Expression And Significance Of Bsep And Mrp2 After Hepatic Ischemia-reperfusion: Experiment With Rats

Objective: To detect the expression and location of Bsep and Mrp2 after hepatic ischemia-reperfusion: experiment with rats,and to investigate the mechanism of hyperbilirubinemia after rat hepatic ischemia-reperfusion.Methods: Sixty healthy MaleSprague-Dawley(S-D) rats were randomly divided into 2 groups(thirty ras in each group):Group A(the control group),rats were not subjected to hepatic ischemia reperfusion.Group B,rats were subjected to 30 min of total hepatic ischemia.At 1h,6h,12h,1d,3d after operation checkpoint was set. Routine biochemistry method detect TBA,DBIL, ALT levels of plasm.The expression of Bsep and Mrp2 mRNA was determined by Real Time reverse transcriptase PCR .The expression of Bsep and Mrp2 protein was detected by immunohistochemical SP method .Results:①TBA,DBIL,ALT:There were no significant differents in levels of serum TBA,DBIL,ALT in control group from 1h-3d after operation(P>0.05)。In group B the expression of TBA,DBIL,ALT were significantly higher compared with group A at 1h,6h,12h,1d after hepatic ischemia-reperfusion(P<0.05),and on significant differences at 3d(P>0.05).②HE:Pathological examination showed that there were only amid inflammation in the liver tissues that had undergone ischemia-reperfusion in Group B,and no necrosis of hepatocytes was seen in both group.③RT-PCR:There were no significant differences in the levels of serum Bsep and Mrp2 from 1h-3d in Group A(P>0.05). The expressions of serum Bsep at 1h,6h,12h,1d after hepatic ischemia-reperfusion were significant differences compared with Group A(P<0.05),and there was no significant differences at 3d(P>0.05).At 1h,6h,12h after hepatic ischemia-reperfusion in rats of Group B were significant lower than Group A(P<0.05),and there were no significant at 1d,3d.(P>0.05).④IHC:The protein of Bsep and Mrp2 were regularly expressed in hepatocyte membrane in both group. At 1h-1d after reperfusion ,the protein of Bsep was significant derease in Group B compared with Group A,no location in hyalomitome,and there were no significan at 3d. The localization of Mrp2 continuously altered at 1h-1d,not only expression in hepatocyte membrane,but also can be seen in hyalomitome,and there were no significan at 3d.Conclusion: Down-regulation of Bsep and absence of Mrp2 localization in hepatocytes membrane may be the mechanism of serum bilirubin after hepatic ischemia-reperfusion.