American Journal of Pharmacological Sciences
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American Journal of Pharmacological Sciences. 2015, 3(3), 79-86
DOI: 10.12691/ajps-3-3-5
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Hepatoprotective Effect of Ethanolic Leaf Extract of Vernonia amygdalina and Azadirachta indica against Acetaminophen-Induced Hepatotoxicity inSprague-Dawley Male Albino Rats

Momoh Johnson1, , Longe Adeteju Olufunmilayo1, Damazio Olanrewaju Anthony2 and Eleyowo Oluwole Olusoji3

1Department of Science Laboratory Technology (Biochemistry units), School of Pure and Applied Sciences, Lagos State Polytechnic, Ikorodu, Lagos, Nigeria

2Department of Science Laboratory Technology (Chemistry units), School of Pure and Applied Sciences, Lagos State Polytechnic, Ikorodu, Lagos, Nigeria

3Department of Science Laboratory Technology (Environmental Biology units), School of Pure and Applied Sciences, Lagos State Polytechnic, Ikorodu, Lagos, Nigeria

Pub. Date: July 05, 2015

Cite this paper:
Momoh Johnson, Longe Adeteju Olufunmilayo, Damazio Olanrewaju Anthony and Eleyowo Oluwole Olusoji. Hepatoprotective Effect of Ethanolic Leaf Extract of Vernonia amygdalina and Azadirachta indica against Acetaminophen-Induced Hepatotoxicity inSprague-Dawley Male Albino Rats. American Journal of Pharmacological Sciences. 2015; 3(3):79-86. doi: 10.12691/ajps-3-3-5


Acetaminophen (paracetamol) is a commonly and widely used analgesic and antipyretic agent, but at high dose it leads to undesirable side effects, such as hepatotoxicity. The study investigate the hepatoprotective effect of ethanolic leaf extract of Vernonia amygdalina and ethanolic leaf extract of Azadirachta indica against acetaminophen-induced hepatotoxicity in Sprague -Dawley male albino rats. Male albino rats were randomly divided into six groups each consisting of five albino rats. Group A rats served as the normal control and were given water daily for a period of 14 days. Hepatotoxicity was induced in-vivo to all animals of Groups B, C, D, E, and F orally by administering 2g /kg body weight of paracetamol once a day for a period of 14 days. Group C, D, E and F were orally administered silymarin (100 mg/kg B.W), Vitamin C (100 mg/kg B.W), ethanolic leaf extract of V. amygdalina (300 mg/kg B.W) and A. indica (300 mg/kg B.W) respectively daily for a period of 14 days. Group B animals served as the paracetamol control and they were not treated. The result of this study shows that animals treated with silymarin, V. amygdalina and A. indica extracts significantly (P<0.05) have reduced WBC count compared to paracetamol control group. HGB, RBC and HCT values in all the groups administered with silymarin, Vitamin C, V. amygdalina and A. indica extracts were significantly (P<0.05) increased when compared to the paracetamol- intoxicated animals without treatment. Oral administration of acetaminophen caused marked liver damage as noted by the significant increased (P<0.05) in activities of plasma AST, ALT, ALP and GGT as well as the level of cholesterol, triglyceride and a reduction in plasma total protein. The drug also resulted to a significant increase (P<0.05) in liver MDA content, decrease in liver GSH content, decrease in SOD and CAT activities. Treatment with silymarin, Vitamin C, V. amygdalina and A. indica extracts showed effective hepatoprotective effect as evidence in the decrease in the plasma levels of liver biomarker enzymes and reduction in oxidative stress parameters. Histopathological evaluation of the liver architecture also revealed that all the treated animals have reduced the incidence of paracetamol- induced liver lesions.

acetaminophen antioxidant Azadirachta indica haematological parameters hepatoprotective effect histopathology liver biomarker enzymes and Vernonia amygdalina

