TRAF6对结核分枝杆菌感染后巨噬细胞自噬的调控作用及机制研究

结核分枝杆菌感染巨噬细胞后可激活TLR样信号通路并诱发细胞自噬，而自噬在结核病的发生、发展及预后阶段发挥着不可或缺的作用，该过程极其复杂并受到多种基因调控。TRAF6作为TLR样信号通路的关键基因参与了结核分枝杆菌感染后的炎性及免疫等多种生命过程，但是TRAF6是否参与结核分枝杆菌感染后自噬的调控过程尚不明晰。. 基于以上研究背景，本项目利用siRNA敲低小鼠RAW264.7细胞系的TRAF6表达量，探讨TRAF6对结核分枝杆菌感染后细胞自噬的调控作用；并利用Rapamycin来研究TRAF6对自噬的调控是否依赖于mTOR途径；随后构建Beclin1和RAB7的过表达载体，探明TRAF6对自噬启始和膜融合环节的调控作用；并用结核分枝杆菌感染TRAF6的基因敲除小鼠，通过体内外试验最终阐明TRAF6对结核分枝杆菌感染后细胞自噬的调控机制，以期为结核病的预防及靶向药物研制提供理论依据。

Mycobacterium tuberculosis (Mtb) can activate the TLR- signal pathway and autophagy of macrophages. Autophagy which is regulated by lots of genes plays a vital role in the pathogenesis and outcome of tuberculosis. The mechanism of autophagy has not been well elucidated. TRAF6, a key gene of TLR signal pathway, is involved in the process of inflammatory and immune in target cells infected with Mtb. However, whether TRAF6 is involved in regulating the autophagy of macrophage infected with Mtb remains unknown. . In the present study, we use small interfering RNA of TRAF6 to explore the regulation of TRAF6 on the autophagy of macrophage RAW264.7 infected with Mtb. Then, Rapamycin is employed to examine whether the mTOR-dependent pathway will involve in the autophagy process regultaed by TRAF6. Subsequently , over expressed vector of Beclin1 and RAB7 are constructed to examine the impact of TRAF6 on initiation and membrane fusion of autophagy. In the end, we use TRAF6 KO mice infected with Mtb to confirm the function of TRAF6 on autophagy. Our datas will reveal the mechanism of TRAF6 on the autophagy mechanism of the macrophages infected with Mtb and provide a new insight into the theory basis for development of targeted drugs of tuberculosis .