Effects Of Qucertin On MRNA Expresion Of CPT-1 And PPARγ In Human L-02 Steatosis Hepatocyte

Object: To observe the effects of qucertin on TG levels , the mRNA expresion of PPARγand CPT-1 in human L-02 steatosis hepatocyte.Method: we incubate the normal human L-02 hepatocyte in RPMI-1640 complete medium with 10% (V/V) fetal bovine serum plus 10% (V/V) medical fat emulsion injection for 24 hours. Then add qucertin of varies concentration into the medium and incubate for another 24 hours. The concentration of qucertin was respectively 20μmol/l, 40μmol/l and 80μmol/l. 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 PPARγand CTP-1 of the hepatocytes of each group. Then expose the cell to the qucertin of 40μmol/l for 6, 12, 24 and 48 hours respectively, observe the above-mentioned parameters and determine the best duration for qucertin to decrease intracellular triglyceride levels, and draw out the duration-effect relation curve. All the data was analyzed by SPSS11.0 statistical software.Result: After incubate in fat emulsion injection for 24 hours, the features of hepatocyte microsteatosis in the cells were observed in human L-02 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. After exposed to different concentrations of qucertin for 24 hours, by oil red O stain the intracellular lipid droplets of steatosis hepatocytes were much less than before, the intracellular TG levels decreased remarkablely with concentration dependently. After exposed to the qucertin of 40μmol/l for 6, 12, 24 and 48 hours, the decrease of TG levels in the foam cell by qucertin was found to be dependent on the duration. The semi quantitative RT-PCR to detect the mRNA levels of PPARγand CTP-1 of the hepatocytes of each group, CTP-1 mRNA expression was found out to be much higher in the group treated with qucertin than in the normal and model group, while the PPARγmRNA expression in the group of treatment of qucertin was found out to be much decreased than in the model group, but higher than the normal.Conclusion:1. The quantity of trigelyceride increase in the fatty-changed hepatocytes,2. Qucertin can decrease the quantity of trigelyceride in the fatty-changed hepatocytes3. The possible mechanisms of qucertin decreasing trigelyceride are related to upregulation of CTP-1 and downregulation of PPARγ.