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[1]  Vitaglione P, Morisco F, Caporaso N, Fogliano V. Dietary antioxidant compounds and liver health. Crit. Rev. Food Sci. Nutr, 2004; 44: 575-586.
[2]  Cristovao FL, Manuel FF, Cristina PW. Drinking of Salvia officinalis tea increases CCl4-induced hepatotoxicity in mice. Food Chem. Toxicol, 2007; 45: 456-464.
[3]  Aruoma, O.I. Nutrition and health aspects of free radicals and antioxidants. Food Chem. Toxicol. 1994; 32: 671-683.
[4]  Wu, J, Danielsson, A, Zern, M.A. Toxicity of hepatotoxins: new insights into mechanisms and therapy. Expert Opin. Investig. Drugs 8, 1999; 585-607.
[5]  Bansal, A.K., Bansal, M., Soni, G., Bhatnagar, D. N-nitrosodiethylamine induced oxidative stress in rat liver. Chem. Biol. Interact. 2005; 156: 101-111.
[6]  James LP, Mayeux PR, Hinson JA. Acetaminophen-induced hepatotoxicity. Drug Metab Dispos. 2003; 31: 1499 -1506.
[7]  Jollow DJ, Mitchell JR, Potter WZ, Davis DC, Gillette JR, Brodie BB. Acetaminophen induced hepatic necrosis. II. Role of covalent binding in vivo. J Pharmacol Exp Ther. 1973; 187: 195-202.
[8]  Tiwari BK, Khosa RL. Hepatoprotective and antioxidant effect of Sphaeranthus indicus against acetaminophen–induced hepatotoxicity in rats. The Internet Journal of Tropical Medicine. 2010; 6: 1540-2681.
[9]  Kshirsagar A, Ingawale D, Ashok P, Vyawahare N. Silymarin: A Comprehensive Review. Pharmacognosy Reviews . 2009; 3: 116-121.
[10]  Dehmlow C, Erhard J, de Groot H. Inhibition of Kupffer cell functions as an explanation for the hepatoprotective properties of silibinin. Hepatology. 1996; 23: 749-754.
[11]  Jeong DH, Lee GP, Jeong WI, Do SH, Yang HJ, Yuan DW, et al. Alterations of mast cells and TGF-beta1 on the silymarin treatment for CCl4- induced hepatic fibrosis World J Gastroenterol. 2005; 11: 1141-1148.
[12]  Fraschini F, Demartini G, Esposti D. Pharmacology of Silymarin. Clin Drug Inv 2002; 22: 51-65.
[13]  Halliwell, B. Role of free radicals in the neurodegenerative diseases: therapeuticimplications for antioxidant treatment. Drugs Ageing, 2001; 18(9): 685-716.
[14]  Woollard, KJ, Loryman CJ, Meredith E, Bevan R, Shaw JA, Lunec J and Griffith HR. Effects of oral vitamin C on monocyte: Endthelial cell adhension in healthy subjects. Biochem Biophys Res. Commun, 2002; 294: 1161-1168.
[15]  Asiley SB, Elliot RM, Archer DB and Southon S. Evidence that dietary supplementation with carotenoids-rich food modulates the DNA damage:Repair balance in human lymphocytes Br. J. Nutr, 2004; 91: 63-72.
[16]  McDowell LR. Vitamins in Animal Nutrition- Comparative Aspects to Human Nutrition: Vitamin C. Acad. Press, London. 1989; 10-52, Pp 93-131,
[17]  Parola M, Leonarduzzi G, Biasi F, Albano E, Biocca ME, Poli G, Dianzani MU. Vitamin E dietary supplementation protects against carbon tetrachlorideinduced chronic liver damage and cirrhosis. Hepatology, 1992; 16: 1014-1021.
[18]  Sies H, Stahi KL and Sundquist AR. Antioxidant functions of vitamin E and C, $ carotene and other carotenoids. Annals. New York Acad. Sci, 1992; 669: 7-20.
[19]  Argheore EM., Makkar HPS, Becker K. Feed value of some browse plants from central zone of Delta State of Nigeria. Trop Sci, 1998; 38: 97-104.
[20]  Egedigwe C A. "Effects of Dietary Incorporation of Vernonia amygdalina and Vernonia colorata on Blood Lipid Profile and Relative Organ Weights in Albino Rats", M.Sc. Dissertaton, Department of Biochemistry, Michael Opara University of Agriculture Umudike, Nigeria. 2010.
