leagues [92] worked on the antioxidant antibacterial andantifungal activities of essential oils and water extracts of Bpilosarsquos leaves and flowers Table 16 summarizes the resultsobtained from DPPH free radical scavenging assay

It can be inferred fromTable 16 that essential oils from theleaves possessed the highest activity It is reported elsewherethat monoterpenes present in essential oils such that of Bpilosa have protective effects and antioxidant properties [92]Beta-carotene bleaching method was also performed Leavesessential oils and aqueous extracts of the leaves and flowersshowed higher activity than the flower essential oils This isdue to the volatility of the flower essential oils The activityexhibited by the aqueous extracts was accounted to thepresence of phenolic compounds that are reported to donatea hydrogen atom to free radicals such that the propagation ofthe chain reaction during lipid oxidation is terminated [92]Overall essential oils and phenolics present in B pilosa canbe thought of as major antioxidant compounds

35 Immunomodulatory Activity B pilosa is thought to bean immunomodulatory plant and is reported to be effectivein the treatment of immune disorders such as allergy [75]arthritis [75] and T1D [48 50 53]

As pointed out in the discussion of its anti-diabeticactivities (Section 33) a combination of phytochemicalsmethod and T-cell activation assays was used to studyimmunomodulatory properties of B pilosa IFN-120574 is a keycytokine released by T and NK cells that mediates immune

cells and sustains immunity against pathogens Defects inIFN-120574 expression regulation and activation result in vulner-ability to diseases caused by bacteria and viruses [69] Anelegant study performed byChang and colleagues using IFN-120574 promoter-driven luciferase reporter construct in JurkatT cells showed that hot water crude extracts of B pilosaincreased IFN-120574 promoter activity two-fold [69] Out of thesubfractions of this extract the butanol fraction but not thewater or ethyl acetate fractions increased IFN-120574 promoteractivity six-fold Centaurein (110) and centaureidin (109)were identified from the butanol fraction and were stated tocause a four-fold increase in IFN-120574 promoter activity withEC50

values of 75 120583gmL and 09120583gmL respectively Themechanism of action of centaurein was determined usingtranscription factors such as AP-1 NFAT and NF120581B whichare reported to bind to IFN-120574 promoter and regulate IFN-120574transcription Unlike with the activity of the positive controlPHA centaurein caused a four-fold increase in NFAT a3-fold increase in NF120581B and had little if any effect onAP-1 enhancer activities (23-fold three-fold and ten-foldincreases respectively were seenwith PHA) [69]The authorsconcluded that centaureinmodulates IFN-120574 expression by theNFAT and NF120581B pathways The article only determined themechanism of action of centaurein Its aglycone centaureidinmay act through the samemechanism though this conclusionneeds to be further verified

B pilosa extract and its compounds are reported to inhibitdifferentiation of naıve CD4+ helper T (Th0) cells into Th1cells [49] Using theThcell differentiation assay as a screeningplatform 3 polyynes 2-120573-D-glucopyranosyloxy-1 -hydroxytrideca-57911-tetrayne (53) 3-120573-D-glucopyranosyl-1-hydroxy-6(E)- tetradecene-81012-triyne (69) and 2-120573-D-glucopyranosyloxy-1-hydroxy-5(E)- tridecene-7911-triyne(49) were discovered from B pilosa [49 53] The data showsthat cytopiloyne and other two polyynes suppressed thedifferentiation of type 1 helper T (Th1) cells and productionof Th1 cytokines and promoted that of type 2 helper T (Th2)cells and production of Th2 cytokines thus explaining theimmunomodulatory and anti-inflammatory effects of Bpilosa and its polyynes

Chang and colleagues were the first to report the effect ofthe butanol extract of B pilosa on the autoimmune diabetesand airway inflammation inmice [48] Imbalance in the levelsofTh1 andTh2 and of various cytokines leads to autoimmunediseases T1D and other autoimmune diseases (rheumatoidarthritis Crohnrsquos disease among others) are exacerbated byan increase Th1 levels (specifically CD4+ Th1 cells) whileTh2 cells antagonize this effect [19] Moreover Th2 cellsmediate asthma in ovalbumin-induced hypersensitivity inBALBc mice [48] In their work [48] Chang and colleaguesshowed that 10mgkg butanol extracts with 15 (ww)compound 69 and 11 (ww) compound 49 (Table 11)ameliorated the development of Th1-mediated diabetes inNOD mice through inhibition of 120573 cell death and leukocyteinfiltration At the same dosage the butanol extracts alsoexacerbated ovalbumin-induced pulmonary inflammationin BALBc mice with an increase in the infiltration ofeosinophils and mast cells into the airway of the mice [48]Despite the different outcomes both mouse models proved

44 Evidence-Based Complementary and Alternative Medicine

the concept that control over the Th1Th2 shift is associatedwith autoimmune diseases and that the B pilosa butanolextract can shift the differentiation of Th0 cells to Th2 cells[48 49]

An extended study presented by Chiang and colleaguesshowed that compound 53 modulates T-cell functions [50]Using CD4+ T cells from BALBc mice they demonstratedthat cytopiloyne decreased levels of IFN-120574 producing cells(Th1) by 122 (from 72 to 598) Since Th1 and Th2 celldifferentiation is antagonistic it was expected that compound53 increased the percentage of mouse IL-4 producing cells(Th2) by 72 (from 237 to 309) Subsequent assess-ment of effect of compound 53 on the modulation of thetranscription of IL-4 and IFN-120574 showed that cytopiloyne asexpected decreased the splenocyte levels of IFN-120574 mRNAand increasing that of IL-4 in a dose-dependent manner Inthe range of 01 to 3 120583gmL of compound 53 the effects werenot attributed to its cytotoxicity Consequently using 3120583gmLcytopiloyne for 72 and 96 hours the protein concentration ofIFN-120574 decreased to 186 and 444 respectively Under thesame conditions cytopiloyne increased IL-4 concentrationsto 1985 and 2470 (for 72 and 96 hours resp) Thismodulation of T-cell differentiation exhibited by compound53 was used to explain its anti-diabetic activity [50]

The anti-diabetic role of cytopiloyne was extensivelydiscussed above (Section 33 anti-diabetic activity) Themolecular basis of the regulation of cytokine expression bycytopiloyne has been described Cytopiloyne directly elevatedthe expression level of IL-4 via GATA-3 upregulation inT cells [53] However reduction of IFN-120574 expression in Tcells seemed to come from the indirect opposing effect IL-4cytokine because the expression level of T-bet was unaltered[53] In this way cytopiloyne skewedTh1 polarization into theTh2 state conferring protection against T1D in NOD miceAside frompolarization ofThcell differentiation cytopiloynealso activated the expression of Fas ligand in pancreaticb cells this increase leading to the partial depletion of Tcells and reduction of immune response in local areas suchas the pancreas Of note cytopiloyne also inhibited T-cellproliferation and activation By targeting T cells from threeimmunomodulatory actions cytopiloyne protects againstT1D and probably other Th1-mediated autoimmune diseases[19]

The phytochemical constituents of B pilosa exert theirfunctions on different immune cells to modulate immuneresponse It is possible that some of the compounds mayhave agonistic or antagonistic effects on immune responseImmune function of B pilosa may depend on its compo-sition and amount of compounds which could explain theapparently conflicting report of B pilosa butanol extractaggravating allergy in mice [48] while cellulosine-treatedextract ameliorated allergy [48 75] IFN-120574 promoter reporterassays and T-cell differentiation assays were used to isolate2 flavonoids [69] and 3 polyynes [49 50] as immunomod-ulatory compounds from the butanol fraction of B pilosaInterestingly the flavonoids promote IFN-120574 expression inNK and T cells In marked contrast the polyynes promoteIL-4 expression and indirectly inhibit IFN-120574 expression indifferentiating T cells Sensitivity appears to be the key to

identifying structure- and bioactivity-related phytochemicalsfrom medicinal plants

36 Antimalarial Activity The use of chemical drugs againstpathogens has resulted in drug-resistant mutants Examplesof drug resistance can be found in the species of thePlasmodium that cause malaria It is important to search fornew compounds to combat Plasmodium parasites [47] Astudy of the anti-malarial activity of the leaf extracts of Bpilosa using a combination of phytochemistry and bioassaysshowed that compound 49 (Table 11) showed activity againsta malaria parasite (P falciparum NF54 strain) with an IC

50

value of 60 120583gmL [77] In addition compound 49 isolatedfrom the aerial parts of B pilosa inhibited growth of the Pfalciparum FCR-3 strain with an IC

50value of 035 120583gmL

This compound was tested for its in vivo effect in miceinfected with P berghei NK-65 strain Results showed thatcompound 49 decreased the average parasitemia in the redblood cells by 207 (from 328 of that of the control to121) after an intravenous injection of 08mgkg BWday forfour days [47] Further studies addressing the anti-malarialmechanism underlying both polyynes and clinical studies areneeded

37 Antibacterial Activity Emergence of multiple antibiotic-resistantmicrobes is becoming a global threat to public healthand a challenge to disease treatment For instance penicillinis commonly used to combat a food-borne intracellularbacteriumListeria however penicillin-resistant bacteria havebeen discovered recently [69] Chang and coworkers iso-lated centaurein (110) and centaureidin (109) from B pilosaextract [69 70] Centaurein enhances expression of IFN-120574a key cytokine for macrophage activation and consequentlyenhances bactericidal activity in macrophages [69 70] Inagreement with observed in vitro effects centaurein wasreported to prevent and treat Listeria infection in C57BL6Jmice [91] Mechanistic studies confirmed that centaureinexerted antilisterial action via IFN-120574 expression in wild-typemice but not IFN-120574 knockout mice [91] Further in vitrostudies on centaurein showed that this compound increasedIFN-120574 expression by 13 (from 17 to 20) 20 (from 21to 41) and 11 (from 6 to 17) in CD4+ T cells CD8+Tcells and NK cells respectively That is to say there wasan increase in IFN-120574 producing immune cells As expectedcentaurein also enhanced the expression level of T-bet a keynuclear factor for IFN-120574 expression Consistently centaureinaugmented the serum IFN-120574 levels inC57BL6Jmice and thisaugmentation peaked 24 hours after compound injectionThe quantity of mouse serum IFN-120574 was sufficient to activatemacrophages in vitro and eradicated GFP-producing Listeriainside macrophages However the entry of Listeria intomacrophages was not affected by centaurein-treated mousesera Centaurein treatment at 20120583g per mouse rescued 30of the mice infected with a lethal dose of Listeria (2 times106 CFU) It is noteworthy that in the presence of ampicillin(5 120583gmouse) centaurein (20 120583gmouse) rescued 70 of themice suggesting an additive effect between ampicillin andcentaurein [91] Despite lower abundance centaureidin was

Evidence-Based Complementary and Alternative Medicine 45

30 times more active than centaurein in terms of IFN-120574production [91]

Aside from the indirect antibacterial action mentionedabove extract andor compounds of B pilosa also showeddirect bacteriostatic andor bactericidal action One studyreported that essential oils and leafflower extracts of Bpilosa could suppress the growth of gram positive and gramnegative bacteria as evidenced by zone of inhibition assaysIn this study antibacterial activity of the essential oils fromthe leaves and flowers of B pilosa was determined in anattempt to identify natural products as food preservatives forprevention of microbial multiplication and food oxidationThe essential oils and extracts of B pilosa leaves and flow-ers showed moderate but different extents of antibacterialactivity (Table 17) In general essential oils had higherantibacterial activity than crude extracts One explanation forthis could be that monoterpenes in the essential oils destroycellular integrity and subsequently inhibit the respirationand ion transport processes The presence of antibacterial120573-caryophyllene could be another explanation as reportedelsewhere [92] Another study reported that the methanoland acetone extract of B pilosa roots displayed antibacterialactivities against the bacteria listed in Table 18 [93] andmethanol extracts from the roots seemed to be the mosteffective

Another study indicated that the polyyne (R)-12-dihydroxytrideca-357911-pentayne (49) from this plantalso suppressed bacterial growth as shown by the minimuminhibitory concentration required to inhibit 50 bacterialgrowth (MIC

50) in Table 19 This compound was highly

effective against several Gram positive and Gram negativebacteria including the drug-resistant bacteria Staphylococcusaureus N315 (MRSA) and Enterococcus faecalis NCTC12201(VRE) [47] Strikingly compound 49 had a similar MIC

50

value to antibiotics (ampicillin tetracycline norfloxacin andamphotericin B) in most of the bacteria tested

Antibacterial activity of B pilosa extracts and compo-nents expressed as MIC

50and the mean zone of inhibition

is tabulated in Tables 17 18 and 19The zone of inhibition forAmpicilline (positive control) ranges from 153 plusmn 03mm to443 plusmn 02mm [92]

38 Antifungal Activity B pilosa has traditionally been usedto treat microbial infection Recently different parts of Bpilosa have been tested for antifungal activities Deba andcolleagues first evaluated the antifungal effect of the hotwater extracts of the B pilosa roots stems and leaves againstCorticium rolfsii Fusarium solani and Fusarium oxysporumThey discovered that C rolfsii was most suppressed bytreatment with B pilosa as its growth was reduced at almostall the tested doses followed by F oxysporum and F solani[99] However the fungicidal activities of the stems androots were greater than the leaves [99] Moreover the samegroup assessed the antifungal activity of the essential oilsand aqueous extracts from B pilosa flowers and leaves [92]They showed that the extracts and oils had antifungal activityagainst C rolfsii F solani and F oxysporum Essential oils

appeared to have better fungicidal activity thanwater extractsas summarized in Table 20

Another study by Ashafa and colleagues showed thatacetone methanol and water extracts of the B pilosa rootsshowed antifungal activities against Aspergillus niger Aflavus and Penicillium notatum using the agar dilutionmethod The results are tabulated in Table 21 [93] Negativecontrols showed 0 growth inhibitionThe methanol extractof the B pilosa roots at 10mgmL was also effective againstCandida albicans [93]Of note B pilosa obtained fromPapuaNew Guinea had no activity against A niger and C albicans[126] but the SouthAfrican (EasternCape) ecotype exhibitedmoderate activity against C albicans [93] This discrepancymay depend on extraction solvents extraction procedureassay techniques different plant parts and abundance ofactive compounds

B pilosa produces a variety of secondary metabolitessuch as flavonoids phenylacetylenes alkaloids sterols ter-penoids and tannis [92 93] However none of them havebeen confirmed as active compounds against fungi Furtherinvestigation of active compounds from B pilosa is necessaryto further understand the antifungal efficacy of this plant

39 Hypotensive and Vasodilatory Activities In early studiesDimo and colleagues used three rat models normotensiveWistar rats (NTR) salt-loading hypertensive rats (SLHR)and spontaneous hypertensive rats (SHR) to investigate thehypotensive effect of the methanol crude extract of B pilosaleaves [28 127] The extract lowered systolic blood pressurein hypertensive rats (SLHR and SHR) to a greater degreethan NTR [28 127] In addition a decrease in urinarysodium ions and an increase in urinary potassium ions wereobserved after treatment with the methanol extract of Bpilosa leaves although neither differences were statisticallysignificant [28 127] Taking the data together the studyproposed that B pilosa leaf extract reduced blood pressurevia vasodilation [28 127] The same group continued totest the antihypertensive effect of aqueous and methylenechloride extracts of B pilosa leaves in a hypertensive ratmodel [28 127] To establish a fructose-induced hypertensionmodel male Wistar rats were given 10 fructose solutionto drink ad libitum for three weeks In addition to freeaccess to 10 fructose the rats were treated with the aqueous(150mgkg) or methyl chloride (350mgkg) extracts of Bpilosa for additional three weeks [28 127] Both extracts ofB pilosa leaves had a hypotensive effect on rats Howeverneither extracts reversed the elevation of serum insulin infructose-fed rats Therefore B pilosa lowered blood pressureirrespective of insulin [28 127] To better understand thehypotensive mechanism the authors investigated the effectof a neutral extract of B pilosa (NBP) a mixture of methanolandmethylene chloride (1 1) extract after neutralization withNaOH and HCl on the heart and the blood pressure ofNTR and SHR [110] This study showed that an intravenousinjection of the NBP resulted in a biphasic reduction insystolic blood pressure In addition one intravenous dose ofthe extract at 10 20 and 30mgkg BW decreased systolicblood pressure in normal rats by 183 425 and 30

46 Evidence-Based Complementary and Alternative Medicine

respectively and the same doses reduced the blood pressurein hypertensive rats by 258 389 and 286 respectivelyOnly the highest dose (30mgkg) affected the force of thecontraction of the heart Atropine and propranolol were usedto interfere with the hypotensive action of the NBP Atropinereduced the initial phase of the hypotensive response in NBPand completely abolished the second phase of hypotensiveresponse in NBP In contrast propranolol increased thefirst hypotensive response but partially abolished the secondhypotensive response provoked by the NBP [110]This mech-anistic study suggested that B pilosa invokes the biphasichypotensive responses via targeting cardiac pump efficiencyduring the first phase and vasodilation at the second phase[110]

A further study was performed to investigate the relaxingeffect of a neutral extract of B pilosa (NBP) on rat aorta con-tractedwithKCl (60mM) andnorepinephrine (01mM) [94]Cumulative addition of NBP relaxed the rat aorta previouslycontracted by KCl in a dose-dependent manner The EC

50

value of theNBP for vasorelaxationwas 032mgmLThedataalso showed that the NBP reduced the contraction of aortapreviously contracted by KCl irrespective of the presence ofaortic endothelium [94]

Pretreatment with glibenclamide an ATP-dependent K+channel blocker did not considerably affect the relaxant effectof the NBP on KCl-induced contraction suggesting thatthe vasodilatory effect of B pilosa was not related to theopening of this ATP-dependent K+ channel [94] On theother hand in the presence of indomethacin or pyrilaminemaleate the relaxant response induced by the plant extractwas significantly inhibited at the lower concentrations Theplant extract was able to reduce the aorta resting tone inhibitthe KCl-induced contractions by 90 at 15mgmL and theCaCl2-induced contractions by 95 at 075mgmL These

results demonstrate that B pilosa can act as a vasodilatorprobably via acting as a calcium antagonist [94]

However no specific compound for the above activityhas been identified from B pilosa to date A bioactivity-guided identification approach may be adopted to identifythe active compounds in B pilosa that possess hypotensiveand vasodilatory effects and understand their mechanism ofaction

310 Wound Healing Activity B pilosa has been traditionallyused to treat tissue injury inCameroon Brazil andVenezuela[26] Hassan and colleagues investigated the wound healingpotential of B pilosa in Wistar rats [128] Mirroring thepositive control neomycin sulfate the ethanol extract of Bpilosa had faster wound closure than control rats 3 6 and 9days after topical application Histological examination alsorevealed better collagenation angiogenesis and organizationof wound tissue seven days after application Epithelializationand total healing time in B pilosa-treated rats were compara-ble to those of neomycin sulfate Together these data suggestthat B pilosa may be a viable alternative to neomycin lotionfor the treatment of wounds

In addition to studying the wound healing effect of Bpilosa on external ulcers Tan and colleagues also examined

the effect of methanol cyclohexane and methyl chlorideextracts of B pilosa on gastric ulcers in Wistar rats fedwith 1mL HClethanol gastric necrotizing solution (150mMHCl in 60 ethanol) and macroscopically visible lesionswere scored [26] Among the three extracts methylene chlo-ride extracts exhibited the highest activity showing 464inhibition of lesion formation at a dose of 500mgkg BWand complete inhibition at 750mgkg [26] The efficacy ofthe ethylene chloride extract was followed by that of themethanol extracts which had inhibition ranging from 304to 822 at concentrations of 500mgkg and 1000mgkgBW respectively [26] The cyclohexane extracts showed thelowest activity against gastric ulcers in rats with 13340 and 797 inhibition at 500 750 and 1000mgkg BWrespectively [26] To better understand the mode of actionof the methylene chloride extract of B pilosa rats werepretreated with indomethacin a COX-2 inhibitor involved inprostaglandin synthesis Pretreatment significantly reducedthe protection against HClethanol-induced ulcers to 313inhibition at 750mgkg BW suggesting a link between theantiulcerative activity of B pilosa and prostaglandin syn-thesis Unexpectedly the methylene chloride extract of Bpilosa showed little gastric mucosal protection against gastriclesions induced by 95 ethanol (1mL) [26] Absolute alcoholis known to cause mucosalsubmucosal tissue destruction viacellular necrosis and the release of tissue-derived mediators(histamine and leukotrieneC4)Thus these data imply thatBpilosa did not prevent the generation or the necrotic action ofthese mediators on the gastric microvasculature In additionpylorus ligation can increase gastric acid secretion withoutan alteration of mucosal histamine content The methylenechloride extract of B pilosa did not possess antisecretoryactivity On the contrary it was observed that increases in thedose of the extract led to elevated gastric juice acidity [26]Results with both absolute ethanol and pylorus ligation ratmodels suggested the possibility that the ineffectiveness of Bpilosa against gastric ulcers was due to lack of antihistaminicactivity in the plant In summary overall the data suggest thatB pilosa protects against HCLethanol-mediated ulcers viainhibition of prostaglandin biosynthesis

Previous phytochemical studies showed that a groupof flavonoids acyclichalcones are present in B pilosa[129 130] and the chalcones were proposed to have anti-ulcerative activity [131] Moreover nine hydroxychalcones 10were reported to possess gastric cytoprotective effects with24-dihydroxychalcone being the most active [132] Sincemethylene chloride extracts appear to be the most active Bpilosa extracts next the specific anti-ulcerative phytochemi-cals in the methylene chloride extracts of B pilosa and theirmodes of action needs to be probed

Despite the claims listed inTable 3 relatively few scientificstudies have been conducted in vitro and in vivo to address thetraditional ethnomedical uses of B pilosa Information aboutthe use of B pilosa as a botanical therapy recorded so far isfar from complete Studies conducted thus far only serve asa starting point for further investigation of B pilosa and theultimate efficacious use of the herb in clinical applications

