Orlistat is the deriva tive selleck bio of lipstatin, a potent natural inhibitor of pancre atic lipases isolated from bacterium. Orlistat promotes body weight loss and reduces the incidence of dia betes by nearly 40% in obese people. However, it has serious side effects, such as steatorrhea, stomach pain, irregular menstrual periods, and headaches. Polygonum cuspidatum has been used Inhibitors,Modulators,Libraries clinically for the treatment of consti pation, gallstones, hepatitis, and inflammation in East Asian countries such as Korea, China, Taiwan, and Japan, little work has been carried out re garding the effects on anti obesity. In the present study, to elucidate activities of P. cuspidatum on anti obesity, we screened candidates for lipase inhibi tory activity from the P. cuspidatum fractions and investigated the effects of butanol fraction of the ethanol extract of P.
cuspidatum on the regulation of adipocyte differentiation. Our findings suggest for the first time that POCU1b inhibits adi pocyte differentiation Inhibitors,Modulators,Libraries through the attenuation of lipid accumulation. Methods Preparation of Polygonum cuspidatum extract Radix of P. cuspidatum were purchased from a com mercial supplier in Jung dong, Daejeon, Korea in November 2008 and identified by Prof. J. H. Kim in the Department of Life Science, Gachon University. A voucher specimen was deposited at the Herbarium of Diabetic Complication Research Team, Korea Institute of Oriental Medicine. The dried plant ma terial was extracted with ethanol Inhibitors,Modulators,Libraries by maceration at room temperature for 3 days and the extracts were concentrated in vacuo at 40 C.
The concentrated extract was diluted in water and then partitioned successively with n hexane, ethyl acetate, n butanol, and water, re spectively. The percentage yield of the lyophi lized butanol Inhibitors,Modulators,Libraries fraction of the ethanol extract of P. cuspidatum was 1. 9%. Inhibitors,Modulators,Libraries For water ex tract, the dried plant was extracted with boiling water and the extract was in vacuo at 40 C. The percentage yield of water extract was 24. 8%. Pancreatic lipase activity of Polygonum cuspidatum extract and its fractions The method for measuring pancreatic lipase activity was modified from that of Kim and colleagues. Briefly, an enzyme buffer was prepared by the addition of a solution of porcine pancreatic lipase to 169 ul of Tris buffer.
Then, either 20 ul of the plant extracts and fractions at the test concentra tion, or orlistat, was mixed with 20 ul of the enzyme buf fer and incubated for 15 min at 37 C with 5 ul of the substrate solution. The enzymatic reactions were allowed to proceed for 30 min at 37 C. Lipase ac tivity was determined by measuring http://www.selleckchem.com/products/XL184.html the hydrolysis of p NPB to p nitrophenol at 405 nm using an ELISA reader. Culture and differentiation The 3 T3 L1 preadipocyte cell line was purchased from the American Type Culture Collection. The cells were cultured in 4.
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