Effect of Ethanol Extract of Indigofera tinctoria Linn (Fabaceae) on Lithium/Pilocarpine-Induced Status Epilepticus and Oxidative Stress in Wistar Rats

Purpose: Indigofera tinctoria Linn. of the Fabaceae family is claimed to be useful in the treatment of a variety of epileptic disorders in Indian traditional system of medicine. The present study was designed to verify this claim.Methods: Status epilepticus was induced in male albino rats of Wistar strain by administration of pilocarpine (30 mg/kg, i.p.) 24 h after injection of lithium chloride (3 mEq/kg, i.p.). Different doses of the ethanol extract of Indigofera tinctoria were administered orally 1 h before the injection of pilocarpine. The severity of status epilepticus was observed and recorded every 15 min for 90 min and thereafter every 30 min for another 90 min, using Racine scoring system. In-vivo lipid peroxidation of rat brain tissue was measured in terms of the thiobarbiturate-reactive substances (TBARS). Both in-vitro free radical nitric oxide (NO) and 2-diphenyl-2-picryl hydrazyl (DPPH) scavenging activities of the extract were also determined.Results: The severity of status epilepticus was significantly (p < 0.01) reduced following oral administration of the extract at 500 and 1000 mg/kg doses. No test animal group exhibited stage 4 seizure. The extract also exhibited both in-vivo and in-vitro antioxidant activities. Conclusion: The ethanol extract of Indigofera tinctoria was found to be useful in controlling lithium/pilocarpine-induced status epilepticus in albino rats.Keywords: Indigofera tinctoria, Free radical scavenging, Status epilepticus, Antioxidant, Albino rats, Polyphenols

DEVELOPMENT AND VALIDATION OF BIOANALYTICAL HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF CILNIDIPINE AND NEBIVOLOL IN HUMAN PLASMA

Objective: To develop and validate a modified isocratic reversed-phase high performance liquid chromatographic (RP-HPLC) method for determination of cilnidipine and nebivolol in human plasma to be used for pharmacokinetic studies.Methods: The drug was extracted from plasma samples by direct protein precipitation technique using acetonitrile. Amlodipine was used as internal standard (IS). Samples were analyzed on BDS C18 column (250 x 4.6 mm, 5 µm), applying ortho phosphoric acid (0.1%): Acetonitrile, at a ratio of 45:55 v/v in isocratic mode as a mobile phase at a flow rate of 1 ml/min to attain adequate resolution. Separations were performed at room temperature and monitored at a wavelength of 260 nm after injection of 50μl samples into the HPLC system. The analytical method was validated according to FDA bioanalytical method validation guidance. The method was applied for pharmacokinetic study of cilnidipine and nebivolol tablets-10 mg and 5 mg were administered as a single dose to 6 healthy male rabbits under fasting condition. Twelve blood samples were withdrawn from each rabbit over 24 h periods. From the plasma concentration-time data of each individual, the pharmacokinetic parameters; Cmax, Tmax, AUC0-t and AUC0-∞ were calculated.Results: A peak area was obtained for cilnidipine and nebivolol at 3.943 and 4.719 min retention time respectively. Linearity was established at a concentration range of 0.20-20 μg/ml (r2=0.999, n=8) for cilnidipine and 0.02-2 μg/ml (r2=0.999, n=8) for nebivolol. The lower limit of quantitation (LLOQ) was identifiable and reproducible at 0.2μg/ml for cilnidipine and 0.02 μg/ml for nebivolol. The coefficients of variation (%cv) of the intra-day and inter-day precision of cilnidipine at 600, 1000 and 1600ng/ml levels were found to be 6.90%, 6.19%, 5.22%; and 7.74%, 6.54%, 5.77%, respectively, which are lower than the accepted criteria limits (15-20 %). The mean recovery (%) cilnidipine at 600, 1000, and 1600ng/ml was found to be 101.03%, 99.27% and 104.87%, and for nebivolol 60, 100, and 160 ng/ml was found to be 106.13%, 107.03% and 98.06% respectively. Stability at different conditions and in autosampler was also established. The mean pharmacokinetic parameters; Cmax, Tmax, AUC0-t and AUC0-∞ were 6 ng/ml, 2 hr, 96.76 mg. hr/ml, 63.45 mg. hr/ml for cilnidipine and 5.8ng/ml, 2hr, 74.78 mg. hr/ml, 100.25 mg. hr/ml for nebivolol respectively.Conclusion: The present analytical method was found to be specific, sensitive, accurate and precise for quantification of cilnidipine and nebivolol in human plasma. It can be successively applied for pharmacokinetics, bioavailability and bioequivalence studies

