Effect Of Hovenia Dulcis Thunb Extracted Liquid On The Expression Of Adipor-2 Genes In The Fatty Degeneration Of Hepatic Cells Induced By Ethanol In Vitro

Purpose:To establish the models of the different concentration of alcohol-induced steatosis in L-02 liver cells. To explore the effect of different ethanol concentration on the lipogenesis and the releasing dose of alanine amino transferase(ALT), aspartate amino transferase(AST) of L-02 liver cells, and to investigate whether hovenia dulcis thumb extracted liquid has therapeutical effect on the models of alcohol-induced steatosis in L-02 liver cells,and to investigate the mechanism involving in it.Methods. The concentrations of alcohol inducing hepatocyte steatosis,were detected by MTT assay (0.3%,0.6%,1.2%,2.4%). The models of fatty degeneration L-02 liver cells were treated with the optimum concentration of hovenia dulcis thumb extracted lipuid for 48h.The accumulation of lipid droplets were observed by light inverted microscopy after red oil-0 staining. An estimation of Releasing dose of alanine amino transferase(ALT), aspartate amino transferase(AST) in the culture was detected through automatic biochemical analyzer, and the intracelluer triglyceride(TG) level would be determined by the kit. The semiquantitative RT-PCR was used to detect the mRNA levels of The concentrations of alcohol inducing hepatocyte steatosis,were detected by MTT assay (0.3%,0.6%,1.2%,2.4%). The models of fatty degeneration L-02 liver cells were treated with the optimum concentration of hovenia dulcis thumb extracted lipuid for 48h..Results:1.After incubated in culture with ethanol more than two days, numerous lipiddroplets were gathered in the cytolymph of human L-02 hepatocyte observed by inverted microscopy, which could be dyed to red with red oil-0 dying.48h later after hovenia dulcis thumb extracted lipuid treated, lipiddroplets made considerabale reduction.2. Compared with the normal group, the releasing dose of alanine amino transferase(ALT), aspartate amino transferase(AST) in the culture and the intracellular TG levels increased in the ethanol groups(p<0.05). And they decreased if treated with hovenia dulcis thumb extracted lipuid for 48 hours(p<0.05).3. Compared with the normal group, the expression of mRNA of adipor2 increased in 0.3% ethanol group but decreased in the rest of ethanol groups.(P<0.05)。After hovenia dulcis Thunb extracted liquid treated, the expression of mRNA of adipor2 had no significant change in normal group and 2.4% ethanol group(P> 0.05),but increased in the rest of ethanol groups(P< 0.05).Conclusion:1. The models of fatty degeneration L-02 liver cells can be successfully established by Ethanol.And the AST, ALT levels in vitro increase in the models.2.The expression of mRNA of adipor2 depress as the ethanol concentration ascending, and is related to the hepatocyte steatosis.3. Hovenia dulcis thumb extracted liquid has therapeutical effect on the fatty changed L-02 cells which are induced by alcohol in vitro. It also can decrease the quantity of TG in the fatty changed human L-02 hepatocytes. The possible mechanisms of hovenia dulcis thumb decreasing the intracellur TG are related to the upregulation of the expression of the mRNA of adipor2 in hepatocytes,which have preventive and therapeutic effect on alcoholic fatty liver.