Ife Journal of Science vol. 16, no. 3 (2014)

NEPHROCURATIVE EFFECTS OF AQUEOUS STEM BARK EXTRACT OF BOSWELLIA PAPYRIFERA (DEL.) AGAINST ACETOMINOPHEN-INDUCED

KIDNEY DAMAGE IN RATS

1* 1 2 2 2 Abdulmumin Y. , Matazu K. I. , Wudil A. M. , Alhassan A. J. , Imam A. A.

1*Department of Science Laboratory Technology, Collage of Science and Technology,Hussaini Adamu Federal Polytechnic, Kazaure, P.M.B 5004, Jigawa State, Nigeria

2 Department of Biochemistry, Faculty of Science, Bayero University, Kano, P.M.B 3011, Kano, Nigeria.1*Corresponding author: yabdulmumin@yahoo.com.

(Received: 30th June, 2014; Accepted: 6th October, 2014)

The properties of Boswellia plants have been exploited for years in traditional medicines of Africa; the advent of synthetic drugs has obscured the pharmaceutical use of Boswellia until it was reported that an ethanolic extract exerts anti-inflammatory and antiarthritis effects. This paper investigated the curative effect of aqueous stem bark extract of Boswellia papyrifera on acetaminophen-induced kidney damage. Three different doses (50, 100 and 150 mg/kg) of the extract were administered daily to the different groups of rats for two-and four-week periods after inducing kidney damage using acetaminophen at a dose of 800mg/kg (po). The serum creatinine, urea, sodium, potassium, chloride and bicarbonate were found not to be different from that of normal control group even at two weeks; the histopathology examination of the kidney after four weeks showed normal kidney architecture. The nephrocurative effects of aqueous stem bark extract of Boswellia papyrifera oral administration was found to be dose- and time- dependent.

Key words: Boswellia papyrifera, curative effect, acetaminophen and kidney damage.

ABSTRACT

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INTRODUCTIONTraditional medicine was a source of discovering many important medical pharmaceuticals (Gilani and Atta-ur-rahman, 2005). Recently, however, plant-derived compounds offer potential source of new anti- microbial, anticancer and anti-HIV agents among other pharmaceuticals (Gurib-Fakim et al., 2005). Traditional medicine is rooted in a philosophical understanding which engages a multitude of healing methods that have been preserved for generations through stories, oral teachings, ceremonies, and through written documentation.. Community is central to the practice of traditional medicine (Maydell, 1986). In some Asian and African countries, up to 80% of the population relies on traditional medicine for their primary health care needs. When adopted outside of its traditional culture, traditional medicine is often called complementary and alternative medicine.

Boswellia papyrifera (Del.) Hochst (Ararrabi) belong to family of tropical plants called Bruceraceae (Fitchl and Admasu, 1994) which is distinguished by the presence of resin ducts in the bark (Groom, 1981). B. papyrifera is a deciduous tree which can be as tall as 12 m, with a rounded crown and a straight, regular bole. The bark is whitish to pale-brown,

peeling off in large flakes; slash is red-brown and it exudes a fragrant resin. The bark contains schizogenous olea-gum-resin pockets (Verghese 1988). Various plant parts and products are utilized for traditional and medicinal purposes. The leaves and roots of the species are used against lymphadenopathy while the resin is used as a febrifuge (Fitchl and Admasu, 1994). Frankincense from Boswellia papyrifera was considered as a stimulant and was once used to treat leprosy in China (Tucker, 1986). The bark is chewed to treat stomach disturbances. It is burnt as a mosquito repellent in the tropics and also chewed by lowlanders to quench thirst during hot days (Tilahun, 1997). Limited information exists on the medicinal uses of B. papyrifera. Nevertheless, the traditional medicinal uses by the local people and experiences with other species of the genus (for example, B. serrata) suggests the potential use of B. papyrifera for medicinal purposes as well. Boswellic acid (which constitutes about 50-70% of the oil) extracted from B. serrata in India is commercially used as a fumigative agent, an anti-arthritic, anti-inflammatory and pharmacological agent (Anon, 2000). Those extracts from B. serrata were found to be more beneficial, less toxic, and more potent than

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standard anti-inflammatory drugs. Boswellia has long been used in Ayurvedic medicine. The boswellic acids that are a component of the resin it produces have shown some promise as a treatment for asthma and various inflammatory conditions (Gupta et al, 1998).