[21]  Girish K. and Shankara BS, Neem A. Green Treasure. Electronic Journal of Biology, 2008; 4(3), 102-111.
[22]  Allameh AMR, Abyaneh MA, Shams MB, Rezaee, Jaimand K. Effects of neem leaf extract on production of aflatoxins and activities of fatty acid synthetase, isocitrate dehydrogenase and glutathione-stransferasein Aspergillusparasiticus. Mycopathologia, 2002; 54:79-84.
[23]  Mossini SA, Oliveira KP, KemmelmeierC. Inhibition of patulinproduction By peniculliumexpansun culture with neem (Azadirachta indica) leaf extracts. Basic Microbiol, 2004; 44:106-113.
[24]  Niehaus WG, Samuelsson B. Formation of malondialdehyde from phosopholipid arachidonate during microsomal lipid peroxidation. Eur J. Biochem, 1968; 6: 126-130.
[25]  Magwere T, Naiks YS, Hasler JA. Effect of Chloroquine treatment on antioxidant enzymes in rat liver and kidney Free. Rad. Biol. Med, 1997; 22: 321-327.
[26]  Sinha KA. Colorimetric assay of catalase. Anal Biochem, 1972; 47: 389-394.
[27]  Ellman GL. Tissue sulphydryl groups. Arch Biochem Biophys, 1959; 82: 70-77.
[28]  Galighor AE, Kozloff EN. Essentials of practical microtechnique 2nd edn, Lea and Febiger, NewYork. 1976.
[29]  Momoh J, Odetunde SK, Longe A O. Phytochemical analysis, in vitro evaluation of antioxidant and antimicrobial activity of methanolic leaf extract of Vernonia amygdalina (bitter leaf) against Staphylococcus aureus and Pseudomonas aeruginosa. Int.J.Curr.Microbiol.App.Sci, 2015; 4(5): 411-426
[30]  Momoh J, Iwalokun BA, Longe OA, Babalola OM. "Antimicrobial Susceptibility patterns of methanolic leaf extract of Azadirachta Indica and some selected antibiotics and Plasmid Profiles of Escherichia Coli Isolates Obtained from Different Human Clinical Specimens in Lagos- Nigeria", Science Journal of Microbiology. 2014; 6 Pages.
[31]  Al- Mamary M, Al-Meeri A, Al-Habori M. Antioxidant activities and total phenolics of different types of honey. Nutrition Research, 2002; 22: 1041-1047.
[32]  Gupta VK, Sharma SK. Plants as natural antioxidants. Natural products radiance. 2006; 5: 326-334.
[33]  Sexena DP, Shukla SK, K, Kumar R. Efficacy studies of In vitro screening of antiplasmodial activity by crude extracts of Diospyros melanoxylem Res. J. Med. Plant. 2011; 5: 312-320.
[34]  Ohaeri CC, Eluwa MC. Abnornal biochemical and hematological indices in trypanosomiasis as a threat to herd production. Vet. Parasitol. 2011;177: 199-202.
[35]  Momoh J, Damazio OA. Hepatotoxicity of House Hold Kerosene (HHK) on liver enzyme markers and its effect on hematological and oxidative stress parameters on wistar albino rats. Science Journal of Medicine and Clinical Trials. 2014: 7 pages.
[36]  Mahesh A, Shaheetha J, Thangadurai D, Rao DM. Protective effect of Indian honey on acetaminophen induced oxidative stress and liver toxicity in rat. Biologia. 2009, 64: 1225-1231.
[37]  Nkosi CZ, Opoku AR, Terblanche SE. Effect of pumpkin seed (Cucurbita pepo) protein isolate on the activity levels of certain plasma enzymes in CCl4-induced liver injury in low-protein fed rats. Phytother Res. 2005; 19: 341-345.
[38]  Bairwa NK, Sethiya, Mishra S. Protective effect of stem bark of Ceiba pentandra linn. against paracetamol-induced hepatotoxicity in rats. Pharmacognsy research. 2010; 2: 26-30.
[39]  Kuriakose GC, Kurup MG. Hepatoprotective effect of Spirulina lonar on paracetamol induced liver damage in rats. Asian J Exp Biol Sci. 2010; 1: 614-662.