Evidence-Based Complementary and Alternative Medicine 47

4 Toxicology

Despite its use as an ingredient in food for human con-sumption studies on systemic toxicity (eg acute subacutechronic and subchronic toxicities) of B pilosa in humansand animals are still inadequate and insufficient So faracute andor subchronic toxicities have been evaluated inrats and mice Oral acute and 28-day toxicities of waterextract of B pilosa leaves were evaluated in Wistar rats[133] An oral dose of water extract of B pilosa leaves at10 gkg BW showed no obvious mortality or changes inthe appearance in rats [134] The same extract at 08 gkgBWday once a day showed no obvious sub-chronic toxicityin rats over 28 days as measured by survival rate bodyweight and gross examination of organs [134] These dataare consistent with our data indicating that oral deliveryof the water extract of the B pilosa whole plant at 1 gkgBWday once a day is safe in rats over 28 days (unpublisheddata) Taken together these studies suggest that ingestion ofB pilosa aqueous extract at up to at 1 gkg BWday oncea day is highly safe in rats In addition the acute toxicityof aqueous and ethanol extracts of B pilosa in mice havebeen reported [134] Five- to six-week-old mice with weightsbetween 28 and 35 g received a peritoneal injection of bothextracts at the different doses The LD

50 the dose that causes

50 lethality of the aqueous and ethanol extracts in micewas 1230 gkg BW and 615 gkg BW respectively [134] Acomplete toxicological study has not been completed forhumans Furthermore the drug interactions of B pilosa withother drugs are unknown Further safety verification andclinical trials should be performed before B pilosa can beconsidered for medicinal use

5 Conclusions

B pilosa is an erect perennial plant with green leaves whiteor yellow flowers and tiny black seeds As it is distributedworldwide and is widely used as a folk remedy B pilosacan be thought of as an extraordinary source of food andmedicine However a comprehensive up-to-date review ofresearch on B pilosa has hitherto been unavailable In thisarticle scientific studies on B pilosa have been summarizedand critically discussed from the perspectives of botanyethnomedicine phytochemistry pharmacology and toxicol-ogy B pilosa is claimed to treat more than 40 disordersand 201 compounds have been identified from this plantThe medicinal utility of B pilosa and its modes of actionin relation to its known phytochemicals were discussedherein Polyynes flavonoids phenylpropanoids fatty acidsand phenolics are the primary bioactive compounds of Bpilosa and they have been reported to be effective in thetreatment of tumors inflammationimmune modulationdiabetes viruses microbes protozoans gastrointestinal dis-eases hypertension and cardiovascular diseases Cautionshould be exercised in the therapeutic use of B pilosa forhypoglycemia hypotension bleeding and allergy

Acknowledgments

The authors thank their laboratory members for construc-tive suggestions and the authors whose publications theycited for their contributions The authors also thank MsMiranda Loney of Academia Sinica for English editing of thispaper This work was supported by Grants 99-CDA-L11 and101S0010056 from Academia Sinica Taiwan Arlene P Bar-tolome is a recipient of MECO-TECO Sandwich ScholarshipProgram between Taiwan and the Philippines

References

[1] T Shen G H Li X N Wang and H X Lou ldquoThe genusCommiphora a review of its traditional uses phytochemistryand pharmacologyrdquo Journal of Ethnopharmacology vol 142 no2 pp 319ndash330

[2] C Long P Sauleau B David et al ldquoBioactive flavonoids ofTanacetum parthenium revisitedrdquo Phytochemistry vol 64 no2 pp 567ndash569 2003

[3] P O Karis and O Ryding Asteraceae Cladistics and Classifica-tion Bremer K Eds pp 559ndash569 Timber press Portland OreUSA 1994

[4] O N Pozharitskaya A N Shikov M N Makarova et alldquoAnti-inflammatory activity of a HPLC-fingerprinted aqueousinfusion of aerial part of Bidens tripartita LrdquoPhytomedicine vol17 no 6 pp 463ndash468 2010

[5] F Q Oliveira V Andrade-Neto A U Krettli and MG L Brandao ldquoNew evidences of antimalarial activity ofBidens pilosa roots extract correlated with polyacetylene andflavonoidsrdquo Journal of Ethnopharmacology vol 93 no 1 pp 39ndash42 2004

[6] Agriculture USDA ldquoPlants databaserdquo 2013in Natural Resources Conservation Servicehttpwwwnrcsusdagovwpsportalnrcssitenationalhome

[7] Y Zhou and X Z Yan ldquoExperimental study of qi deficiencysyndrome and Codonopsis pillosulae and Astragalus injectionon the immune response in micerdquo Zhong Xi Yi Jie He Za Zhivol 9 no 5 pp 286ndash262 1989

[8] S C Chien P H Young Y J Hsu et al ldquoAnti-diabeticproperties of three common Bidens pilosa variants in TaiwanrdquoPhytochemistry vol 70 no 10 pp 1246ndash1254 2009

[9] M J Alcaraz and M J Jimenez ldquoFlavonoids as anti-inflammatory agentsrdquo Fitoterapia vol 59 no 1 pp 25ndash38 1988

[10] FAO Agriculture Food and Nutrition for AfricamdashA ResourceBook for Teachers of Agriculture Publishing ManagementGroup FAO Information Division Rome Italy 1997

[11] M B Rokaya Z Munzbergova B Timsina and K R BhattaraildquoRheum australe D Don a review of its botany ethnobotanyphytochemistry and pharmacologyrdquo Journal of Ethnopharma-cology vol 141 no 3 pp 761ndash774 2012

[12] P H Young Y J Hsu and W C Yang ldquoBidens pilosa L andits medicinal userdquo in Recent Progress in Medicinal Plants DrugPlant II A S AwaadVK Singh and JNGovil Eds StandiumPress Houston Tex USA 2010

[13] C Ge ldquoCytologic study of Bidens bipinnata LrdquoZhongguo ZhongYao Za Zhi vol 15 no 2 pp 72ndash125 1990

[14] L C Chiang J S Chang C C Chen L T Ng and C CLin ldquoAnti-herpes simplex virus activity of Bidens pilosa and

48 Evidence-Based Complementary and Alternative Medicine

Houttuynia cordatardquoAmerican Journal of ChineseMedicine vol31 no 3 pp 355ndash362 2003

[15] N P Rybalchenko V A Prykhodko S S Nagorna et alldquoIn vitro antifungal activity of phenylheptatriyne from Bidenscernua L against yeastsrdquo Fitoterapia vol 81 no 5 pp 336ndash3382010

[16] K Redl W Breu B Davis and R Bauer ldquoAnti-inflammatoryactive polyacetylenes from Bidens campylothecardquo Planta Med-ica vol 60 no 1 pp 58ndash62 1994

[17] M Ayyanar and S Ignacimuthu ldquoTraditional knowledge ofKani tribals inKouthalai of Tirunelveli hills TamilNadu IndiardquoJournal of Ethnopharmacology vol 102 no 2 pp 246ndash255 2005

[18] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant intraditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[19] AKAbbas KMMurphy andA Sher ldquoFunctional diversity ofhelper T lymphocytesrdquo Nature vol 383 no 6603 pp 787ndash7931996

[20] J H Cano and G Volpato ldquoHerbal mixtures in the traditionalmedicine of Eastern Cubardquo Journal of Ethnopharmacology vol90 no 2-3 pp 293ndash316 2004

[21] J Namukobe J M Kasenene B T Kiremire et al ldquoTraditionalplants used for medicinal purposes by local communitiesaround the Northern sector of Kibale National Park UgandardquoJournal of Ethnopharmacology vol 136 no 1 pp 236ndash245 2011

[22] C Lans ldquoComparison of plants used for skin and stomachproblems in Trinidad and Tobago with Asian ethnomedicinerdquoJournal of Ethnobiology andEthnomedicine vol 3 article 3 2007

[23] S Dharmananda ldquoA Popular Remedy Ecapes Notice ofWesternPractitionersrdquo 2013 httpwwwitmonlineorgartsbidenshtm

[24] Y J Hsu T H Lee C L T Chang Y T Huang andW C YangldquoAnti-hyperglycemic effects and mechanism of Bidens pilosawater extractrdquo Journal of Ethnopharmacology vol 122 no 2 pp379ndash383 2009

[25] E Noumi F Houngue and D Lontsi ldquoTraditional medicinesin primary health care plants used for the treatment ofhypertension in Bafia Cameroonrdquo Fitoterapia vol 70 no 2 pp134ndash139 1999

[26] P V Tan T Dimo and E Dongo ldquoEffects of methanolcyclohexane and methylene chlo ride extracts of Bidens pilosaon various gastric ulcer models in ratsrdquo Journal of Ethnophar-macology vol 73 no 3 pp 415ndash421 2000

[27] R L C Pereira T Ibrahim L Lucchetti A J R Da Silva and VL G De Moraes ldquoImmunosuppressive and anti-inflammatoryeffects of methanolic extract and the polyacetylene isolatedfrom Bidens pilosa Lrdquo Immunopharmacology vol 43 no 1 pp31ndash37 1999

[28] T Dimo J Azay P V Tan et al ldquoEffects of the aqueous andmethylene chloride extracts of Bidens pilosa leaf on fructose-hypertensive ratsrdquo Journal of Ethnopharmacology vol 76 no 3pp 215ndash221 2001

[29] CWiartMedicinal Plants of Southeast Asia Pelanduk SelabgorDarul Ehsan Malaysia 2000

[30] F L Silva D C H Fischer J F Tavares M S Silva P FDe Athayde-Filho and J M Barbosa-Filho ldquoCompilation ofsecondary metabolites from Bidens pilosa LrdquoMolecules vol 16no 2 pp 1070ndash1102 2011

[31] A H Chen S R Lin and C H Hong ldquoPhytochemical studyon Bidens pilosa L varminorrdquo Huaxue pp 38ndash42 1975

[32] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[33] M C Zulueta M Tada and C Y Ragasa ldquoA diterpene fromBidens pilosardquo Phytochemistry vol 38 no 6 pp 1449ndash14501995

[34] J Wang H Yang Z W Lin and H D Sun ldquoFlavonoids fromBidens pilosa var radiatardquo Phytochemistry vol 46 no 7 pp1275ndash1278 1997

[35] A Zhao Q Zhao L Peng et al ldquoA new chalcone glycoside fromBidens pilosardquo Acta Botanica Yunnanica vol 26 no 1 pp 121ndash126

[36] P Gao Y Wu S Cui and Y Zou ldquoSynthesis and absoluteconfiguration of the 7-phenylhepta-4 6-diyne-1 2-diol isolatedfrom Bidens pilosardquo Synthesis no 13 pp 2131ndash2135 2011

[37] S Wang B Yang D Zhu D He and L Wang ldquoActivecomponents of Bidens pilosa Lrdquo Zhongcaoyao vol 36 pp 20ndash21

[38] F Bohlmann T Burkhardt and C Zdero Naturally OccuringAcetylenes Academic Press New York NY USA 1973

[39] T M Sarg A M Ateya N M Farrag and F A AbbasldquoConstituents and biological activity of Bidens pilosa L grownin EgyptrdquoActa Pharmaceutica Hungarica vol 61 no 6 pp 317ndash323 1991

[40] H A L Valdes and H P Rego ldquoBidens pilosa Linnerdquo RevistaCubana de Plantas Medicinales vol 6 pp 28ndash33

[41] F Bohlmann H Bornowski and K M Kleine ldquoNew polyynesfrom the tribe Heliantheaerdquo Chemische Berichte vol 97 pp2135ndash2138

[42] RWang Q XWu and Y P Shi ldquoPolyacetylenes and flavonoidsfrom the aerial parts ofBidens pilosardquo PlantaMedica vol 76 no9 pp 893ndash896 2010

[43] L W Wu Y M Chiang H C Chuang et al ldquoA novelpolyacetylene significantly inhibits angiogenesis and promotesapoptosis in human endothelial cells through activation of theCDK inhibitors and caspase-7rdquo Planta Medica vol 73 no 7 pp655ndash661 2007

[44] L W Wu Y M Chiang H C Chuang et al ldquoPolyacetylenesfunction as anti-angiogenic agentsrdquo Pharmaceutical Researchvol 21 no 11 pp 2112ndash2119 2004

[45] H L Yang S C Chen N W Chang et al ldquoProtection fromoxidative damage using Bidens pilosa extracts in normal humanerythrocytesrdquo Food and Chemical Toxicology vol 44 no 9 pp1513ndash1521 2006

[46] H Q Wang S J Lu H Li and Z H Yao ldquoEDTA-enhancedphytoremediation of lead contaminated soil by Bidens maxi-mowiczianardquo Journal of Environmental Sciences vol 19 no 12pp 1496ndash1499 2007

[47] S TobinagaMK SharmaWG L Aalbersberg et al ldquoIsolationand identification of a potent antimalarial and antibacterialpolyacetylene from Bidens pilosardquo Planta Medica vol 75 no 6pp 624ndash628 2009

[48] C L T Chang H K Kuo S L Chang et al ldquoThe distincteffects of a butanol fraction of Bidens pilosa plant extract onthe development of Th1-mediated diabetes and Th2-mediatedairway inflammation in micerdquo Journal of Biomedical Sciencevol 12 no 1 pp 79ndash89 2005

[49] S L Chang C L T Chang Y M Chiang et al ldquoPolyacetyleniccompounds and butanol fraction from Bidens pilosa can mod-ulate the differentiation of helper T cells and prevent autoim-mune diabetes in non-obese diabetic micerdquo Planta Medica vol70 no 11 pp 1045ndash1051 2004

Evidence-Based Complementary and Alternative Medicine 49

[50] Y M Chiang C L T Chang S L Chang W C Yang and LF Shyur ldquoCytopiloyne a novel polyacetylenic glucoside fromBidens pilosa functions as a T helper cell modulatorrdquo Journal ofEthnopharmacology vol 110 no 3 pp 532ndash538 2007

[51] R PUbillas CDMendez S D Jolad et al ldquoAntihyperglycemicacetylenic glucosides from Bidens pilosardquo Planta Medica vol66 no 1 pp 82ndash83 2000

[52] L Alvarez S Marquina M L Villarreal D Alonso E Arandaand G Delgado ldquoBioactive polyacetylenes from Bidens pilosardquoPlanta Medica vol 62 no 4 pp 355ndash357 1996

[53] C L T Chang S L Chang Y M Lee et al ldquoCytopiloyne apolyacetylenic glucoside prevents type 1 diabetes in nonobesediabetic micerdquo Journal of Immunology vol 178 no 11 pp 6984ndash6993 2007

[54] G Kusano A Kusano and Y Seyama Novel Hypoglycemicand Anti-Inflammatory Polyacetylenic Compounds Their Com-positions Bidens Plant Extract Fractions and CompositionsContaining the Plant or Fraction JPO Tokyo Japan 2004

[55] M G L Brandao A U Krettli L S R Soares C G CNery and H C Marinuzzi ldquoAntimalarial activity of extractsand fractions from Bidens pilosa and other Bidens species(Asteraceae) correlated with the presence of acetylene andflavonoid compoundsrdquo Journal of Ethnopharmacology vol 57no 2 pp 131ndash138 1997

[56] A U Krettli V F Andrade-Neto M D G L Brandao andW M S Ferrari ldquoThe search for new anti-malarial drugs fromplants used to treat fever and malaria or plants ramdomlyselected a reviewrdquoMemorias do Instituto Oswaldo Cruz vol 96no 8 pp 1033ndash1042 2001

[57] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[58] C K Wat R K Biswas E A Graham L Bohm G H NTowers and E R Waygood ldquoUltraviolet-mediated cytotoxicactivity of phenylheptatriyne from Bidens pilosa Lrdquo Journal ofNatural Products vol 42 no 1 pp 103ndash111 1979

[59] Y Sashida K Ogawa M Kitada H Karikome Y Mimakiand H Shimomura ldquoNew aurone glucosides and new phenyl-propanoid glucosides from Bidens pilosardquo Chemical and Phar-maceutical Bulletin vol 39 no 3 pp 709ndash711 1991

[60] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens pilosa L (TFB) on animal liver injury andliver fibrosisrdquo Journal of Ethnopharmacology vol 116 no 3 pp539ndash546 2008

[61] B Hoffmann and J Holzl ldquoAcylated compounds from Bidenspilosardquo Planta Medica vol 55 pp 108ndash109

[62] B Hoffmann and J Holzl ldquoA methylated chalcone glucosidefrom Bidens pilosardquo Phytochemistry vol 27 no 11 pp 3700ndash3701 1988

[63] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[64] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[65] B Hoffmann and J Holzl ldquoWeitere acylierte chalkone ausBidens pilosardquo Planta Medica vol 54 pp 450ndash451 1988

[66] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant in11traditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[67] V V Pham V V T K P V L Hoang and V K Phan ldquoFlavonoidcompounds from the plant Bidens pilosa L (Asteraceae)rdquo TapChi Duoc Hoc vol 50 pp 48ndash53

[68] S D Sarker B Bartholomew R J Nash and N Robinson ldquo5-O-methylhoslundin an unusual flavonoid from Bidens pilosa(Asteraceae)rdquo Biochemical Systematics and Ecology vol 28 no6 pp 591ndash593 2000

[69] S L Chang Y M Chiang C L T Chang et al ldquoFlavonoidscentaurein and centaureidin from Bidens pilosa stimulate IFN-120574 expressionrdquo Journal of Ethnopharmacology vol 112 no 2 pp232ndash236 2007

[70] Y M Chiang D Y Chuang S Y Wang Y H Kuo P WTsai and L F Shyur ldquoMetabolite profiling and chemopreventivebioactivity of plant extracts from Bidens pilosardquo Journal ofEthnopharmacology vol 95 no 2-3 pp 409ndash419 2004

[71] A Kusano Y Seyama E Usami et al ldquoStudies on the antioxi-dant active constituents of the dried powder from Bidens pilosaL var radiata SchrdquoNaturalMedicines vol 57 no 3 pp 100ndash1042003

[72] I Doleckova L Rarova J Gruz et al ldquoAnti-proliferative andanti-angiogenic effects of flavone eupatorin an active con-stituent of chloroform extract ofOrthosiphon stamineus leavesrdquoFitoterapia vol 83 no 6 pp 1000ndash1007

[73] K Bairwa R Kumar R J Sharma and R K Roy ldquoAn updatedreview on Bidens pilosa Lrdquo Der Pharma Chemica vol 2 no 3pp 325ndash337

[74] Q Xia Y Liu and Y Li ldquoDetermination of hyperoside indifferent parts and different species of Herba Bidens by RP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol 50pp 48ndash53

[75] M Horiuchi and Y Seyama ldquoImprovement of the antiinflam-matory and antiallergic activity of Bidens pilosa L var radiataSCHERFF treated with enzyme (Cellulosine)rdquo Journal of HealthScience vol 54 no 3 pp 294ndash301 2008

[76] M G L Brandao C G C Nery M A S Mamao and AU Krettli ldquoTwo methoxylated flavone glycosides from Bidenspilosardquo Phytochemistry vol 48 no 2 pp 397ndash400 1998

[77] P Kumari K Misra B S Sisodia et al ldquoA promising anticancerand antimalarial component from the leaves of Bidens pilosardquoPlanta Medica vol 75 no 1 pp 59ndash61 2009

[78] G Seelinger I Merfort U Wolfle and C M Schempp ldquoAnti-carcinogenic effects of the flavonoid luteolinrdquoMolecules vol 13no 10 pp 2628ndash2651 2008

[79] G Seelinger I Merfort and C M Schempp ldquoAnti-oxidantanti-inflammatory and anti-allergic activities of luteolinrdquoPlanta Medica vol 74 no 14 pp 1667ndash1677 2008

[80] C C Yit and N P Das ldquoCytotoxic effect of butein on humancolon adenocarcinoma cell proliferationrdquo Cancer Letters vol82 no 1 pp 65ndash72 1994

[81] J A Beutler E Hamel A J Vlietinck et al ldquoStructure-activityrequirements for flavone cytotoxicity and binding to tubulinrdquoJournal of Medicinal Chemistry vol 41 no 13 pp 2333ndash23381998

[82] M R Kviecinski K B Felipe T Schoenfelder et al ldquoStudy ofthe antitumor potential of Bidens pilosa (Asteraceae) used inBrazilian folkmedicinerdquo Journal of Ethnopharmacology vol 117no 1 pp 69ndash75 2008

[83] P Sundararajan A Dey A Smith A G Doss M Rajappanand S Natarajan ldquoStudies of anticancer and antipyretic activityof Bidens pilosawhole plantrdquo African Health Sciences vol 6 no1 pp 27ndash30 2006

[84] YMChiang C P Lo Y P Chen et al ldquoEthyl caffeate suppressesNF-120581B activation and its downstream inflammatory mediatorsiNOS COX-2 and PGE2 in vitro or in mouse skinrdquo BritishJournal of Pharmacology vol 146 no 3 pp 352ndash363 2005

50 Evidence-Based Complementary and Alternative Medicine

[85] J S Kim H J Lee M H Lee J Kim C Jin and J HRyu ldquoLuteolin inhibits LPS-stimulated inducible nitric oxidesynthase expression in BV-2 microglial cellsrdquo Planta Medicavol 72 no 1 pp 65ndash68 2006

[86] P A Ruiz and D Haller ldquoFunctional diversity of flavonoidsin the inhibition of the proinflammatory NF-120581B IRF and Aktsignaling pathways in murine intestinal epithelial cellsrdquo Journalof Nutrition vol 136 no 3 pp 664ndash671 2006

[87] A Xagorari A Papapetropoulos A Mauromatis MEconomou T Fotsis and C Roussos ldquoLuteolin inhibitsan endotoxin-stimulated phosphorylation cascade andproinflammatory cytokine production in macrophagesrdquoJournal of Pharmacology and Experimental Therapeutics vol296 no 1 pp 181ndash187 2001

[88] N Yoshida T Kanekura Y Higashi and T Kanzaki ldquoBidenspilosa suppresses interleukin-1120573-induced cyclooxygenase-2expression through the inhibition of mitogen activated proteinkinases phosphorylation in normal human dermal fibroblastsrdquoJournal of Dermatology vol 33 no 10 pp 676ndash683 2006

[89] M Muchuweti C Mupure A Ndhlala T Murenje and M AN Benhura ldquoScreening of antioxidant and radical scavengingactivity of Vigna ungiculata Bidens pilosa and Cleome gynan-drardquo American Journal of Food Technology vol 2 no 3 pp 161ndash168 2007