Nephrotoxicity is a poisonous effect of some substances notably toxic chemicals and drugs, on the kidneys (Galley, 2000). It is the most common kidney problem and occurs when the body is exposed to a drug or toxin that causes damage to the kidneys. When kidney damage occurs, the body is unable to get rid of excess urine, and wastes. The blood electrolytes such as potassium, sodium, chloride and magnesium will be elevated (Abeloff et al; 2000). Nephrotoxicity can be temporary with a temporary elevation of blood urea nitrogen (BUN) and/or creatinine (Berg, 2001).

The underlying characteristics of the offending agent also play an important role in the development of nephrotoxicity Acetaminophen is most widely used in the world as an analgesic and antipyretic drug that is safe at therapeutic dosages. However, it is also known to cause hepatic necrosis and renal failure in humans (Hengy et al, 2009) and animals (Ghosh et al.,2010) in overdoses. In the human, acetaminophen represents a growing cause of renal failure in current medical practice. Acetaminophen-induced renal insufficiency is consistent with acute tubular necrosis, an increase in the plasma creatinine level and a decrease in the glomerular f i l trat ion rate (GFR). The development of acute renal failure (ARF) in rats within 24hrs by administration of a single dose of acetaminophen (750 mg/kg, po) was reported (Pelani et al., 2009).

The World Health Organisation notes, that "inappropriate use of traditional medicines or practices can have negative or dangerous effects" and that "further research is needed to ascertain the efficacy and safety" of several of the practices and medicinal plants used by traditional medicine systems.

This paper is aimed at determining the effect of aqueous bark extract of Boswellia papyrifera on kidney function indices (serum creatinine, urea

+ + - -and electrolytes (Na , K , Cl and HCO ) on 3

acetaminophen--induced kidney damaged rats in order to give an insight on the possible nephrocurative effects of the extract on damaged kidney.

MATERIAL AND METHODS

Plant Material and Preparation of ExtractThe plant (stem bark) was collected in September, 2012 from Zabainawa town of Dawakin tofa Local Government, Kano State, Nigeria. The stem bark collected was washed in clean water and

Oit was then dried at room temperature of 27 C after which it was pulverized to coarse powder

3 ,100g was soaked in 500cm distilled water in a 1 Litre Conical flask. The extract was then obtained after 48 hours by filtration using Whatman No1 filter paper and concentrated using vacuum evaporator. The concentration of the aqueous

3stem bark extract was found to be 50mg/cm . The dose of the extract was administered based on the weight of the rats and doses administered

Experimental Design A total of 24 white male albino rats were used in the study. The rats were divided into eight groups of five rats each. The rats in group I were not induced with kidney damage, they served as normal control. Group II to VIII were administered with 800mg/kg acetaminophen according to Pelani et al. (2009) prior to oral administration of aqueous stem bark extract of Boswellia papyrifera to groups III to VIII. Groups III to V were administered with 50, 100, 150mg/kg of the extract for two weeks while groups VI to VIII were also administered with 50, 100, 150mg/kg for four weeks. All the rats were sacrificed by decapitation. The blood samples collected were allowed to clot and serum was obtained for the determination of serum creatinine, urea, potassium, sodium, chloride and bicarbonate level.

Biochemical AnalysisSerum was obtained and analyzed for serum creatinine level by the method of Bartels and Bohmer (1972); serum urea level by Berthelot's reaction (Weatherburn, 1967); serum sodium level analysis was carried out by Maruna (1958) method; serum potassium level by Henry (1974);

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serum chloride level by the method of Schales (White,1970) and serum bicarbonate level was carried out by the method of (Forrester et al., 1976).

Histological AnalysisThe liver of the rats for all the groups were removed immediately after sacrifice and kept in 10% normal saline for histological examination and photomicrography using Nickon Eclipse-E-200, Photomicrograph system (Auwiro, 2010).

