The Expression And Effect Of IL-17on Rat Acute Hepatic Allograft Rejection

Part1:Establishment of the Models of Rat Acute Hepatic Allograft RejectionObjective:To establish the stable models of rat acute rejection of allograft in inbred rats.Methods:120cases of rat OLT were performed by modified Kamada’s two-cuff technique in closed colony SD rats, following by30cases of Lewis to BN in inbred rat. The grades of acute rejection in allografts were judged according to Banff scheme.Results:120cases of rat OLT were divided into three parts, every part contains40cases. Of the aftermost40cases, the successful rates of operation and1w survive rates were92.5%and82.5%respectively, the duration of anhepatic time was17.23±2.32min. Rejection activity index were rised on day1,3,5,7,9postoperation.Conclusion:The rat OLT models were performed by modified Kamada’s two-cuff technique. Lewis to BN rat strain combination is a stable acue rejection model. rejectionPart2:IL-23Promotes NKT Cell Production of IL-17during Acute Rejection after Rat Orthotopic Liver TransplantationObjective:(1) To study the possibility of NKT cell can produces IL-17during acute rejection after rat orthotopic liver transplantation.(2) To study whether IL-23promotes NKT cell production of IL-17during acute rejection or not.Methods:Experimental animal were divided into two groups(n=18), orthotopic liver transplantation were performed in each group. Group A: Lewâ†'Lew. Group B:Lewâ†'BN.6rats were sacrificed and random divided into two groups on day3,7and9postoperation, the levels of IL-17and IL-23in liver allografts were determined by ELISA. IL-17-positive cells in CD1d CD4+cells of grafts were detected by flow cytometry on day7postoperation.Results:High expression of IL-17and IL-23was observed in liver allografts. The ratios of NKT cells was dramatically increased in group B compared with group A(P<0.01). In vitro, blockage of IL-23using anti-IL-23antibody can inhibit increasing expression of IL-17(P<0.01).Conclusion:NKT cell contribute to production of IL-17mediated by IL-23on rat acute allograft rejection model of orthotopic liver transplantation. Part3:Expression and Effect of IL-17on Rat Acute Hepatic Allograft RejectionObjective:To study the expression and effect of IL-17in acute allograft rejection after rat orthotopic liver transplantation.Methods:Experimental animal were divided into three groups(n=6), orthotopic liver transplantation were performed in each group. Group A: Lewâ†'Lew. Group B:Lewâ†'BN. Group C:Lewâ†'BN, and blockage of IL-17using anti-IL-17antibody on day1,2and3postoperation. Sacrificed6rats of each group on day7postoperation, serum levels of ALT, AST and TBIL were measured; Part of the liver tissues were made into paraffin-embedded specimens to detect rat liver histological change; The serum levels of IL-17in each group were measured by ELISA; IL-17, INF-γ, ICAM-1and MIP-2mRNA in each group were measured by RT-PCR.Results:On day7postoperation, serum levels of ALT, AST and TBIL in group B were obviously increased than group A (P<0.05); Acute rejection was not emerged in group A, but was significant on group B. Serum levels of IL-17in group B were significantly increased expression than group A(P<0.05). The expression of IL-17, INF-γ, ICAM-1and MIP-2mRNA in group B were all increased expression than group A(P<0.05). Compared with group B, serum levels of ALT, AST and TBIL in group C were obviously depression(P<0.05); Acute rejection were moderated in group C, and RAI was depression(P<0.05); Serum levels of IL-17in group C were significantly depressed expression (P<0.05); The expression of IL-17, INF-γ, ICAM-1and MIP-2mRNA in group C were all significantly depressed expression(P<0.05).Conclusion:IL-17emerged high expression in acute rejection after rat orthotopic liver transplantation and the cytokine levels, such as INF-γ, ICAM-1and MIP-2were all arose at the same time.