同种异基因大鼠肝移植急性排斥反应模型的建立

Establishment of acute rejection models in allogeneic rats following liver transplantation

背景:移植后急性排斥反应是制约国内肝移植发展的主要障碍之一。目的:探索同种异基因大鼠肝移植急性排斥反应模型的建立方法。设计、时间及地点:病理学水平的实验方法改进,于2007-06/2008-11在中南大学实验动物学部完成。材料:供体选用SD大鼠,雌雄不拘;受体选用雄性Lewis大鼠,供受体各120只。方法:采用改良Kamada’s二袖套法,并在此基础上加以改良:肝上下腔静脉重建采用改良连续缝合;支架法重建肝动脉,肝脏灌注采用全自动静脉输液泵控制,建立同种异基因大鼠肝移植急性排斥反应模型。共实施大鼠肝移植120例,其中定型手术80例。主要观察指标:移植手术时间,移植成功率,大鼠精神状态,对刺激的反应,活动及进食情况,急性排斥反应Banff评分。结果:80例定型手术中,供肝热缺血时间为0～2min,冷缺血时间为80min,无肝期平均为16～21min,手术成功率为85%(68/80)。急性排斥反应出现于移植后第3天,表现为食欲不佳、精神萎靡;移植后第5天出现局部黄染;移植后第7天黄疽进行性加重,并出现明显腹水和死亡。移植后第3,5,7天的急性排斥反应Banff评分分别为3.98,4.96及6.03。结论:同种异基因大鼠采用改良Kamada’s二袖套法、重建肝动脉、改进肝上下腔静脉缝合及肝脏灌注方法可建立稳定的大鼠肝移植急性排斥反应模型。

BACKGROUND： Acute rejection following transplantation was one of primary obstruction for liver transplantation in China. OBJECTIVE： To study the technique of establishing models of acute rejection following orthotopic liver transplantation （OLT） in allogeneic rats. DESIGN, TIME AND SETTING： A pathological empirical method improvement. Experiments were performed at the Department of Laboratory Animal Science, Central South University from June 2007 to November 2008. MATERIALS： Sprague Dawley rats of both genders were considered as donors （n=120）, while male Lewis rats served as recipients （n=-120）, METHODS： Modified Kamada＇s cuff technique was used and modified with hepatic arterial reconstruction and modified suture for suprahepatic inferior vena cava. Hepatic perfusion method was used to create models of acute rejection after ischemia reperfusion injury in allogeneic OLT rats using full-automatic venous transfusion pump. A total of 120 rat models of OLT were generated, and 80 cases formal operations were practiced. MAIN OUTCOME MEASURES： The following parameters were measured： transplantation time, success rate of transplantation, rat mental state, reaction to simulation, activity and eating, acute rejection Banff score, RESULTS： 80 cases formal operations were practiced. The anhepatic time was 16 21 minutes and the successful rate was 85% （68/80） after the liver grafts undergoing 0 2 minutes of warm ischemia and 80 minutes of cold ischemia. Acute rejection was emerged in 3 days post-operation, with the presence of poor appetite and depressed. Local stained yellow appeared at day 5 following transplantation. The appearance was most typical in 7 days post-operation, with the significant presence of ascites and death. Banff scores were 3.98, 4.96 and 6.03 at days 3, 5 and 7 following transplantation. CONCLUSION： The allogeneic rat＇s OLT models were established using Kamada＇s cuff technique with hepatic arterial reconstruction and modified suture for superior vena. Hepatic perfusion method was reliable to study acute rejection after ischemia reperfusion injury.