| BackgroundLiver transplantation is the most effective treatment for end-stage liver disease.With the development of the technique and perioperative management,the 1-year survival rate after liver transplantation has reached more than 90%.Liver transplantation is suitable for cirrhosis,acute/chronic liver failure and liver malignancy.Due to the number of available organs is limited,the shortage of donor liver has become an important factor restricting the operation of liver transplantation.Donation after brain death is a major component of organ donor source.Brain death causes hemodynamic changes mainly because the release of catecholamine instantaneous.The donor heart rate increased,and the blood vessel constriction resulted in short supply of blood.The study showed that the incidence of biliary tract complications after liver transplantation was 19%.ITBI is the main cause of biliary complications after transplantation.In this paper,we hope to explore the mechanism of ischemic biliary tract injury through animal experiment and cell experiment.Objective1.To explore whether brain death can aggravate ischemic-type biliary injury after liver transplantation.2.To explore the molecular mechanism of brain death for ischemic biliary injury after liver transplantation.Methods1.To construct a model of rat brain death donor liver transplantation: the experimental was divided into three groups: control group,normal rat donor liver transplantation group and rat brain death donor liver transplantation group.The rats in the control group did not intervene,and the normal rat donor liver transplantation group was that received the normal rat liver in situ and transplanted into another rat.The rat brain death donor liver transplantation group was used to obtain the liver after 6h of the successful rat model of brain death,and transplanted it into another rat.2.After 1 month the model was builded,collecting three groups of bile and liver tissue,using the method of ELISA tou detect the bile duct function markers including GGT/LDH.Using the method of HE dyeing to observe bile duct changes in morphology;Using the method of Western Blotting to detect EMT related markers in liver tissue,and the relationship between EMT and brain death in ischemic-type biliary injury was studied.3.By extracting the primary bile duct epithelial cells of normal rats and establishing the model of ischemia/reperfusion of the primary bile duct epithelial cells to simulate the brain death process.Using the method of Western Blotting to detect the expression of EMT related markers after ischemia/reperfusion of the primary bile duct epithelial cells,to speculate the possible mechanism of brain death for ischemic biliary injury after liver transplantation.Results1.By comparing the HE staining from three groups showed that the liver cells and bile duct form are normal in liver tissues of normal rats.During the normal rat liver transplantation group,the liver cells were slightly swollen,new capillary bile duct increased,intrahepatic bile duct mild dilation,the biliary wall thickened and the surrounding inflammatory cells infiltrated.Hepatic cell fragment necrosis was observed in liver tissue of brain death donor liver transplantation group.Inflammatory cell infiltration,a large number of intrahepatic bile duct hyperplasia obvious,structural disorders,cholestasis bile duct expansion.By comparing three groups of indicators of bile biochemical marker GGT and LDH,it was found that the contents of GGT and LDH were significantly higher than those in normal rats and normal rat donor liver transplantation group.2.The expression of EMT related proteins in three groups of liver tissues was detected by Western blotting.With the increasing of biliary injury in the liver transplantation group,EMT related protein E-cad was down-regulated,and the interstitial markers of N-cad and FSP-1 were up-regulated.3.The ischemia/reperfusion model include control group,ischemia reperfusion group 1h,ischemia reperfusion group 6h.The ischemia/reperfusion model was detected by Western Blotting.It was found that the expression of EMT related markers increased during the occurrence of ischemia/reperfusion injury in bile duct epithelial cells.Meanwhile,the expression of JunD also increased significantly.Conclusion1.Brain death leads to ischemic-type biliary injury after liver transplantation;2.The molecular mechanism might be the that the development of rats brain death activation of JunD,caused the EMT happen and induced the ischemic-type biliary injury of rats after liver transplantation;... |
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