[90] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens bipinnata L against carbon tetrachloride-induced liver fibrosis in ratsrdquo Journal of Pharmacy and Pharma-cology vol 60 no 10 pp 1393ndash1402 2008

[91] S L Chang H H Yeh Y S Lin Y M Chiang T K Wuand W C Yang ldquoThe effect of centaurein on interferon-120574 expression and Listeria infection in micerdquo Toxicology andApplied Pharmacology vol 219 no 1 pp 54ndash61 2007

[92] F Deba T D Xuan M Yasuda and S Tawata ldquoChemical com-position and antioxidant antibacterial and antifungal activitiesof the essential oils from Bidens pilosa Linn var Radiatardquo FoodControl vol 19 no 4 pp 346ndash352 2008

[93] A O T Ashafa and A J Afolayan ldquoScreening the root extractsfrom Biden pilosa L var radiata (Asteraceae) for antimicrobialpotentialsrdquo Journal of Medicinal Plant Research vol 3 no 8 pp568ndash572 2009

[94] T B Nguelefack T Dimo E P Nguelefack Mbuyo P V TanS V Rakotonirina and A Kamanyi ldquoRelaxant effects of theneutral extract of the leaves of Bidens pilosa linn on isolated ratvascular smooth musclerdquo Phytotherapy Research vol 19 no 3pp 207ndash210 2005

[95] M T Grombone-Guaratini K L Silva-Brandao V N SolferiniJ Semir and J R Trigo ldquoSesquiterpene and polyacetyleneprofile of the Bidens pilosa complex (Asteraceae Heliantheae)from Southeast of Brazilrdquo Biochemical Systematics and Ecologyvol 33 no 5 pp 479ndash486 2005

[96] L L Lin C Y Wu H C Hsiu M T Wang and H ChuangldquoStudies on diabetes mellitus I The hypoglycemic activity ofphytosterin on alloxan diabetic ratsrdquo Taiwan Yi Xue Hui Za Zhivol 66 no 2 pp 58ndash65 1967

[97] M A N Benhura and I C Chitsiku ldquoThe extractable 120573-carotene content of Guku (Bidens pilosa) leaves after cookingdrying and storagerdquo International Journal of Food Science andTechnology vol 32 no 6 pp 495ndash500 1997

[98] J S Chang L C Chiang C C Chen L T Liu K C Wangand C C Lin ldquoAtileukemic activity of Bidens pilosa l varminor (blume) sherff andHouttuynia cordata thunbrdquo AmericanJournal of Chinese Medicine vol 29 no 2 pp 303ndash312 2001

[99] F Deba T D Xuan M Yasuda and S Tawata ldquoHerbicidal andfungicidal activities and identification of potential phytotoxinsfrom Bidens pilosa L var radiata Scherff research paperrdquoWeedBiology and Management vol 7 no 2 pp 77ndash83 2007

[100] J K Kumar and A K Sinha ldquoA new disubstituted acetylacetonefrom the leaves ofBidens pilosa LinnrdquoNatural Product Researchvol 17 no 1 pp 71ndash74 2003

[101] K Ogawa and Y Sashida ldquoCaffeoyl derivatives of a sugarlactone and its hydroxy acid from the leaves of Bidens pilosardquoPhytochemistry vol 31 no 10 pp 3657ndash3658 1992

[102] Q Xia Y Liu and Y Li ldquoDetermination of gallic acid fromdifferent species and different medical parts of Herba Bidens byRP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol24 no 03 pp 308ndash310

[103] W J Lee L F Wu W K Chen C J Wang and T HTseng ldquoInhibitory effect of luteolin on hepatocyte growth fac-torscatter factor-induced HepG2 cell invasion involving bothMAPKERKs and PI3K-Akt pathwaysrdquo Chemico-BiologicalInteractions vol 160 no 2 pp 123ndash133 2006

[104] J A Beutler J H Cardellina C M Lin E Hamel G MCragg andM R Boyd ldquoCentaureidin a cytotoxic flavone fromPolymnia fruticosa inhibits tubulin polymerizationrdquo BioorganicandMedicinal Chemistry Letters vol 3 no 4 pp 581ndash584 1993

[105] A Verma A Su A M Golin B OrsquoMarrah and J K AmorosaldquoThe lateral view a screening method for knee traumardquo Aca-demic Radiology vol 8 no 5 pp 392ndash397 2001

[106] J Corren M Lemay Y Lin L Rozga and R K RandolphldquoClinical and biochemical effects of a combination botanicalproduct (ClearGuard) for allergy a pilot randomized double-blind placebo-controlled trialrdquo Nutrition Journal vol 7 no 1article 20 2008

[107] M S Gachet J S Lecaro M Kaiser et al ldquoAssessment of anti-protozoal activity of plants traditionally used in Ecuador in thetreatment of leishmaniasisrdquo Journal of Ethnopharmacology vol128 no 1 pp 184ndash197 2010

[108] W C Yang M Ghiotto B Barbarat and D Olive ldquoThe role ofTec protein-tyrosine kinase in T cell signalingrdquo The Journal ofBiological Chemistry vol 274 no 2 pp 607ndash617 1999

[109] S Tewtrakul H Miyashiro N Nakamura et al ldquoHIV-1 inte-grase inhibitory substances from Coleus parvifoliusrdquo Phytother-apy Research vol 17 no 3 pp 232ndash239 2003

[110] T Dimo T B Nguelefack P V Tan et al ldquoPossible mechanismsof action of the neutral extract from Bidens pilosa L leaves onthe cardiovascular system of anaesthetized ratsrdquo PhytotherapyResearch vol 17 no 10 pp 1135ndash1139 2003

[111] T S C Li Chinese and Related North American Herbs CRCPress New York NY USA 2002

[112] H Wu H Chen X Hua Z Shi L Zhang and J ChenldquoClinical therapeutic effect of drug-separated moxibustion onchronic diarrhea and its immunologic mechanismsrdquo Journal ofTraditional Chinese Medicine vol 17 no 4 pp 253ndash258 1997

[113] S W Wright R R Harris J S Kerr et al ldquoSynthesis chemicaland biological properties of vinylogous hydroxamic acids dualinhibitors of 5-lipoxygenase and IL-1 biosynthesisrdquo Journal ofMedicinal Chemistry vol 35 no 22 pp 4061ndash4068 1992

[114] W R Almiron and M E Brewer ldquoClassification of immaturestage habitats of Culicidae (Diptera) collected in CordobaArgentinardquo Memorias do Instituto Oswaldo Cruz vol 91 no 1pp 1ndash9 1996

[115] N L Wang J Wang X S Yao and S Kitanaka ldquoTwo newmonoterpene glycosides and a new (+)-jasmololone glucoside

Evidence-Based Complementary and Alternative Medicine 51

from Bidens parvifloraWilldrdquo Journal of Asian Natural ProductsResearch vol 9 no 5 pp 473ndash479 2007

[116] P Champagnat ldquoRole of the terminal bud in the actionexercised by the cotyledon of Bidens pilosus L var radiatus onits axillary budrdquo Comptes Rendus des Seances et Memoires de laSociete de Biologie vol 145 no 17-18 pp 1374ndash1376 1951

[117] S F Nielsen S B Christensen G Cruciani A Kharazmiand T Liljefors ldquoAntileishmaniai chalcones statistical designsynthesis and three-dimensional quantitative structure-activityrelationship analysisrdquo Journal of Medicinal Chemistry vol 41no 24 pp 4819ndash4832 1998

[118] Y C Hwang J J H Chu P L Yang W Chen and M V YatesldquoRapid identification of inhibitors that interfere with poliovirusreplication using a cell-based assayrdquo Antiviral Research vol 77no 3 pp 232ndash236 2008

[119] V F Andrade-Neto M G L Brandao F Q Oliveira et alldquoAntimalarial activity of Bidens pilosa L (Asteraceae) ethanolextracts fromwild plants collected in various localities or plantscultivated in humus soilrdquo Phytotherapy Research vol 18 no 8pp 634ndash639 2004

[120] R J Marles and N R Farnsworth ldquoAnti-diabetic plants andtheir active constituentsrdquo Phytomedicine vol 2 no 2 pp 137ndash189

[121] M Habeck ldquoDiabetes treatments get sweet help from naturerdquoNature Medicine vol 9 no 10 p 1228 2003

[122] H W Lin G Y Han and S X Liao ldquoStudies on the activeconstituents of the Chinese traditional medicine Polygonatumodoratum (Mill) DrucerdquoActa Pharmaceutica Sinica vol 29 no3 pp 215ndash222 1994

[123] C L T Chang Y C Chen H M Chen N S Yang andW C Yang ldquoNatural cures for type 1 diabetes a reviewof phytochemicals biological actions and clinical potentialrdquoCurrent Medicinal Chemistry vol 20 no 7 pp 899ndash907 2013

[124] L Dey A S Attele and C S Yuan ldquoAlternative therapies fortype 2 diabetesrdquo Alternative Medicine Review vol 7 no 1 pp45ndash58 2002

[125] C L T ChangH Y Liu T F Kuo et al ldquoAnti-diabetic effect andmode of action of cytopiloynerdquo Evidence-Based Complementaryand Alternative Medicine vol 2013 Article ID 685642 13 pages2013

[126] M R Khan M Kihara and A D Omoloso ldquoAnti-microbialactivity of Bidens pilosa Bischofia javanica Elmerillia papuanaand Sigesbekia orientalisrdquo Fitoterapia vol 72 no 6 pp 662ndash6652001

[127] T Dimo T B Nguelefack P Kamtchouing E Dongo ARakotonirina and S V Rakotonirina ldquoHypotensive effects ofmethanol extract from Bidens pilosa Linn on hypertensive ratsrdquoComptes Rendus de lrsquoAcademie des Sciences Serie III vol 322no 4 pp 323ndash329 1999

[128] A K Hassan O Deogratius J F Nyafuono O Francis and OP Engeu ldquoWound healing potential of the ethanolic extracts ofBidens pilosa and Ocimum suaverdquo African Journal of Pharmacyand Pharmacology vol 5 no 2 pp 132ndash136 2011

[129] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[130] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[131] C Kandaswami and EMiddleton ldquoFree radical scavenging andantioxidant activity of plant flavonoidsrdquo Advances in Experi-mental Medicine and Biology vol 366 pp 351ndash376 1994

[132] K Yamamoto H Kakegawa H Ueda et al ldquoGastric cytopro-tective anti-ulcerogenic actions of hydroxychalcones in ratsrdquoPlanta Medica vol 58 no 5 pp 389ndash393 1992

[133] J O C Ezeonwumelu A K Julius C N Muhoho et alldquoBiochemical and histological studies of aqueous extract ofBidens pilosa leaves from Ugandan Rift valley in ratsrdquo BritishJournal of Pharmacology and Toxicology vol 2 no 6 pp 302ndash309

[134] L Frida S Rakotonirina A Rakotonirina and J P SavineauldquoIn vivo and in vitro effects of Bidens pilosa L (Asteraceae)leaf aqueous and ethanol extracts on primed-oestrogenized ratuterine musclerdquo African Journal of Traditional Complementaryand Alternative Medicines vol 5 no 1 pp 79ndash91 2008

52 Evidence-Based Complementary and Alternative Medicine

Composition Comments

1 Please provide valid postal code to the address if

possible

2 We made the highlighted change in Table 4 for the sake

of correctness Please check

3 Please check the correctness of the highlighted part in

Table 5

4 We considered the highlighted part in Table 11 as

reference Please check similar cases throughout

5 Please check the format of Table 18

6 We reformatted Table 20 Please check

7 We made the highlighted change in Table 4 for the sake

of consistency Please check similar cases throughout the

paper

8 We made the highlighted change(s) for the sake

of consistency Please check simila highlighted cases

throughout

9 We made the highlighted change in Table 9 for the sake

of clarity Please check similar cases through out

10 We made the highlighted change(s) for the sake of

correctness Please check

11 Comment on ref [66] This reference is a repetition

of [18] Please check similar cases throughout

Author(s) Name(s)

Author 1

Author 2

Author 3

14 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

81

1-[2-(120573-D

-Glucopyrano

syloxy)-

4-hydroxyphenyl]-2-

hydroxy-3-(3-

hydroxyphenyl)-2-

prop

en-1-on

e

Chalcone120572

32101584041015840-

tetrahydroxy-

21015840-O

-120573-D

-glucop

yranosyl

HO

O

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[35]

821-(24-Dihydroxyph

enyl)-

3-(34-dihydroxyph

enyl)-

2-prop

en-1-on

eBu

tein

O

OH

OH

HO

OH

Aeria

l(Ch

ina)

[35]

833-(34-D

ihydroxyph

enyl)-

1-(234-trihydroxyph

enyl)-

2-prop

en-1-on

eOkanin

O

HO

HO

OH

OH

OH

Leaves

(China)

[60]

84

3-(34-D

ihydroxyph

enyl)-

1-[3-(120573-D

-glucop

yranosyloxy)-

24-dihydroxyphenyl]-2-

prop

en-1-on

e

Okanin

31015840-O

-120573-D

-glucosid

e

O

HO

OH

OH

OH

OO

HO

H

HO

H

H

HOH

H

OH

Leaves

(Germany)

flowers(Germany)

[61ndash63]

85

3-(34-D

ihydroxyph

enyl)-

1-[4-(120573-D

-glucop

yranosyloxy)-

23-dihydroxyphenyl]-

2-prop

en-1-on

e

Okanin

41015840-O

-120573-D

-glucosid

e

OOH

OH

OH

HOO

OHO

H

HO

H

H

HOH

H

OH

Flow

ers(Germany)

leaves

(Japan)

[5963]

Evidence-Based Complementary and Alternative Medicine 15

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

86Okanin41015840-O

-120573-D

-(610158401015840-

O-acetylglucosid

e)

O

O

OH

OH

OH

OHO

H

HO

H

H

HOH

H

HO

O

O

Flow

ers(Germany)

[63]

87

1-[4-[(46-di-O

-Acetyl-

120573-D

-glucopyrano

syl)-oxy]-

23-dihydroxyphenyl]-3-

(34-dihydroxyph

enyl)-2-

prop

en-1-on

e

Okanin

41015840-O

-120573-D

-(410158401015840610158401015840-diacetyl)-

glucop

yranoside

O

O

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Aeria

l(Ch

ina)

[34]

88Okanin41015840-O

-120573-D

-(210158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[64]

89Okanin41015840-O

-120573-D

-(310158401015840410158401015840610158401015840-tr

iacetyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

O

OAe

rial(Ch

ina)

[34]

16 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

90

1-[23-Dihydroxy-4-

[[6-O-[3-(4-hydroxyph

enyl)-

1-oxo-2-prop

enyl]-120573-D

-glucop

yranosyl]oxy]-ph

enyl]-

3-(34-dihydroxyph

enyl)-

2-prop

en-1on

e

Okanin

41015840-O

-120573-D

-(610158401015840-tr

ans-

p-coum

aroyl)-

glucoside

O

O

OH

OH

OH

OHOH

HO

H

H

HOH

H

HO

OO

OH

Leaves

(Germany)

[64]

91Okanin41015840-O

-120573-D

-(410158401015840-acetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HOH

H

HO

OO

O

Leaves

(Germany)

[65]

92

Okanin

41015840-O

-120573-D

-(210158401015840410158401015840-diacetyl-

610158401015840-tr

ans-p

-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OH

OO

H

HO

H

H

HO

H

HO

OO

O

O

Leaves

(Germany)

[65]

Evidence-Based Complementary and Alternative Medicine 17

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

93

Okanin

41015840-O

-120573-D

-(310158401015840410158401015840-diacetyl-610158401015840-

trans-p-cou

maroyl)-

glucoside

O

O

OH

OH

OH

OO

H OH

H

HOH

H

HO

OO

OH

O O

Leaves

(Germany)

[65]

94

Okanin

41015840-O

-[120573-D

-glucopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

OOH

OH

OH

HO

OH

HO

HHO

H

HOH

HO

OH

OH

HO

HHO

H

HOH

HO

Flow

er(G

ermany)

[63]

95Okanin

3101584041015840-di-O

-120573-D

-glucosid

e

OOH

OH

OH

O OO

H HO

HHOH

HOH

HHO

OHO

H

HO

H

H

HOH

H

OH

Flow

er(G

ermany)

[63]

18 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

96

1-[3-(120573-D

-Glucopyrano

syloxy)-

24-dihydroxyphenyl]-

3-(3-hydroxy)-

4-metho

xyph

enyl)-

2-prop

en-1-on

e

Okanin

4-methylether-

31015840-O

-120573-D

-glucop

yranoside

OOH

O

OH

HO

O

H HO

HHOH

HOH

H

OHO

Leaves

(Germany)aerial

(China)

[3562]

97

Okanin4-methylether-

3101584041015840-di-O

-120573-(41015840101584061015840101584041015840101584010158406101584010158401015840-

tetraacetyl)-

glucop

yranoside

OOH

OOH

O

HH

OH

HO

HOH

H

OO

O

Ac

Ac

AcAc

OO

H

HO

H

H

HOH

H

O

Aeria

l(Ch

ina)

[42]

98Ch

alcone210158404101584061015840-tr

imetho

xy-

4-O-D

-glucopyrano

syl-

dihydro

Not

foun

dLeaves

(China)

[60]

992-(34-D

ihydroxyph

enyl)-23-

dihydro-78

-dihydroxy-

4H-1-

benzop

yran-4-one

Okanin

isoOO

OH

HO

OH

OH

Leaves

(China)

[60]

100

2-(34-D

ihydroxyph

enyl)-

23-dihydro-8-hydroxy-

7[(246-tr

i-O-acetyl-120573

-D-

glucop

yranosyl)oxy]-4H

-1-b

enzopyran-4-on

eOO

OH

OH

OH

OH

O

HHO

H

HO

HO

OAc

Ac

Ac

Aeria

l(Ch

ina)

[34]

Evidence-Based Complementary and Alternative Medicine 19

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

101

57-Dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

OO

OH

HO

OH

Aeria

l(TanzaniaC

hina)

[4266]

102

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Apigenin

7-O-glucopyrano

side

OO

OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Tanzania)

[66]

103

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-4H

-1-benzop

yran-4-one

Luteolin

OO

HO

OH

OH

OH

Aeria

l(TanzaniaC

hina

Vietnam)

[343542

6667]

104

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5hydroxy-4H-1-

benzop

yran-4-one

Luteolin

7-O-120573-D

-glucop

yranoside

OOOH

OH

OH

OO

HO

H

HO

H

H

HOH

OH H

Aeria

l(Tanzania)

[66]

105

57-Dim

etho

xy-6-

(5-m

etho

xy-6-m

ethyl-

4-oxo-4H

-pyran-3-yl)-

2-ph

enyl-4H-1-

benzop

yran-4-one

5-O-M

ethylhosludin

O OOO

O

O

O

Aeria

l(Ugand

a)[68]

20 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

106

3-(120573-D

-Glucopyrano

syloxy)-

57-dihydroxy-2-

(4-hydroxyph

enyl)-

4H-1-

benzop

yran-4-one

Astragalin

Kaem

pferol-3-O

-120573-D

-glucopyrano

side

HO

OH

O O

OH

O

O

HO

HHO

H

H

HOH

OH

H

Aeria

l(Ch

ina)

[35]

107

Kaem

pferol3-

(23-di-E

-p-cou

maroyl-

120572-L-rhamno

pyrano

side)

Not

foun

dAe

rial(Vietnam)

[67]

108

2-(34-D

ihydroxyph

enyl)-

7-(120573-D

-glucopyrano

syloxy)-

5-hydroxy-36-dimetho

xy-

4H-1-

benzop

yran-4-one

Axillarosid

eO O

OH

OH

OO

OH

OO

HO

H

HO

H

H

HOH

H

OH

Aeria

l(Ch

ina)

[42]

109

57-Dihydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaureidin

O

O

O

O

O

HO

OH

OH

Who

le(Taiwan)

[69]

110

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Centaurein

O O

OO

O OH

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan)w

hole

(Taiwan)

[69ndash

71]

Evidence-Based Complementary and Alternative Medicine 21

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

111

5-Hydroxy-2-

(3-hydroxy-4-

metho

xyph

enyl)-

67-dimetho

xy-4H-

chromen-4-one

Eupatorin

iso

OO O

OH

O

O OH

Not

foun

d(C

hina)

[3772]

112

2-(34-D

imetho

xyph

enyl)-7-

(120573-D

-glucopyrano

syloxy)-35-

dihydroxy-

8-metho

xy-4H-1-

benzop

yran-4-one

O O

OH

O O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

113

7-(120573-D

-Glucopyrano

syloxy)-

5hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-38-

dimehtoxy-4H-1-

benzop

yran-4-one

O O

O

OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

O

Not

foun

d(Ja

pan)

[73]

114

Isorhamnetin

3-[O

-120572-

L-rham

nopyrano

syl-

(1-2)-120573-D

-glucop

yranoside]

Not

foun

dAe

rial(Vietnam)

[67]

115

7-[(6-deoxy-120572-L-

Manno

pyrano

syl)o

xy]-

3-120573-D

-glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-3-

metho

xyph

enyl)-4H

-1-benzop

yran-4-one

Luteoside

O O

OH

O

OH

OH

OH

HOH

H

H HO

HO

HO

OH

HO

H

HO

OH

OH

HH

Aeria

l(Ch

ina)

[42]

22 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

116

Luteolin

3-O-120573-D

-Glucopyrano

side

O O

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

HO

Aeria

l(Tanzania)

[66]

117

57-Dihydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

Quercetagetin

3631015840-tr

imethylether

OOO

O

O OH

HO

OH

Aeria

l(Ch

ina)

[42]

118

6-((2S3S4S5S)-

Tetrahydro-345-tr

ihydroxy-

6-(hydroxymethyl)-2H

-pyran-2-yloxy)-5-hydroxy-

2-(4-hydroxy-3-

metho

xyph

enyl)-

37-dimetho

xy-4H-

chromen-4-one

Quercetagetin

3731015840-

trim

ethylether-6-O

-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Ch

ina)

[42]

119

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-36-

dimetho

xy-4H-1-

benzop

yran-4-one

JaceinQ

uercetin

3631015840-

trim

ethyl

ether-7-O-120573-glucosid

eOO

O

O OH

O

OH

OH

HO

HHO

H

HOH

HO

OH

Aeria

l(Japan

China)

who

le(Taiwan)

