结核分枝杆菌分泌性酸性磷酸酶(SapM)抑制小鼠巨噬细胞自噬  

Secretory acid phosphatase of Mycobacterium tuberculosis inhibits the autophagy of murine macrophages

作　　者：胡东[1,2] 王婉[1] 赵润鹏[1] 徐雪维 邢应如[3] 徐从景[3] 张荣波[1,2] 吴静[1,2] 

机构地区：[1]安徽理工大学医学检验中心 [2]安徽理工大学医学院医学免疫学与检验教研室 [3]安徽理工大学附属肿瘤医院,安徽淮南232001

出　　处：《细胞与分子免疫学杂志》2016年第6期726-729,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81302524;81202294;81172778);安徽省自然科学基金(KJ2013A105)

摘　　要：目的研究结核分枝杆菌分泌性酸性磷酸酶(Sap M)对小鼠巨噬细胞自噬的影响。方法分别用Sap M、渥曼青霉素、饥饿处理GFP-LC3-Raw264.7细胞。荧光显微镜下观察自噬体的形成,Western blot法检测微管相关蛋白1轻链3(LC3)Ⅱ水平;再用Sap M处理GFP-LC3-Raw264.7细胞后加氯喹,Western blot法检测LC3Ⅱ水平。结果 Sap M和饥饿均导致GFP-LC3斑点增多和LC3Ⅱ水平增高,在Sap M处理的细胞中加入氯喹并没有引起LC3Ⅱ的进一步升高。结论 Sap M可阻断自噬体与溶酶体的融合,从而抑制小鼠巨噬细胞自噬。Objective To investigate the effect of secretory acid phosphatase as a virulence factor of Mycobacterium tuberculosis（ Sap M） on the autophagy of murine macrophages. Methods GFP-LC3-Raw264. 7 cells were treated with Sap M,wortmannin,or starvation. Then the formation of autophagosomes was observed under a fluorescence microscope. The level of microtubule-associated protein 1 light chain 3（ LC3） Ⅱ was detected using Western blotting. After chloroquine was added in the Sap M-treated cells,LC3Ⅱ level was again tested by Western blotting. Results Both starvation and Sap M increased the number of GFP-LC3 puncta and the level of LC3Ⅱ. There was no further increase of LC3Ⅱlevel in Sap M-treated cells after chloroquine addition. Conclusion Sap M can block autophagosome-lysosome fusion and inhibit autophagy of murine macrophages.

关 键 词：结核分枝杆菌分泌性酸性磷酸酶(SapM) 自噬 融合 Raw264.7细胞 微管蛋白相关轻链3(LC3) 

分 类 号：R378.911[医药卫生—病原生物学] R392.33[医药卫生—基础医学]