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Journal of Chinese Integrative Medicine ›› 2011, Vol. 9 ›› Issue (12): 1373-1379.doi: 10.3736/jcim20111215

• Original Experimental Research • Previous Articles     Next Articles

Effects of combined leaf extract of Vernonia amygdalina and Azadirachta indica on hepatic morphology and hepatotoxicity markers in streptozotocin-induced diabetic rats

Oluwole Busayo Akinola1(), Gabriel Olaiya Omotoso1, Oluwafunmike Sharon Akinola1, Olufunke Olubusola Dosumu2, Esther Tomi Adewoye1   

  1. 1. Department of Anatomy, Faculty of Basic Medical Sciences, College of Health Sciences, University of Ilorin, Ilorin, Nigeria
    2. Department of Anatomy, College of Medicine, University of Lagos, Lagos, Nigeria
  • Received:2011-07-05 Accepted:2011-09-28 Online:2011-12-20 Published:2011-11-15

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Objective: In this work, we studied liver morphology, markers of hepatic oxidative stress and some liver enzymes in diabetic rats treated with the combined leaf extract (CLE) of Vernonia amygdalina (bitter leaf) and Azadirachta indica (neem).
Methods: Diabetes was induced in fasted male Wistar rats with intraperitoneal injection of streptozotocin (STZ). Oral CLE (500 mg/kg body weight) and metformin (150 mg/kg body weight) were administered to different groups of diabetic rats for eight weeks. Blood glucose and change in body weight were estimated weekly. All animals were sacrificed under anaesthesia after eight weeks. Hepatic sections were stained with periodic acid-Schiff. Liver samples were homogenized and assayed for contents of malondialdehyde (MDA) and glutathione peroxidase (GPx), while the plasma was assayed for contents of alanine aminotransferase (ALT) and aspartate aminotransferase (AST).
Results: Metformin and CLE treatment produced normoglycaemia in the diabetic rats in the course of the treatment period. Significant increases in body weight were observed in the treatment groups compared with the diabetic control rats (P<0.05). In the control and treatment groups, light microscopic study showed intact hepatic histology. Plasma ALT and AST were not significantly different from the control values in the CLE-treated rats. In addition, from week four onwards, blood glucose concentrations in the CLE-treated rats were not different from the normal control (P>0.05). Besides, hepatic MDA (P<0.05) significantly decreased in the CLE-treated rats compared with the normal control.
Conclusion: These findings suggest that CLE ameliorates hyperglycemia and hepatic oxidative stress when administered to diabetic rats as a chronic regimen, and there was no morphologic or biochemical evidence of liver damage at the dose tested.

Key words: diabetes mellitus, experimental, Vernonia, Azadirachta, plant extracts, hepatotoxicity, oxidative stress, rats

Figure 1

Blood glucose concentrations of control and treatment groups over a period of eight weeks Data are presented as mean±SEM, n=8, *P<0.05, vs normal control group; △P<0.05, vs diabetic control group. SEM: standard error of mean; STZ: streptozotocin; CLE: combined leaf extract of Vernonia amygdalina and Azadirachta indica."

Figure 2

Changes in body weights of the rats after eight weeks of treatment Data are presented as mean±SEM, n=8, *P<0.05, vs normal control group; △P<0.05, vs diabetic control group. SEM: standard error of mean; STZ: streptozotocin; CLE: combined leaf extract of Vernonia amygdalina and Azadirachta indica."

"

Group n Plasma ALT (IU/L) Plasma AST (IU/L) Hepatic MDA (μmol/mg protein) Hepatic GPx (U/mg protein)
Normal control 8 36.3±5.1 68.1±6.2 1 650±150 1.20±0.15
Diabetic control 8 37.0±3.2 88.3±7.1* 1 410±132 1.22±0.30
STZ plus metformin 8 45.2±4.8* 89.8±6.5* 1 305±98 1.60±0.41*
STZ plus CLE 8 28.6±3.1 30.1±2.8△ 903±69*△ 0.62±0.05
CLE control 8 25.1±5.5 30.1±3.1△ 1 906±161△ 1.50±0.35*

Figure 3

Hepatic histology after eight weeks of treatment (Light microscopy, ×400) A: Normal control; B: Diabetic control; C: STZ plus CLE; D: STZ plus metformin; E: CLE alone. In each of the groups, hepatic sections are periodic acid-Schiff-negative; besides, no notable differences exist in hepatic morphology of the control and treated rats. Thick white arrows point to the Kupffer cells; thick black arrows point to the sinusoid; thin arrows point to the hepatocyte nucleus; C means the central vein. STZ: streptozotocin; CLE: combined leaf extract of Vernonia amygdalina and Azadirachta indica."