[425470]

120

2-(34-D

ihydroxyph

enyl)-

357-trihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

OO

HO

OH

OH

OH

OH

Aeria

l(Ch

ina)leaves

(China)who

le(C

hina)

[356074]

Evidence-Based Complementary and Alternative Medicine 23

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

121

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-galactopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-4-one

Quercetin

3-O-120573-D

-galactosidehyperin

hyperosid

e

O O

HO

OH

OH

OH

OOHH

HOH

H

H HO

HO

OH

Aeria

l(TanzaniaJapan)

Leaves

(China)Who

le(C

hina)

[376066

717475]

122

2-(34-D

ihydroxyph

enyl)-

3-(120573-D

-glucopyrano

syloxy)-

57-dihydroxy-

4H-1-

benzop

yran-

4-on

e

Quercetin

3-O-120573-

D-glucopyrano

side

O O

HO

OH

OH

OH

OH

OH

HOH

H

H HO

HO

HO

Aeria

l(TanzaniaC

hina

Japan)L

eaves(Japan)

[355966

75]

123

2-(34-D

ihydroxyph

enyl)-

57-dihydroxy-

4-oxo-4H

-1-benzop

yran-3-yl-

120573-D

-glucopyrano

siduron

icacid

Quercetin

3-O-120573-D

-glucuron

opyranoside

O O

HO

OH

OH

OH

OHOH

HOH

H

H HO

HO

OHO

Aeria

l(TanzaniaJapan)

[6671]

124

3-[[6-O-(6-

Deoxy-120572-L-m

anno

pyrano

syl)-

120573-D

-galactopyrano

syl]o

xy]-

2-(34-dihydroxyph

enyl)-57-

dihydroxy-4H

-1-benzop

yran-4-one

Quercetin

3-O-rob

inob

iosid

e

O O

HO

OH

OH

OH

O

HOHH OH

HO

H HH

OO

OHH

HOH

H

H HO

HO

Aeria

l(Japan)W

hole

(Taiwan)

[7071]

24 Evidence-Based Complementary and Alternative Medicine

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

125

7-(120573-D

-Glucopyrano

syloxy)-

5-hydroxy-2-(4-hydroxy-

3-metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

OH

Roots(Brazil)

[55676]

126

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-

120573-D

-glucopyrano

syl]o

xy]-

5-hydroxy-2-

(4-hydroxy-3-

metho

xyph

enyl)-

3-metho

xy-4H-1-

benzop

yran-4-one

Quercetin

331015840-

dimethylether

7-O-120572-

L-rham

nopyrano

syl-

(1rarr

6)-120573-D

-glucop

yranoside

O OOH

OH

O

O

OH

HO

HHO

H

HOH

HO

O

HHO HHO

HO H

HH

ORo

ots(Brazil)

[5676]

127

7-[[6-O-(6-Deoxy-120572-L-

manno

pyrano

syl)-120573-D

-glucop

yranosyl]oxy]-5-

hydroxy-2-(3-hydroxy-4-

metho

xyph

enyl)-3-metho

xy-

4H-1-

benzop

yran-4-one

Quercetin

341015840-

dimethyl

ether-7-O-rutinoside

O OOH

OH

O

O

OH O

HOH

H

HHO

H

O

HHO HHO

HO H

HH

O

HO

Aeria

l(Ch

ina)

[3435]

128

2-(34-D

ihydroxyph

enyl)-3-

(120573-D

-glucofurano

syloxy)-57-

dihydroxy-4H

-1-benzop

yran-

4-on

e

Isoq

uercitrin

O OOH

OH

OHO

H

OHH

OH

HO

H O HOH

OH

Aeria

l(Japan

China)

[3571]

Evidence-Based Complementary and Alternative Medicine 25

Table5Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art

(Cou

ntry)

References

129

Quercetin

3-O-120573-D

-galactop

yranoside

O

OH

HO

OH OH

O

O

HO

OH

OHOH

OAe

rial(no

tstated)

[18]

130

Quercetin

3-O-rutinoside

O

O

R

OO

OH

OHO

H

O

OH

OH

H3C

H3C

H3C

OH

HO

Who

le(Taiwan)

[70]

26 Evidence-Based Complementary and Alternative Medicine

Table 6 Terpenoids isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(Country) References

131371111-Tetramethylbicyclo[810]undeca-

26-dieneBicyclogermacrene Leaves

(Brazil) [95]

13241111-Trimethyl-8-

methylenebicyclo[720]undec-4-ene

E-Caryophyllene Leaves(Brazil) [95]

1331-Methyl-5-methylene-8-

(1-methylethyl)-16-cyclodecadiene

Germacrene D Leaves(Brazil) [95]

1344-(15-Dimethyl-4-hexen-1-ylidene)-1-methyl-cyclohexene

Z-120574-Bisabolene Leaves(Brazil) [95]

135Decahydro-114-trimethyl-7-methylene-1H-cycloprop[e]-

azulene120573-Gurjunene Leaves

(Brazil) [95]

136 2669-Tetramethyl-148-cycloundecatriene

120572-Humulene120572-caryophyllene

Leaves(Brazil) [95]

13712344a568a-Octahydro-1-isopropyl-7-methyl-4-methylenenaphthalene

120575-Muurolene Leaves(Brazil) [95]

138

12344a568a-Octahydro-4a8-dimethyl-2-(1-methylethylidene)-

naphthalene

Selina-37(11)-diene

Leaves(Brazil) [95]

139(2E7R11R)-371115-

Tetramethyl-2-hexadecen-1-ol

Phytol

HO

Whole(Taiwan) [57]

Evidence-Based Complementary and Alternative Medicine 27

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

140 371115-Tetramethyl-2-hexadecenoic acid Phytanic acid

O

OH

Whole(Taiwan) [57]

141371115-Tetramethyl-2-

hexadecenyl ester-heptanoicacid

Phythylheptanoate O O

Leaves(Not stated) [33]

142

(3S10R13R)-17-((2R5R)-5-Ethyl-6-methylheptan-2-yl)-

2347891011121314151617-tetradecahydro-

1013-dimethyl-1H-cyclopenta[a]phenanthren-

3-ol

Campestrol

HO

Aerial(Tanzania) [18]

143 Not found Phytosterin-B Not foundNot found(TaiwanEgypt)

[39 96]

144 Stigmast-5-en-3-ol 120573-sitosterol

HOH

H

H

Aerial(Tanzania)

whole(Taiwan)

[18 31 57]

145

1314151617-Tetradecahydro-1013-

dimethyl-1H-cyclopenta[a]phenanthren-

3-yloxy)-tetrahydro-6-(hydroxymethyl)-2H-pyran-345-triol

120573-Sitosterolglucoside

H

H

HOH

HO

HHO

H

HOHH O

OH Not found(Egypt) [39]

146 5120572-Stigmasta-7-en-3120573-ol

HO

Whole(Taiwan) [57]

147 5120572-Stigmasta-722t-dien-3120573-ol

HO

Whole(Taiwan) [57]

148 Stigmasta-522-dien-3-ol Stigmasterol

HOH

H

H

Not found(Taiwan)aerial

(Tanzania)leaves (notstated)whole

(Taiwan)

[18 31 3357]

28 Evidence-Based Complementary and Alternative Medicine

Table 6 Continued

S N IUPAC names Common names Structure Plant part(Country) References

149 Lup-20(29)-en-3-ol Lupeol

H

HH

HOH

Not found(Egypt) [39]

150 Lup-20(29)-en-3-ol acetate Lupeol acetate

H

HH

O

OH

Not found(Egypt) [39]

151 Olean-12-en-3-ol 120573-amyrin

H

HOH

Not found(Egypt) [39]

152 5913-Trimethyl-242526-trinoroleanan-3-ol Friedelan-3120573-ol

HO

Aerial(Tanzania) [18]

153 5913-Trimethyl-242526-trinoroleanan-3-one

Friedelinfriedelan-3-one

O

Aerial(Tanzania) [18]

1542610151923-

Hexamethyl-2610141822-tetracosahexaene

Squalene

Aerial(Tanzania)Leaves (Notstated)Whole(Taiwan)

[18 33 57]

155 120573120573-Carotene 120573-Carotene Leaves (Notstated) [97]

Evidence-Based Complementary and Alternative Medicine 29

studies [83] Hot water extracts of B pilosa varminor Sheriffwere also assessed for its antileukemic effects on leukemiccell lines L1210 U937 K562 Raji and P3HR1 using XTT-based colorimetric assays The extract inhibited the five celllines with IC

50values ranging from 145120583gmL to 586120583gmL

L1210 K562 Raji and P3HR1 were more sensitive to B pilosaextract with IC

50values below 200120583gmL [98]

Consistent with the antitumor activities of B pilosaextracts and fractions some of its phytochemicals alsoshowed anticancer activity as outlined in Table 11 Amongthem luteolin (103) a well-studied flavonoid with multiplebioactivities was more effective against tumor cell prolifera-tion than its derivatives with IC

50values ranging from 3 120583M

to 50120583M in cells and 5 to 10mgkg in animals Luteolin wasalso found to fight cancer as a food additive at concentrationsof 50 to 200 ppm [78] and prevent skin cancer [78] and cancer3invasion [103] Lee and colleagues reported that luteolinprevents cancer by inhibiting cell adhesion and invasion[103] Significant inhibition concentration was reported to be5 120583M and complete inhibition concentration was reported tobe 40120583M Moreover luteolin was reported to inhibit hepa-tocyte growth factor (HGF)-induced cell scattering as wellas cytoskeleton changes such as filopodia and lamellipodiawhichwas determined using phase-contrast and fluorescencemicroscopy Furthermore luteolin also inhibited the HGF-induced phosphorylation of c-Met ERK 12 and Akt aswell as the MAPKERK and P13K-Akt pathways [78 103]Other mechanisms underlying the anticancer activities ofluteolin are the inhibition of topoisomerase I and II whichinhibits cell replication and DNA repair thus promotingapoptosis regulation of PI-3-KinaseAktMAPKERKJNKactivation of apoptosis in the mitochondrial pathway byactivating caspase 9 and caspase 3 which were found inmalignant cells but not in normal human peripheral bloodmononuclear cells death receptor-induced apoptosis and acell-cycle arrest mechanism inhibition of fatty acid synthasewhich is upregulated in many cancer cells and sensitizationto chemotherapywhereby luteolin increases the susceptibilityof cancer cells to chemotherapy [78] Butein (82) is anotherflavonoid that showed a cytotoxic effect on human colonadenocarcinoma cell proliferation with a reported IC

50value

of 175 120583M Butein at 2 120583M affected the incorporation of[14C]-labeled leucine thymidine and uridine which cancause the inhibition of DNA RNA and protein synthesis ofhuman colon cancer cells Moreover butein also exhibitednoncompetitive inhibition of 1-chloro-24-dinitrobenzene(CDNB) in glutathione S-transferase (GST) activity Tumorresistance was correlated with high levels of GST thus buteininhibited proliferation of cancer cells [80] Another flavonoidpresent in B pilosa centaureidin (109) also showed anti-cancer activity in B lymphoma cells Centaureidin isolatedfrom Polymnia fruticosa inhibited tubulin polymerization invitro and induced mitotic figure formation in CA46 Burkittlymphoma cells Using turbidimetric assay the IC

50value of

centaureidin in inhibition of mitosis was 3120583M [104] Cyto-toxicity of centaureidin was further analyzed using AmericanNational Cancer Institute (NCI) 60 human tumor cell linesThe cytotoxicity potency of centaureidin expressed as GI

50

(50 growth inhibition in the NCI tumor line panel) was024120583M [81] These data mark centaureidin as a promisingantimitotic agent for tumor therapy

In addition to anti-tumor flavones polyynes found in B 4pilosa have also been shown to possess anti-tumor proper-ties Based on a bioactivity-directed isolation approach Wuand colleagues identified two polyyne aglycones from theethyl acetate fraction of B pilosa [44] 12-Dihydroxytrideca-57911-tetrayne (48) and 13-Dihydroxy-6(E)- tetradecene-81012-triyne (46) exhibited significant anticell prolifera-tion activity in primary human umbilical vein endotheliumcells (HUVEC) with IC

50values of 125 120583M and 173 120583M

respectively They also decreased angiogenesis and pro-moted apoptosis in human endothelial cells Their anti-angiogenic and cytotoxic effects correlated with activationof the CDK inhibitors and caspase-7 [44] In addition 12-Dihyroxy-5(E)-tridecene-7911-triyne (45) showed antian-giogenic effects in HUVECs with an IC

50value of 124 120583M as

evidenced by a decrease in the tube formation and migrationof HUVECs [43] The IC

50value of compound 45 in the

inhibition of basic fibroblast growth factor-induced HUVECgrowth was 282120583M However it had higher IC

50values

than those for lung carcinoma cells and keratinocytes Thiscompound could also inhibit cell proliferation of HUVECslung carcinoma A549 cells and HACAT keratinocytes Themechanism by which compound 45 inhibits HUVEC growthand angiogenesis is complicated and includes decreasing theexpression of cell cycle regulators (CDK4 cyclins D1 and Aretinoblastoma (Rb) and vascular endothelial growth factorreceptor 1) caspase-mediated activation of CDK inhibitorsp21 (Cip1) and p27 (Kip) upregulation of Fas ligand expres-sion downregulation of Bcl-2 expression and activation ofcaspase-7 and poly (ADP-ribose) polymerase [43]

32 Anti-Inflammatory Activity B pilosa is commonly usedto treat inflammatory disorders The anti-inflammatory phy-tochemicals present in B pilosa are listed in Table 11 5Cyclooxygenase-2 (COX-2) is a physiologically importantenzyme that converts arachidonic acid to prostaglandin(PGE2) Its expression is induced by a wide variety of external

stimuli indicating its involvement in inflammatory diseasesand it is used as an inflammatory marker [88] Yoshida andcolleagues studied the effects of the aqueous extracts of Bpilosa aerial parts in the production of COX-2 and PGE

2as

well as on the activation of mitogen activated protein kinases(MAPKs) in normal human dermal fibroblasts (HDFs) inresponse to inflammatory cytokine IL-1120573 This work showedthat IL-1120573 activated MAPKs such as ERK12 p38 and JNKto different extents and induced COX-2 expression TheCOX-2 expression in HDFs was regulated mainly by p38following IL-1120573 stimulation Consistently the p38 inhibitorSB203580 blocked this expression Using this cell platformB pilosa extracts were tested for inhibition of inflammationThe extract dose-dependently suppressed the activation ofp38 and JNK and moderately suppressed ERK12 as wellas suppressing COX-2 expression and PGE

2production

30 Evidence-Based Complementary and Alternative Medicine

Table7Ph

enylprop

anoids

isolatedfro

mB

pilosa

[30]

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

156

3-(4-H

ydroxyph

enyl)-2-prop

enoica

cid

p-Cou

maricacid

OHO

OH

Who

le(Ja

pan)

[99]

157

2-Metho

xy-4-(2-prop

en-1-yl)-ph

enol

Eugeno

lOOH

Leaves

androots(Japan)

[99]

158

3-(4-H

ydroxy-3-m

etho

xyph

enyl)-

2-prop

enoica

cid

Ferulic

acid

O

OH

O

HO

Who

le(Ja

pan)

[99]

159

3-(34-D

ihydroxyph

enyl)-2-prop

enoic

acid

Caffeicacid

O

HO

OH

OH

Who

leandaeria

l(Japan)

[7599]

160

3-Prop

yl-3[(245-trim

etho

xyph

enyl)-

metho

xy]-24-pentanedione

3-Prop

yl-3-(245-

trim

etho

xy)benzyloxy-

pentan-24-dione

O

O

O

O

O

O

Leaves

(India)

[100]

161

3-(34-D

ihydroxyph

enyl)-2-

prop

enoica

cidethyleste

rCa

ffeateethyl

O

O

OH

OH

Not

foun

d(Taiwan)

who

le(Taiwan)

[434584]

Evidence-Based Complementary and Alternative Medicine 31Ta

ble7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

162

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

d-Erythron

icacid

2-O-caffeoyl-2-C

-methyl

O

HO

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

163

2-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-

34-dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

2-O-caffeoyl-2-C

-methyl

O

O

OH

HO

O

O

OH

OH

Leaves

(Japan)

[101]

164

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

enyl]oxy]-24-

dihydroxy-2-methyl-b

utanoica

cid

methyleste

r

d-Erythron

atemethyl

3-O-caffeoyl-2-C

-methyl

O

O

HO

HO

O

O

OH

OH

Leaves

(Japan)

[101]

165

4-(A

cetyloxy)-3-[[3-(34-

dihydroxyphenyl)-

1-oxo-2-propen-1-y

l-]oxy]-2-hydroxy-

2-methyl-b

utanoica

cid

O

HO

OH

O OHO

HO

OO

Not

foun

d(Ja

pan)

[73]

166

3-(34-D

ihydroxyph

enyl)-tetrahydro-4-

hydroxy-4-

methyl-5

-oxo-3-fu

ranyl

ester-2-prop

enoica

cid

3-O-C

affeoyl-2-C

-methyl-

D-erythrono

-14-la

cton

eHO

HO

O

OO

OOH

Leaves

(Japan)

[101]

167

3-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]-oxy]-145-

trihydroxy-cyclohexanecarboxylicacid

Chlorogenica

cid

O

HO

HO

OH

O

OH

O

OH

HO

Aeria

l(Japan)w

hole

(Taiwan)

[707175]

32 Evidence-Based Complementary and Alternative Medicine

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

168

4-[[3-(34-D

ihydroxyph

enyl)-1-o

xo-2-

prop

en-1-

yl]oxy]-

135-tr

ihydroxy-cyclohexanecarboxylic

acid

4-O-C

affeoylqu

inicacid

OHO

HO

OH

OH

O

O

OH

OH

Aeria

l(Japan)

[71]

169

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-15-

dihydroxy-cyclo

hexanecarboxylicacid

34-Di-O

-caffeoylqu

inic

acid

HO

HO

O

O

O

O

OH

OH

HO

OH O

H

O

Aeria

l(Japan)W

hole

(Taiwan)

[45497071]

170

34-bis[[2(E)-3-(34-D

ihydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-oxy]-14-dihydroxy-

cyclo

hexanecarboxylicacid

35-Di-O

-caffeoylqu

inic

acid

HO

HO

O O

HO

O

HO

OH OO

OHOH

Aeria

l(Japan)w

hole

(Taiwan)

[45497071]

Evidence-Based Complementary and Alternative Medicine 33

Table7Con

tinued

SN

IUPA

Cnames

Com

mon

names

Structure

Plantp

art(coun

try)

References

171

34-bis[[(2E

)-3-(34-D

ihydroxyph

enyl)-1-

oxo-2-prop

en-1-

yl]oxo]-15

-dihydroxy-

cyclo

hexanecarboxylicacid

45-Di-O

-caffeoylqu

inic

acid

OHO

OH

HO

O

O

OH

OH

O

OH

HO

OWho

le(Taiwan)

[454970]

172

3-[4-[[-6-O-[3-(4-H

ydroxyph

enyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]-phenyl]-2-prop

enoica

cid

120573-D

-p-C

oumaricacid4-O

-(6-O

-p-caoum

aroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HOH

HO

O

O

Leaves

(Japan)

[59]

173

3-[4-[[2-O

-Acetyl-6

-O-[3-(4-

hydroxyphenyl)-

1-oxo-2-propen-1-y

l]-120573-D

-glucop

yranosyl]-

oxy]ph

enyl]2-propeno

icacid

120573-D

-p-C

oumaricacid4-O

-(2-O

-acetyl-6

-O-p-

caou

maroyl-

glucop

yranosyl)

HO

O

OH

HO

HHO

H

HO

HO

O

O

Leaves

(Japan)aerial

(China)

[3459]

174

67-Dihydroxy-2-chrom

enon

eEsculetin

cicho

rigenin

OHO HO

O

Not

foun

d(Egypt)

[39]

34 Evidence-Based Complementary and Alternative Medicine

Table 8 Aromatic compounds isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

175 12-Benzenediol PyrocatechinHO

HOWhole(Japan) [99]

176 4-Ethyl-12-benzenediol Pyrocatechol

OH

OH

Whole(Japan) [99]

177 Dimethoxyphenol26-dimethoxyphenol

O

HO

O

Roots (Japan) [99]

178 4-Ethenyl-2-methoxy-phenol p-Vinylguaiacol

O

OHWhole(Japan) [99]

179 2-Hydroxy-6-methylbenzaldehyde

6-Methyl-salicylaldehyde O

OH

Whole(Japan) [99]

180 Benzene-ethanol 2-Phenyl-ethanolHO

Whole(Japan) [57]

181 4-Hydroxy-3-methoxy-benzaldehyde Vanillin O

O

HOAerial(Japan) [99]

182 3-Hydroxy-4-methoxy-benzaldehyde Vanillin iso

OO

OH

Leaves(Japan) [99]

183 4-Hydroxybenzoic acid p-Hydroxybenzoicacid

O

HOOH Whole

(Japan) [99]

184 2-Hydroxybenzoic acid Salicylic acidO

HO

HO

Stem androots (Japan) [99]

185 34-Dihydroxybenzoicacid Protocatechuic acid

O

HOOH

OH

Whole(Japan) [99]

1864-Hydroxy-

3methoxybenzoicacid

Vanillic acid

OOH

O

OH

Aerial(Uganda)

Roots (Japan)[68 99]

Evidence-Based Complementary and Alternative Medicine 35

Table 8 Continued

S N IUPAC names Common names Structure Plant part(country) References

187 345-Trihydroxybenzoicacid Gallic acid

O

OHHO

HO

HO

Whole(China) [102]

[88] This work supports the use of B pilosa as an anti-inflammatory agent however no compounds responsible forthe anti-inflammatory activity of B pilosa were identified

