The Effects Of MethyL Ferulic Acid On Ethanol-induced Oxidative Damage In Human Hepatocytes

Objective: Establish the hepatocyte injury model by ethanol,to study the effect and possible mechanism of methyl ferulic acid(MFA)extracted from cicada on the anti-hepatocyte oxidative damage.Methods: The methyl ferulic acid monomer was extracted from the rhizome of the celestilla.L-02 cells were cuLtured in vitro and passaged.The optimum concentration of MFA and ethanol was screened by MTT method.The optimum time of ethanol was screened by MTT method.Cell model was estabLished with 5% ethanol stimulation.The cells were divided into 5 groups: control group(DMEM medium);model group(5% ethanoL group);positive drug group(VC,15mg/mL);MFA10μg/mL dose group;MFAμg/mL dose group.The apoptosis of L-02 cells was detected by flow cytometry,the content of ROS in L-02 cells was detected by fluorescence probe method.Alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in each cell supernatant were measured by enzyme activity method,the contents of superoxide dismutase(SOD),catalase(CAT)and intracellular gLutathione peroxidase(GSH-px)in cells were measured by spectrophotometry.The expression levels of Bax mRNA in each group were detected by PCR.The expression of Bax protein was detected by Western blotting.Results: EthanoL significantly promote the apoptosis of L-02 cells,enhance the level of oxidative stress,decrease the levels of SOD,CAT and intracellular GSH-px,and increase the levels of ALT and AST in the supernatant,increase the expression of Bax mRNA and the level of Bax protein.MFA could significantly reduce the level of oxidative stress,increase the levels of SOD,CAT and GSH-px of cell,while decrease the levels of ALT,and AST in cell supernatant,reduce the expression of Bax mRNA and the level of Bax protein,compared with the control group,the difference was significant(P <0.05).Conclusions:(1)MFA can inhibit the damage of ethanol to L-02 cells,improve the survivaL rate,a certain range of MFA can increase the ethanol-induced L-02 cell injury survival rate.(2)MFA can inhibit ethanol damage L-02-induced apoptosis.(3)MFA can improve the level of oxidative stress after ethanol injury.(4)The protective mechanism of MFA on L-02 cells may be through down-reguLation of Bax gene and protein expression to play an antioxidant injury.