STATISTICAL ANALYSISThe data were analysed using one way Analysis of Variance (ANOVA) by Graphpad instat 3 software (2000) version 3.05 by Graphpad Inc.

RESULTS AND OBSERVATIONSTable 1 summarizes the results for serum level of creatinine, urea, sodium, potassium, chloride and

bicarbonate for groups I normal and II test control 24hours after orally administered with 800mg/kg of acetaminophen while groups III to V present results of rat orally administered with acetaminophen and various doses of aqueous stem bark extract (ASBE) of Boswellia papyrifera for two weeks. Table 2 presents the results for groups VI, VII and VIII rats orally administered with various doses of aqueous stem bark extract (ASBE) of Boswellia papyrifera for four weeks. There was significant difference (p<0.05) between group I and II in the parameters analysed. Similarly, significant decreases (p<0.05) in serum level creatinine, urea, sodium, potassium and chloride was observed while bicarbonate increases in group III to V and VI to VIII after two and four weeks respectively.

I

GROUP

/Treatment CREATININE

(µmol/L) UREA

(mmol/L) HCO3

(mmol/L) Na

(mmol/L) K

(mmol/L) Cl

(mmol/L) NO Acetaminophen

Administered

(NORMAL)

41.33 ± 6.03 2.43 ± 0.78 24.3 ±4.51

115.00 ±5.00 3.83 ± 0.55

89.00 ±7.21

II

80 mg/kg

of Acetaminophen

administered

(TEST CONTROL RATS)

66.00 ± 6.25* 5.54 ± 1.42*

10.67±1.16*

148.33±5.77*

7.93 ±1.40*

128.33 ± 7.64*

III 50mg/kg

ASBE of B.papyrifera

Administered

45.33 ± 9.87n 3.03 ± 0.21o 12.00 ±2.65p 128.33 ± 7.64q 4.43 ±0.15r 106.33±11.85s

IV

100mg/kg

ASBE of B.papyrifera

Administered

47.67±4.93 n 2.63 ± 0.55

o 13.67±4.16 p 114.00 ± 5.29

q 4.30 ± 0.10 r 99.33 ±5.13 s

V 150 mg/kg ASBE of B.papyrifera Administered

44.00 ±3.61 n 2.20 ± 0.26 o 16.67±4.93 p 113.33±2.89 q 3.93 ± 0.38 r 110.67±11.37

Results are expressed as mean ± SD,n = 5Values with asterisk are significantly different at p<0.05 when compared with the normal rats.Values in the same column with the same superscript are significantly different at p<0.05 when compared with the test control HCO - Bicarbonate Na - sodium, K- potassium, Cl- chloride3 ,

Table 1: Serum Kidney Function Indices of Acetaminophen-induced Kidney Damage Rats after the Two Weeks Oral Administration of Aqueous Stem Bark Extract of Boswellia papyrifera.

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HISTOPATHOLOGIC FINDING OF THE KIDNEYHistopathology evaluation of the rat's kidney section from the normal group I (Plate 1) showed normal kidney architecture with the cortex containing the glomerulus and the medulla containing the renal tubules while the test control (group II) showed distortion of the normal kidney architecture with dilation of the renal tubules and collapsed of bowman's capsule (Plate 2). This is

due to the damage caused by acetaminophen oral administration. However, following oral administration of 50, 100, 150 mg/kg of aqueous stem bark extract of B. papyrifera for two and four weeks respectively, the kidney morphology returned to normal with glomerular area, mesengial space, podocyte, proximal and distal tubule similar to that of the normal ( Plates 3 to 8); this may be due to curative effects of the B. papyrifera aqueous stem bark extract.