A further study also reported the anti-inflammatoryactivity as well as the antiallergic activity of B pilosa [75] Inthis study dried powder of the aerial part of B pilosa whichhad been pretreated with the enzyme cellulosine was usedfor further tests The results showed that oral administrationof the cellulosine-treated B pilosa lowered the level of serumIgE in mice 10 days after immunization with DNP (24-dintrophenyl)-Ascaris as an antigen This treatment alsoreduced dye exudation in skin induced by passive cutaneousanaphylaxis and production of inflammatory mediators his-tamine and substance P in rats [75] Phytochemical analysisshowed that cellulosine treatment increased the percentage ofcaffeic acid and flavonoids This study suggests that B pilosaand its phenolics have anti-inflammatory functions

Phenolics and polyynes are major anti-inflammatoryphytochemicals present in B pilosa (Table 11) Unsurpris-6ingly phenolics such as luteolin (103) and ethyl caffeate(161) that are major constituents of B pilosa have alsobeen reported to possess anti-inflammatory activity Lute-olin was reported to exhibit anti-inflammatory activity inmacrophages Xagorari and colleagues showed that luteolininhibited the release of inflammatory cytokines TNF-120572 andinterleukin-6 in RAW 2647 cells following LPS stimulation[87] It inhibited TNF-120572 production with an IC

50value

of 1120583M The underlying anti-inflammatory mechanism ofluteolin was reported to be the inactivation of Akt andNF-120581B activation [87] In addition luteolin was reportedto confer anti-inflammatory activity through inhibition ofLPS-stimulated iNOS expression in BV-2 microglial cellsIt inhibited LPS-activated microglia in a dose-dependentmanner with an IC

50value of 69 120583MMoreover immunoblot

and RT-PCR data proved that luteolin suppressed I120581B-120572degradation and iNOS expression in LPS-activated microglia[85] Kim and colleagues stated that luteolin may havebeneficial effects on inflammatory neural diseases throughinhibition of iNOS expression [85] A related study revealedthat luteolin decreased the transcriptional activity of NF-120581BRelA via partial inhibition of TNF-mediated NF-120581B DNAbinding activity Luteolin also inhibited Akt phosphorylationand induced degradation of a transcription factor interferonregulatory factor (IRF) [86]

Chiang and colleagues showed that ethyl caffeate(161) significantly inhibited NO production in mousemacrophages RAW 2647 cells [84] Based on MTT assays

they concluded that this inhibition was not due to thecytotoxicity of ethyl caffeate The IC

50value of ethyl caffeate

in the inhibition of NO production was 55 120583gmL slightlylower than curcumin (positive control) which has anIC50

value of 65120583gmL They demonstrated that ethylcaffeate exerted anti-inflammatory activity via the reducedtranscription and translation of iNOS (inducible nitric oxidesynthase) in RAW 2467 cells In addition this compoundalso suppressed COX-2 expression in RAW 2467 cellsand MCF-7 cells The in vivo anti-inflammatory effectof ethyl caffeate was verified by testing in TPA-treatedmouse skin Like celecoxib the positive control ethylcaffeate significantly abolished COX-2 expression in adose-dependent manner Ethyl caffeate at 1mg200120583Lsite(24mM) inhibited COX-2 expression at a level comparableto celecoxib at 1mg200120583Lsite (13mM) Remarkably ethylcaffeate at 48mM (2mg200120583Lsite) was more effectivethan celecoxib at 131mM (10mg200120583Lsite) Moreoverthis compound inhibited the activation of nuclear factor-120581B(NF-120581B) by LPS via the prevention of NF-120581B binding toDNA [84] In addition 3 ethyl caffeate analogs (ethyl 34-dihydroxyhydrocinnamate ethyl cinnamate and catechol)also showed different degrees of NF-120581B binding to DNA aslisted in Table 12 Ethyl cinnamate lacks a catechol moietywhich results in ineffective inhibition of NF-120581B binding toDNA [84]

Pereira and colleagues assessed the anti- 7inflammatory and immunomodulatory activities ofB pilosa methanol extract as well as one polyyne2-O-120573-glucosyltrideca-11(E)-en-3 579-tetrayn-12-diol(54) in T lymphocytes and a zymosan-induced arthritismouse model [27] They first examined the in vitro effect of 8the B pilosa extract and compound 54 on cell proliferation ofhuman T cells stimulated with 5120583gmL phytohemagglutinin(PHA) or 100 nM 12-O-tetradecanoyl phorbol-13-acetate(TPA) plus 15 120583M ionomycin and on cell proliferation ofmouse T cells stimulated with 5 120583gmL concanavalin A(Con A) The data demonstrated that both methanol extractand compound 54 suppressed T-cell proliferation in adose-dependent manner The estimated IC

50values of the B

pilosa extract against human T cells stimulated with 5120583gmLPHA and 100 nM TPA plus 15120583M ionomycin were 125 and25 120583gmL respectively In comparison with the methanolextract compound 54 showed 10-fold more inhibition ofhuman T-cell proliferation with an estimated IC

50value of

15 120583gmL Accordingly the B pilosa extract and compound54 dose-dependently suppressed mouse T-cell proliferation

36 Evidence-Based Complementary and Alternative Medicine

Table 9 Porphyrins isolated from B pilosa [30]

S N IUPAC names Common names Structure Plant part(country) References

188

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(15S16S)-10-ethenyl-5-ethyl-1161820-tetrahydro-6111522-tetramethyl-1820-dioxo-15H-912-imino-212-

metheno-471714-dinitrilopyrano[43-

b]azacyclononadecine-16-propanoic

acid

AristophyllC

NH N

HNN

O

O OOOPhytyl

O

Leaves(Taiwan) [100]

189

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -20b-(methoxycarbonyl)-9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-dinitrilo-12-

dioxeto-[310158404101584034]-cyclo-pent[1 2]azacyclononadecine-

19-propanoic acid

Bidenphytin A

NH N

HNN

OPhytylO O O

O O

Leaves(Taiwan) [100]

190

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(2S18S19S20bR)-13-

ethenyl-8-ethyl-2a1819 20b-tetrahydro -2a-

hydroxy-20b-(methoxycarbonyl)-

9141824-tetramethyl-4H-1215-imino-35-metheno-7102017-

dinitrilo-12-dioxeto[310158404101584034]-cyclo-pent[12-b]-

azacyclononadecine-19-propanoic acid

Bidenphytin B

NH N

HNN

OPhytylO O O

O O

OH

Leaves(Taiwan) [100]

191

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO O

HOOO

Leaves(Taiwan) [100]

192

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-21-

hydroxy-21-(methoxycarbonyl)-4891318-pentamethyl-20-oxo-

3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin A

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

Evidence-Based Complementary and Alternative Medicine 37

Table 9 Continued

S N IUPAC names Common names Structure Plant part(country) References

193

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21R)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132R)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

Leaves(Taiwan) [100]

194

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3R4S21S)-14-ethyl-13-

formyl-21-hydroxy-21-(methoxycarbonyl)-48918-

tetramethyl-20-oxo-3-phorbinepropanoic acid

(132S)-132-Hydroxypheophytin B

NH N

HNN

OPhytylO OHO

OO

O

Leaves(Taiwan) [100]

195

(2E7R11R)-371115-Tetramethyl-2-hexadecen-1-ylester-(3S4S21R)-9-ethenyl-14-ethyl-21-(methoxycarbonyl)-

481318-tetramethyl-20-oxo-3-phorbinepropanoic acid

Pheophytin A

NH N

HNN

PhytylO

O

OO O

Leaves(Taiwan) [100]

with estimated IC50values of 30 and 25 120583gmL respectively

Taken together the data indicate that the B pilosa extractand compound 54 act on human and mouse T cells Totest the in vivo effect of the B pilosa extract and compound54 a zymosan-induced arthritis mouse model was usedThis model was established from B10ASgSnJ mice with aninjection of zymosan (015mg) The zymosan-injected mice9received an intraperitoneal injection of the B pilosa extract(1 5 or 10mg) at one dose a day for 5 days Popliteal lymphnode (PLN) weight was monitored to check the developmentof arthritis The results revealed that 10mg of the methanolextract of B pilosa extract could significantly diminishinflammation as evidenced by PLN weight [27] This worksuggests that B pilosa (and compound 54) can suppressimmune response and inflammation

33 Antidiabetic Activity Anti-diabetic agents are primarilydeveloped from plants and other natural resources [8 24 4951 53] B pilosa is one of 1200 plant species that have beeninvestigated for antidiabetic activity [120 121]B pilosa is usedas an anti-diabetic herb in America Africa and Asia [51 120

122] Many studies have indicated that B pilosa could treattype 1 diabetes (T1D) and type 2 diabetes (T2D) in animals

Etiologically speaking T1D is caused by autoimmune-mediated destruction of pancreatic 120573 cells leading to insulindeficiency hyperglycemia and complications Currentlythere is no cure for T1D Polarization ofTh cell differentiationcontrols the development of T1D Suppression of Th1 celldifferentiation and promotion of Th2 cell differentiationameliorate T1D [123] One study showed that the butanolfraction of B pilosa inhibited T-cell proliferation decreasedTh1 cells and cytokines and increased Th2 cells andcytokines leading to prevention of T1D in nonobesediabetic (NOD) mice [49] Based on a bioactivity-directedisolation strategy 3 polyynes 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne (53) also known ascytopiloyne 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D- Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne (50) were identifiedfrom B pilosa [49 53] The IC

50value of the butanol fraction

was 200120583gmL This inhibition was reported to be partiallyattributed to cytotoxicity because the butanol fraction at

38 Evidence-Based Complementary and Alternative Medicine

Table 10 Other compounds isolated from B pilosa [30]

SN IUPAC names Common names Structure Plant part(country) References

19637-Dihydro-137-

trimethyl-1H-purine-26-dione

CaffeineN

NN

NO

O

Aerial(Uganda) [68]

197

1-((2R4S5R)-Tetrahydro-4-hydroxy-5-(hydroxymethyl)furan-2-yl)-5-

methylpyrimidine-24(1H3H)-dione

ThymidineHO

HO

ON

O

NHO Not found

(China) [37]

198 1-(2-Thienyl)-ethanone 2-Acetyl-thiophene

S

O

Roots(Germany) [41]

199

(2R3S4S5S)-2-(Heptan-2-yloxy)-tetrahydro-6-

((tetrahydro-34-dihydroxy-5-

(hydroxymethyl)furan-2-yloxy)methyl)-

2H-pyran-345-triol

Heptanyl2-O-120573-xylofuranosyl-

(1rarr 6)-120573-glucopyranoside

O

OOH

OH

HO

OHO

H

HO

H

H

HOHH

O

Whole(Taiwan) [70]

200

2-[(3R7R11R)-3-Hydroxy-371115-

tetramethylhexadecyl]-356-trimethyl-

25-cyclohexadiene-14-dione

120572-Tocopherylquinone

OO

HO

Whole(Taiwan) [32]

2017-O-(410158401015840610158401015840-Diacetyl)-

120573-D-glucopyranoside

Not found Leaves(China) [60]

180 120583gmL could cause 50 death of Th1 cells Moreoverthis study suggested that the butanol fraction may preventdiabetes in NOD mice in vivo via downregulation of Th1cells or upregulation Th2 cells that have effects which areantagonistic of those of Th1 cells [49] This was proven byintraperitoneal injection of the butanol fraction at a doseof 3mgkg BW 3 times a week to NOD mice from 4 to 27weeks This dosage resulted in lower incidence of diabetes(33) At a dose of 10mgkg the butanol fraction of Bpilosa totally eliminated (0) the initiation of the diseaseTo further support this result assessment of IgG2a and IgEproduction was performed in the serum of NOD mice Asin vivo results obtained from intracellular cytokine stainingexperiments were not very conclusive levels of IgG2a and IgE

were measured since Th1 cytokine IFN120574 and Th2 cytokineIL-4 favor the production of IgG2a and IgE respectively Asexpected high levels of IgE and some decline in the levels ofIgG2a were observed in the serum Profiling of the butanolextract revealed five compounds 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne (69) 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne(50) 45-Di-O-caffeoyl- quinic acid 35-Di-O-caffeoylquinicacid and 34-Di-O-caffeoylquinic acid Only the first twocompounds showed similar effects on the prevention ofdiabetes in NOD mice as the B pilosa butanol fractionMoreover compound 50 showed greater activity thancompound 69 in terms of enhancement (by 34 comparedto 8) of differentiation of Th0 to Th2 at 15 120583gmL (both

Evidence-Based Complementary and Alternative Medicine 39

Table 11 Chemical constituents of B pilosa and their biological activities

SN Name Classification Molecularformula Biological activities

109 Centaureidin [91] Flavonoid C18H16O8Anti-listerial [69 91]

Cytotoxic [10]

110 Centaurein [91] Flavonoid C24H26O13

Anti-listerial [69 91]Cytotoxic [10]Anti-viral [105]

103 Luteolin [106] Flavonoid C15H10O6

Anti-viral [107 108]Cytotoxic [77]

Anti-inflammatory [109]Anti-allergic [109]

82 Butein [110] Flavonoid C15H12O5Anti-leishmanial [111]

Cytotoxic [78]48 12-Dihydroxytrideca-57911-tetrayne [112] Polyyne C13H12O2 Anti-angiogeneic [112]46 13-Dihyroxy-6(E)-tetradecene-81012-triyne [112] Polyyne C14H16O2 Anti-angiogeneic [112]

45 12-Dihyroxy-5(E)-tridecene-7911-triyne [113] Polyyne C13H14O2Anti-angiogeneic [113]Anti-proliferative [113]

64 1-Phenylhepta-135-triyne [114] Polyyne C13H8

Anti-microbial [115]Anti-malarial [3]Cytotoxic [3]Antifungal [15]

27 Linoleic acid [79] Fatty acid C18H32O2Anti-viral (100)Cytotoxic [17]

161 Ethyl caffeate [84] Phenylpropanoid C11H12O4 Anti-inflammatory [84]

54 2-O-120573-Glucosyltrideca-11(E)-en-3579-tetrayn-12-diol [26] Polyyne C19H20O7Immunosuppressive andAnti-inflammatory [26]

53 2-120573-D-Glucopyranosyloxy-1-hydroxytrideca-57911-tetrayne [50] Polyyne C19H22O7Anti-diabetic [116]

Anti-inflammatory [50]

69 3-120573-D-Glucopyranosyl-1-hydroxy-6(E)-tetradecene-81012-triyne [28] Polyyne C20H26O7Anti-diabetic [28]

Anti-inflammatory [94]

50 2-120573-D-Glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7911-triyne [28] Polyyne C19H24O7

Anti-diabetic [28]Anti-inflammatory [94]

Anti-malarial andantibacterial [47]

129 Quercetin 3-O-120573-D-galactopyranoside [18] Flavonoid C21H20O12 Anti-inflammatory [117]

170 35-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

171 45-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

169 34-Di-O-caffeoylquinic acid [94] Phenylpropanoid C25H24O12Anti-viral [57]Antioxidant [70]

126 Quercetin 331015840-dimethyl ether7-O-120572-L-rhamnopyranosyl-(1rarr 6)-120573-D-glucopyranoside [118] Flavonoid C21H20O11 Anti-malarial [119]

125 Quercetin 331015840-dimethyl ether-7-O-120573-D-glucopyranoside [118] Flavonoid C21H20O12 Anti-malarial [119]70 1-Phenyl-13-diyn-5-en-7-ol-acetate [27] Polyyne C15H12O2 Anti-malarial [27]199 Heptanyl 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside [70] Miscellaneous C18H44O10 Antioxidant [70]124 3-O-Rabinobioside [70] Saccharide C27H38O15 Antioxidant [70]130 Quercetin 3-O-rutinoside [70] Flavonoid C27H38O15 Antioxidant [70]167 Chlorogenic acid [70] Phenolic C16H26O9 Antioxidant [70]119 Jacein [70] Flavonoid C24H26O13 Antioxidant [70]

49 (R)-12-dihydroxytrideca-357911-pentayne [47] Polyyne C13H8O2Anti-malarial andAntibacterial [47]

40 Evidence-Based Complementary and Alternative Medicine

Table 12 Structure and activity relationship studies of ethyl caffeateusing in vitro NF-120581BDNA binding assays [84]

Compound Concentration(120583M)

NF-120581BDNA binding

Ethyl caffeate 50 100inhibition

Ethyl 34-dihydroxyhydrocinnamate 100 100inhibition

Catechol 400 100inhibition

Ethyl cinnamate 400 Noinhibition

Table 13 Apoptosis in cocultures of T cells and pancreatic 120573 cells[53]

Cellmedium apoptosis andnecrosis

CD4+ T cellscontrol medium lt4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 2

CD4+ T cellswith cytopiloyne-treated 120573 cells 18CD4+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 7

CD8+ T cellscontrol medium 4CD4+ T cellswith PBS-treated 120573 cellsof NOD-SCID mice 4

CD8+ T cellswith cytopiloyne-treated 120573 cells 4CD8+ T cellswith cytopiloyne-treated 120573 cells inthe presence of 120572-FasL antibody 4

Table 14 Radical scavenging activities of B pilosa extracts [70]

Extractscontrol DPPH assayIC50 (120583gmL)

NBThypoxanthinesuperoxide assayIC50 (120583gmL)

Quercetin 198 15Ascorbic acid 634 Not determined120572-tocopherol 897 Not determinedEthyl acetateextract 1383 597

Butanol extract 1669 114Water extract gt100 gt100

compounds) and inhibition (by 40 compared to 10) ofdifferentiation toTh1 at the same concentration [49]

Among the three polyynes found in B pilosa cytopiloyne(53) had the most potent anti-T1D activity [53] To test the invivo effect of cytopiloyne NODmice received intraperitonealor intramuscular injection of cytopiloyne at 25120583gkg BW3 times per week Twelve-week-old NOD mice started todevelop T1D and 70 of NOD mice aged 23 weeks andover developed T1D Remarkably 12- to 30-week-old NODmice treated with cytopiloyne showed normal levels of bloodglucose (lt200mgdL) and insulin (1-2 ngmL) Consistent

Table 15 Radical scavenging activity of secondarymetabolites fromB pilosa [70]

Metabolite (Table 11)control DPPH assay IC50 (120583gmL)199 Not determined124 53130 68167 105169 33171 38119 Not determined110 Not determinedQuercetin 256Caffeic acid 890

Table 16 Antioxidant activity of the essential oils and water extractsfrom B pilosa [92]

Extract IC50 (120583gmL)Leaf essential oils 47Flower essential oils 50Leaf extract 61Flower extract 172

with T1D incidence cytopiloyne delayed and reduced theinvasion of CD4+ T cells into the pancreatic islets [53]

In vitro study showed that cytopiloyne (53) inhibited thedifferentiation of naıveTh (Th0) cells (ie CD4+ T cells) intoTh1 cells and promoted differentiation of Th0 cells into Th2cells [50] The in vitro data are consistent with the in vivoresults indicating that cytopiloyne reducedTh1differentiationand increased Th2 differentiation as shown by intracellularcytokine staining and FACS analysis [53] In line with theskewing of Th differentiation the level of serum IFN-120574 andIgG2c decreased while that of serum IL-4 and serum IgEincreased compared to the negative controls (PBS-treatedmice) Cytopiloyne also enhanced the expression of GATA-3 a master gene for Th2 cell differentiation but not theexpression of T-bet a master gene forTh1 cell differentiationfurther supporting its role in skewingTh differentiation [53]

Also importantly cytopiloyne partially depleted CD4+rather than CD8+ T cells in NOD mice [53] As shown inTable 13 coculture assays showed that the depletion of CD4+T cells was mediated through the induction of Fas ligandexpression on pancreatic islet cells by cytopiloyne leading toapoptosis of infiltrating CD4+ T cells in the pancreas via theFas and Fas ligand pathways However cytopiloyne did notinduce the expression of TNF-120572 in pancreatic islet cells andthus had no effect on CD8+ T cells [53]

In addition Chang and colleagues showed that cytopi-loyne dose-dependently inhibited T-cell proliferation stimu-lated by IL-2 plus Con A or anti-CD3 antibody using [3H]thymidine incorporation assay [53]

Overall the mechanism of action of cytopiloyne andprobably its derivatives in T1D includes inhibition of T-cellproliferation skewing of Th cell differentiation and partialdepletion of Th cells Due to the anti-diabetic mechanisms

Evidence-Based Complementary and Alternative Medicine 41

Table 17 Antibacterial activity of essential oils and flower extracts from B pilosa [92]

Strain Mean zone of inhibition (mm)Leaf essential oil Flower essential oil Leaf extract Flower extract

Micrococcus flavus 127 plusmn 03 87 plusmn 03 102 plusmn 02 108 plusmn 03Bacillus subtilis 173 plusmn 19 117 plusmn 02 109 plusmn 02 103 plusmn 02Bacillus cereus 190 plusmn 14 112 plusmn 03 118 plusmn 04 185 plusmn 10Bacillus pumilus 123 plusmn 07 108 plusmn 02 105 plusmn 04 77 plusmn 02Escherichia coli 137 plusmn 04 203 plusmn 07 102 plusmn 11 140 plusmn 13Pseudomonas ovalis 125 plusmn 08 137 plusmn 15 102 plusmn 06 125 plusmn 06

Table 18 Antibacterial activity of root extracts from B pilosa [93]

Strain MIC50 (mgmL)Methanol extract Acetone extract

Bacillus cereus 10 mdashEscherichia coli 5 5Klebsilla pneumonia 5 10Micrococcus kristinae 10 mdashPseudomonas aeruginosa 10 10Staphylococcus aureus 5 10Sraphylococcus epidermidis 5 5Serratia marcescens 10 mdashShigelea flexneri 10 mdashStreptococcus faecalis 10 mdash

Table 19 Antibacterial activity of B pilosa of compound 29 [47]

Strain MIC50 (120583gmL)Escherichia coli NIHJ 1Escherichia coli ATCC25922 1Klebsiella pneumoniae ATCC700603 128Serratia marcescens ATCC13880 16Pseudomonas aeruginosa ATCC27853 8Staphylococcus aureus FDA209P 05Staphylococcus aureus ATCC29213 025Staphylococcus aureus N315 (MRSA) 05Enterococcus faecalis ATCC29212 2Enterococcus faecalis NCTC12201 (VRE) 1Bacillus subtilis ATCC6633 05Candida albicans ATCC10231 025

of action it was hypothesized that cytopiloyne protectsNOD mice from diabetes by a generalized suppression ofadaptive immunity To evaluate this hypothesis ovalbumin(Ova) was used as a T-cell dependent antigen to primeNOD mice which had already received cytopiloyne orPBS vehicle Ova priming boosted similar anti-Ova titersin cytopiloyne-treated mice and PBS-treated mice but adifference in immunoglobulin isotype was observed in thetwo groups Thus it was concluded that cytopiloyne is animmunomodulatory compound rather than an immunosup-pressive compound [50 53]

