| PART I The relationship between different ischemia time of non-heart beating liver graft and bile ductObjective:At present, the use of multi-liver transplant cardiac death (DCD) donor, and is vigorously promoting the hospital DCD donor donated work. Therefore, DCD donor is a major organ of the current and future sources of clinical transplantation, namely the inevitable presence of its unique warm ischemia injury problems learning. Among them, the warm ischemia injury caused by liver transplantation biliary disease plagued a major problem, seriously affecting the long-term survival and quality of life of patients. Safety studies on liver warm ischemia time focused on the impact of multi-liver cells and liver function, and less research on biliary warm ischemia injury, especially on warm ischemia time DCD donor rats with biliary tract disease Science studies the relationship between change and its complications, yet little has been reported that oral arguments, which for further study DCD donor warm ischemia injury and predict the incidence of biliary complications after liver transplantation has important significance. This study was designed to investigate the relationship between the different rat DCD donor warm ischemia time and biliary injury.Experimental groups:no-heparin rat model of DCD donor liver transplantation, according to donor warm ischemia time is divided into0min (WO),10min (w10),15min (W15),30min (W30)4groups of20rats. At specified time period taken periportal pathology testing, and inferior vena cava blood biochemical detection. OUTCOME MEASURES:Dynamic biliary pathology observed changes in rat liver function indicators.Statistical methods for analysis of all data:SPSS13.0statistical software Measurement data test with t.Results:When donor warm ischemia less than10min, biliary pathology lighter and reversible change; When donor warm ischemia time was30min, heavy and biliary pathology is irreversible change; Conclusion Heparin DCD liver of rats with warm ischemia time exceeds30min, bile duct injury after transplantation significantly, can lead to irreversible change. PART II The Innuence of Warm Ischemia-reperfusion Injury on Damage and Repair of Bile Ducts in rat liVer transplantation received DCDMATERIALS AND METHODS:Donor without heparin (because it is more in line with the characteristics of cardiac death donor sources of addition, the use of foreign experience DCD donor liver transplantation showed that the removal of life support systems before intravenous heparin certain amount of energy. effectively reduce biliary stricture). Use of cardiac vascular clamp clamp method to establish the base of warm ischemia model of cardiac death. Experimental animals were randomly divided into three groups:SO group, sham operation group, as normal controls; WO group that no donor warm ischemia group; W10group, namely liver warm ischemia time was10minutes. W30groups, namely liver warm ischemia time was30min. In liver transplant graft reperfusion3h,12h, Id,3d, respectively liver tissue specimens and serum samples after7d. Each group at each time point six pairs of rats Comparison of two surgical cases, and postoperative complications. Serum ALT, GGT and TBIL level. HE staining, light microscopy periportal inflammatory cell infiltration and the extent of structural damage to the bile duct, and semi-quantitative ratings and comparisons. Statistical periportal bile duct number, number of blood vessels, blood vessels associated with bile duct number, not the number of blood vessels associated with vascular and biliary several isolated.Results:The two groups each time the main steps needed surgery there is no statistical difference. W10incidence of hepatic artery thrombosis and biliary obstruction than WO increased. W30hepatic artery thrombosis and a higher incidence of biliary obstruction. WO and W1O after liver transplantation group ALT peaked at6hours after surgery and returned to normal within weeks after1. However, GGT and TBIL W10group reached the highest value in the first three days after surgery, respectively, to30and14after the first genius to normal. W30group ALT, GGT, TBIL showed a sustained rise. W10group structure of the extent of damage and bile duct infiltration of inflammatory cells was significantly increased compared with w0 and lasts longer. W30group bile duct discontinuous endothelium, endothelial cell swelling significantly, showing irreversible damage. HE staining showed that after the first seven days W10group periportal fibrosis was significantly increased compared with WO. W30group showed significant fibrosis proliferation, scar tissue formation. Electron microscopy showed:W10group0hours in addition to visible biliary epithelial cells and endothelial cells mild swelling of mitochondria, but also shows the formation of micro-thrombi within the PBP. Swelling of endothelial cells after reperfusion, dissolution, necrosis, apoptosis gradually increased, especially intraluminal thrombosis micro vascular endothelial cells damage is more serious. Early postoperative vascular WO number periportal no significant reduction in the number of vessels not associated with the highest values in the bile duct after24hours, after which the portal area with the formation of blood vessels, the blood vessels are not accompanied by decreasing the number of bile ducts. The early postoperative vascular W10portal area and the number of the number of bile ducts associated with vascular (ie biliary/vascular units) compared to WO group and normal liver were significantly reduced. W10group24hours after liver transplantation periportal bile duct hyperplasia, periportal biliary number gradually increased, but until7days after the number of blood vessels in portal area has been increased slowly. W10group after24hours and the number of blood vessels after the first three days periportal WO group were significantly less than the corresponding time point and the number of blood vessels in normal liver portal area.7days after the vascular portal area only to restore normal liver tissue with a similar level. This leads to a number of periportal bile ducts associated with vascular consistently lower than the corresponding time points wO group and normal liver tissue level, rather than the number of blood vessels associated with bile duct hyperplasia has continued to increase. The number of postoperative vascular W30periportal significantly reduced,3d small amount of periportal bile duct hyperplasia, but not until after seven days periportal bile duct hyperplasia slowly.Conclusion1. BEC is susceptible to warm ischemia-reperfusion injury in a cell population, warm ischemia-reperfusion injury leads to liver transplantation increased incidence of biliary obstruction after planting.2. The PBP micro-thrombosis, increased vascular endothelial cell damage and loss of function, and eventually suffered again BEC warm ischemia, increased BEC damage.3. Warm ischemia-reperfusion injury repair process has lagged far behind in renewable PBP bile duct continuous regeneration in the regenerative ischemia, can not play a normal physiological role.4. Bile duct susceptible warm ischemia-reperfusion injury may be due to ischemia and reperfusion PBP hot micro thrombosis, caused by increased vascular endothelial cell injury, leading to biliary microcirculation and regeneration PBP has lagged far behind in the bile duct regeneration. |
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