去端肽胶原介导小干扰RNA抑制大鼠静脉移植物内膜增生  

Atelocollagen-mediated small interfering RNA delivery for effective gene silencing in rat vein grafts

作　　者：邱雪峰[1] 董念国[1] 孙宗全[1] 苏伟[1] 史嘉玮[1] 

机构地区：[1]华中科技大学附属协和医院心血管外科,武汉430022

出　　处：《中华外科杂志》2009年第13期1028-1031,共4页Chinese Journal of Surgery

基　　金：湖北省卫生厅科研基金资助项目（JX2816）;国家自然科学基金资助项目（30571839）

摘　　要：目的评价局部应用组织因子小干扰RNA（siRNA）抑制静脉移植物内膜增生的效果。方法sD大鼠120只，体重260～300g，建立颈外静脉-颈总动脉移植模型后随机分成4组，每组30只。A组：去端肽胶原-TF Stealth^TM Select RNAi组，B组：去端肽胶原-TF Stealth^TM RNAi阴性对照组，C组：去端肽胶原组，D组：空白对照组。使用去端肽胶原-siRNA混合物涂抹于静脉移植物周围，免疫印迹检测术后1、3、7、14、28d管壁组织因子蛋白表达，免疫组织化学法检测术后3d管壁组织因子表达，同时检测术后14d管壁增殖细胞核抗原指数，计算术后28d新生内膜厚度。去端肽胶原-BLOCK—iT^TM荧光寡核苷酸平行实验组12只，分别于术后3、7d检测管壁BLOCK—iT^TM寡核苷酸荧光确认其稳定性和转染效率。结果平行实验组静脉移植物可观察到BLOCK—iT^TM绿色荧光并持续到术后7d。术后3dA组siRNA有效抑制管壁TF蛋白表达，术后14 dA组与其他组比较增殖细胞核抗原指数明显减少（P〈0．05），术后28dA组新生内膜厚度明显低于其他组（P〈0．05）。结论去端肽胶原携带siRNA可有效下调静脉移植物组织因子表达，抑制内膜增生。Objective To evaluate the efficacy of using small interfering RNA targeting TF as a therapy for vein graft failure. Methods External jugular vein to carotid artery interposition vein grafts, which were applied to a low flow condition, were made in 120 Sprague-Dawley rats weighing 260 to 300 g. These rats were randomly divided into 4 groups, 30 rats each group. Group A was atelocollagen-TF StealthTM Select RNAi group. Group B was atelocollagen-TF StealthTM RNAi group. Group C was ateloeollagen group. Group D was control group. Small interfering RNA mixed with atelocollagen was administrated to the external wall of grafted veins. The TF protein expression of vein grafts was analyzed by Western blot at 1, 3, 7, 14, and 28 d postoperatively, and by immunochemistry at 3 d postoperatively. The proliferation index was determined at 14 d postoperatively. Neointimal hyperplasia was evaluated at 28 d postoperatively. BLOCK- iTTM fluorescent oligo was used to confirm its stability and successful transfer into the vein graft wall at 3 and 7 d postoperatively for another group （ n = 12 ）. Results Fluorescence of BLOCK-iTTM fluorescent oligo could be detected in the graft wall even at 7 d postoperatively. Knockdown of the TF expression was achieved by perivascular application of siRNA using ateloeollagen. Compared with control group, the intima thickness at 28 d after grafting was significantly reduced （ P 〈 0. 05 ）. This phenomenon was preceded by significant reduction of cell proliferation in siRNA-treated grafts at 14 d postoperatively （P 〈0.05 ）. Conclusion The expression of TF in vein grafts can be effectively inhibited by specific siRNAs using a atelocollagen-based nonviral delivery approach in vivo, so that the neointimal thickening can be prevented.

关 键 词：血管内膜 移植物 靶向治疗 RNA于扰 

分 类 号：R686[医药卫生—骨科学]