siRNA干扰OX40/OX40L通路对小鼠移植心脏的免疫保护作用  

The immunoprotection of transplant mouse heart by interfering OX40/OX40L pathway with siRNA

作　　者：李军良[1] 马毅[1] 何晓顺[1] 王国栋[1] 胡安斌[1] 朱晓峰[1] 

机构地区：[1]中山大学附属第一医院器官移植中心,广州510080

出　　处：《中华实验外科杂志》2010年第1期93-95,共3页Chinese Journal of Experimental Surgery

基　　金：广东省科技计划资助项目（200811030301296）;广东省自然科学基金资助项目（9151008901000052）

摘　　要：目的 观察真核表达载体介导siRNA对小鼠移植心脏的免疫保护作用.方法 在C57/BL6J→BALB/C组合[主要组织相容性复合体（MHC）不合]、DBA/2→BALB/C组合[次要组织相容性复合体（mHC）不合]中分别于小鼠心脏移植术后第1、4天胸腺注射生理盐水,pRNAT-CTL阴性对照载体和pRNAT-OX40 siRNA,观察移植心脏存活情况.术后第7天取移植心脏行病理学检查,取血清检测心肌酶谱,取脾脏行逆转录.聚合酶链反应（RT-PCR）和Western blot检测OX40mRNA表达和OX40膜蛋白表达.结果 在C57/BL6J→BALB/C组合中（MHC不合）,pRNAT-OX40siRNA组移植心脏生存时间较生理盐水组和阴性对照组无明显延长,移植心脏病理学检查无明显改善.而在DBA/2→BALB/C组合中（mHC不合）,pRNAT-OX40 siRNA组小鼠移植心脏生存时间较生理盐水组和阴性对照组显著延长,病理改变显著减轻,乳酸脱氢酶（LDH）、谷草转氨酶（AST）、肌钙蛋白（Tn）以及脾脏OX40 mRNA表达和OX40膜蛋白表达与空白对照组和阴性对照组比较明显下降.结论 在DBA/2→BALB/C组合中（mHC不合）,单独阻断OXdO/OX40L通路对小鼠移植心脏有保护作用.在C57/BL6J→BALB/C的组合中（MHC不合）,单独阻断OX40/OX40L通路未发现对移植心脏的保护作用.Objective To investigate the immunoprotection of monse transplant heart mediated by eukaryotic expression vector of siRNA technique.Methods Sodium chloride,pRNAT-CTL negative control vector and pRNAT-OX40 siRNA were injected into thymus on the day I and 4 after heart transplantation using strains representing both major histocompatibility complex （MHC）（C57/BL6J→BALB/C） and minor histocompatibility complex（mHC）（DBA/2→BALB/C）antigen mismatches.Transplant heart,bloodserum and spleen were harvested on the day 7 after operation for pathological examination,cardiac muscle zymogram,RT-PCR and Western blot assays,respectively.Results In C57/BL6J→BALB/C group（MHC incompatible）.the survival time of transplant heart in pRNAT-OX40 siRNA was not prolonged and the transplant heart pathological changes were not improved as compared with sodium chloride and pRNAT-CTL negative control vector.In DBA/2→BALB/C group（mHC incompatible）,the survival time of tramplant heart in pRNAT-OX40 siRNA was significantly prolonged.the transplant heart pathological changes were notablely improved,and lactate dehydrogenase,glutamic oxalacetic transaminase,troponin,spleen OX40 mRNA expression and OX40 membrane protein expression were obviously depressed as compaired with sodium chloride and pRNAT-CTL negative control vector.Conclusion Blockade of OX40/OX40L pathway prevents reiection of mHC-incompatible（DBA/2→BALB/C）but not MHC incompatible（C57/BL6J→BALB/C）cardiac allografts.

关 键 词：心脏移植 siRNA 真核表达载体 OX40 免疫保护 

分 类 号：R654.2[医药卫生—外科学]