TRPC6基因启动子区-254(C>G)SNP通过NF-κB调控足细胞功能的机制研究

细胞内钙离子内流增加是导致足细胞损伤并最终产生蛋白尿的主要原因之一。业已证明，作为足细胞表面唯一的钙离子通道蛋白，TRPC6基因突变或表达异常与细胞内钙离子内流增加密切相关，但其具体调控机制尚不清楚。我们前期研究首次发现TRPC6基因启动子区-254(C>G)是调控该基因表达的重要单核苷酸多态性(SNP)，而该变异位点可能与核转录因子NF-κB结合参与调控足细胞功能，但其确切分子机制有待阐明。因此本课题以人类足细胞株为研究对象，应用EMSA、基因克隆、基因转染和免疫共沉淀等技术：（1）明确-254(C>G)通过NF-κB 介导并调控TRPC6 基因启动子活性的分子机制；（2）进一步分析TRPC6钙离子通道及其相关的钙离子信号通路对足细胞结构和功能的影响，从而阐明-254(C>G)SNP通过NF-κB调控足细胞功能的可能机制，为蛋白尿发生的分子机制提供新的理论依据。

Increased Ca2+ influx is the one of the major causation in the development of proteinuria. Moreover, TRPC6, as the only calcium channel protein in podocyte, involved in the major procedure of podocyte injury and repair. Gene mutations or abnormal expression of TRPC6 gene have been approved most associated with increased Ca2+ influx, however, the specific molecular mechanism remains unclear. Our previous study firstly demonstrated that in the promoter region -254(C>G) is an important SNP to regulate TRPC6 expression. Besides, the gene variant may create a binding site for NF-κB, however, the exact molecular mechanism remains to be made clear. Therefore, by the study of human podocyte cell line, apply the technology of EMSA, gene clone, gene transfection, and co-immunoprecipitation etc.,this research intended to: (1) Confirm the molecular mechanism of NF-κB regulating -254(C>G) in promoter region can alter the promoter activation of TRPC6. (2) Investigate the effection of TRPC6 associated Ca2+ signal pathway on the cytoarchitecture and function of podocyte. Accordingly, the research can specify the molecular mechanism of NF-κB regulating the podocyte function through -254(C>G) SNP in TRPC6 promoter, and provide new theoretical basis for the molecular mechanism of proteinuria.