Evaluation of antidiabetic and antioxidative efficacy of ethyl acetate fraction of methanolic extract of Camellia sinensis (green tea) leaves in streptozotocin induced diabetic albino rat
Keywords:
Streptozotocin, C.sinensis, Antioxidant, Hexokinase-I, Bax, Bcl-2Abstract
Objective: To investigate the antidiabetic and antioxidative potentiality of ethyl-acetate fraction of methanolic extract of Camellia sinensis (Green tea) leaves in streptozotocin induced diabetic rat. Methods: Streptozotocin induced diabetic state was confirmed by increased level of fasting blood glucose, decreased level of serum insulin along with inhibition in carbohydrate metabolomics. Oxidative stress was assessed by measuring antioxidative enzyme activities of hepatic and skeleto-muscular tissue. Hepatic Hexokinase-I, pro-apoptotic Bax and anti-apoptotic Bcl-2 gene expression patterns were noted by qRT-PCR technique. Results: After treatment with ethyl-acetate fraction of methanolic extract of Camellia sinensis (Green tea) leaves at a dose of 100 mg/kg body weight/day to diabetic rats for 28 days, a significant (p < 0.05) recovery was noted in fasting blood glucose level, serum insulin level along with activities of carbohydrate metabolic enzymes in hepatic tissue in respect to the vehicle treated diabetic group. This fraction also resulted a significant (p < 0.05) recovery in the activities of antioxidative enzymes in hepatic and skeleto - mascular tissue. In streptozotocin induced diabetic rat the low level of expression of Hexokinase-I, anti-apoptotic Bcl-2 and high level of expression of pro-apoptotic Bax gene were observed in hepatic tissue in respect to vehicle treated control. There were recovered significantly after the treatment with the said fraction. Conclusion: From the results, it may be concluded that ethyl-acetate fraction of methanolic extract of leaves of C. sinensis has a promising anti-diabetic and antioxidative activities for the management of streptozotocin induced diabetic state.
References
.Namita P, Mukesh R, Vijay KJ. Camellia sinensis (Green tea): A Review. Global J Pharmacol, 2012; 6: 52-59. [2].Chatterjee K, Ali KM, De D, Mallick C, Ghosh D. Antihyperglycemic, antioxidative activities of a formulated polyherbal drug MTEC (Modified) in streptozotocin-induced diabetic rat. J Med Plants Res, 2009; 3: 468-480. [3].Sekiguchi N, Inoguchi T, Kobayashi K, Nawata H. Effect of erythropoietin on endothelial cell apoptosis induced by high glucose. Diabetes Res Clin Prac, 2004; 66: 103-107. [4].EL-Hela A, Abdulla A. Antioxidant and antimicrobial activity in relation to polyphenolic content,J Applied Seires, 2000; 6: 683-689. [5].De D, Ali KM, Chatterjee K, Bera TK, Ghosh D. Protective effect of aqueous extract of seed of Psoralea corylifolia (Somraji) and seed ofTrigonella foenum-graecum L. (Methi) in streptozotocin-induced diabetic rat: A comparative evaluation. Trop J Pharm Res, 2009; 7 : 1-10. [6].WHO. World Health Organization Expert committee on Diabetes Mellitus. Second report, Technical report series, Geneva. 1980; 646: 1-80. [7].Research Council of the National Academies, Guide for the Care and Use of Laboratory Animals, 8th ed, Committee for the the upated of the guide for the care and use of laboratory animals, Publication no. N01- OD-4-2139 #188, The National Academies Press, NW, Washing DC, 2011; 1-246. [8].Ali KM, Chatterjee K, De D, Bera TK, Ghosh D. Efficacy of aqueous extract of seed of Holarrhena antidysenterica for the management of diabetes in experimental model rat: A correlative study with antihyperlipidemic activity. Int J App Res Nat Product, 2009; 2: 13-21. [9].Lenzen S. The mechanisms of alloxan and streptozotocin-induced diabetes. Diabetolog, 2008; 51: 216-226. [10].Mallick C, Chatterjee K, Mandal U, Ghosh D. Antihyperglycemic, antilipidperoxidative and antioxidative effects of extracts of Musa paradisiaca and Coccinia indica in streptozotocin-induced diabetic rat. Ethiop Pharm J, 2007; 25: 9-22. [11].Chou AC, Wilson JE. Carbohydrate