Effects Of Curcumin On MRNA Expression Of Acetyl-CoA Carboxylase In Human Hepatocyte

Objective: To observe the effect of curcumin to triglyceride's disposition and expression of acetyl-CoA carboxylase mRNA in normal and nonalcoholic induced fatty liver cell.Methods: The investigation was conducted by means of randomized controlled trial. To establish a model of hepatocyte steatosis, we incubate the normal human HL-7702 hepatocyte in RPMI-1640 complete medium with 10% (V/V) fetal bovine serum plus 10% (V/V) medical fat emulsion injection for 24 hours. The experiment is divided into 6 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 be12.5,25, 50 ,100μmol/l respectively. After each group is incubated for 24 hours, And observe the formation of cellular morphology and lipid droplets in the cells under oil red O stain and determin the intracellular triglyceride levels through high performance liquid chromatography (HPLC). The semiquantitative RT-PCR was used to detect the mRNA levels of acetyl-CoA carboxylase of the hepatocytes of each group. According to the descendent level of intracellular triglyceride, the dose-effect relationship curve was made out. Then the cells were exposed to curcumin at the 100μmol/L concentration for 6hr, 12hr, 24hr , 48hr respectively, observe the above-mentioned parameters and determine the best duration for curcumin to decrease intracellular triglyceride levels, and draw out the duration-effect relation curve.Using±s to represent experimental data, using SPSS10.0 statistical software to analyze the data, a P value of <0.05 is considered significantly important. Results: After incubate in fat emulsion injection for 24 hours, the features of hepatocyte microsteatosis in the cells were observed in human HL-7702 hepatocyte under oil red O stain, and through measurement of HPLC intracellular TG levels were found out to be increase dramatically. Thus the model of steatosis hepatocytes was established successfully. Then expose the fatty changed hepatocytes to different concentrations of curcumin for 24 hours. The result displayed: oil red O stain showed, the intracellular lipid droplets were less than before exposure. After HL-7702 liver cell were turned into fatty liver cell , the RT-PCR showed that acetyl-CoA carboxylase mRNA's expression increased obviously and the HPLC show triglyceride's disposition increased clearly. When we gave curcumin to fatty liver cell, triglyceride's disposition decreased and the variation was positive correlated with curcumin ' s concentration. the RT- PCR showed that acetyl-CoA carboxylase mRNA' s expression decreased than model group. Then after exposed to the curcumin of 100μmol/l with the fatty changed hepatocytes for 6, 12, 24hours, the results said: the decrease of TG in the cells by curcumin was dependent on the duration , at the same time the decrease of acetyl-CoA carboxylase mRNA's expression was dependent on the duration and the intracellular lipid droplets decreases duration dependently.Conclusions:1 Curcumin could lighten hepatocyte steatosis and decrease triglyceride's disposition in the fatty-changed hepatocytes .2. The possible mechanisms of curcumin decreasing triglyceride's disposition in the fatty-changed hepatocytes were related to downregulation of acetyl-CoA carboxylase mRNA.