SMAD7调控内皮间质转化可预防跟腱损伤后的异位骨化

发布时间：2018-07-21 22:17

【摘要】：背景:现有的研究结果证明,内皮细胞向间质细胞转化(End MT)在异位骨化病程中起主要作用。目的:基于SMAD7基因在阻止肌成纤维细胞转化等内皮-间质转化进展中的潜能,验证其是否是异位骨化潜在的治疗靶点。方法:构建了过表达SMAD7的慢病毒颗粒,并在大鼠主动脉内皮细胞中测定了其最佳滴度和转染效率。随后,将该病毒颗粒注入构建的大鼠跟腱损伤模型中,对照组注射生理盐水。使用qPCR和蛋白印迹实验分析了损伤处内皮和间质标志物的表达,采用X射线和组织染色观察异位骨化病程转化。结果与结论:(1)局部注射转染SMAD7基因的病毒载体可引起内皮细胞标志物(CD31,VE-cadherin)发生上调,同时间质细胞标志物(N-cadherin,vimentin)发生下调,提示SMAD7的局部高表达阻断了内皮-间质转化改变。这种表达差异在造模后10周时比在造模后6周时更明显;(2)X射线片和组织染色显示,相比对照组,注入表达SMAD7的慢病毒后,肌腱组织中骨化结构缺失;(3)因此认为,促进局部SMAD7的高表达可用来预防术后异位骨化形成,而不影响正常的伤口愈合过程。
[Abstract]:Background: current studies have demonstrated that endothelial cell to mesenchymal cell transformation (end MT) plays a major role in the course of ectopic ossification. Aim: to investigate whether SMAD7 gene is a potential therapeutic target for ectopic ossification based on the potential of SMAD7 gene in preventing the development of endothelium-mesenchymal transformation such as myofibroblast transformation. Methods: the lentivirus granules expressing SMAD7 were constructed and the optimal titer and transfection efficiency were determined in rat aortic endothelial cells. Then, the virus particles were injected into the model of Achilles tendon injury in rats, and the control group was injected with normal saline. The expression of endothelial and interstitial markers was analyzed by qPCR and Western blot. The course of ectopic ossification was observed by X-ray and tissue staining. Results and conclusion: (1) Local injection of SMAD7 gene vector could induce up-regulation of endothelial cell marker (CD31- VE-cadherin) and down-regulation of N-cadherin- vimentin, suggesting that local overexpression of SMAD7 blocked the change of endothelium-interstitial transformation. The difference in expression was more significant at 10 weeks than at 6 weeks after modeling. (2) X-ray and tissue staining showed that the ossification structure of tendon tissue was missing after injecting lentivirus expressing SMAD7 in the control group; (3) the results showed that the ossification structure of the tendon was absent after injection of the lentivirus expressing SMAD7. The high expression of SMAD7 can be used to prevent the formation of ectopic ossification without affecting the normal wound healing process.
【作者单位】：解放军第二军医大学附属长海医院;上海交通大学附属第六人民医院;
【基金】：上海市卫生局青年基金(20134y110)~~
【分类号】：R686

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