Comparative Evaluation of Analgesic Activity of ... · Comparative Evaluation of Analgesic Activity of Oleogumresin and Stem bark extract of Guggulu in Albino rats. Malviya Reeta1*,

Malviya Reeta et al, 2015/ Comparative evaluation of analgesic activity of Oleogumresin

International Research Journal of Pharmaceutical and Biosciences (IRJPBS) 2 (5) 01- 09 1

RESEARCH ARTICLE

International Research Journal of Pharmaceutical and Biosciences

Pri -ISSN: 2394 - 5826 http://www.irjpbs.com e-ISSN: 2394 - 5834

Comparative Evaluation of Analgesic Activity of Oleogumresin and Stem bark extract of Guggulu in Albino rats.

Malviya Reeta1*, Sharma Ravindra2, Sharma Tarun,3 Ramamurthy A,4 and Kotecha Mita5

*1Dept. of Dravya Guna, Mahatma Jyotiba Fule Ayurvedic college, chomu, Jaipur, Rajasthan. 2Dept. of P.G. Studies in Dravya Guna, National Institute of Ayurveda, Jaipur, Rajasthan. 3Dept. of P.G. Studies in Dravya Guna, National Institute of Ayurveda, Jaipur, Rajasthan. 4Dept. of P.G. Studies in Dravya Guna. National Institute of Ayurveda, Jaipur, Rajasthan. 5Dept. of P.G. Studies in Dravya Guna. National Institute of Ayurveda, Jaipur, Rajasthan.

Article info Abstract

Article history:

Received 25 AUG 2015

Accepted 02 SEP 2015

*Corresponding author:

[email protected]

Copyright 2015 irjpbs

Background: Guggulu (commiphora wightii Bhandari) is a very important plant widely used for its antihyperlipidaemic, analgesic and anti- inflammatory actions by both Ayurvedic contemprory medicinal practionars. The Oleo-gumresin is a very widely used raw material of this medicinal plant and it is commonly used in Vata roga and Medo roga in Ayurveda. Objective: The current research paper provides a detailed report of comparative study the Analgesic activity of Guggulu resin & its stem bark extract in albino rats. Methods: An artery clip is applied to the root of the tail of mice and the reaction time is noted. Male mice with a weight between 18 and 25 g are used. The Control Group, Standard Group and Test Group consist of 6 mice in each. The test compounds are administered orally to fasted animals. The drug is administered 15, 30 or 60 min prior testing. An artery clip is applied to the root of the tail (approximately 1 cm from the body) to induce pain. The animal quickly responds to these noxious stimuli by biting the clip or the tail near the location of the clip. A stopwatch in 1/10 seconds increments measures the time between stimulation onset and response. A cut – off time will determine by taking the average reaction time plus 3 times the standard deviation of combined latencies of the control mice at all times periods. Any reaction time of the best animals which greater. Then the cut-off time is called a positive response indicative of analgesic activity. The length of time until response indicates the period of greatest activity after dosing. An ED50 value is calculated at the peak time of drug activity. Results: All three test sample provided significant effect as Analgesic agent. Conclusion: It can be concluded that although aqueous extract and methanolic extract of stem bark provided significant effect as analgesic agent they haven’t provided better effect or, same effect as compared crude resin extract.

Keywords: Guggulu resin, Guggulu stems bark, analgesic activity, Methanolic extract.

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International Research Journal of Pharmaceutical and Biosciences (IRJPBS) 2 (5) 01 - 09 2

INTRODUCTION Guggulu (commiphora wightii Bhandari) is a very important plant widely used for its antihyperlipidaemic, analgesic and anti- inflammatory actions by both Ayurvedic contemprory medicinal practionars. The Oleo-gumresin is a very widely used raw material of this medicinal plant and it is commonly used in Vata roga and Medo roga in Ayurveda. Unfortunately the plant Commiphora wightii has become endangered because of its slow

growing nature, poor seed setting, and lack of cultivation, poor seed germination rate and

excessive and unscientific tapping for its gum resin by the pharmaceutical industries and

religious prophets. This plant is incorporated in Data Deficient category of IUCN’s Red Data

list. The plant Commiphora wightii has become endangered because of its slow growing

nature, poor seed setting, and lack of cultivation, poor seed germination rate and excessive

and unscientific tapping for its gum resin by the pharmaceutical industries and religious

purposes. In case of C. wightii, stem cuttings offer several advantages over seeds. They save

time and labor, and produce genetically superior and uniform planting materials from

superior parentstocks. Stem cuttings are also inexpensive and easier to practice than other

vegetative -propagation methods, such as tissue culture. In addition, stem cuttings can

continuously supply planting stocks throughout the year for reforestation activities.

