受体胸腺内注射Fas抗体对急性移植排异的作用及机制

Effect and mechanism of recipient intrathymic injection of anti-Fas monoclonal antibody on reject reaction after heart transplantation

目的:探讨胸腺内注射死亡受体(Fas)抗体对心脏移植急性排异、半胱氨酸基天冬氨酸特异性蛋白酶(Caspase-3)表达及心肌细胞凋亡的影响。方法:应用大鼠腹腔心脏移植模型(供体:Wistar大鼠,受体:SD大鼠),随机分为实验组、环孢素组和对照组。心脏移植前24h,向实验组受体胸腺内注射Fas抗体(0.1mg/kg)1次,腹腔注射生理盐水2mL1次/d;向环孢素组受体胸腺内注射磷酸缓冲液(PBS)(1mL)1次,腹腔注射环孢素(8mg/kg)1次/d;向对照组受体胸腺内注射PBS(1mL)1次,腹腔注射生理盐水(2mL)1次/d。术后5d取移植心脏,进行病理学检查,排斥分级;半定量逆转录聚合酶链反应(RT-PCR)检测移植心脏Caspase-3 mRNA表达;免疫组化检测Caspase-3蛋白表达;原位末端标记法(TUNEL)检测凋亡细胞。结果:1.排斥级别:实验组2.13±0.64低于对照组3.75±0.46高于环饱素组1.25±0.68(P<0.05);2.Caspase-3 mRNA水平:实验组0.62±0.05低于对照组0.87±0.12,高于环孢素组0.59±0.08(P<0.05);3.Caspase-3蛋白阳性细胞数:实验组113±12.5低于对照组223±4.80高于环孢素组95±4.50(P<0.05);4.凋亡指数:实验组7.32±0.58低于对照组11.07±1.18高于环孢素组5.67±0.43(P<0.05)。结论:受体胸腺内注射Fas抗体减轻心脏移植急性排异程度。机理是下调移植心脏Caspase-3基因mRNA及蛋白表达、减少心肌细胞凋亡。

Objective：To investigate the impact on the expression of Caspase-3 and myocardia apoptosis of intrathymic injection of anti-fas monoclonal antibody on reject reaction after rat heart transplantation. Method：Using rat allograft heterotopic heart transplantation models,Wistar rat hearts were transplanted to SD rats and randomly divided into three groups.Study group： （intrathymic injection of anti-fas monoclonal antibody 0.1 mg/kg once and abdominal injection of NS 2 mL o.d.）; CsA group（intrathymic injection of PBS 1 mL once and abdominal injection of CsA 8 mg/kg o.d. ）Control group： （intrathymic injection of PBS 1 mL once and abdominal injection of NS 2 mL o.d.）. Animals were given above drugs one days before operations after operation. Myocardial pathological damage and rejection grade were measured, the express of Caspase-3 mRNA were measured by RT-PCR and the active Caspase-3 protein by immunohistochemistry and apoptosis by TUNEL. Result：1.Rejection grade of Study group 2.13±0.64 were lower than control group 3.75±0.46 （P〈0.05）, and higher than CsA group 1.25±0.68（P〈0.05）. 2.Expression of Caspase-3 mRNA on the transplanted heart were lower in study group 0.62±0.05 than control group 0.87±0.12（P〈0.05）,and higher than CsA group 0.59±0.08（P〈0.05）. 3.The expression of active Caspase-3 protein were lower in study group 113±12.5 than control group 223±4.80 （P〈0.05）,and higher than CsA group 95±4.50 （P〈0.05）. 4.The AI were lower in study group 7.32±0.58 than the control group 11.07±1.18 （P〈0.05）,and higher than CsA group 5.67±0.43 （P〈0.05）.Conclusion：Recipient intrathymic injection of anti-Fas monoclonal antibody can lessen the rejection grade after heart transplantation and reduce expression of Caspase-3 mRNA and active Caspase-3 protein, cardiomyocyte apoptosis in the experimental transplanted heart.