姜黄素对人肝L-02细胞胆固醇合成及转运蛋白表达的影响-消化内科专业论文.docxVIP

PAGE PAGE 10 Effects of Curcumin on Cholesterol Synthesis and Transport in human L-02 hepatocyte ABSTRACT Object: investigate the formation of lipid droplets and expression of cholesterol rate-limiting enzyme－HMG-CoA reductase and transported protein－ caveolin-1,detect the amount of cholesterol on human L-02 hepatocyte under the control of curcumin, to find out the effects of curcumin on cholesterol synthesis and transport. Methods: Use L-02 hepatocyte of normal human and the hepatocyte under fatty change after exposure to 50% fetal bovine serum for 24 hours as study objects. The experiment is divided into 8 groups: The first group is normal hepatocytes; the second group is fatty change hepatocytes;from group 3 to group 6, different amounts of curcumin is added to the fatty changed hepatocytes, to make the concentration of curcumin to be 0, 12.5, 25, 50 ,100 respectively; Group 7 is the positive control group, Probucol is added to the hepatocyte with fatty change to make the concentration to be 50; In group 8, curcumin is added to the normal hepatocyte to make the concentration to be 50. After each group is incubated for 24 hours, observe the formation of lipid droplets in the cells under oil red O dye.Detect the intracellular amount of total cholesterol(TC) 、 free cholesterol(FC)and cholesterol ester(CE) through high performance liquid chromatography (HPLC) and calculate the ratio of intracellular CE to TC and the amount of FC in the medium. Then detect the HMG-CoA reductase and caveolin-1 mRNA ‘s expression of the hepatocytes of each group and decide the best concentration of curcumin that decreases the amount of intracellular cholesterol and make out the amount – effect relation curve. And expose the cell to curcumin at the best concentration for 0hr, 6hr, 12hr, 24hr respectively, observe the formation of intracellular lipid droplets under oil red O dye.Detect the intracellular amount of TC and CE and calculate the ratio of intracellular CE to TC. Then