Osteogenic and Anti I-Obesity Potential of Cissus Quadrangularis: Elucidation of Differentiation Process in Mouse Pre-Osteoblast and Pre-Adipocyte Cell Lines

Abstract

Traditional (herbal) medicine has been utilized to treat human ailments since ancient times. Compared to modern medicine, plant and animal-based medicines have less side effects to human health. Cissus quadrangularis (Haddjod) is a medicinal herb reported in ayurveda literature for its diverse pharmacological properties including healing bone disorders and obesity. It has previously been established in this lab that CQ crude ethanolic extract enhances mineralization of MC3T3-E1 cell line. With an aim to isolate and identify the main active constituent, CQ-E was fractionated into four solvent fractions i.e. ethyl acetate (CQ-E), butanol (CQ-B), hexane (CQ-H) and dichloromethane (CQ-D). The solvent fractions were subjected to various biochemical tests to identify the phytochemical constituents of each fraction and GC-MS to predict the phytochemical constituents of the fractions. Afterwards the osteogenic potential of these solvent fractions on MC3T3-E1 - mouse pre osteoblast cell line - was investigated. Moreover, the effect of these solvent fractions on osteoclastogenesis and adipogenesis has also been studied using mouse macrophage Raw264.7 and mouse pre-adipocyte 3T3-L1 cell lines. For this, viability, growth parameters, proliferation and metabolic activity of the cells were analyzed to assess the effect of these solvent fractions on the cells. Effect on growth parameters were studied by plotting growth curve. Cells were treated with mitogenic concentrations of each solvent fraction during differentiation and expression profiles of osteoblast, osteoclast and adipocyte marker genes were also plotted. The findings of this study indicated that CQ-H fraction has the most osteoblastogenic, anti-osteoclastogenic and anti-adipogenic potential with 10ng/ml as the most efficacious dosage with EC50 of 168µg/ml. Moreover, CQ-H was more effective anti-osteoporotic and anti-obesity agent than resveratrol. xxiv The bioactivity of CQ has been attributed to its phytochemical constituents. Among numerous other phytochemicals, resveratrol and its metabolites have been reported as one of abundant phytochemical constituents of CQ therefore it was used as a standard for analyzing osteoporotic and anti-obesity potential of CQ. The results revealed resveratrol as an effective anti-osteoporotic and anti-obesity agent with 1µM (0.02µg/ml) as efficacious dose of resveratrol with EC50 of 150µM (3.48µg/ml). The results of this study provide in vitro analyses of two important reported properties of CQ i.e. anti-osteoporosis and anti-obesity, strengthening or antagonizing the previously reported results, thus indicating the potential of CQ to be used as an alternative medicine for osteoporosis and weight management