T2D is a chronic metabolic disease with serious com-plications resulting from defects in either insulin secretion

insulin action or both [124] A study by Ubillas et alshowed that the aqueous ethanol extract of the aerial partof B pilosa at 1 gkg body weight (BW) lowered bloodglucose in dbdb mice a T2D mouse model [51] Basedon a bioactivity-guided identification compounds 69 and50 were identified Further the mixture of the compounds(69 50) in a 2 3 ratio significantly decreased blood glucoseconcentration and reduced food intake on the second dayof treatment when administered at doses of 250mgkg twicea day to C5BLKs-dbdb mice When tested at 500mgkga more substantial drop in blood glucose level as wellas the stronger anorexic effect (food intake reduced from58 gmouseday to 25 gmouseday) was observed [51] Inthis study it was suggested that the blood glucose lower-ing effect of B pilosa was caused in part by the hungersuppressing effect of its polyynes [51] However the hungersuppressing effect of the ethanol extract of B pilosa wasnot found in the studies described below In another study[24] water extracts of B pilosa (BPWE) were used indiabetic dbdb mice aged 6ndash8 weeks with postprandialblood glucose levels of 350 to 400mgdL Like oral anti-diabetic glimepiride which stimulates insulin release onesingle dose of BPWE reduced blood glucose levels from374 to 144mgdL The antihyperglycemic effect of BPWEwas inversely correlated to an increase in serum insulinlevels suggesting that BPWE acts to lower blood glucose viaincreased insulin production However BPWE had differentinsulin secretion kinetics to glimepiride [24] One flaw incurrent anti-diabetics is their decreasing efficacy over timeThe authors investigated the long term anti-diabetic effectof BPWE in dbdb mice BPWE reduced blood glucoseincreased blood insulin improved glucose tolerance andreduced the percentage of glycosylated hemoglobin (HbA1c)Both long-term and one-time experiments strongly supportthe anti-diabetic action of BPWE [24] In sharp contrast toglimepiride BPWE protected against islet atrophy in mousepancreas The investigators further evaluated anti-diabeticproperties of 3 B pilosa varieties B pilosa L var radiate(BPR) Bpilosa L var pilosa (BPP) and B pilosa L varminor(BPM) in dbdb mice [8] One single oral dose (10 50 and250mgkg body weight) of BPR BPP or BPM crude extractsdecreased postprandial blood glucose levels in dbdb micefor up to four hours and the reduction of glucose levels inthe blood appeared to be dose-dependent Comparing thethree variants BPR extract resulted in a higher reduction inblood glucose levels when administered at the same dose as

42 Evidence-Based Complementary and Alternative Medicine

Table 20 Antifungal activity of B pilosa [92]

Partextract Concentration (ppm) Strain InhibitionCortiicum rolfsii Fusarium solani Fusarium oxysporum

Leaves

Essentialoils

100 857 plusmn 09 682 plusmn 0 745 plusmn 17250 960 plusmn 08 779 plusmn 18 879 plusmn 04

AqueousExtracts

100 446 plusmn 17 605 plusmn 21 716 plusmn 07250 942 plusmn 03 689 plusmn 07 824 plusmn 19

Flowers

Essentialoils

100 604 plusmn 09 892 plusmn 04 869 plusmn 05250 894 plusmn 12 980 plusmn 03 949 plusmn 06

AqueousExtracts

100 331 plusmn 11 714 plusmn 07 573 plusmn 22250 661 plusmn 14 912 plusmn 0 900 plusmn 07

Table 21 Antifungal activity of B pilosa root extracts [93]

Strain LC50 (mgmL)Acetoneextracts

Methanolextracts

Waterextracts

Aspergillus niger 014 006 007Aspergillus flavus 1091 658 0Penicillium notatum 005 005 005

the other two varieties In terms of serum insulin levels a doseof 50mgkg of each extract was used and the BPR extracttogether with the three polyynes significantly increased theserum insulin level in dbdb mice Long-term experiments(28-day treatment) were then conducted using diabetic micewith postprandial glucose levels from 370 to 420mgdL andglimepiride was used as positive controlThe range of dosagesapplied was from 10mgkg BW to 250mgkg BW Resultsshowed that the positive control as well as the crude extractsof the three varieties lowered the blood glucose levels indbdb mice However only BPR extract containing a higherpercentage of cytopiloyne (53) reduced blood glucose levelsand augmented blood insulin levelsmore than BPP andBPMThe percentage of glycosylated hemoglobin A1c (HbA1c) wasalso measured and found to be 79 plusmn 05 in mice aged 10ndash12 weeks and 66 plusmn 02 61 plusmn 03 and 62 plusmn 03 inthe blood of age-matchedmice following treatment with BPRcrude extract (50mgkg) glimepiride (1mgkg) and com-pound 53 (05mgkg) respectively [8] Among the polyynesfound in B pilosa cytopiloyne was the most effective againstT2D Hence cytopiloyne was used for further study on anti-diabetic action and mechanism [125] The data confirmedthat cytopiloyne reduced postprandial blood glucose lev-els increased blood insulin improved glucose tolerancesuppressed HbA1c level and protected pancreatic islets indbdb mice Nevertheless cytopiloyne failed to decreaseblood glucose in streptoztocin (STZ)-treated mice whose bcells were already destroyed Additionally cytopiloyne dose-dependently increased insulin secretion and expression in bcells as well as calcium influx diacylglycerol and activationof protein kinase C120572 Collectively the mechanistic studiessuggest that cytopiloyne treats T2D via regulation of insulin

production involving the calciumDAGPKC120572 cascade in bcells

The studies detailed above point to the conclusion thatcytopiloyne and related polyynes (compounds 69 and 49)are anti-diabetics in animal models The data uncover a newbiological action of polyynes It should be noted that likeall anti-diabetic drugs cytopiloyne failed to prevent or curediabetes completely but reduced diabetic complications [125]Intriguingly 34 polyynes have been found in B pilosa so farIt remains to be seen whether all the polyynes present in thisplant have anti-diabetic activities

34 Antioxidant Activity Free radicals can damage cellularcomponents via a series of chemical reactions [89] leadingto development and progression of cardiovascular diseasecancer neurodegenerative diseases and ageing [70] Freeradicals nitric oxide (NO) and superoxide anions can beproduced in macrophages to kill microbes However anexcessive generation of the free radicals under pathologicalconditions is associated with a wide range of illnessesPlants are known to be rich in antioxidant phytochemicalsChiang and colleagues evaluated the free radical scaveng-ing activity of crude extract fractions and compounds ofB pilosa using 11-diphenyl-2-picrylhydrazyl (DPPH) andhypoxanthinexanthine oxidase assays [70] Using DPPHand hypoxanthinexanthine oxidase assays they found thatthe B pilosa crude extract and the ethyl acetate butanoland water fractions had free radical scavenging activityNine compounds Heptyl- 2-O-120573-xylofuranosyl-(1rarr 6)-120573-glucopyranoside (199) 3-O-Rabinobioside (124) Quercetin3-O-rutinoside (130) Chlorogenic acid (167) 34-Di-O-caffeoylquinic acid (169) 35-Di-O-caffeoylquinic acid (170)45-Di-O-caffeoylquinic acid (171) Jacein (119) and Cen-taurein (110) had DPPH radical scavenging activity [70]The IC

50values of the B pilosa crude extractfractions and

compounds are summarized in Tables 14 and 15 respectivelyMeasurement of free radical scavenging activities is one

way of assessing the antioxidant activities of B pilosa andits fractions and compounds It is interesting that the ethylacetate and butanol fractions are more active than the waterfraction and B pilosa crude extract [70] Of the secondarymetabolites only phenolic compounds 124 130 167 169 and171 showed significant DPPH-radical scavenging activities

Evidence-Based Complementary and Alternative Medicine 43

Further analysis of the structure-activity relationship of thecompounds suggested that substitution of the C3 hydroxylgroup with glycosides increased the activity approximately2-fold (for example in compounds 124 and 130) relative toquercetin (-OH in its C3) [70] Further modification of thestructures of the active compounds needs to be performed totest the effects of various substituents on activity The reasonwhy most of the antioxidant compounds contain phenolmoieties in their structure could be that the reduction-oxidation (redox) properties of phenols allow them to actas reducing agents singlet oxygen quenchers and hydrogendonors

A complementary study by Muchuweti and colleaguesdetermined phenolic content antioxidant activity and thephenolic profile of B pilosa methanol extract [89] Theyestimated that the phenolic content of the methanol extractof B pilosawas 11028plusmn22mgg [89] Vanillin hydroxyben-zaldehyde caffeic acid coumaric acid and ferulic acid werefound in this extractThe B pilosa extract also showed DPPHradical scavenging activity Furthermore the antioxidantactivity of the flavonoids found in B pilosa was correlatedwith its hepatoprotective effects through their inhibitionof NF-120581B activation which may lessen the oxidative stresscaused by the production of free radicals during liver injury[60]This activity might also be due to the anti-inflammatoryeffects of the aqueous extracts of B pilosa aerial parts on theinhibition of COX-2 and PGE

2production [88]

Essential oils from B pilosa flowers and leaves are alsoreported to possess antioxidant activity With the aim ofreplacing chemically synthesized additives Deba and col-leagues [92] worked on the antioxidant antibacterial andantifungal activities of essential oils and water extracts of Bpilosarsquos leaves and flowers Table 16 summarizes the resultsobtained from DPPH free radical scavenging assay

It can be inferred fromTable 16 that essential oils from theleaves possessed the highest activity It is reported elsewherethat monoterpenes present in essential oils such that of Bpilosa have protective effects and antioxidant properties [92]Beta-carotene bleaching method was also performed Leavesessential oils and aqueous extracts of the leaves and flowersshowed higher activity than the flower essential oils This isdue to the volatility of the flower essential oils The activityexhibited by the aqueous extracts was accounted to thepresence of phenolic compounds that are reported to donatea hydrogen atom to free radicals such that the propagation ofthe chain reaction during lipid oxidation is terminated [92]Overall essential oils and phenolics present in B pilosa canbe thought of as major antioxidant compounds

35 Immunomodulatory Activity B pilosa is thought to bean immunomodulatory plant and is reported to be effectivein the treatment of immune disorders such as allergy [75]arthritis [75] and T1D [48 50 53]

As pointed out in the discussion of its anti-diabeticactivities (Section 33) a combination of phytochemicalsmethod and T-cell activation assays was used to studyimmunomodulatory properties of B pilosa IFN-120574 is a keycytokine released by T and NK cells that mediates immune

cells and sustains immunity against pathogens Defects inIFN-120574 expression regulation and activation result in vulner-ability to diseases caused by bacteria and viruses [69] Anelegant study performed byChang and colleagues using IFN-120574 promoter-driven luciferase reporter construct in JurkatT cells showed that hot water crude extracts of B pilosaincreased IFN-120574 promoter activity two-fold [69] Out of thesubfractions of this extract the butanol fraction but not thewater or ethyl acetate fractions increased IFN-120574 promoteractivity six-fold Centaurein (110) and centaureidin (109)were identified from the butanol fraction and were stated tocause a four-fold increase in IFN-120574 promoter activity withEC50

values of 75 120583gmL and 09120583gmL respectively Themechanism of action of centaurein was determined usingtranscription factors such as AP-1 NFAT and NF120581B whichare reported to bind to IFN-120574 promoter and regulate IFN-120574transcription Unlike with the activity of the positive controlPHA centaurein caused a four-fold increase in NFAT a3-fold increase in NF120581B and had little if any effect onAP-1 enhancer activities (23-fold three-fold and ten-foldincreases respectively were seenwith PHA) [69]The authorsconcluded that centaureinmodulates IFN-120574 expression by theNFAT and NF120581B pathways The article only determined themechanism of action of centaurein Its aglycone centaureidinmay act through the samemechanism though this conclusionneeds to be further verified

B pilosa extract and its compounds are reported to inhibitdifferentiation of naıve CD4+ helper T (Th0) cells into Th1cells [49] Using theThcell differentiation assay as a screeningplatform 3 polyynes 2-120573-D-glucopyranosyloxy-1 -hydroxytrideca-57911-tetrayne (53) 3-120573-D-glucopyranosyl-1-hydroxy-6(E)- tetradecene-81012-triyne (69) and 2-120573-D-glucopyranosyloxy-1-hydroxy-5(E)- tridecene-7911-triyne(49) were discovered from B pilosa [49 53] The data showsthat cytopiloyne and other two polyynes suppressed thedifferentiation of type 1 helper T (Th1) cells and productionof Th1 cytokines and promoted that of type 2 helper T (Th2)cells and production of Th2 cytokines thus explaining theimmunomodulatory and anti-inflammatory effects of Bpilosa and its polyynes

Chang and colleagues were the first to report the effect ofthe butanol extract of B pilosa on the autoimmune diabetesand airway inflammation inmice [48] Imbalance in the levelsofTh1 andTh2 and of various cytokines leads to autoimmunediseases T1D and other autoimmune diseases (rheumatoidarthritis Crohnrsquos disease among others) are exacerbated byan increase Th1 levels (specifically CD4+ Th1 cells) whileTh2 cells antagonize this effect [19] Moreover Th2 cellsmediate asthma in ovalbumin-induced hypersensitivity inBALBc mice [48] In their work [48] Chang and colleaguesshowed that 10mgkg butanol extracts with 15 (ww)compound 69 and 11 (ww) compound 49 (Table 11)ameliorated the development of Th1-mediated diabetes inNOD mice through inhibition of 120573 cell death and leukocyteinfiltration At the same dosage the butanol extracts alsoexacerbated ovalbumin-induced pulmonary inflammationin BALBc mice with an increase in the infiltration ofeosinophils and mast cells into the airway of the mice [48]Despite the different outcomes both mouse models proved

44 Evidence-Based Complementary and Alternative Medicine

the concept that control over the Th1Th2 shift is associatedwith autoimmune diseases and that the B pilosa butanolextract can shift the differentiation of Th0 cells to Th2 cells[48 49]

An extended study presented by Chiang and colleaguesshowed that compound 53 modulates T-cell functions [50]Using CD4+ T cells from BALBc mice they demonstratedthat cytopiloyne decreased levels of IFN-120574 producing cells(Th1) by 122 (from 72 to 598) Since Th1 and Th2 celldifferentiation is antagonistic it was expected that compound53 increased the percentage of mouse IL-4 producing cells(Th2) by 72 (from 237 to 309) Subsequent assess-ment of effect of compound 53 on the modulation of thetranscription of IL-4 and IFN-120574 showed that cytopiloyne asexpected decreased the splenocyte levels of IFN-120574 mRNAand increasing that of IL-4 in a dose-dependent manner Inthe range of 01 to 3 120583gmL of compound 53 the effects werenot attributed to its cytotoxicity Consequently using 3120583gmLcytopiloyne for 72 and 96 hours the protein concentration ofIFN-120574 decreased to 186 and 444 respectively Under thesame conditions cytopiloyne increased IL-4 concentrationsto 1985 and 2470 (for 72 and 96 hours resp) Thismodulation of T-cell differentiation exhibited by compound53 was used to explain its anti-diabetic activity [50]

The anti-diabetic role of cytopiloyne was extensivelydiscussed above (Section 33 anti-diabetic activity) Themolecular basis of the regulation of cytokine expression bycytopiloyne has been described Cytopiloyne directly elevatedthe expression level of IL-4 via GATA-3 upregulation inT cells [53] However reduction of IFN-120574 expression in Tcells seemed to come from the indirect opposing effect IL-4cytokine because the expression level of T-bet was unaltered[53] In this way cytopiloyne skewedTh1 polarization into theTh2 state conferring protection against T1D in NOD miceAside frompolarization ofThcell differentiation cytopiloynealso activated the expression of Fas ligand in pancreaticb cells this increase leading to the partial depletion of Tcells and reduction of immune response in local areas suchas the pancreas Of note cytopiloyne also inhibited T-cellproliferation and activation By targeting T cells from threeimmunomodulatory actions cytopiloyne protects againstT1D and probably other Th1-mediated autoimmune diseases[19]

The phytochemical constituents of B pilosa exert theirfunctions on different immune cells to modulate immuneresponse It is possible that some of the compounds mayhave agonistic or antagonistic effects on immune responseImmune function of B pilosa may depend on its compo-sition and amount of compounds which could explain theapparently conflicting report of B pilosa butanol extractaggravating allergy in mice [48] while cellulosine-treatedextract ameliorated allergy [48 75] IFN-120574 promoter reporterassays and T-cell differentiation assays were used to isolate2 flavonoids [69] and 3 polyynes [49 50] as immunomod-ulatory compounds from the butanol fraction of B pilosaInterestingly the flavonoids promote IFN-120574 expression inNK and T cells In marked contrast the polyynes promoteIL-4 expression and indirectly inhibit IFN-120574 expression indifferentiating T cells Sensitivity appears to be the key to

identifying structure- and bioactivity-related phytochemicalsfrom medicinal plants

36 Antimalarial Activity The use of chemical drugs againstpathogens has resulted in drug-resistant mutants Examplesof drug resistance can be found in the species of thePlasmodium that cause malaria It is important to search fornew compounds to combat Plasmodium parasites [47] Astudy of the anti-malarial activity of the leaf extracts of Bpilosa using a combination of phytochemistry and bioassaysshowed that compound 49 (Table 11) showed activity againsta malaria parasite (P falciparum NF54 strain) with an IC

50

value of 60 120583gmL [77] In addition compound 49 isolatedfrom the aerial parts of B pilosa inhibited growth of the Pfalciparum FCR-3 strain with an IC

50value of 035 120583gmL

This compound was tested for its in vivo effect in miceinfected with P berghei NK-65 strain Results showed thatcompound 49 decreased the average parasitemia in the redblood cells by 207 (from 328 of that of the control to121) after an intravenous injection of 08mgkg BWday forfour days [47] Further studies addressing the anti-malarialmechanism underlying both polyynes and clinical studies areneeded

37 Antibacterial Activity Emergence of multiple antibiotic-resistantmicrobes is becoming a global threat to public healthand a challenge to disease treatment For instance penicillinis commonly used to combat a food-borne intracellularbacteriumListeria however penicillin-resistant bacteria havebeen discovered recently [69] Chang and coworkers iso-lated centaurein (110) and centaureidin (109) from B pilosaextract [69 70] Centaurein enhances expression of IFN-120574a key cytokine for macrophage activation and consequentlyenhances bactericidal activity in macrophages [69 70] Inagreement with observed in vitro effects centaurein wasreported to prevent and treat Listeria infection in C57BL6Jmice [91] Mechanistic studies confirmed that centaureinexerted antilisterial action via IFN-120574 expression in wild-typemice but not IFN-120574 knockout mice [91] Further in vitrostudies on centaurein showed that this compound increasedIFN-120574 expression by 13 (from 17 to 20) 20 (from 21to 41) and 11 (from 6 to 17) in CD4+ T cells CD8+Tcells and NK cells respectively That is to say there wasan increase in IFN-120574 producing immune cells As expectedcentaurein also enhanced the expression level of T-bet a keynuclear factor for IFN-120574 expression Consistently centaureinaugmented the serum IFN-120574 levels inC57BL6Jmice and thisaugmentation peaked 24 hours after compound injectionThe quantity of mouse serum IFN-120574 was sufficient to activatemacrophages in vitro and eradicated GFP-producing Listeriainside macrophages However the entry of Listeria intomacrophages was not affected by centaurein-treated mousesera Centaurein treatment at 20120583g per mouse rescued 30of the mice infected with a lethal dose of Listeria (2 times106 CFU) It is noteworthy that in the presence of ampicillin(5 120583gmouse) centaurein (20 120583gmouse) rescued 70 of themice suggesting an additive effect between ampicillin andcentaurein [91] Despite lower abundance centaureidin was

Evidence-Based Complementary and Alternative Medicine 45

30 times more active than centaurein in terms of IFN-120574production [91]

Aside from the indirect antibacterial action mentionedabove extract andor compounds of B pilosa also showeddirect bacteriostatic andor bactericidal action One studyreported that essential oils and leafflower extracts of Bpilosa could suppress the growth of gram positive and gramnegative bacteria as evidenced by zone of inhibition assaysIn this study antibacterial activity of the essential oils fromthe leaves and flowers of B pilosa was determined in anattempt to identify natural products as food preservatives forprevention of microbial multiplication and food oxidationThe essential oils and extracts of B pilosa leaves and flow-ers showed moderate but different extents of antibacterialactivity (Table 17) In general essential oils had higherantibacterial activity than crude extracts One explanation forthis could be that monoterpenes in the essential oils destroycellular integrity and subsequently inhibit the respirationand ion transport processes The presence of antibacterial120573-caryophyllene could be another explanation as reportedelsewhere [92] Another study reported that the methanoland acetone extract of B pilosa roots displayed antibacterialactivities against the bacteria listed in Table 18 [93] andmethanol extracts from the roots seemed to be the mosteffective

Another study indicated that the polyyne (R)-12-dihydroxytrideca-357911-pentayne (49) from this plantalso suppressed bacterial growth as shown by the minimuminhibitory concentration required to inhibit 50 bacterialgrowth (MIC

50) in Table 19 This compound was highly

effective against several Gram positive and Gram negativebacteria including the drug-resistant bacteria Staphylococcusaureus N315 (MRSA) and Enterococcus faecalis NCTC12201(VRE) [47] Strikingly compound 49 had a similar MIC

50

value to antibiotics (ampicillin tetracycline norfloxacin andamphotericin B) in most of the bacteria tested

Antibacterial activity of B pilosa extracts and compo-nents expressed as MIC

50and the mean zone of inhibition

is tabulated in Tables 17 18 and 19The zone of inhibition forAmpicilline (positive control) ranges from 153 plusmn 03mm to443 plusmn 02mm [92]

