Bioassay Guided Isolation for Anti-inflammatory and Antioxidant Components from Celastrus paniculatus Seeds. (#29)
Aim: Celastrus species are used as a folk remedy in India for the treatment of various inflammatory conditions. In order to identify the components responsible for pharmacological activity, in vivo anti-inflammatory activity of Celastrus paniculatus seeds were investigated using carrageenan induced rat paw edema as a screening model.
Methods: The methanolic extract of CP was fractionated sequentially with petroleum ether (PCP), chloroform (CCP), ethyl acetate (ECP) and n-butanol (BCP) solvents. Active PCP fraction was subjected to fractionation through successive column chromatography on silica gel, eluted with a gradient of petroleum ether: ethyl acetate to afford five sub-fractions (C1 – C5). The activity of C2 subfraction was evaluated against oxidative stress induced by carrageenan in rat paw tissues by studying the levels of antioxidant enzymes viz. superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH) and myeloperoxidase (MPO). Characterization of sub-fraction (C2) was performed by spectral techniques to identify active phytoconstituents.
Results: The results exhibited varying degree of anti-inflammatory activities in different fractions, whereas petroleum ether fraction of CP (PCP) showed significant reduction in paw edema volume. Among the PCP sub-fractions (C1 – C5), sub-fraction C2 exhibited significant anti- inflammatory effect. Also, PCP sub-fraction C2 improved the levels of SOD, CAT, GSH and MPO in rat paw tissues. Bioassay-guided fractionation of CP led to the isolation and identification of β-sitosterol by GC-MS, IR, 1H and 13C NMR from the active PCP sub-fraction C2.
Conclusion: The presence of β-sitosterol in CP provides a scientific basis for the utilization of Celastrus paniculatus as traditional medicine in treatment of inflammatory conditions.