Effect Of Liquid Extracted From Hovenia Dulcis Thunb On The Fatty Degeneration Of Hepatic Cells Induced By Ehanol In Vitro

Objective: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 different concentration of alcohol-induced steatosis in L-02 liver cells.Method: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 for same time with The optimum concentration of hovenia dulcis thumb extracted lipuid.The accumulation of lipid droplets were observed by light inverted microscopy after red oil-O 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 SREBP-1c in hepatocytes of each group.Result:1.When the ethanol concentration in the culture is 0.1% and 0.2%,the hepatocytes would have higher survival rate than in the normal culure. The survival rate will decrease as the ethanol concentration more than 0.3%,and the most significant decline of survival rate happened when the ethanol concentration was 2.4%(p<0.05).2.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 can be dyed to red with red oil-O dying.48h later after hovenia dulcis thumb extracted lipuid treated, lipiddroplets make considerabale reduction.3.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).4.The expression of mRNA of SREBP-lc in hepatocytes of each ethanol group increased gradually as the concentration of ethanol ascend(p<0.05), which can be depressed by hovenia dulcis thumb extracted liquid(p<0.05).Conclusion:1.Low concentration of ethanol can promote the viability of hepatocytes within a certain range. On the contrary,it will promote the mortality of the hepatocytes if the concentration of ethanol beyond a certain range.2.Ethanol could establish the models of fatty degeneration L-02 liver cells successfully and increase the AST,ALT levels in vitro with enough time, which have positive relationship with the ethanol concentration.3.hovenia dulcis thumb extracted liquid has therapeutical effect on the fatty changed L-02 cells which were 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 downregulation of the expression of the mRNA of SREBP-1c in hepatocytes,which have preventive and therapeutic effect on alcoholic fatty liver.