小鼠心脏移植中阻断 OX40信号通路诱导免疫保护的实验研究

摘要

目的：为了探讨真核表达载体介导siRNA在小鼠移植心脏术后的免疫保护作用。方法我们选用次要组织相容性复合体不合的小鼠分成三组作心脏移植并于移植术后第1天、第4天对三组移植小鼠的胸腺分别注射生理盐水、pRNAT-CTL 阴性对照载体和 pRNAT-OX40siRNA。在小鼠移植心脏术后第7天取移植小鼠心脏、血清、脾脏分别进行病理学免疫组化检查、心肌酶谱检测以及OX40mRNA和OX40膜蛋白表达的检测。结果 pRNAT-OX40siRNA组较生理盐水组和阴性对照组，小鼠移植心脏生存时间显著延长，病理改变显著减轻，谷草转氨酶（AST）、肌钙蛋白（Tn）、乳酸脱氢酶（LDH）以及OX40mRNA和OX40膜蛋白表达与空白对照组和阴性对照组相比明显下降，差异具有显著性。结论小鼠次要组织相容性复合体不合的心脏移植中阻断OX40／OX40L免疫通路能够诱导免疫保护效应。%Objective To value the immunoprotection of transplant heat that was mediated by eukaryotic expression vector of siRNA technique. Methods First, we did mouse heart transplantation using strains representing minor histocompatibility complex antigen mismatches (mHC), and all the experimental animals were divided into three groups. Then we injected respectively sodium chloride ,pRNAT-CTL negative control vector and pRNAT-OX40siRNA into the three groups of mouse thymus at the 1st day and the fourth day after heart transplantation..At the 7th day of post-transplantation, we collected the transplant heart,.blood serum and spleen for pathological examination, besides cardiac muscle zymogram,..RT-PCR and Western blot tests,.respectively..Results The survival time of transplant heart in pRNAT-OX40siRNA was significantly prolonged,..and the transplant heart pathological examination was significantly bettered and lactate dehydrogenase ,.glutamic oxalacetic transaminase, troponin, spleen OX40mRNA expression and OX40 membrane protein expression were significantly lowered compared with those in mice with sodium chloride and pRNAT-CTL negative control vector..Conclusion Our results clearly show that blockade OX40/OX40L pathway has immunoprotection for heart transplantation of mouse with mHC-incompatible.