GROUP CREATININE UREA HCO3 Na K Cl /Treatment (µmol/L) (mmol/L) (mmol/L) (mmol/L) (mmol/L) (mmol/L)

I NO Acetaminophen 41.33 ± 6.03 2.43 ± 0.78 24.3 ±4.51 115.00 ±5.00 3.83 ± 0.55 89.00 ±7.21 Administered (NORMAL) II 80 mg/kg of acetaminophen 66.00 ± 6.25* 5.54 ± 1.42* 10.67 ± 1.16* 148.33 ± 5.77* 7.93 ± 1.40* 128.33 ± 7.64* (TEST CONTROL RATS) VI 50mg/kg ASBE of B.papyrifera 45.33 ± 2.52s 2.73 ± 0.21t 19.67 ± 1.528u 101.67 ± 10.41v 3.70 ± 0.61w 101.00±7.81x Administered VII 100mg/kg ASBE of B.papyrifer 42.33 ± 6.43 s 2.67 ± 0.65 t 20.33 ± 2.31 u 111.00 ±11.53 v 3.47 ± 0.35 w 98.00 ±1.00 x Administered VIII 150 mg/kg ASBE of B.papyrifera 40.00 ± 1.00 s 2.27 ±0.83 t 19.67 ± 5.03 u 114.33± 9.02 v 3.53 ± 0.65 w 98.33±2.08 x Administered

Results are expressed as mean ± SD,n = 3 Values with asterisk are significantly different at p<0.05 when compared with the normal rats. Values in the same column with the same superscript are significantly different at p<0.05 when compared with the test control HCO3 - Bicarbonate , Na - sodium, K- potassium, Cl- chloride

Table 2: Serum Kidney Function Indices of Acetaminophen Induced Kidney Damage Rats after the Four Weeks Oral Administration of Aqueous Stem Bark Extract of Boswellia Papyrifera.

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PLATE 3: Histopathology Lesion, photomicrograph of transverse section of acetaminophen induced rats kidney, group III, the kidney showing no significant pathology after two weeks oral administration of 50mg/kg B. papyrifera aqueous stem bark extract, the kidney showed normal architecture with glomerular area, mesengial space, podocyte proximal and distal tubule similar to that of the normal H & E stain X 10.

P L AT E 2 : H i s t o p a t h o l o g y L e s i o n , Photomicrograph of transverse section of test control rats kidney, group II, showing distortion of the normal kidney architecture with dilation of the renal tubules and collapsed of bowman's capsule due to damage caused by acetaminophen administered to the rats. H & E Stain X 10

P l a t e 1 ; H i s t o p a t h o l o g y L e s i o n , photomicrograph of transverse section of normal rats kidney group I , showing normal kidney architechure with the cortex containing the glomerulus and the medulla containing the renal tubules H & E STAIN X 10

PLATE 4: Histopathology Lesion, Photomicrograph of transverse section of acetaminophen induced rats kidney group IV, the kidney showing no significant pathology after two weeks oral administration of 100mg/kg B. papyrifera aqueous stem bark extract, the kidney showed normal architecture with glomerular area, mesengial space, podocyte proximal and distal tubule similar to that of the normal H & E stain X 10.

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PLATE 5: Histopathology Lesion, photomicrograph of transverse section of acetaminophen induced rats kidney, group V the kidney showing no significant pathology after two weeks oral administration of 150mg/kg of B. papyrifera aqueous stem bark extracts, the kidney showed normal architecture with glomerular area, mesengial space, podocyte proximal and distal tubule similar to that of the normal H & E stain X 10.

PLATE 6: Histopathology Lesion, photomicrograph of transverse section of acetaminophen induced rats kidney group VI the kidney showing no significant pathology a f t e r four weeks o ra l administration of 50mg/kg B. papyrifera aqueous stem bark extract , the k idney showed nor ma l architecture with glomerular area, mesengial space, podocyte proximal and distal tubule similar to that of the normal H & E stain X 10.

PLATE 7: Histopathology Lesion, photomicrograph of transverse section of acetaminophen induced rats kidney group VII the kidney showing no significant pathology after four weeks oral administration of 100mg/kg B. papyrifera aqueous stem bark extract, the kidney showed normal architecture with glomerular area, mesengial space, podocyte proximal and distal tubule similar to that of the normal H & E stain X 10.

PLATE 8: Histopathology Lesion, photomicrograph of transverse section of acetaminophen induced rats kidney group VIII the kidney showing no significant pathology after four weeks oral administration of 1 50mg/kg B. papyrifera aqueous stem bark extract, the kidney showed normal architecture with glomerular area, mesengial space, podocyte proximal and distal tubule similar to that of the normal H & E stain X 10.