[40]  Randle LE, Sathish JG, Kitteringham NR, Macdonald I, Williams DP, Park BK. Alpha(1)-adrenoceptor antagonists prevent paracetamol-induced hepatotoxicity in mice. Br J Pharmacol. 2008; 153: 820-830.
[41]  Rasool MK, Sabina EP, Ramya SR, Preety P, Patel S, Mandal N, et al. Hepatoprotective and antioxidant effects of gallic acid in paracetamolinduced liver damage in mice. J Pharm Pharmacol. 2010; 62: 638-643.
[42]  Catala A. Lipid peroxidation of membrane phospholipids generates hydroxy-alkenals and oxidized phospholipids active in physiological and/or pathological conditions. Chemistry and Physics of Lipids. 2009; 157: 1-11.
[43]  Peter B, Wartena M, Kampinga HH, Konings AW. Role of lipid peroxidation and DNA damage in paraquat toxicity and the interaction of paraquat with ionizing radiation. Biochem Pharmacol. 1992; 43: 705-715.
[44]  Bors W, Saran M. Radical scavenging by flavonoids antioxidants. Free Radic Res Commun. 1987; 2: 289-294.
[45]  Manna SK, Mukhopadhyay A, Van NT, Aggarwal BB. Silymarin suppresses TNF-induced activation of NF-kappa B, c-Jun N-terminal kinase, and apoptosis. J Immunol. 1999; 163: 6800-6809.
[46]  Hiscott J, Marois J, Garoufalis J, D’Addario M, Roulston A, Kwan I, et al. Characterization of a functional NF-kappa B site in the human interleukin 1 beta promoter: evidence for a positive autoregulatory loop. Mol Cell Biol. 2006; 13: 6231-6240.
[47]  Hwang D, Jang BC, Yu G, Boudreau M. Expression of mitogeninducible cyclooxygenase induced by lipopolysaccharide: mediation through both mitogen-activated protein kinase and NF-kappaB signalling pathways in macrophages. Biochem Pharmacol. 1997; 54: 87-96.
[48]  Soon KJ, Young JJ, Song-Kyu P, Kyu-Hwan Y, Mook KH. Protection against lipopolysaccharide-induced sepsis and inhibition of interleukin-1b and prostaglandin E2 synthesis by silymarin. Biochemical Pharmacology. 2004; 67: 175-181.
[49]  Videla LA, Valenzuela A. Alcohol ingestion, liver glutathione and lipoperoxidation: metabolic interrelations and pathological implications. Life Sci. 1982; 31: 2395-2407.
[50]  Tirmenstein MA, Nelson SP. Sub cellular binding and effects on calcium homeostasis produced by acetaminophen and a non-hepatotoxic regioisomer 3-hydroxyacetanilide in mouse liver. J Biol Chem, 1989; 264: 9814-9819.
[51]  Vermeulen NPE, Bessems JGM, Van DE, Straat R. Molecular aspects of paracetamol-induced hepatotoxicity and its mechanism based prevention. Drug Metab Rev, 1992; 24: 367-407.
[52]  Bailey B, Amre DK, Gaudreault P. Fulminant hepatic failure secondary to acetaminophen poisoning: a systematic review and meta-analysis of prognostic criteria determining the need for liver transplantation. Crit Care Med. 2003; 31: 299-305.
[53]  Lee WM. Acetaminophen and the U.S. Acute Liver Failure Study Group: lowering the risks of hepatic failure. Hepatology. 2004; 4: 6-9.
[54]  Moore M, Thor H, Moore G, Nelson S, Moldeus P, Orrenius S. The toxicity of acetaminophen and N-acetyl-p-benzoquinone imine in isolated hepatocytes is associated with thiol depletion and increased cytosolic Ca2+. J Biol Chem. 1985; 260: 13035-13040.
[55]  Oz HS, McClain CJ, Nagasawa HT, Ray MB, De Villiers WJ, Chen TS. Diverse antioxidants protect against acetaminophen hepatotoxicity. J Biochem Mol Toxicol. 2004; 18: 361-368.
[56]  Volmar B, Menger MD. The hepatic microcirculation: mechanistic contributions and therapeutic targets in liver injury and repair. Physiol Rev. 2009; 89: 1269-1339.