EVALUATION OF PROTECTIVE EFFECT OF BASSIA MALABARICA LEAVES AGAINST CISPLATININDUCED NEPHROTOXICITY AND DOXORUBICIN-INDUCED CARDIOTOXICITY IN RATS

Objective: The objective of the current investigation is to study the effect of ethanolic extract of Bassia malabarica leaves (EBML) against cisplatin-induced nephrotoxicity and doxorubicin (DOX)-induced cardiotoxicity in healthy adult male Wistar rats.Methods: Nephrotoxicity was induced by cisplatin (7 mg/kg, i.p) and cardiotoxicity was induced by DOX (15 mg/kg, i.p). In both the models, EBML (150 mg/kg and 300 mg/kg, p.o) was administered for 15 days to assess the prophylactic and curative effect. Urinary parameters, biochemical parameters, and in vivo antioxidants were monitored for nephroprotective effect. For assessing cardioprotective effect serum parameters, cardiac ATPases and in vivo antioxidants were measured. A statistical significance was set at p&lt;0.05 which was analyzed by one-way analysis of variance followed by Tukey's multiple comparison test.Results: The study results show that the EBML has significantly (p&lt;0.05) restored the urinary parameters, serum parameters of cisplatin-induced nephrotoxic rats, and serum parameters of DOX -induced cardiotoxic rats. A significant decrease (p&lt;0.05) in levels of malondialdehyde and increase in reduced glutathione and catalase were seen in both nephrotoxic and cardiotoxic rats. Ca+2 ATPase was significantly decreased and Na+ K+ ATPase was significantly increased (p&lt;0.05) in the treatment groups when compared to DOX disease control group.Conclusion: EBML showed a protective effect against cisplatin-induced nephrotoxicity and DOX -induced cardiotoxicity

Effect of alcoholic extract of roots of Dichrostachys cinerea Wight & Arn. against cisplatin-induced nephrotoxicity in rats

12-18The alcoholic extract of roots of Dichrostachys cinerea Wight & Arn. (200 and 400 mg/kg, p.o.) was studied for its protective effect against cisplatin-induced renal injury in rats. Cisplatin (6mg/kg, i.p.) significantly elevated serum markers level, increased urinary protein excretion, reduced urine to serum creatinine ratio and creatinine clearance. In curative regimen, the alcoholic extract exhibited dose dependent protection. Animals which received prophylactic treatment also showed partial protection against cisplatin-induced effects. Histopathological studies substantiated the above results. Further, the alcoholic extract showed marked nitric oxide scavenging effect and reducing power suggesting an antioxidant property. A triterpenoid, fatty acid and a steroid were isolated from the n-hexane, ethyl acetate fractions of alcoholic extract. The results suggested that the roots of D. cinerea showed protective effect against cisplatin-induced nephrotoxicity which may probably be mediated by its antioxidant property

IN SILICO STUDIES ON FUNCTIONALIZED AZAGLYCINE DERIVATIVES CONTAINING 2, 4-THIAZOLIDINEDIONE SCAFFOLD ON MULTIPLE TARGETS

Objective: The 2, 4-thiazolidinedione containing compounds could lead to most promising scaffolds with higher efficiency toward the targets recognized for its antidiabetic activity when combined with azaglycine moiety. The objective of the present work was to merge functionalized aza glycines with 2, 4-thiazolidinediones, perform in silico evaluation by molecular properties prediction and undertake the molecular docking studies with targets relevant to diabetes, bacterial and viral infections using Swiss Dock programme for unraveling the target identification which can be used for further designing.Methods: (i) In silico studies were performed using Molinspiration online tool, Swiss ADME website and Swiss Target Prediction websites to compute the physicochemical descriptors, oral bioavailability and brain penetration. (ii) Molecular docking studies were performed using Swiss Dock web service for enumeration of binding affinities and assess their biological potentiality.Results: The results predicted good drug likeness, solubility, permeability and oral bioavailability for the compounds. All the compounds showed good docking scores as compared to the reference drugs. The N-oleoyl functionalized aza glycine derivative demonstrated superior binding properties towards all the studied target reference proteins, suggesting its significance in pharmacological actions.Conclusion: The binding interactions observed in the molecular docking studies suggest good binding affinity of the oleoyl functionalized aza glycine derivative, indicating that this derivative would be a promising lead for further investigations of anti-viral, anti-inflammatory and anti-diabetic activities