metabolism. In: Wood WA, (ed.) Methods in Enzymol. Vol. XIII. New York, Academic Press 1975; 20-21. [12].Swanson MA. Glucose-6-phosphatase from liver. In: Colowick SP, Kaplan NO (eds.) Methods in Enzymol: Vol. II. Academic Press, New York 1955; 541-543. [13].Langdon RG, Glucose-6-phosphate dehydrogenase from erythrocytes. In: Wood WA (ed.): Methods in Enzymol. Vol. IX. Academic Press, New York 1966; 126-131. [14].Beers RF, Sizer IW. Spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Bio Chem, 1952; 195: 133-140. [15].Marklund S, Marklund G. Involvement of superoxide anion in autooxidation of pyrogallol and a convenient assay of superoxide dismutase. Europ J Biochem, 1974; 47: 469-474. [16].Jonas JC, Sharma A, Hasenkamp W, Ilkova H, Patane G, Laybutt R. Chronic hyperglycemia triggers loss of pancreatic beta cell differentiation in an animal model of diabetes. J Biol Chem, 1999; 274: 14112-14121. [17].Henry RJ, Chiamori M, Golub OJ, Berkman S. Revised spectrophotometric methods for the determination of glutamate-oxaloacetic transaminase, glutamate-pyruvate transaminase and lactic acid dehydrogenase. Am J Clinl Pathol, 1960; 34: 381-398. [18].Sokal RR, Rohle FJ. Introduction to Analysis of Variance. In: Biometry, Sokal RR, Rohle FJ (eds) WH Freeman and Company, New York. 1997; 179-206. [19].Kamalakkanan N, Prince PSM. Antihyperglycaemic and antioxidant effect of rutin, a polyphenolic flavonoid, in streptozotocin induced diabetic Wistar rats. Basic Clin Pharmacol Toxicol, 2006; 98: 97-103. [20].Murali B, Upadhaya UM, Goyal RK. Effect of chronic treatment with Enicosistemma littorale in non-insulin-dependent diabetic (NIDDM) rats. J Ethnopharmacol, 2002; 81: 199-204. [21].Chauhan S, Nath N, Tule V. Antidiabetic and antioxidant effects of Picrorhiza kurrooa rhizome extracts in diabetic rats. Ind J Clin Biochem, 2008; 23: 238-242. [22].Mallick C, Surva M, Baric B, Bhattacharya A, Ghosh D. Protection of testicular dysfunction by METC, a formulated herbal drug in STZ induced diabetic rat. Biol Pharm Bull, 2007; 30: 84-90. [23].Jana K, Chatterjee K, Bera TK, Maiti S, De D, Ali KM, Ghosh D. Antihyperglycemic and antihyperlipidemic effects of hydro-methanolic extract of seed of Caesalpinia bonduc in streptozotocin induced diabetic male albino rat. Int J Pharm Tech Res, 2010; 2: 2234-2242. [24].Pari L, Murugan P. Effect of tetrahydrocurcumin on blood glucose, plasma insulin and hepatic key enzymes in streptozotcin induced diabetic rats. J Basic Clin Physiol Pharmacol, 2005; 16: 257-274. [25].Murphy ED, Anderson JW. Tissue glycolytic and gluconeogenic enzymes activities in mildly and moderate diabetic rats. Influence of tolbutamide administration. Endocrinol, 1974; 94: 27-34. [26].Bera TK, De D, Chatterjee K, Ali KM, Ghosh D. Effect of Diashis, a polyherbal formulation, in streptozotocin-induced diabetic male albino rats. Int J Ayurveda Res, 2010; 1: 18-24. [27].Saxena AK, Srivastava P, Kale RK, Baquer NZ. Impaired antioxidant status in diabetic rat liver. Effect of vanadate. Biochem Pharmacol, 1993; 45: 539-542. [28]. Panda DK, Ghosh D, Bhat B, Talwar SK, Jaggi M, Mukherjee R. Diabetic therapeutic effects of ethyl acetate fractions from the roots of Musa Paradisiaca and seeds of Eugenia jambolana in streptozotocin-induced male diabetic rats. Methods Find Exp Clin Pharmacol, 2009; 31: 571-584. [29].Griffin LD, Gelb BD, Adams V, McCabe ERB. Developmental expression of hexokinase I in the rat. Biochim Biophys Acta, 1992; 1129: 309-317. [30].Jana K, Chatterjee K, Ali KM, De D, Bera TK, Ghosh D. Antihyperglycemic and antioxidative effect of the hydro-methanolic extract of the seeds of the Caesalpinia bonduc on streptozotocin-induced diabetes in male albino rats. Pharm Res, 2012; 4: 57-62. [31].Jaeschke H. Reactive oxygen and mechanisms of inflammatory liver injury. J Gastroen Hepatol, 2000; 15: 718–724. [32].Ghosh S, Suryawanshi SA. Effect of Vinca rosea extracts in treatment of alloxan diabetes in male albino rats. Ind J Exp Biol, 2001; 39: 748-759.
