大鼠异位辅助性肝移植术后PFP mRNA的表达

Expression of Perforin mRNA of rat heterotopic auxiliary liver transplantation

目的建立稳定的脾窝异位辅助性肝移植大鼠模型,观察术后移植肝病理学变化,并应用RT-PCR法,研究外周血中PFP mRNA的表达与急性排斥反应的关系。方法采用健康清洁成年Wistar和SD大鼠。实验分为3组:A组:空白对照组n=30;B组:肝移植术后免疫抑制组,n=30;C组:肝移植术后非免疫抑制组,n=30。各组在术后第1、3、5、7、10d随机取6只动物肝脏和外周血,将肝脏作病理学检查,并应用RT-PCR方法监测外周血的穿孔素mRNA的表达。结果外周血PFP mRNA表达能随病理变化,以数值的形式反应术后的急性排斥反应变化过程。通过检测未发现A组PFP mRNA表达,而B、C组均有表达,对比各时间观察的穿孔素表达水平发现B组与C组比较有显著差异(P<0.05)。结论 (1)通过各时间点的监测,发现外周血PFPmRNA表达水平与移植肝的病理变化趋同,因而应用半定量RT-PCR检测外周血穿孔素mRNA表达可作为早期诊断肝移植急性排斥反应一种较好的方法。(2)免疫抑制剂的应用,可以有效地减少炎性细胞的浸润,降低穿孔素mRNA的表达,对减轻急性排斥反应作用明显。但仍无法较完全地控制肝细胞坏死和出血。

Objective To establish a stable rat model of heterotopic auxiliary liver transplantation in spleen fossa with splenectomy.To observate the pathological changes of graft liver after operation.useing semi-quantitative RT-PCR to monitor the expression of Perforin mRNA in peripheral blood.To explore the relationship between the expression of perforin mRNA and acute hepatic allograft rejection.Methods This design of experiment use adult healthy clean rats.They was divide into three groups：Group A：blank group n=30;Group B： allograft transplantation group（graft liver came form Wistar rats,host liver came form SD rats）and treated with cyclosporine n=30;Group C,allograft transplantation group and untreated controls n=30.The samples were obtained at different times,which were timed on the 1st,3rd,5th,7th,10th day postoperative.To select six rats at every observation point.semi-quantitative was used to measure the expression of perforin mRNA in peripheral blood,and the pathological changes of the graft were observed as well.Results Postoperative acute rejection could be reflected through the pathological changes of graft liver.During acute rejection in rat auxiliary liver transplantation,The pathological changes of graft liver was similar to the expression of Perform mRNA in peripheral blood.The result show it found the expression of Perforin mRNA in peripheral blood both Group B and gooup C,but Group A.Compared the Group B with the group C,there were significantly statistical differences between different time points（P0.05）.Conclusion（1）According to monitor the changes of the expression of perform mRNA in peripheral blood in different time point,we find the change was paralleled between the pathological changes of graft liver and it.While acute rejection in rat liver transplantation was occurring,the expression levels of perform mRNA between grafts and peripheral blood,which was coherent with the level of the severity degree of histologic damage.（2）Cyclosporine A can decrease the number of infiltrating cells and degrade the expression levels of perform mRNA.It has palpable effect to reduce acute rejection.but can not change hepatocyte necrosis and hemorrhage.