Pain is a subjective symptom and a major health problem throughout the globe now days as

it leaves major negative effects of physical and psychological health of the people. The gap

between the sophisticated knowledge of pain and its treatment and the effective application

of that knowledge is large and widening. Both acute and chronic pain is often poorly

managed for a wide variety of reasons.

In India 30% of people, women and elderly in particular and other adults in general are

affected with chronic pain. About 20-25% of this population suffer pain from

musculoskeletal and joint disorders. The back pain accounts for another 25-30% people.

Headache and migraine accounts for about 5% among the cases of persistent pain. In only 1-

2 % of adults with chronic pain result from cancer. Trigeminal neuralgia, post hepatic

neuralgia and other neuropathic pain syndromes constitute another 5-7% of patients

suffering from chronic pain. Now interventional pain management procedures like trigger

point injections, percutaneous disectomy and nerve blocks etc. however with no good

results. Hence an attempt has been made to find out simple and safe remedies from

Ayurveda.

Mode of Action

Guggulu can counteract upon Shita and reduce pain sansationsa by having direct effect on

Vata. Hence Guggulu have been described as best Vathara drug of Ayurveda. Vagbhat has

mentioned Guggulu as “xqXxqyq esnks·fuykgjk.kke~”A in Agrya Prakarana. And it pacifies

the Vata by its Vatashamak action releiving pain.



A dominant Analgesic activity of Commiphora wightii (guggul) plant, earlier known as

Commiphora mukul (family-Burseraceae) add impetus to this research approach. Oleogum

resin, known as guggul or gum guggul, is obtained from C. mukul found in India, Bangladesh

and Pakistan (Dev, 1987). One of the most revered plant Guggul, Commiphora wightii, has

been used in Ayurvedic medicine to treat a variety of diseases since several thousand years

(Satyavati, 1988). It is used extensively for the treatment of different ailments including

dysmenorrhea, endometritis, hypercholesterolemia, hypertension, bronchitis, inflammation,

arthritis, cancer and cardiovascular disorders (Shishodia et al., 2008). The phytochemical

constituents of guggul like Z- and E-isomers of guggulsterone and its related guggulsterols I-

IV (Zhu et al., 2001). It possess a wide array of pharmacological activities including anti-

lipidemic (Satyavati et al., 1969), anti-apoptotic (Macha et al., 2010), anti-inflammatory

(Francis et al., 2004), anti-thyroid (Panda and Kar, 2005; Tripathi et al., 1998), hypolipidemic

(Chander et al., 1996) and hepatoprotective (Gowrishankar et al., 2008) has been reported.

MATERIAL AND METHODS

The animal experiment was approved by Institutional Animal Ethics Committee (IAEC) of

Pinnacle Biomedical Research Institute (PBRI) Bhopal (Reg No. 1283/c/09/CPCSEA). Protocol

Approval Reference No. PBRI/13/IAEC/PN-337.

Present study was undertaken in order to

Animal used and their Weight

Healthy Adult Male Wistar Albino Rats (200±30gm) Swiss Albino Male Mice (25±5gm).

Route of administration - P.O.

Housing Condition

Animals were housed in a group of four in separate cages under controlled conditions of

temperature (22 ± 2°C). All animals were given standard diet (Golden Feed, New Delhi) and

water regularly. Animals were further divided in eight groups with four animals in each

group.

Procedure

Animal

Male Swiss albino mice with a weight between 25±5gm.

Induction of Algesia (Pain)

An artery clip is applied to the root of the tail (approximately 1 cm from the body) to induce

pain. The animal quickly responds to these noxious stimuli by biting the clip or the tail near

the location of the clip. The time between stimulation onset and response is measured by a

stopwatch in 1/10 seconds increments. The drug is administered 30 min prior testing. The

animals were divided into four groups of six rats each.



Group-I (Control): Administer 5 ml of water orally.

Group-II (Standard): Administer 500 Mg/Kg. of Aqueous Extract of Commiphora

wightii Bhandari. Resin orally.

Group-III (Test A): Administer 500 Mg/Kg. of Aqueous Extract of Commiphora wightii

Bhandari. Stem bark orally.

Group-IV (Test B): Administer 500 Mg/Kg. of Methanol Extract of Commiphora

wightii Bhandari. Stem Bark orally

A cut-off time is determined by taking the average reaction time after the drug

administration at time interval 0, 30 and 60 min.

Fig.1. Pictures of Commiphora Wightii (Bhandari)



Fig.2. T.S. of the Leave, Portion of the Cortex of Midrib Region Of Commiphora

Wightii (Bhandari).

Fig.3. T.S. of Mature Stem of Commiphora Wightii

(Bhandari).

Fig.4. T.S. of Mature Stem of Commiphora Wightii

(Bhandari).



