Carbon tetrachloride (CCl4), riboflavin, deoxyribose, carrageenan and silymarin were purchased from Sigma Chemicals, USA. Serum glutamate pyruvate transaminase (SGPT), Serum Glutamate Oxaloacetate Transaminase (SGOT), Serum Alkaline Phosphatase (ALP) and Serum Total Bilirubin (T. Bil) assay kits were purchased from Span diagnostics Ltd, Gujarat, India. All other chemicals used were of analytical Crizotinib price grade. Adult albino Wistar rats (National Institute of Nutrition, Hyderabad, India) of either sex weighing 150–200 g were used in the studies.
The animals were maintained under standard laboratory conditions at an ambient temperature of 23 ± 2 °C having 50 ± 5% relative humidity with 12 h light and dark cycle. The use and care of the animals in the experimental protocol has been approved by the local Institutional Animal Ethics Committee (Regd. No. 516/01/A/CPCSEA) following the guidelines of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Government of India.
The acute toxicity study was conducted for hydroalcoholic, methanolic, ethyl acetate and hexane extracts of ABT737 G. gynandra as per OECD (Organisation for Economic Co-operation and Development) guidelines 420 (OECD.2001). Total phenolic content was determined using the Folin–Ciocalteu reagent7 using standard gallic acid calibration curve, measure the concentration of phenolic content in gallic acid total equivalents using unit’s mg/gm (GAE).8 The Fazel Shamsa et al, 2008 method using atropine calibration curve, measure the concentration of alkaloid content in atropine equivalents using unit’s mg/g.9 Superoxide scavenging activity10 of the plant extracts was determined by McCord & Fridovich method, which depends on light induced superoxide generation by riboflavin and the corresponding reduction of nitroblue tetrazolium.11 Hydroxyl radical scavenging activity was measured by
studying the competition between deoxyribose and the extracts for hydroxyl radicals generated from the Fe2+/EDTA/H2O2 system (Fenton reaction). The hydroxyl radical attacks deoxyribose, which CYTH4 eventually results in the formation of thiobarbituric acid reacting substances (TBARS).12 The scavenging activity for DPPH free radicals was measured according to the procedure described by Braca et al, 2003.13 and 14 In the present work hepatoprotective activity of different extracts of G. gynandra were tested against carbon tetrachloride (CCl4) induced hepatotoxicity by measuring biochemical enzymes (SGOT, SGPT, ALP and T. BIL). An increase in the enzymes levels of these biochemical parameters is a sensitive index of hepatic damage. The standard and test group animals were treated with 50 mg/kg dose of silymarin and 100, 200, 400 mg/kg doses of different extracts of G. gynandra for 6 days. On 6th day, 1 h after treatment with standard drug and selected plant extracts, the animals were intoxicated with CCl4 in liquid paraffin (1:1 v/v, 0.75 ml of CCl4/kg, i.p.).