| Objective Warm ischaemia reperfusion injury(IRI)plays a key role in biliary complication after liver transplantation,especially for donation after cardiac death donors.The excessive inflammatory response is the characteristic of early physiopathological changes of liver IRI.S-Adenosylmethionine(SAM)is an important metabolic intermediate that modulate inflammatory reactions,which can inhibit cytokines production.However,its role in liver warm IRI is not known.This study was designed to investigate the protective effects of SAM on bile duct early Warm ischemia reperfusion injury after liver transplantation and its mechanism in rats.Method 96 pure SPF level male SD rats,body weight was 300-320 g.64 of them was established the rat orthotopic autologous liver transplantation model with bile ducts warm ischaemia reperfusion injury.The cold perfusion time was set to same 15 min,while the anhepatic time was unified to 30 min.The 64 rats were randomly divided into two groups.The SAM group were treated with SAM(170 ?mol/kg body weight)immediately after operation,while the IRI group was treated with same volume of normal saline.Another 32 rats were sham group with exploratory laparotomy.The histopathologic structure of bile duct and was analyzed.Simultaneously,the oxidative stress and inflammatory response were assessed.Furthermore,the apoptosis of biliary epithelial cells was tested with TUNEL.The active caspase-3 expression in bile duct cells was detedted with immunohistochemistry.The date was analyzed to investigate the influence of SAM on bile duct IRI.Result1.Histological changes and serum enzymesHE-staining shown that bile duct injury of intra-hepatic and extra-hepatic in the IRI group was more serious than that in the SAM group.The number of edematous,necrotic and deciduous biliary epithelial cells and inflammatory cells filling in SAM group was less than IRI group at 24 h after liver transplantation.The BDISS scores of bile duct histology of SAM group were significantly lower than that of IRI group(P<0.05).The ALT,ALP and DBIL levels for evaluating bile duct function in the IRI group were much higher than that in the SAM group at all indicated time points(P<0.05).2.The oxidative stress after transplantationRats in IRI group had higher hydrogen peroxide(H2O2)and malondialdehyde(MDA)levels in the liver compared to mice in SAM group(P<0.05).The reduced and oxidized glutathione ratio(GSH/GSSG)in liver tissues of IRI group was much lower than that of SAM group(P<0.05).3.The inflammatory response after reperfusionThe TNF-? mRNA and protein expression in liver tissues was rapidly up-regulated in a time-dependent way after transplantation in the IRI group and significantly higher than that in the SAM group at the same time point(P<0.05).At the same time,the gene and protein expression of IL-1 were markedly down-regulated by SAM administration compared with IRI group(P<0.05).The ratio of p-JNK/JNK protein expression in IRI group was rapidly increased in a time-dependent manner after liver transplantation and much higher than that in the SAM group at the observation period(P<0.05).The cAMP concentration in bile duct tissues was up-regulated by SAM compared to that in the IRI group(P<0.05).4.Apoptosis of biliary epithelial cellsTUNEL staining shown that the number of TUNEL-stained intra-hepatic and extra-hepatic bile duct cells in the IRI groups was much more than the SAM group(P<0.05).The active casepase-3 positive bile duct cells in the SAM group were much less than that in the IRI group at 24 h after operation.Conclusion This study used rats orthotopic autologous liver transplantation model simulated the process of bile duct IRI.Compared the change in histopathologic structure,enzyme indicator level,oxidative stress level,inflammatory response,and apoptosis of biliary epithelial cells.The outcome shown that SAM can attenuated bile duct early warm ischemia reperfusion injury after rat liver transplantation by inhibiting oxidative stress and inflammatory response. |
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