paniculata and S. see more chirayita at the dose of 200 mg/kg b.w. orally daily for 16 days respectively. Vehicle, extract and standard drug administered 1 h before CCl4 administration. After 24 h of last dose, blood collected from overnight fasted rats of each group by cardiac puncture, for estimation of serum biochemical parameters. Then the rats sacrificed after 24 h after induction by cervical dislocation for the study of liver biochemical and histopathological parameters.
After 24 h of last dose the animals were dissected under ether anesthesia. Blood was collected from overnight fasted rats of each group by cardiac puncture and collected in previously labeled centrifuging tube stand and allowed to clot for 30 min at room temperature. Serum was separated by centrifugation at 3000 rpm for 15 min. The separated serum was used for the estimation of some biochemical parameters, 10% liver portion was homogenate and used for liver biochemical evaluation. Serum was analyzed for various serum biochemical parameters i.e. serum glutamine oxaloacetate transaminase (SGOT or AST), serum glutamine pyruvate transaminase (SGPT or ALT),13 serum alkaline
phosphatase (SALP),14 serum total bilirubin (TB),15 γ-glutamate transpeptidase (GGTP)16 and total protein (TP)17 content using reported method with the help of commercially available kits (SPAN Diagnostics). The homogenate portions of liver used SAHA HDAC supplier for the estimation of various biochemical parameters like level of lipid peoxidation (LPO)18 and expressed as nM/mg protein of liver tissue. The reduced glutathione (GSH) content of liver tissue was determined as per reported method19 and expressed as mM/gm of liver tissue. The catalase (CAT) activities in liver tissue were assayed as per the methods described20 and expressed in terms of U/mg protein of liver tissue. The superoxide dismutases (SOD)21 level also estimated according to the prescribed methods. In histopathological study, liver from each animal removed after dissection and preserved immediately in 10% formalin, dehydrated
in ethanol (50–100%). Then representative blocks of liver tissues from each lobe taken and processes for paraffin embedding using the standard microtechnique. Montelukast Sodium Sections (5 μm) of livers stained with hematoxylin and alcoholic eosin dye for photo-microscopic observation for histopathological studies. All results were expressed as the mean ± standard error of mean (SEM). The results were analyzed for statistical significance One-way Analysis of Variance (ANOVA) followed by Dunnett’s post hoc multiple comparison tests using Graph Pad Prism software, P < 0.01 was considered as statistically significant. The extracts were found non-toxic up to the dose of 2000 mg/kg b.w. Neither mortality nor any significant behavioral changes were observed, thus 2000 mg/kg was considered as NOAEL and 1/10th of these doses is oral LD50 in both A. paniculata and S. chirayita plant was 200 mg/kg b.w.