38 Antifungal Activity B pilosa has traditionally been usedto treat microbial infection Recently different parts of Bpilosa have been tested for antifungal activities Deba andcolleagues first evaluated the antifungal effect of the hotwater extracts of the B pilosa roots stems and leaves againstCorticium rolfsii Fusarium solani and Fusarium oxysporumThey discovered that C rolfsii was most suppressed bytreatment with B pilosa as its growth was reduced at almostall the tested doses followed by F oxysporum and F solani[99] However the fungicidal activities of the stems androots were greater than the leaves [99] Moreover the samegroup assessed the antifungal activity of the essential oilsand aqueous extracts from B pilosa flowers and leaves [92]They showed that the extracts and oils had antifungal activityagainst C rolfsii F solani and F oxysporum Essential oils

appeared to have better fungicidal activity thanwater extractsas summarized in Table 20

Another study by Ashafa and colleagues showed thatacetone methanol and water extracts of the B pilosa rootsshowed antifungal activities against Aspergillus niger Aflavus and Penicillium notatum using the agar dilutionmethod The results are tabulated in Table 21 [93] Negativecontrols showed 0 growth inhibitionThe methanol extractof the B pilosa roots at 10mgmL was also effective againstCandida albicans [93]Of note B pilosa obtained fromPapuaNew Guinea had no activity against A niger and C albicans[126] but the SouthAfrican (EasternCape) ecotype exhibitedmoderate activity against C albicans [93] This discrepancymay depend on extraction solvents extraction procedureassay techniques different plant parts and abundance ofactive compounds

B pilosa produces a variety of secondary metabolitessuch as flavonoids phenylacetylenes alkaloids sterols ter-penoids and tannis [92 93] However none of them havebeen confirmed as active compounds against fungi Furtherinvestigation of active compounds from B pilosa is necessaryto further understand the antifungal efficacy of this plant

39 Hypotensive and Vasodilatory Activities In early studiesDimo and colleagues used three rat models normotensiveWistar rats (NTR) salt-loading hypertensive rats (SLHR)and spontaneous hypertensive rats (SHR) to investigate thehypotensive effect of the methanol crude extract of B pilosaleaves [28 127] The extract lowered systolic blood pressurein hypertensive rats (SLHR and SHR) to a greater degreethan NTR [28 127] In addition a decrease in urinarysodium ions and an increase in urinary potassium ions wereobserved after treatment with the methanol extract of Bpilosa leaves although neither differences were statisticallysignificant [28 127] Taking the data together the studyproposed that B pilosa leaf extract reduced blood pressurevia vasodilation [28 127] The same group continued totest the antihypertensive effect of aqueous and methylenechloride extracts of B pilosa leaves in a hypertensive ratmodel [28 127] To establish a fructose-induced hypertensionmodel male Wistar rats were given 10 fructose solutionto drink ad libitum for three weeks In addition to freeaccess to 10 fructose the rats were treated with the aqueous(150mgkg) or methyl chloride (350mgkg) extracts of Bpilosa for additional three weeks [28 127] Both extracts ofB pilosa leaves had a hypotensive effect on rats Howeverneither extracts reversed the elevation of serum insulin infructose-fed rats Therefore B pilosa lowered blood pressureirrespective of insulin [28 127] To better understand thehypotensive mechanism the authors investigated the effectof a neutral extract of B pilosa (NBP) a mixture of methanolandmethylene chloride (1 1) extract after neutralization withNaOH and HCl on the heart and the blood pressure ofNTR and SHR [110] This study showed that an intravenousinjection of the NBP resulted in a biphasic reduction insystolic blood pressure In addition one intravenous dose ofthe extract at 10 20 and 30mgkg BW decreased systolicblood pressure in normal rats by 183 425 and 30

46 Evidence-Based Complementary and Alternative Medicine

respectively and the same doses reduced the blood pressurein hypertensive rats by 258 389 and 286 respectivelyOnly the highest dose (30mgkg) affected the force of thecontraction of the heart Atropine and propranolol were usedto interfere with the hypotensive action of the NBP Atropinereduced the initial phase of the hypotensive response in NBPand completely abolished the second phase of hypotensiveresponse in NBP In contrast propranolol increased thefirst hypotensive response but partially abolished the secondhypotensive response provoked by the NBP [110]This mech-anistic study suggested that B pilosa invokes the biphasichypotensive responses via targeting cardiac pump efficiencyduring the first phase and vasodilation at the second phase[110]

A further study was performed to investigate the relaxingeffect of a neutral extract of B pilosa (NBP) on rat aorta con-tractedwithKCl (60mM) andnorepinephrine (01mM) [94]Cumulative addition of NBP relaxed the rat aorta previouslycontracted by KCl in a dose-dependent manner The EC

50

value of theNBP for vasorelaxationwas 032mgmLThedataalso showed that the NBP reduced the contraction of aortapreviously contracted by KCl irrespective of the presence ofaortic endothelium [94]

Pretreatment with glibenclamide an ATP-dependent K+channel blocker did not considerably affect the relaxant effectof the NBP on KCl-induced contraction suggesting thatthe vasodilatory effect of B pilosa was not related to theopening of this ATP-dependent K+ channel [94] On theother hand in the presence of indomethacin or pyrilaminemaleate the relaxant response induced by the plant extractwas significantly inhibited at the lower concentrations Theplant extract was able to reduce the aorta resting tone inhibitthe KCl-induced contractions by 90 at 15mgmL and theCaCl2-induced contractions by 95 at 075mgmL These

results demonstrate that B pilosa can act as a vasodilatorprobably via acting as a calcium antagonist [94]

However no specific compound for the above activityhas been identified from B pilosa to date A bioactivity-guided identification approach may be adopted to identifythe active compounds in B pilosa that possess hypotensiveand vasodilatory effects and understand their mechanism ofaction

310 Wound Healing Activity B pilosa has been traditionallyused to treat tissue injury inCameroon Brazil andVenezuela[26] Hassan and colleagues investigated the wound healingpotential of B pilosa in Wistar rats [128] Mirroring thepositive control neomycin sulfate the ethanol extract of Bpilosa had faster wound closure than control rats 3 6 and 9days after topical application Histological examination alsorevealed better collagenation angiogenesis and organizationof wound tissue seven days after application Epithelializationand total healing time in B pilosa-treated rats were compara-ble to those of neomycin sulfate Together these data suggestthat B pilosa may be a viable alternative to neomycin lotionfor the treatment of wounds

In addition to studying the wound healing effect of Bpilosa on external ulcers Tan and colleagues also examined

the effect of methanol cyclohexane and methyl chlorideextracts of B pilosa on gastric ulcers in Wistar rats fedwith 1mL HClethanol gastric necrotizing solution (150mMHCl in 60 ethanol) and macroscopically visible lesionswere scored [26] Among the three extracts methylene chlo-ride extracts exhibited the highest activity showing 464inhibition of lesion formation at a dose of 500mgkg BWand complete inhibition at 750mgkg [26] The efficacy ofthe ethylene chloride extract was followed by that of themethanol extracts which had inhibition ranging from 304to 822 at concentrations of 500mgkg and 1000mgkgBW respectively [26] The cyclohexane extracts showed thelowest activity against gastric ulcers in rats with 13340 and 797 inhibition at 500 750 and 1000mgkg BWrespectively [26] To better understand the mode of actionof the methylene chloride extract of B pilosa rats werepretreated with indomethacin a COX-2 inhibitor involved inprostaglandin synthesis Pretreatment significantly reducedthe protection against HClethanol-induced ulcers to 313inhibition at 750mgkg BW suggesting a link between theantiulcerative activity of B pilosa and prostaglandin syn-thesis Unexpectedly the methylene chloride extract of Bpilosa showed little gastric mucosal protection against gastriclesions induced by 95 ethanol (1mL) [26] Absolute alcoholis known to cause mucosalsubmucosal tissue destruction viacellular necrosis and the release of tissue-derived mediators(histamine and leukotrieneC4)Thus these data imply thatBpilosa did not prevent the generation or the necrotic action ofthese mediators on the gastric microvasculature In additionpylorus ligation can increase gastric acid secretion withoutan alteration of mucosal histamine content The methylenechloride extract of B pilosa did not possess antisecretoryactivity On the contrary it was observed that increases in thedose of the extract led to elevated gastric juice acidity [26]Results with both absolute ethanol and pylorus ligation ratmodels suggested the possibility that the ineffectiveness of Bpilosa against gastric ulcers was due to lack of antihistaminicactivity in the plant In summary overall the data suggest thatB pilosa protects against HCLethanol-mediated ulcers viainhibition of prostaglandin biosynthesis

Previous phytochemical studies showed that a groupof flavonoids acyclichalcones are present in B pilosa[129 130] and the chalcones were proposed to have anti-ulcerative activity [131] Moreover nine hydroxychalcones 10were reported to possess gastric cytoprotective effects with24-dihydroxychalcone being the most active [132] Sincemethylene chloride extracts appear to be the most active Bpilosa extracts next the specific anti-ulcerative phytochemi-cals in the methylene chloride extracts of B pilosa and theirmodes of action needs to be probed

Despite the claims listed inTable 3 relatively few scientificstudies have been conducted in vitro and in vivo to address thetraditional ethnomedical uses of B pilosa Information aboutthe use of B pilosa as a botanical therapy recorded so far isfar from complete Studies conducted thus far only serve asa starting point for further investigation of B pilosa and theultimate efficacious use of the herb in clinical applications

Evidence-Based Complementary and Alternative Medicine 47

4 Toxicology

Despite its use as an ingredient in food for human con-sumption studies on systemic toxicity (eg acute subacutechronic and subchronic toxicities) of B pilosa in humansand animals are still inadequate and insufficient So faracute andor subchronic toxicities have been evaluated inrats and mice Oral acute and 28-day toxicities of waterextract of B pilosa leaves were evaluated in Wistar rats[133] An oral dose of water extract of B pilosa leaves at10 gkg BW showed no obvious mortality or changes inthe appearance in rats [134] The same extract at 08 gkgBWday once a day showed no obvious sub-chronic toxicityin rats over 28 days as measured by survival rate bodyweight and gross examination of organs [134] These dataare consistent with our data indicating that oral deliveryof the water extract of the B pilosa whole plant at 1 gkgBWday once a day is safe in rats over 28 days (unpublisheddata) Taken together these studies suggest that ingestion ofB pilosa aqueous extract at up to at 1 gkg BWday oncea day is highly safe in rats In addition the acute toxicityof aqueous and ethanol extracts of B pilosa in mice havebeen reported [134] Five- to six-week-old mice with weightsbetween 28 and 35 g received a peritoneal injection of bothextracts at the different doses The LD

50 the dose that causes

50 lethality of the aqueous and ethanol extracts in micewas 1230 gkg BW and 615 gkg BW respectively [134] Acomplete toxicological study has not been completed forhumans Furthermore the drug interactions of B pilosa withother drugs are unknown Further safety verification andclinical trials should be performed before B pilosa can beconsidered for medicinal use

5 Conclusions

B pilosa is an erect perennial plant with green leaves whiteor yellow flowers and tiny black seeds As it is distributedworldwide and is widely used as a folk remedy B pilosacan be thought of as an extraordinary source of food andmedicine However a comprehensive up-to-date review ofresearch on B pilosa has hitherto been unavailable In thisarticle scientific studies on B pilosa have been summarizedand critically discussed from the perspectives of botanyethnomedicine phytochemistry pharmacology and toxicol-ogy B pilosa is claimed to treat more than 40 disordersand 201 compounds have been identified from this plantThe medicinal utility of B pilosa and its modes of actionin relation to its known phytochemicals were discussedherein Polyynes flavonoids phenylpropanoids fatty acidsand phenolics are the primary bioactive compounds of Bpilosa and they have been reported to be effective in thetreatment of tumors inflammationimmune modulationdiabetes viruses microbes protozoans gastrointestinal dis-eases hypertension and cardiovascular diseases Cautionshould be exercised in the therapeutic use of B pilosa forhypoglycemia hypotension bleeding and allergy

Acknowledgments

The authors thank their laboratory members for construc-tive suggestions and the authors whose publications theycited for their contributions The authors also thank MsMiranda Loney of Academia Sinica for English editing of thispaper This work was supported by Grants 99-CDA-L11 and101S0010056 from Academia Sinica Taiwan Arlene P Bar-tolome is a recipient of MECO-TECO Sandwich ScholarshipProgram between Taiwan and the Philippines

References

[1] T Shen G H Li X N Wang and H X Lou ldquoThe genusCommiphora a review of its traditional uses phytochemistryand pharmacologyrdquo Journal of Ethnopharmacology vol 142 no2 pp 319ndash330

[2] C Long P Sauleau B David et al ldquoBioactive flavonoids ofTanacetum parthenium revisitedrdquo Phytochemistry vol 64 no2 pp 567ndash569 2003

[3] P O Karis and O Ryding Asteraceae Cladistics and Classifica-tion Bremer K Eds pp 559ndash569 Timber press Portland OreUSA 1994

[4] O N Pozharitskaya A N Shikov M N Makarova et alldquoAnti-inflammatory activity of a HPLC-fingerprinted aqueousinfusion of aerial part of Bidens tripartita LrdquoPhytomedicine vol17 no 6 pp 463ndash468 2010

[5] F Q Oliveira V Andrade-Neto A U Krettli and MG L Brandao ldquoNew evidences of antimalarial activity ofBidens pilosa roots extract correlated with polyacetylene andflavonoidsrdquo Journal of Ethnopharmacology vol 93 no 1 pp 39ndash42 2004

[6] Agriculture USDA ldquoPlants databaserdquo 2013in Natural Resources Conservation Servicehttpwwwnrcsusdagovwpsportalnrcssitenationalhome

[7] Y Zhou and X Z Yan ldquoExperimental study of qi deficiencysyndrome and Codonopsis pillosulae and Astragalus injectionon the immune response in micerdquo Zhong Xi Yi Jie He Za Zhivol 9 no 5 pp 286ndash262 1989

[8] S C Chien P H Young Y J Hsu et al ldquoAnti-diabeticproperties of three common Bidens pilosa variants in TaiwanrdquoPhytochemistry vol 70 no 10 pp 1246ndash1254 2009

[9] M J Alcaraz and M J Jimenez ldquoFlavonoids as anti-inflammatory agentsrdquo Fitoterapia vol 59 no 1 pp 25ndash38 1988

[10] FAO Agriculture Food and Nutrition for AfricamdashA ResourceBook for Teachers of Agriculture Publishing ManagementGroup FAO Information Division Rome Italy 1997

[11] M B Rokaya Z Munzbergova B Timsina and K R BhattaraildquoRheum australe D Don a review of its botany ethnobotanyphytochemistry and pharmacologyrdquo Journal of Ethnopharma-cology vol 141 no 3 pp 761ndash774 2012

[12] P H Young Y J Hsu and W C Yang ldquoBidens pilosa L andits medicinal userdquo in Recent Progress in Medicinal Plants DrugPlant II A S AwaadVK Singh and JNGovil Eds StandiumPress Houston Tex USA 2010

[13] C Ge ldquoCytologic study of Bidens bipinnata LrdquoZhongguo ZhongYao Za Zhi vol 15 no 2 pp 72ndash125 1990

[14] L C Chiang J S Chang C C Chen L T Ng and C CLin ldquoAnti-herpes simplex virus activity of Bidens pilosa and

48 Evidence-Based Complementary and Alternative Medicine

Houttuynia cordatardquoAmerican Journal of ChineseMedicine vol31 no 3 pp 355ndash362 2003

[15] N P Rybalchenko V A Prykhodko S S Nagorna et alldquoIn vitro antifungal activity of phenylheptatriyne from Bidenscernua L against yeastsrdquo Fitoterapia vol 81 no 5 pp 336ndash3382010

[16] K Redl W Breu B Davis and R Bauer ldquoAnti-inflammatoryactive polyacetylenes from Bidens campylothecardquo Planta Med-ica vol 60 no 1 pp 58ndash62 1994

[17] M Ayyanar and S Ignacimuthu ldquoTraditional knowledge ofKani tribals inKouthalai of Tirunelveli hills TamilNadu IndiardquoJournal of Ethnopharmacology vol 102 no 2 pp 246ndash255 2005

[18] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant intraditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[19] AKAbbas KMMurphy andA Sher ldquoFunctional diversity ofhelper T lymphocytesrdquo Nature vol 383 no 6603 pp 787ndash7931996

[20] J H Cano and G Volpato ldquoHerbal mixtures in the traditionalmedicine of Eastern Cubardquo Journal of Ethnopharmacology vol90 no 2-3 pp 293ndash316 2004

[21] J Namukobe J M Kasenene B T Kiremire et al ldquoTraditionalplants used for medicinal purposes by local communitiesaround the Northern sector of Kibale National Park UgandardquoJournal of Ethnopharmacology vol 136 no 1 pp 236ndash245 2011

[22] C Lans ldquoComparison of plants used for skin and stomachproblems in Trinidad and Tobago with Asian ethnomedicinerdquoJournal of Ethnobiology andEthnomedicine vol 3 article 3 2007

[23] S Dharmananda ldquoA Popular Remedy Ecapes Notice ofWesternPractitionersrdquo 2013 httpwwwitmonlineorgartsbidenshtm

[24] Y J Hsu T H Lee C L T Chang Y T Huang andW C YangldquoAnti-hyperglycemic effects and mechanism of Bidens pilosawater extractrdquo Journal of Ethnopharmacology vol 122 no 2 pp379ndash383 2009

[25] E Noumi F Houngue and D Lontsi ldquoTraditional medicinesin primary health care plants used for the treatment ofhypertension in Bafia Cameroonrdquo Fitoterapia vol 70 no 2 pp134ndash139 1999

[26] P V Tan T Dimo and E Dongo ldquoEffects of methanolcyclohexane and methylene chlo ride extracts of Bidens pilosaon various gastric ulcer models in ratsrdquo Journal of Ethnophar-macology vol 73 no 3 pp 415ndash421 2000

[27] R L C Pereira T Ibrahim L Lucchetti A J R Da Silva and VL G De Moraes ldquoImmunosuppressive and anti-inflammatoryeffects of methanolic extract and the polyacetylene isolatedfrom Bidens pilosa Lrdquo Immunopharmacology vol 43 no 1 pp31ndash37 1999

[28] T Dimo J Azay P V Tan et al ldquoEffects of the aqueous andmethylene chloride extracts of Bidens pilosa leaf on fructose-hypertensive ratsrdquo Journal of Ethnopharmacology vol 76 no 3pp 215ndash221 2001

[29] CWiartMedicinal Plants of Southeast Asia Pelanduk SelabgorDarul Ehsan Malaysia 2000

[30] F L Silva D C H Fischer J F Tavares M S Silva P FDe Athayde-Filho and J M Barbosa-Filho ldquoCompilation ofsecondary metabolites from Bidens pilosa LrdquoMolecules vol 16no 2 pp 1070ndash1102 2011

[31] A H Chen S R Lin and C H Hong ldquoPhytochemical studyon Bidens pilosa L varminorrdquo Huaxue pp 38ndash42 1975

[32] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[33] M C Zulueta M Tada and C Y Ragasa ldquoA diterpene fromBidens pilosardquo Phytochemistry vol 38 no 6 pp 1449ndash14501995

[34] J Wang H Yang Z W Lin and H D Sun ldquoFlavonoids fromBidens pilosa var radiatardquo Phytochemistry vol 46 no 7 pp1275ndash1278 1997

[35] A Zhao Q Zhao L Peng et al ldquoA new chalcone glycoside fromBidens pilosardquo Acta Botanica Yunnanica vol 26 no 1 pp 121ndash126

[36] P Gao Y Wu S Cui and Y Zou ldquoSynthesis and absoluteconfiguration of the 7-phenylhepta-4 6-diyne-1 2-diol isolatedfrom Bidens pilosardquo Synthesis no 13 pp 2131ndash2135 2011

[37] S Wang B Yang D Zhu D He and L Wang ldquoActivecomponents of Bidens pilosa Lrdquo Zhongcaoyao vol 36 pp 20ndash21

[38] F Bohlmann T Burkhardt and C Zdero Naturally OccuringAcetylenes Academic Press New York NY USA 1973

[39] T M Sarg A M Ateya N M Farrag and F A AbbasldquoConstituents and biological activity of Bidens pilosa L grownin EgyptrdquoActa Pharmaceutica Hungarica vol 61 no 6 pp 317ndash323 1991

[40] H A L Valdes and H P Rego ldquoBidens pilosa Linnerdquo RevistaCubana de Plantas Medicinales vol 6 pp 28ndash33

[41] F Bohlmann H Bornowski and K M Kleine ldquoNew polyynesfrom the tribe Heliantheaerdquo Chemische Berichte vol 97 pp2135ndash2138

[42] RWang Q XWu and Y P Shi ldquoPolyacetylenes and flavonoidsfrom the aerial parts ofBidens pilosardquo PlantaMedica vol 76 no9 pp 893ndash896 2010

[43] L W Wu Y M Chiang H C Chuang et al ldquoA novelpolyacetylene significantly inhibits angiogenesis and promotesapoptosis in human endothelial cells through activation of theCDK inhibitors and caspase-7rdquo Planta Medica vol 73 no 7 pp655ndash661 2007

[44] L W Wu Y M Chiang H C Chuang et al ldquoPolyacetylenesfunction as anti-angiogenic agentsrdquo Pharmaceutical Researchvol 21 no 11 pp 2112ndash2119 2004

[45] H L Yang S C Chen N W Chang et al ldquoProtection fromoxidative damage using Bidens pilosa extracts in normal humanerythrocytesrdquo Food and Chemical Toxicology vol 44 no 9 pp1513ndash1521 2006

[46] H Q Wang S J Lu H Li and Z H Yao ldquoEDTA-enhancedphytoremediation of lead contaminated soil by Bidens maxi-mowiczianardquo Journal of Environmental Sciences vol 19 no 12pp 1496ndash1499 2007

[47] S TobinagaMK SharmaWG L Aalbersberg et al ldquoIsolationand identification of a potent antimalarial and antibacterialpolyacetylene from Bidens pilosardquo Planta Medica vol 75 no 6pp 624ndash628 2009

[48] C L T Chang H K Kuo S L Chang et al ldquoThe distincteffects of a butanol fraction of Bidens pilosa plant extract onthe development of Th1-mediated diabetes and Th2-mediatedairway inflammation in micerdquo Journal of Biomedical Sciencevol 12 no 1 pp 79ndash89 2005

