TGF-β/Smads信号通路介导基质金属蛋白酶在三角帆蚌伤口修复中的作用研究

Hyriopsis cumingii is one of the most important freshwater mussels for pearl production in China.The damage in the process of insertion nucleus will lead to infection and inflammation. TGF-β/Smads signaling pathway mediated matrix metalloproteinases plays a vital role in the process of wound repair. The purposes of the project are to clone the key genes of TGF-β/Smads signaling pathway and repair relevant target genes-gelatinase (MMP-2, 9), to compare and analysis their sequence features and gene polymorphism and evolution mechanism；to clone promoters of those genes and analysis their activities and effect factors.The inserted nuclear damage model of Hyriopsis cumingiiwill be constructed. Regulation mechanism of TGF-β/Smads signal pathway on target genes and their role in wound repair will be studied by RNAi and specific inhibitor blocking pathway technique.Effects of these genes on the pathway will be studied by constructing the eukaryotic vector transfected cell line, and the recombinant protein functions will be further studied in vitro.The completion of this project will not only help to understand the molecular mechanisms of wound repair of invertebrates, the results can also provide a theoretical basis for disease control and pearl culture.

三角帆蚌是我国主要的淡水育珠蚌之一。插核手术创伤将增加手术蚌的感染和炎症发生。TGF-β/Smads介导的基质金属蛋白酶(MMPs)在创伤修复过程中起着至关重要的作用。本项目拟克隆三角帆蚌TGF-β/Smads信号通路关键基因及修复相关的靶基因MMPs家族的主要成员-明胶酶（MMP-2,9）以及基因的启动子，分析基因的序列特征、多态性及其进化机制；检测启动子活性和影响因素。构建三角帆蚌插核损伤模型。利用RNAi和特异性抑制剂阻断技术研究TGF-β/Smads信号通路对靶基因的调控机制及其在伤口修复中的作用。通过构建真核表达载体转染外源细胞系，研究信号通路中各基因对信号通路的影响。探索重组蛋白的体外修复功能。这不仅有助于理解无脊椎动物的损伤修复的分子机制，还可以为育珠蚌的病害防治和养殖提供理论基础。

三角帆蚌是我国主要的淡水育珠蚌之一。插核手术创伤将增加手术蚌的感染和炎症发生。TGF-β/Smads介导的基质金属蛋白酶(MMPs)在创伤修复过程中起着至关重要的作用。本项目克隆了三角帆蚌TGF-β/Smads信号通路关键基因(TGF-β,TGF-βRI, Smad3/4/5) 及修复相关的靶基因(MMP1,MMP19, TIMP1和TIMP2)，分析了 基因的序列特征、多态性及其进化机制； 构建了三角帆蚌插核损伤模型, 利用RNAi和特异性抑制剂阻断技术研究TGF-β/Smads信号通路对靶基因的调控机制及其在伤口修复中的作用。通过构建真核表达载体转染外源细胞系，研究了信号通路中各基因对信号通路的影响。探索了重组蛋白的体外功能。研究结果不仅有助于理解无脊椎动物的损伤修复的分子机制，还可以为育珠蚌的病害防治和养殖提供理论基础。