大鼠肝移植后颗粒酶B mRNA的表达与急性排斥反应的关系

Relationship between expression of granzyme B mRNA and acute rejection after liver transplantation in rats

目的探讨大鼠肝移植后肝组织、外周血和胆汁中颗粒酶B m RNA的表达变化与急性排斥反应的关系,评价术后检测颗粒酶B m RNA的表达对早期诊断急性排斥反应发生的价值。方法首先建立大鼠原位肝移植胆道T管引流模型。实验动物分为4组,A组:正常对照组,n=20;B组:同种同基因组,n=30;C组:异基因移植+Cs A组(SD→Wistar大鼠),术后给予免疫抑制剂Cs A 5mg·kg-1·d-1,n=30;D组:急性排斥反应组(SD→Wistar大鼠),n=30。应用RT-PCR方法分别检测各组术后第1、3、5、7和10d外周血和胆汁中颗粒酶B mRNA的表达水平,同时行肝脏病理组织学观察。结果大鼠肝移植后D组肝组织和外周血自术后3d起可以检测到颗粒酶B mRNA的表达,颗粒酶B mRNA的表达在术后第3、5、7、10d与其他组相比有显著统计学差异。各组在胆汁中均未检测到颗粒酶B mRNA的表达。D组急性排斥表现与病理学检查相吻合。结论肝移植术后检测颗粒酶B mRNA的表达,可以为早期发现急性排斥反应的发生提供诊断依据,而从外周血中检测其表达可以提供一种急性排斥反应早期诊断和检测的方法。

Objective To investigate relationship between acute rejection and expression of granzyme B mRNA in liver tissue,peripheral blood and bile after liver transplantation in the rat.To evaluate the value of early judging acute rejection by examining expression of granzyme B mRNA. Methods To establish a stable orthotopic liver transplantation model with biliary extra-drainage by mini T-tube in rats.Experimental animals were divided into four groups. Group A:the Wistar rats were accepted a bile drainage;group B:both donor and receptor were Wistar rats(Wistar→Wistar rats);group C:acute rejection group(SD→Wistar rats) with immunosuppressive therapy;group D:acute rejection group(SD →Wistar rats). Animals were executed four or six on 1,3,5,7,10 d after surgery and collected peripheral blood,liver tissue and bile.The expression of granzyme B mRNA was examined by reverse transcription polymerase chain reaction. Results Granzyme B mRNA could be detected in liver tissue and peripheral blood since the third day after surgery in group D,and there were significantly statistical differences compared with other groups on1,3,5,7,10 d after operation.Expression of granzyme B mRNA was not detected in each sample of bile.Histological findings confirmed the occurrence of acute rejection in group D.Conclusion An early diagnosis methods to examine the occurrence of acute rejection can be provided by means of monitoring the expression of granzyme B mRNA from peripheral blood.