[49] S L Chang C L T Chang Y M Chiang et al ldquoPolyacetyleniccompounds and butanol fraction from Bidens pilosa can mod-ulate the differentiation of helper T cells and prevent autoim-mune diabetes in non-obese diabetic micerdquo Planta Medica vol70 no 11 pp 1045ndash1051 2004

Evidence-Based Complementary and Alternative Medicine 49

[50] Y M Chiang C L T Chang S L Chang W C Yang and LF Shyur ldquoCytopiloyne a novel polyacetylenic glucoside fromBidens pilosa functions as a T helper cell modulatorrdquo Journal ofEthnopharmacology vol 110 no 3 pp 532ndash538 2007

[51] R PUbillas CDMendez S D Jolad et al ldquoAntihyperglycemicacetylenic glucosides from Bidens pilosardquo Planta Medica vol66 no 1 pp 82ndash83 2000

[52] L Alvarez S Marquina M L Villarreal D Alonso E Arandaand G Delgado ldquoBioactive polyacetylenes from Bidens pilosardquoPlanta Medica vol 62 no 4 pp 355ndash357 1996

[53] C L T Chang S L Chang Y M Lee et al ldquoCytopiloyne apolyacetylenic glucoside prevents type 1 diabetes in nonobesediabetic micerdquo Journal of Immunology vol 178 no 11 pp 6984ndash6993 2007

[54] G Kusano A Kusano and Y Seyama Novel Hypoglycemicand Anti-Inflammatory Polyacetylenic Compounds Their Com-positions Bidens Plant Extract Fractions and CompositionsContaining the Plant or Fraction JPO Tokyo Japan 2004

[55] M G L Brandao A U Krettli L S R Soares C G CNery and H C Marinuzzi ldquoAntimalarial activity of extractsand fractions from Bidens pilosa and other Bidens species(Asteraceae) correlated with the presence of acetylene andflavonoid compoundsrdquo Journal of Ethnopharmacology vol 57no 2 pp 131ndash138 1997

[56] A U Krettli V F Andrade-Neto M D G L Brandao andW M S Ferrari ldquoThe search for new anti-malarial drugs fromplants used to treat fever and malaria or plants ramdomlyselected a reviewrdquoMemorias do Instituto Oswaldo Cruz vol 96no 8 pp 1033ndash1042 2001

[57] C K Lee ldquoThe low polar constituents from Bidens pilosa L varminor (blume) sherffrdquo Journal of the Chinese Chemical Societyvol 47 no 5 pp 1131ndash1136 2000

[58] C K Wat R K Biswas E A Graham L Bohm G H NTowers and E R Waygood ldquoUltraviolet-mediated cytotoxicactivity of phenylheptatriyne from Bidens pilosa Lrdquo Journal ofNatural Products vol 42 no 1 pp 103ndash111 1979

[59] Y Sashida K Ogawa M Kitada H Karikome Y Mimakiand H Shimomura ldquoNew aurone glucosides and new phenyl-propanoid glucosides from Bidens pilosardquo Chemical and Phar-maceutical Bulletin vol 39 no 3 pp 709ndash711 1991

[60] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens pilosa L (TFB) on animal liver injury andliver fibrosisrdquo Journal of Ethnopharmacology vol 116 no 3 pp539ndash546 2008

[61] B Hoffmann and J Holzl ldquoAcylated compounds from Bidenspilosardquo Planta Medica vol 55 pp 108ndash109

[62] B Hoffmann and J Holzl ldquoA methylated chalcone glucosidefrom Bidens pilosardquo Phytochemistry vol 27 no 11 pp 3700ndash3701 1988

[63] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[64] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[65] B Hoffmann and J Holzl ldquoWeitere acylierte chalkone ausBidens pilosardquo Planta Medica vol 54 pp 450ndash451 1988

[66] P Geissberger and U Sequin ldquoConstituents of Bidens pilosa Ldo the components found so far explain the use of this plant in11traditional medicinerdquo Acta Tropica vol 48 no 4 pp 251ndash2611991

[67] V V Pham V V T K P V L Hoang and V K Phan ldquoFlavonoidcompounds from the plant Bidens pilosa L (Asteraceae)rdquo TapChi Duoc Hoc vol 50 pp 48ndash53

[68] S D Sarker B Bartholomew R J Nash and N Robinson ldquo5-O-methylhoslundin an unusual flavonoid from Bidens pilosa(Asteraceae)rdquo Biochemical Systematics and Ecology vol 28 no6 pp 591ndash593 2000

[69] S L Chang Y M Chiang C L T Chang et al ldquoFlavonoidscentaurein and centaureidin from Bidens pilosa stimulate IFN-120574 expressionrdquo Journal of Ethnopharmacology vol 112 no 2 pp232ndash236 2007

[70] Y M Chiang D Y Chuang S Y Wang Y H Kuo P WTsai and L F Shyur ldquoMetabolite profiling and chemopreventivebioactivity of plant extracts from Bidens pilosardquo Journal ofEthnopharmacology vol 95 no 2-3 pp 409ndash419 2004

[71] A Kusano Y Seyama E Usami et al ldquoStudies on the antioxi-dant active constituents of the dried powder from Bidens pilosaL var radiata SchrdquoNaturalMedicines vol 57 no 3 pp 100ndash1042003

[72] I Doleckova L Rarova J Gruz et al ldquoAnti-proliferative andanti-angiogenic effects of flavone eupatorin an active con-stituent of chloroform extract ofOrthosiphon stamineus leavesrdquoFitoterapia vol 83 no 6 pp 1000ndash1007

[73] K Bairwa R Kumar R J Sharma and R K Roy ldquoAn updatedreview on Bidens pilosa Lrdquo Der Pharma Chemica vol 2 no 3pp 325ndash337

[74] Q Xia Y Liu and Y Li ldquoDetermination of hyperoside indifferent parts and different species of Herba Bidens by RP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol 50pp 48ndash53

[75] M Horiuchi and Y Seyama ldquoImprovement of the antiinflam-matory and antiallergic activity of Bidens pilosa L var radiataSCHERFF treated with enzyme (Cellulosine)rdquo Journal of HealthScience vol 54 no 3 pp 294ndash301 2008

[76] M G L Brandao C G C Nery M A S Mamao and AU Krettli ldquoTwo methoxylated flavone glycosides from Bidenspilosardquo Phytochemistry vol 48 no 2 pp 397ndash400 1998

[77] P Kumari K Misra B S Sisodia et al ldquoA promising anticancerand antimalarial component from the leaves of Bidens pilosardquoPlanta Medica vol 75 no 1 pp 59ndash61 2009

[78] G Seelinger I Merfort U Wolfle and C M Schempp ldquoAnti-carcinogenic effects of the flavonoid luteolinrdquoMolecules vol 13no 10 pp 2628ndash2651 2008

[79] G Seelinger I Merfort and C M Schempp ldquoAnti-oxidantanti-inflammatory and anti-allergic activities of luteolinrdquoPlanta Medica vol 74 no 14 pp 1667ndash1677 2008

[80] C C Yit and N P Das ldquoCytotoxic effect of butein on humancolon adenocarcinoma cell proliferationrdquo Cancer Letters vol82 no 1 pp 65ndash72 1994

[81] J A Beutler E Hamel A J Vlietinck et al ldquoStructure-activityrequirements for flavone cytotoxicity and binding to tubulinrdquoJournal of Medicinal Chemistry vol 41 no 13 pp 2333ndash23381998

[82] M R Kviecinski K B Felipe T Schoenfelder et al ldquoStudy ofthe antitumor potential of Bidens pilosa (Asteraceae) used inBrazilian folkmedicinerdquo Journal of Ethnopharmacology vol 117no 1 pp 69ndash75 2008

[83] P Sundararajan A Dey A Smith A G Doss M Rajappanand S Natarajan ldquoStudies of anticancer and antipyretic activityof Bidens pilosawhole plantrdquo African Health Sciences vol 6 no1 pp 27ndash30 2006

[84] YMChiang C P Lo Y P Chen et al ldquoEthyl caffeate suppressesNF-120581B activation and its downstream inflammatory mediatorsiNOS COX-2 and PGE2 in vitro or in mouse skinrdquo BritishJournal of Pharmacology vol 146 no 3 pp 352ndash363 2005

50 Evidence-Based Complementary and Alternative Medicine

[85] J S Kim H J Lee M H Lee J Kim C Jin and J HRyu ldquoLuteolin inhibits LPS-stimulated inducible nitric oxidesynthase expression in BV-2 microglial cellsrdquo Planta Medicavol 72 no 1 pp 65ndash68 2006

[86] P A Ruiz and D Haller ldquoFunctional diversity of flavonoidsin the inhibition of the proinflammatory NF-120581B IRF and Aktsignaling pathways in murine intestinal epithelial cellsrdquo Journalof Nutrition vol 136 no 3 pp 664ndash671 2006

[87] A Xagorari A Papapetropoulos A Mauromatis MEconomou T Fotsis and C Roussos ldquoLuteolin inhibitsan endotoxin-stimulated phosphorylation cascade andproinflammatory cytokine production in macrophagesrdquoJournal of Pharmacology and Experimental Therapeutics vol296 no 1 pp 181ndash187 2001

[88] N Yoshida T Kanekura Y Higashi and T Kanzaki ldquoBidenspilosa suppresses interleukin-1120573-induced cyclooxygenase-2expression through the inhibition of mitogen activated proteinkinases phosphorylation in normal human dermal fibroblastsrdquoJournal of Dermatology vol 33 no 10 pp 676ndash683 2006

[89] M Muchuweti C Mupure A Ndhlala T Murenje and M AN Benhura ldquoScreening of antioxidant and radical scavengingactivity of Vigna ungiculata Bidens pilosa and Cleome gynan-drardquo American Journal of Food Technology vol 2 no 3 pp 161ndash168 2007

[90] L P Yuan F H Chen L Ling et al ldquoProtective effects of totalflavonoids of Bidens bipinnata L against carbon tetrachloride-induced liver fibrosis in ratsrdquo Journal of Pharmacy and Pharma-cology vol 60 no 10 pp 1393ndash1402 2008

[91] S L Chang H H Yeh Y S Lin Y M Chiang T K Wuand W C Yang ldquoThe effect of centaurein on interferon-120574 expression and Listeria infection in micerdquo Toxicology andApplied Pharmacology vol 219 no 1 pp 54ndash61 2007

[92] F Deba T D Xuan M Yasuda and S Tawata ldquoChemical com-position and antioxidant antibacterial and antifungal activitiesof the essential oils from Bidens pilosa Linn var Radiatardquo FoodControl vol 19 no 4 pp 346ndash352 2008

[93] A O T Ashafa and A J Afolayan ldquoScreening the root extractsfrom Biden pilosa L var radiata (Asteraceae) for antimicrobialpotentialsrdquo Journal of Medicinal Plant Research vol 3 no 8 pp568ndash572 2009

[94] T B Nguelefack T Dimo E P Nguelefack Mbuyo P V TanS V Rakotonirina and A Kamanyi ldquoRelaxant effects of theneutral extract of the leaves of Bidens pilosa linn on isolated ratvascular smooth musclerdquo Phytotherapy Research vol 19 no 3pp 207ndash210 2005

[95] M T Grombone-Guaratini K L Silva-Brandao V N SolferiniJ Semir and J R Trigo ldquoSesquiterpene and polyacetyleneprofile of the Bidens pilosa complex (Asteraceae Heliantheae)from Southeast of Brazilrdquo Biochemical Systematics and Ecologyvol 33 no 5 pp 479ndash486 2005

[96] L L Lin C Y Wu H C Hsiu M T Wang and H ChuangldquoStudies on diabetes mellitus I The hypoglycemic activity ofphytosterin on alloxan diabetic ratsrdquo Taiwan Yi Xue Hui Za Zhivol 66 no 2 pp 58ndash65 1967

[97] M A N Benhura and I C Chitsiku ldquoThe extractable 120573-carotene content of Guku (Bidens pilosa) leaves after cookingdrying and storagerdquo International Journal of Food Science andTechnology vol 32 no 6 pp 495ndash500 1997

[98] J S Chang L C Chiang C C Chen L T Liu K C Wangand C C Lin ldquoAtileukemic activity of Bidens pilosa l varminor (blume) sherff andHouttuynia cordata thunbrdquo AmericanJournal of Chinese Medicine vol 29 no 2 pp 303ndash312 2001

[99] F Deba T D Xuan M Yasuda and S Tawata ldquoHerbicidal andfungicidal activities and identification of potential phytotoxinsfrom Bidens pilosa L var radiata Scherff research paperrdquoWeedBiology and Management vol 7 no 2 pp 77ndash83 2007

[100] J K Kumar and A K Sinha ldquoA new disubstituted acetylacetonefrom the leaves ofBidens pilosa LinnrdquoNatural Product Researchvol 17 no 1 pp 71ndash74 2003

[101] K Ogawa and Y Sashida ldquoCaffeoyl derivatives of a sugarlactone and its hydroxy acid from the leaves of Bidens pilosardquoPhytochemistry vol 31 no 10 pp 3657ndash3658 1992

[102] Q Xia Y Liu and Y Li ldquoDetermination of gallic acid fromdifferent species and different medical parts of Herba Bidens byRP-HPLCrdquoWest China Journal of Pharmaceutical Sciences vol24 no 03 pp 308ndash310

[103] W J Lee L F Wu W K Chen C J Wang and T HTseng ldquoInhibitory effect of luteolin on hepatocyte growth fac-torscatter factor-induced HepG2 cell invasion involving bothMAPKERKs and PI3K-Akt pathwaysrdquo Chemico-BiologicalInteractions vol 160 no 2 pp 123ndash133 2006

[104] J A Beutler J H Cardellina C M Lin E Hamel G MCragg andM R Boyd ldquoCentaureidin a cytotoxic flavone fromPolymnia fruticosa inhibits tubulin polymerizationrdquo BioorganicandMedicinal Chemistry Letters vol 3 no 4 pp 581ndash584 1993

[105] A Verma A Su A M Golin B OrsquoMarrah and J K AmorosaldquoThe lateral view a screening method for knee traumardquo Aca-demic Radiology vol 8 no 5 pp 392ndash397 2001

[106] J Corren M Lemay Y Lin L Rozga and R K RandolphldquoClinical and biochemical effects of a combination botanicalproduct (ClearGuard) for allergy a pilot randomized double-blind placebo-controlled trialrdquo Nutrition Journal vol 7 no 1article 20 2008

[107] M S Gachet J S Lecaro M Kaiser et al ldquoAssessment of anti-protozoal activity of plants traditionally used in Ecuador in thetreatment of leishmaniasisrdquo Journal of Ethnopharmacology vol128 no 1 pp 184ndash197 2010

[108] W C Yang M Ghiotto B Barbarat and D Olive ldquoThe role ofTec protein-tyrosine kinase in T cell signalingrdquo The Journal ofBiological Chemistry vol 274 no 2 pp 607ndash617 1999

[109] S Tewtrakul H Miyashiro N Nakamura et al ldquoHIV-1 inte-grase inhibitory substances from Coleus parvifoliusrdquo Phytother-apy Research vol 17 no 3 pp 232ndash239 2003

[110] T Dimo T B Nguelefack P V Tan et al ldquoPossible mechanismsof action of the neutral extract from Bidens pilosa L leaves onthe cardiovascular system of anaesthetized ratsrdquo PhytotherapyResearch vol 17 no 10 pp 1135ndash1139 2003

[111] T S C Li Chinese and Related North American Herbs CRCPress New York NY USA 2002

[112] H Wu H Chen X Hua Z Shi L Zhang and J ChenldquoClinical therapeutic effect of drug-separated moxibustion onchronic diarrhea and its immunologic mechanismsrdquo Journal ofTraditional Chinese Medicine vol 17 no 4 pp 253ndash258 1997

[113] S W Wright R R Harris J S Kerr et al ldquoSynthesis chemicaland biological properties of vinylogous hydroxamic acids dualinhibitors of 5-lipoxygenase and IL-1 biosynthesisrdquo Journal ofMedicinal Chemistry vol 35 no 22 pp 4061ndash4068 1992

[114] W R Almiron and M E Brewer ldquoClassification of immaturestage habitats of Culicidae (Diptera) collected in CordobaArgentinardquo Memorias do Instituto Oswaldo Cruz vol 91 no 1pp 1ndash9 1996

[115] N L Wang J Wang X S Yao and S Kitanaka ldquoTwo newmonoterpene glycosides and a new (+)-jasmololone glucoside

Evidence-Based Complementary and Alternative Medicine 51

from Bidens parvifloraWilldrdquo Journal of Asian Natural ProductsResearch vol 9 no 5 pp 473ndash479 2007

[116] P Champagnat ldquoRole of the terminal bud in the actionexercised by the cotyledon of Bidens pilosus L var radiatus onits axillary budrdquo Comptes Rendus des Seances et Memoires de laSociete de Biologie vol 145 no 17-18 pp 1374ndash1376 1951

[117] S F Nielsen S B Christensen G Cruciani A Kharazmiand T Liljefors ldquoAntileishmaniai chalcones statistical designsynthesis and three-dimensional quantitative structure-activityrelationship analysisrdquo Journal of Medicinal Chemistry vol 41no 24 pp 4819ndash4832 1998

[118] Y C Hwang J J H Chu P L Yang W Chen and M V YatesldquoRapid identification of inhibitors that interfere with poliovirusreplication using a cell-based assayrdquo Antiviral Research vol 77no 3 pp 232ndash236 2008

[119] V F Andrade-Neto M G L Brandao F Q Oliveira et alldquoAntimalarial activity of Bidens pilosa L (Asteraceae) ethanolextracts fromwild plants collected in various localities or plantscultivated in humus soilrdquo Phytotherapy Research vol 18 no 8pp 634ndash639 2004

[120] R J Marles and N R Farnsworth ldquoAnti-diabetic plants andtheir active constituentsrdquo Phytomedicine vol 2 no 2 pp 137ndash189

[121] M Habeck ldquoDiabetes treatments get sweet help from naturerdquoNature Medicine vol 9 no 10 p 1228 2003

[122] H W Lin G Y Han and S X Liao ldquoStudies on the activeconstituents of the Chinese traditional medicine Polygonatumodoratum (Mill) DrucerdquoActa Pharmaceutica Sinica vol 29 no3 pp 215ndash222 1994

[123] C L T Chang Y C Chen H M Chen N S Yang andW C Yang ldquoNatural cures for type 1 diabetes a reviewof phytochemicals biological actions and clinical potentialrdquoCurrent Medicinal Chemistry vol 20 no 7 pp 899ndash907 2013

[124] L Dey A S Attele and C S Yuan ldquoAlternative therapies fortype 2 diabetesrdquo Alternative Medicine Review vol 7 no 1 pp45ndash58 2002

[125] C L T ChangH Y Liu T F Kuo et al ldquoAnti-diabetic effect andmode of action of cytopiloynerdquo Evidence-Based Complementaryand Alternative Medicine vol 2013 Article ID 685642 13 pages2013

[126] M R Khan M Kihara and A D Omoloso ldquoAnti-microbialactivity of Bidens pilosa Bischofia javanica Elmerillia papuanaand Sigesbekia orientalisrdquo Fitoterapia vol 72 no 6 pp 662ndash6652001

[127] T Dimo T B Nguelefack P Kamtchouing E Dongo ARakotonirina and S V Rakotonirina ldquoHypotensive effects ofmethanol extract from Bidens pilosa Linn on hypertensive ratsrdquoComptes Rendus de lrsquoAcademie des Sciences Serie III vol 322no 4 pp 323ndash329 1999

[128] A K Hassan O Deogratius J F Nyafuono O Francis and OP Engeu ldquoWound healing potential of the ethanolic extracts ofBidens pilosa and Ocimum suaverdquo African Journal of Pharmacyand Pharmacology vol 5 no 2 pp 132ndash136 2011

[129] B Hoffmann and J Holzl ldquoChalcone glucosides from Bidenspilosardquo Phytochemistry vol 28 no 1 pp 247ndash250 1989

[130] B Hoffmann and J Holzl ldquoNew chalcones from Bidens pilosardquoPlanta Medica vol 54 no 1 pp 52ndash54

[131] C Kandaswami and EMiddleton ldquoFree radical scavenging andantioxidant activity of plant flavonoidsrdquo Advances in Experi-mental Medicine and Biology vol 366 pp 351ndash376 1994

[132] K Yamamoto H Kakegawa H Ueda et al ldquoGastric cytopro-tective anti-ulcerogenic actions of hydroxychalcones in ratsrdquoPlanta Medica vol 58 no 5 pp 389ndash393 1992

[133] J O C Ezeonwumelu A K Julius C N Muhoho et alldquoBiochemical and histological studies of aqueous extract ofBidens pilosa leaves from Ugandan Rift valley in ratsrdquo BritishJournal of Pharmacology and Toxicology vol 2 no 6 pp 302ndash309

[134] L Frida S Rakotonirina A Rakotonirina and J P SavineauldquoIn vivo and in vitro effects of Bidens pilosa L (Asteraceae)leaf aqueous and ethanol extracts on primed-oestrogenized ratuterine musclerdquo African Journal of Traditional Complementaryand Alternative Medicines vol 5 no 1 pp 79ndash91 2008

52 Evidence-Based Complementary and Alternative Medicine

Composition Comments

1 Please provide valid postal code to the address if

possible

2 We made the highlighted change in Table 4 for the sake

of correctness Please check

3 Please check the correctness of the highlighted part in

Table 5

4 We considered the highlighted part in Table 11 as

reference Please check similar cases throughout

5 Please check the format of Table 18

6 We reformatted Table 20 Please check

7 We made the highlighted change in Table 4 for the sake

of consistency Please check similar cases throughout the

paper

8 We made the highlighted change(s) for the sake

of consistency Please check simila highlighted cases

throughout

9 We made the highlighted change in Table 9 for the sake

of clarity Please check similar cases through out

10 We made the highlighted change(s) for the sake of

correctness Please check

11 Comment on ref [66] This reference is a repetition

of [18] Please check similar cases throughout

Author(s) Name(s)

Author 1

Author 2

Author 3