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DISCUSSIONIn this study, acetaminophen was used to induce nephrotoxicity in rats, followed by treatment with aqueous stem bark extract of B. papyrifera at various doses. A single dose of 800mg/kg body weight of acetaminophen dissolved in normal saline was orally administered to induce nephrotoxicity in rats within 24 hours.

The rats injected with 800mg/kg body weight of acetaminophen (po) showed significant (p<0.05) increase in serum level of creatinine, urea, potassium, sodium, chloride and low serum level of bicarbonate when compared with the normal. This indicated inducement of renal failure in the injected rats. This is consistent with previous studies (Pelani et al; 2009), Cekman et al, (2008) and Adeneye et al, (2008) which demonstrated renal damage in rats following acetaminophen administration. At higher doses, acetaminophen induce renal insufficiency characterized by acute tubular necrosis, increase in serum creatinine level and electrolyte imbalances. Oxidative stress is reported to play roles in the pathogenesis of acetaminophen induced renal damage, whose metabolism occurs via cytochrome p450 enzymes (Bessems and Vermeulen, 2001).

The groups III , IV and V administered with 50, 100, 150 mg/kg of aqueous stem bark extract respectively for two weeks once daily showed insignificant difference ( p>0.05) in serum level of creatinine, urea, potassium, chloride and sodium when compared with the normal but significantly higher (p<0.05) when compared with the test control (Table 1). However, the mean serum level of bicarbonate showed significant decrease (p<0.05) when compared with normal but statistically not significant (p>0.05) when compared with the test control. The decrease in serum kidney indices is pronounced in groups administered daily dose of 100 and 150mg/kg body weight daily. This may be due to serum bioavailability of the aqueous stem bark extract; this showed the possible curative effect of the extract on the damage due to acetaminophen toxicity.

However, groups VI, VII and VIII administered daily doses of 50, 100 and 150mg/kg respectively of B.papyrifera aqueous stem bark extract for four

weeks showed no significant difference (p>0.05) when compared with the normal, but showed significant difference (p<0.05) when compared with the test control (group II; Table 2), This confirmed the curative activities of B. papyrifera aqueous stem bark extract, which may be due to phytochemical constituent of the plant. Flavonoids found in the stem bark extract have potential to be biological response modifiers such as anti-allergic, anti-inflammatory, anti-microbial and anti-cancer activities (Gaby, 1999).

it could therefore be suggested that the phytochemical constituents of aqueous stem bark extract of Boswellia papyrifera such as alkaloids, tannins, flavonoids, saponins and cardiac glycosides previously reported (Abdulmumin et al., 2014) may be responsible for the nephrocurative effect of the aqueous stem bark extract of the plant. They are super antioxidants and free radical scavengers which prevent oxidative cell damage and have strong anti-cancer activity and protects against all stages of carcinogens. Flavonoids in the body are known to reduce the risk of heart diseases (Urquiaga and Leighton, 2000). In terms of anti-cancer activity, they inhibit the initiation, promotion and progression of tumors (Urquiga and Leighton, 2000; Okwu, 2004).

The anti-oxidant property of Boswellia papyrifera could probably be mediated by enhancing superoxide dismutase and glutathione peroxidase activity or even increase catalase content of the kidney tissue (Hirano et al; 1991; Kavutcu et al; 1996; Ozturk et al, 1997). The histopathology of the kidney (Plates 6,7 and 8) four weeks after the administration of B. papyrifera aqueous stem bark extract showed no significant pathology, normal kidney with the cortex containing the glomerulus and the medulla containing the renal tubules; this supports the biochemical result obtained.

CONCLUSIONIn conclusion, Boswellia papyrifera aqueous stem bark extract has shown nephrocurative activity ag a ins t ace taminophen- induced rena l insufficiency. This may be attributed to the phytochemical content of the stem bark extract of the plant. We therefore suggest further

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research on the plant for its nephrocurative potentials.

ACKNOWLEDGEMENTS: This work is supported by Tertiary Education Trust Fund (TETFUND) and Hussaini Adamu Federal Polytechnics, Kazaure, Jigawa State.

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