RESULTS AND DISCUSSION

ANALGESIC ACTIVITY (Haffner’s Tail Clip Method- 1954)

S. No. Treatment Reaction Time (sec.)

Initial Final % Increase

1. Control

(5 ml/kg of water p.o.)

2.23±0.484 2.33±0.367 2.28±0.354

2. Standard (Aq. Extract of CWB

resin – 500 mg/kg p.o.)

2.17±0.543 4.87±0.550 8.43±0.631

3. Test A (Aq. Extract of CWB

stem bark – 500 mg/kg p.o.)

2.37±0.398 3.03±0.288 4.80±0.400

4. Test B (MeOH Extract of CWB

stem bark – 500 mg/kg p.o.)

2.15±0.596 3.35±0.532 6.62±0.637

A) After 0 min

One Way Analysis of Variance

Data source: Data 1 in Notebook 1

Group Name N Missing Mean Std Dev SEM

Row 1 6 0 2.230 0.484 0.198

Row 2 6 0 2.170 0.543 0.222

Row 3 6 0 2.370 0.398 0.162

Row 4 6 0 2.150 0.596 0.243

Source of Variation DF SS M F P

Between Groups 3 0.178 0.0592 0.227 0.876

Residual 20 5.214 0.261

Total 23 5.391

The differences in the mean values among the treatment groups are not great enough to

exclude the possibility that the difference is due to random sampling variability; there is

not a statistically significant difference (P = 0.876).

Power of performed test with alpha = 0.050: 0.049



The power of the performed test (0.049) is below the desired power of 0.800.

Less than desired power indicates you are less likely to detect a difference when one

actually exists. Negative results should be interpreted cautiously.

B) AFTER 30 min




Row 1 6 0 2.330 0.367 0.150

Row 2 6 0 4.870 0.550 0.225

Row 3 6 0 3.030 0.288 0.118

Row 4 6 0 3.350 0.532 0.217

Source of Variation DF SS MS F P

Between Groups 3 20.671 6.890 34.316 <0.001

Residual 20 4.016 0.201

Total 23 24.686

The differences in the mean values among the treatment groups are greater than would

be expected by chance; there is a statistically significant difference (P = <0.001).


All Pairwise Multiple Comparison Procedures (Bonferroni t-test):

Comparison Diff of Means T P P<0.050

Row 2 vs. Row 1 2.540 9.818 <0.001 Yes

Row 2 vs. Row3 1.840 7.112 <0.001 Yes

Row 2 vs. Row4 1.520 5.875 <0.001 Yes

Row 4 vs. Row1 1.020 3.943 0.005 Yes

Row 4 vs. Row3 0.320 1.237 1.000 No

Row 3 vs. Row1 0.700 2.706 0.082 No

C) After 60 min






Row 1 6 0 2.280 0.354 0.145

Row 2 6 0 8.430 0.631 0.258

Row 3 6 0 4.800 0.400 0.163

Row 4 6 0 6.620 0.637 0.260

Source of Variation DF SS MS F P

Between Groups 3 124.161 41.387 151.984 <0.001

Residual 20 5.446 0.272

Total 23 129.607

The differences in the mean values among the treatment groups are greater than would

be expected by chance; there is a statistically significant difference (P = <0.001).


All Pairwise Multiple Comparison Procedures (Bonferroni t-test):

Comparisons for factor

Comparison Diff of Means T P P<0.050

Row 2 vs. Row 1 6.150 20.413 <0.001 Yes

Row 2 vs. Row3 3.630 12.049 <0.001 Yes

Row 2 vs. Row4 1.810 6.008 <0.001 Yes

Row 4 vs. Row1 4.340 14.405 <0.001 Yes

Row 4 vs. Row3 1.820 6.041 <0.001 Yes

Row 3 vs. Row1 2.520 8.364 <0.001 Yes

CONCLUSION

Analgesic activity of crude resin’s extract and stem bark‘s methanolic and aqueous extract

was compared on the basis of Haffner’s tail clip method. Reaction time (sec) was considered

as analysis parameter. At 30th min there was significant difference observed in between

resin extract treated animals and control group (P<0.001). Methanolic extract of stem bark

also showed significant effect (P<0.05) but aqueous extract showed no significant effect at



30th min the same time. At 60th min it was observed that all three treatments provided

significant effect (P<0.001). There was significant difference present in between crude resin

extract treated animals, aqueous extract of stem bark treated animals and methanolic

extract of stem bark treated animals (P<0.001). Hence it can be concluded that although

aqueous extract and methanolic extract of stem bark provided significant effect as analgesic

agent they haven’t provided better effect or, same effect as compared crude resin extract.

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Comparative Evaluation of Analgesic Activity of ... · Comparative Evaluation of Analgesic Activity of Oleogumresin and Stem bark extract of Guggulu in Albino rats. Malviya Reeta1*,