Regulation Of Uncoupling Protein 2 MRNA To The Effect Of Tolerance Of Ischemia Reperfusion Injury On Hepatocytes

Objective: To discuss the influence of hepatocytes'tolerance ability which was given ischemia treatment by regulating the expression of uncoupling protein 2.Methods: Gene UCP-2 eukaryotic expression plasmid and targeting gene UCP-2 siRNA eukaryotic expression plasmid were established, then transfected the both of plasmid to the normal and steatosis LO2 liver cell respectively. Ischemia-reperfusion was performed on the transfected liver cell. Assess the rate of cell apoptosis, necrosis and survival at different time points after reperfusion by flow cytometry(FCM);collect the culture fluid to determine the level of ATP, ROS ,MDA; Use real-time PCR to determine the expression of UCP-2 mRNA.Results:After ischemia treatment,in the ucp-2 mRNA over-expression group: the level of ROS,MDA,ATP are lower than the control groups'obviously on the same time point(P<0.05), and the same is cell survival rate, the rate of cell necrosis is higher than the control group's obviously(P<0.05), the rate of cell apoptosis have no difference (P>0.05); in the ucp-2 mRNA inhibited expression group:the level of ROS,MDA,ATP and the rate of cell apopotisi have no difference compared with control group after reperfusion 6 hours (P>0.05).However, the level of ATP and the rate of cell survive is higher than fatty liver cell control group at the same time point after reperfusion 6 hours (P<0.05).Conclusion : Inhibiting the expression of UCP-2 mRNA can alleviate ischemia reperfusion injury of fatty liver cell. However, it is no sense for normal liver cell to change the expression of UCP-2 mRNA.