miR-9调控VSMC表型转化在脑血管支架植入术后再狭窄中的作用及机制研究

项目名称： miR-9调控VSMC表型转化在脑血管支架植入术后再狭窄中的作用及机制研究

项目编号： No.81471194

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 周振华

作者单位： 中国人民解放军第三军医大学

项目金额： 70万元

中文摘要： 脑血管支架植入术后再狭窄是脑血管病防治中的重点和难点。血管平滑肌细胞（VSMC）表型转化是防治的关键环节。我们前期在脑动脉再狭窄病人及大鼠模型中研究发现TGFβ和miR-9均增高且呈正相关，进一步体外预实验发现miR-9能下调Myocd基因并能促进VSMC表型转化，但具体机制尚不清楚。基于前期研究和文献报道，我们提出假设：TGFβ上调miR-9，后者通过靶蛋白Myocd调控VSMC表型转化，引发脑血管支架植入术后再狭窄。本项目应用荧光素酶报告基因、qPCR、WB和siRNA等技术，在细胞水平与动物模型明确TGFβ和Myocd分别是miR-9的上游调控基因和下游靶基因；观察TGFβ/SMAD信号通路是否可通过上调miRNA-9，抑制Myocd表达，从而促进VSMC表型转化。该研究旨在阐明脑血管支架植入术后miR-9促进VSMC表型转化的功能和分子机制，为支架植入术后再狭窄的防治提供理论依据。

中文关键词： 再狭窄；脑血管病；微小RNA；血管平滑肌细胞

英文摘要： Restenosis after cerebrovascular stent implantation is one of the most important and difficult problem in prevention and treatment of cerebrovacular disease, and the key is phenotype switch of vascular smooth muscle cell ( VSMC ). Our previous study found that the phenotype switch of VSMC is closely related to TGFβ and miR-9 on the cerebrovascular restenosis of patient and rat, but the definite mechanism is unknown and is to be disclosed. According to the latest research and the foundation, We suppose that TGFβ can upregulate miR9, which regulate VSMC phenotype switch by targeting protein Myocd, and then trigger cerebrovascular restenosis after stent implantation. Using the method of luciferase system, this project aims that TGFβ and Myocd are respectively the upstream regulatory gene and the downstream target gene of miR-9. By means of qPCR ,Western blot and siRNA technology, We observe that whether TGFβ /SMAD signaling pathway upregulate miR9, inhibit the expression of Myocd, and promote the differentiation of VSMC phenotype. The purpose is to elucidate the specific mechanism of VSMC phenotype switch after the cerebrovascular stent implantation and the role of regulatory of TGFβ and miR-9, which can provide a new way to inhibit and prevent the clinical restenosis after cerebrovascular stent implantation.

英文关键词： restenosis;cerebralvascular disease;miRNA;vascular smooth muscle cell