Effect Of Hovenia Dulcis Thunb Liquid On The Expression Of UCP2 Gene In Ethanol-induced Superfatted L-02 Cells In Vitro

Objective:To establish the model of alcohol-induced hepatocyte steatosis in L-02 liver cells in vitro; Investigate the relationship between cell damage, UCP2 gene expression and ethanol concentrations in L-02 cells of steatosis; Explore the protective effect of Hovenia Dulcis Thunb extracted liquid; Reveal the molecular mechanisms of AFLD caused by ethanol, and solutions for prevention and cure.Method:The best ethanol concentrations to induce liver L-02 cell steatosis was established by MTT method. Hovenia Dulcis Thunb extracted liquid at the most compatible dosage was applied to the L-02 cell of steatosis. Then L02 cells were stained with red O-oil and observed by light inverted microscope. ALT and AST of each group were detected on the automatic bio-chemical analyzer, intracelluar triglyceride (TG) was analyzed with TG-kit, and UCP2 mRNA was detected by the semiquantitative RT-PCR method.Result:1, MTT method revealed that cell survival rate is less than 50% when ethanol concentration is higher than 5% (P<0.05); When ethanol is 2.4%, cell survival rate increased to more than 80%. Therefore the 0.3%-2.4% was selected as the most compatible concentrations. 2, After cultured for 48-72 hours with 0.3-2.4% ethanol and stained with red O-oil, obvious red fatty drops were seen in ethanol-treated L-02 cells. And the fatty change levels and fatty change cell numbers are increased when the ethanol concentration increased.3, In the steatosis cells, the average values of ALT, AST and TG increased (P<0.05). While treated with Hovenia Dulcis Thunb extracted liquid these values all decreased (P<0.05).4, In steatosis model groups, UCP2mRNA expression level was down regulated (P<0.05). But in the steatosis groups treated with Hovenia Dulcis Thunb extracted liquid, UCP2mRNA levels is obviously increased compared to steatosis group (P<0.05).Conclusion:1, Ethanol has cell toxicity to the L-02 cells. Ethanol can cause the L-02 cell fatty change. The ethanol-induced L-02 cell damage aggravates as the dosage increases.2, Ethanol can down-regulate the UCP2mRNA expression level of L-02 cell in a certain period. The low expression of UCP2mRNA is probably relevant to AFLD occurrence.3, Hovenia Dulcis Thunb can prevent the cell toxicity of ethanol and decrease steatosis level. Hovenia Dulcis Thunb extracted liquid can increase the UCP2mRNAs expression level of the L-02 cells.