[1] Savvidou S, Goulis J, Giavazis I, Patsiaoura K, Hytiroglou P, Arvanitakis C . Herb-induced hepatitis by Teucrium polium L. : report of two cases and review of the literature[J]. Eur Gastroenterol Hepatol. 2007 ; 19(6):507-511.
doi: 10.1097/01.meg.0000252634.26538.eb pmid: 17489062
[2] Freire RB, Borba HR, Coelho CD . Ruta graveolens L. toxicity in Vampirolepis nana infected mice[J]. Indian Pharmacol. 2010; 42(6):345-350.
doi: 10.4103/0253-7613.71898 pmid: 2991689
[3] Alexander GC, Sehgal NL, Moloney RM, Stafford RS . National trends in treatment of type 2 diabetes mellitus, 1994-2007. Arch Intern Med. 2008; 168(19):2088-2094.
doi: 10.1001/archinte.168.19.2088
[4] Akinola OS, Akinola OB, Caxton-Martins EA . Vernonia amygdalina upregulates hepatic enzymes and improves liver microanatomy in experimental diabetes mellitus. Pharmacologyonline. 2009 ; 2 : 1231-1242.
[5] Akinola OB, Caxton-Martins EA, Dini L . Chronic treatment with ethanolic leaf extract of Azadirachta indica ameliorates lesions of pancreatic islets in strepto- zotocin diabetes[J]. Int Morphol. 2010; 28(4):292-302.
[6] Ebong PE, Atangwho IJ, Eyong EU, Egbung GE . The antidiabetic efficacy of combined extracts from two continental plants : Azadirachta indica( A. Juss) 2008; 4(3):239-244.
[7] Matsui Y, Hirasawa Y, Sugiura T, Toyoshi T, Kyuki K, Ito M . Metformin reduces body weight gain and improves glucose intolerance in high-fat diet-fed C57BL/6J mice. Biol Pharm Bull. 2010; 33(6):963-970.
doi: 10.1248/bpb.33.963
[8] Bancroft JD, Stevens A . Theory and practice of histological techniques. 2nd ed. New York: Churchill Livingstone. 1982: 131-135.
[9] Reitman S, Frankel S . A colorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases. Am[J] Clin Pathol. 1957 ; 28(1):56-63.
[10] Ohkawa H, Ohishi N, Yagi K . Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem. 1979; 95(2) : 351-358.
doi: 10.1016/0003-2697(79)90738-3
[11] Paglia DE, Valentine WN . Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase.[J] Lab Clin Med. 1967; 70(1):158-169.
[12] Weichselbaum TE . An accurate and rapid method for the determination of proteins in small amounts of blood serum and plasma[J]. Am Clin Pathol. 1946 ; 10 : 40-49.
[13] Osinubi AA . Effect of Vernonia amygdalina and chlorpropamide on blood glucose[J]. Med Islamic World Acad Sci. 2006; 16:115-119.
[14] Fu S, Korkmaz E, Braet F, Ngo Q, Ramzan I . Influence of kavain on hepatic ultrastructure[J]. World Gastroenterol. 2008; 14(4) : 541-546.
doi: 10.3748/wjg.14.541 pmid: 2681144
[15] Chen GC, Ramanathan VS, Law D, Funchain P, Chen GC, French S, Shlopov B, Eysselein V, Chung D, Reicher S, Pham BV . Acute liver injury induced by weight-loss herbal supplements[J]. World Hepatol. 2010; 2(11) : 410-415.
doi: 10.4254/wjh.v2.i11.410
[16] Tarantino G, Pezzullo MG, Dario di Minno MN, Milone F, Pezzullo LS, Milone M, Capone D . Drug- induced liver injury due to anatural products^ used for weight loss : a case report [J]. World Gastroenterol. 2009; 15(19) : 2414-2417.
doi: 10.3748/wjg.15.2414
[17] Chau T . Drug-induced liver injury: an update. Med Bull. 2008; 13(3):23-26.
[18] Vianna R, Brault A, Martineau LC, Couture R, Arnason JT, Haddad PS . In vivo anti-diabetic activity of the ethanolic crude extract of Sorbus decora CK Schneid. ( Rosacea) : a medicinal plant used by Canadian James Bay Cree Nations to treat symptoms related to diabetes. Evid Based Complement Alternat Med. 2009 Nov 3. [Epub ahead of print]
[19] Olivera-Gonzalez S, de Escalante-Yanguela B, Velilla-Soriano C, Amores-Arriaga B, Martin-Fortea P, Navarro-Aguilar ME . Metformin-associated hepatotoxicity. Med Intensiva. 2010; 34(7) : 483-487.
doi: 10.1016/j.medin.2009.10.006
[20] Calabrese AT, Coley KC, DaPos SV, Swanson D, Rao RH . Evaluation of prescribing practices: risk of lactic acidosis with metformin therapy. Arch Intern Med. 2002; 162(4) : 434-437.
doi: 10.1001/archinte.162.4.434
[21] Large V, Beylot M . Modifications of citric acid cycle activity and gluconeogenesis in streptozotocin-induced diabetes and effects of metformin. Diabetes. 1999 ; 48(6):1251-1257.
doi: 10.2337/diabetes.48.6.1251
[22] McGuinness ME, Talbert RL . Phenformin-induced lactic acidosis : a forgotten adverse drug reaction. Ann Phar- macother. 1993; 27(10):1183-1187.
doi: 10.1177/106002809302701004 pmid: 8251683
[23] Baynes JW . Role of oxidative stress in development of complications in diabetes. Diabetes. 1991; 40(4):405-412.
doi: 10.2337/diabetes.40.4.405 pmid: 2010041
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