The Impact Of Adenovirus Carrier Ad-hIL-10 On Ischemia Reperfusion Injury In Rat's Orthotopic Liver Transplantation

Part 1 Estabishing the Rat's Model of Orthotopic Liver Transplantation(OLT)Objective:To study the operation techniques of rat's orthotopic liver transplantation(OLT)and to provide a reliable model for studying ischemia reperfusion injury of allogene OLT.Methods:120 cases of rats' OLT operations were performed by modified Kamada's cuff technique with hepatic arterial reconstruction and modified suture for suprahepatic inferior vena cava and hepatic perfusing method in order to study the ischemia reperfusion injury of allogene OLT. 40 cases were trial experiments and 80 cases were performed by sterotypia operation.Results:Out of the 80 cases,68 were successful(achievement ratio 85%).The anhepatic time was 15～25min,the warm ischemia was 0～5 min and the cold ischemia was 90 min.The index expressing ischemia reperfusion injury in these cases had obvious changes.Conclusion:The rat's OLT model performed by Kamada's cuff technique with modified suture for superior vena and hepatic perfusing method was reliable to study ischemia reperfusion injury. Part 2 The impact of adenovirus carrier Ad -hIL-10 on ischemia reperfusion injury in Rat's Orthotopic Liver TransplantationObjective:To investigate the impact and mechanism of adenovirus cartier Ad -hIL-10 on ischemia reperfusion injury in Rat's Orthotopic Liver Transplantation.Methods:Sprague-Dawley(SD)rats were used as donors and male SD rats were used as recipients.OLT operations were performed by modified Kamada's cuff technique with modified suture for superior vena and hepatic perfusing method.Four groups were divided by genetic transfection.They were sham group(groupⅠ),control group(groupⅡ), blank carrier control group(groupⅢ)and genetic transfection group (groupⅣ).The animals were killed at 24 hours after graft reperfusion or sham operation respectively.Plasma samples were collected for ALT, AST,AKP and r-GT tests and the degree of ischemia reperfusion injury was assessed.Part of the liver tissue was made into paraffin-embedded specimens to detect rat liver histological change(HE).RT-PCR was used to detect the expressing levels of TNF-a and hIL-10,and TUNEL method was used to detect apoptosis.Result:Compared with the control groups,the liver function(ALT, AST,AKP andγ-GT)in groupⅣwas significantly improved(P＜0.05), the degree of cell apoptosis in groupⅣwas significantly decreased(P＜ 0.05),and the level of hepatic TNF-αmRNA in groupⅣwas significantly decreased(P＜0.05).The gratis of groupⅣrevealed overexpression of hIL-10 mRNA,whereas groupⅠ,ⅡandⅢshowed no expression of hIL- 10 mRNA.Conclusion:Ad -hIL-10 had protective effect to the injury of I/R. The mechanism is that it can inhibit the express of TNF-αmRNA and decrease the degree of cell apoptosis in liver cells.