结核分枝杆菌ESAT6-CFP10融合蛋白对小鼠巨噬细胞自噬功能的影响

The Effects of Mycobacterium tuberculosis ESAT6-CFP10 Fusion Protein on the Function of Autophagy in Mouse Macrophages

目的研究结核分枝杆菌(MTB)ESAT6-CFP10融合蛋白对小鼠巨噬细胞自噬功能的影响。方法H37Rv菌株感染小鼠巨噬细胞后加入纯化的重组ESAT6-CFP10融合蛋白,通过透射电镜检测自噬体的形成。提取细胞总RNA和蛋白,以实时定量RT-PCR及Western blot方法检测自噬相关基因(atg)分子水平和蛋白表达水平。结果ESAT6-CFP10融合蛋白可抑制小鼠巨噬细胞自噬体的形成,并导致atg分子表达水平下降,其中atg8表达量下降最为明显。结论MTB ESAT6-CFP10融合蛋白通过调控atg分子表达水平影响小鼠巨噬细胞自噬功能。

Objective To study the effects of Mycobacterium tuberculosis ESAT6-CFP10 fusion protein on the autophagic function of mouse macrophages.Methods Macrophages of mouse were first infected with M.tuberculosis H37Rv,and then infected with ESAT6-CFP10 fusion protein.Transmission electron microscope was used to detect the formation of autophagosome.Total RNA and protein of infected macrophages were extracted and used to measure the expression levels of autophagy-related genes（atgs） by RT-PCR and Western blot.Results ESAT6-CFP10 fusion protein inhibited the formation of autophagosome in mouse macrophages,and the expression level of atgs also reduced,especially the atg8 had a significant reduction.Conclusion ESAT6-CFP10 fusion protein can inhibit the formation of autophagosome in mouse macrophages by the way of down-regulate the expression level of atgs to control the function of autophagy.