Effect Of Hovenia Dulcis Thunb Liquid On The Injured L-02Cells Induced By Ehanol In Vitro

ObjectiveTo establish the models of the different concentration of alcohol—induced damage in L-02liver cells. To explore the effect of different ethanol concentration on the Oxidative Stress and lipid peroxidation, the releasing dose of alanine amino transferase(ALT), aspartate aminotransferase(AST), Malondialdehyde(MDA), Reduced Glutathione(GSH) of L-02liver cells, the Expression of protein NF-KB1, and to investigate whether hoveniadulcis thumb extracted liquid has therapeutical effect on the models of different concentration of alcohol-induced damage in L-02liver cells.MethodAccording to the MTT experiments, appropriate injured concentrations of alcohol were(0.3%、0.6%、1.2%、2.4%) respectively. The models of injured L-02liver cells were treated for same time with The optimum concentration of hovenia dulcis thumb extracted lipuid. ROS was detected through loading probe in situ.An estimation of Releasing dose of alanine amino transferase(ALT), aspartate amino transferase(AST)in the culture were detected through automatic biochemical analyzer. Theintracelluer Malondialdehyde(MDA), Reduced Glutathione(GSH) level would be determined by the kit. The method of immunocytochemistry was used to test the expression of NF-KB1and observe its intensity or location in L-02liver cells of each group.Result1. When the ethanol concentration in the culture was under0.2%. the hepatocytes revealed higher survival rate than in the normal culure. The survival rate decreased as the ethanol concentration more than0.3%,but the rate still was up on80%. The most significant decline of survival rate happened when the ethanol concentration was over2.4%(p<0.05),so the appropriate injured concentrations were (0.3%、0.6%、1.2%、2.4%).2. After incubated in culture with ethanol, a little fluorescent appeard in the normal group. When increasing the ethanol concentration,the fluorescent was more and more bright in the human L-02hepatocyte which was observed by inverted microscopy. However it is the strongest with the ethanol concentration of2.4%, the cells in contrary decreased. after hovenia dulcis thumb extracted lipuid treated, the fluorescent appeard considerabale reduction.3. Compared with the normal group、the releasing dose of alanine amino transferase(ALT), aspartate amino transferase(AST)in the culture increased in the ethanol groups(p<0.05). However these index decreased if treated with hovenia dulcis thumb extracted lipuid (p<0.05).4. The intracelluer Malondialdehyde(MDA) increased and Glutathione(GSH) decreased with the growing ethanol concentration (p<0.05). After the therepy of hovenia dulcis thumb extracted liquid the above-mentioned index appeard inverse change(p<0.05).5. The normal hepatocytes were detected the expression of NF-KB1in endochylema. the expression of each ethanol group increased gradually as the concentration of ethanol ascended(p<0.05) in endochylema and in karyon, but1.2%group and2.4%group appeard no obvious difference (P>0.05). Each ethanol group Can be depressed by hovenia dulcis thumb extracted liquid(p<0.05).Conclusion1. 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 can establish the models of injured L-02liver cells successfully, and can increase the ROS、LPO、AST、ALT、NF-KB1levels,and can decrease levels the GSH in vitro with enough time, which have positive relationship with the ethanol concentration.3. Hovenia dulcis thumb extracted liquid has therapeutical effect on the injured L-02cells which is induced by alcohol in vitro. Decreasing the Oxidative Stress and lipid peroxidation level, and downregulating the expression of NF-KB1in hepatocytes are the possible mechanisms.