外周血CTL活化基因在大鼠肾移植急性排斥的表达及意义

目的：探讨大鼠同种异体肾移植外周血淋巴细胞（peripheral blood lymphocyte，PBL）穿孔素、颗粒酶B和FasL mRNA表达与急性排斥反应（acute reiection，AR）的关系。方法：采用TdT介导的脱氧核苷酸原位末端标记法（TUNEL）检测移植肾脏中的凋亡细胞。另外同时检测血清肌酐水平。采用逆转录聚合酶链反应（RT-PCR）检测移植肾和PBL中穿孔素、颗粒酶B和FasLmRNA表达。并以同基因肾移植为对照。结果：病理学检查结果显示实验组大鼠在术后第5、7、9～11天分别发生轻、中、重度排斥。对照组无明显排斥现象。实验组于手术后第5、7、9～11天细胞凋亡数明显高于对照组（P〈0．01）：肾小管上皮细胞凋亡程度与血清肌酐水平、病理学检查结果一致。RT-PCR结果显示实验组于术后3～5天起，肾组织和PBL中穿孔素、颗粒酶B和FasLmRNA表达水平即显著升高，于术后7-9天达到高峰。与对照组相比差异有显著性（P〈0．01）。外周血和肾组织中FasL、穿孔素和颗粒酶BmRNA的表达水平平均在急性排斥反应发生前3～5天开始明显上升，其变化趋势早于血清肌酐。结论：定量RT-PCR测定外周血淋巴细胞中穿孔素、颗粒酶B和FasLmRNA表达可以较敏感预测肾移植急性排斥反应的发生，具有临床诊断参考价值。

The experssion of CTL active gene in peripheral blood lymphocytes in acute rat renal allo grafts rejection

Objective: To explore the relationship between the expression of perforin, granzyme B and FasL mRNA in peripheral blood lymphocytes and acute rejection in xenogenic renal transplantation. Methods: Orthotopic left kidney transplantation was performed in inbred SD rat. and inbred Wister rat Apoptosis cells in grafts were monitored by terminal deoxynucleotidy transferase- mediated dUTP nick-end labeling (TUNEL) method and the levec of serum carnine was measured. The perforin, granzyme B and FasL mRNA were detected by using RT-PCR in the renal allografts and peripheral blood lymphocyte PBL. Results: The histology showed mild, moderate and severe acute rejection occurred on day 5, 7, 9 -11 after operation in the experimental group respectively. But none of control group had the histological evidence of acute rejection. The TUNEL showed that the number of apoptosis cells in the experimental group was significantly more than that in the control group (P < 0.01). The apoptosis of tubular epithelial cells were associated with the level of serum creatinine and the severity degree of histologic damage. The RT-PCR showed that the expression of perforin, granzyme B and FasL mRNA in renal allograft and PBL increased significantly in experimental group compared with the control group from the 3th to 5th day after opteration (P < 0.01), and peaked on the 7th to 9th day. The expression of perforin, granzyme B and FasL mRNA in real allograft and PBL increased significantly 3 to5 days before the presence of acute rejection. The expression of perforin, granzyme B and FasL mRNA occurred earlier than the change of blood serum carnine. Conclusion: The level of FasL expression in PBL is sensitive predictor for early diagnosis of acute rejection.