QuantumRNA 18S was used as an internal control. PCR products were technical support analyzed by agarose gel electrophoresis along with DNA molecular markers, stained with ethidium bromide, and visualized under UV light. The intensities of RT PCR products Inhibitors,Modulators,Libraries in the agarose gels were quantified by densitometry. Statistical analysis Data are expressed as the mean S. E. M. from multiple experiments. Comparison between groups was per formed using a one way ANOVA followed by a Tukeys test. Results Lipase inhibitory activity of Polygonum cuspidatum extract and fractions First, we evaluated in vitro the lipase inhibitory activity of natural products and P. cuspidatum was chosen for more detail investigation. Pancreatic lipase inhibition of P. cuspidatum extract and fractions is expressed as per centage and IC50.

Butanol fraction of the etha nol extract of P. cuspidatum exhibited the strongest inhibitory effect on lipase with an IC50 value of 15. 8 ugml. Next, we tested the effect of P. cuspidatum extract and fractions on adipocyte differentiation of 3 T3 L1 preadipocytes. POCU1b was the most effective in the prevention of adipogenesis. Thus, we investigated the anti obesity effects Inhibitors,Modulators,Libraries of POCU1b on adipocyte differentiation of 3 T3 L1 pre adipocytes and the related molecular mechanism of lipid accumulation. Cytotoxicity assay Cytotoxicity assay was performed to evaluate Inhibitors,Modulators,Libraries whether POCU1b had any effects on 3 T3 L1 viability and to de termine Inhibitors,Modulators,Libraries the optimal conditions required. During the dif ferentiation processes, 3 T3 L1 preadipocytes were treated with insulin and POCU1b for 5 and 12 days.

There were no effects on cytotoxicity at low concentra tions of POCU1b and HCA. However, high concentrations of POCU1b altered cell viability after 12 days. Hydroxycitric acid did not alter cytotoxicity after 12 days at concentrations up to 50 ugml. POCU1b inhibits adipocyte differentiation Inhibitors,Modulators,Libraries and GPDH activity To determine the effects of POCU1b on adipogenic con version, 3 T3 L1 preadipocytes were induced to differen tiate in the presence or absence of POCU1b and stained with Oil red O. Lipid droplets in untreated control cells stained very strongly with Oil red O, an indication that the cells accumulated substantial amounts of cytoplas mic triglycerides. Representative images of Oil red O staining in treated cells showed that POCU1b sup pressed nothing both triglyceride accumulation and adipocyte differentiation in a dose dependent manner. HCA treatment of cells also reduced staining with Oil red O, indicative of low triglyceride content. Expression of GPDH is induced the conversion of pre adipocytes to adipocytes. We examined the effect of POCU1b on the activity of GPDH. POCU1b treatment of 3 T3 L1 adipocytes resulted in a marked decrease in GPDH activity in a dose dependent manner.