下调Mcl-1表达对H37Rv感染小鼠模型调控作用研究  

The regulatory role of down-regulated Mcl-1 expression in a mouse model of H37Rv infection

作　　者：王新敏[1] 王小芳[2] 卢洋 杨菩 韩玲 王英姿[2] 张万江[2] 章乐[2] WANG Xin-min;WANG Xiao-fang;LU Yang;YANG Pu;HAN Lin;WANG Ying-zi;ZHANG Wan-jiang;ZHANG Le(Urology,the First Affiliated Hospital of Shihezi University,Shihezi 832002,Xinjiang,China;Department of Pathophysiology,College of Medicine,Shihezi University)

机构地区：[1]石河子大学医学院第一附属医院泌尿外科,新疆石河子832002 [2]石河子大学医学院病理生理学教研室

出　　处：《中国病原生物学杂志》2018年第7期709-713,718,共6页Journal of Pathogen Biology

基　　金：国家自然科学基金资助项目(No.81660330)

摘　　要：目的用不同阻断剂下调调控抗凋亡蛋白Mcl-1表达信号通路,探讨下调Mcl-1表达对结核杆菌感染小鼠腹腔巨噬细胞凋亡和炎症反应的影响。方法制备结核分枝杆菌国际标准强H37Rv菌株菌悬液,感染BALB/c小鼠,再分别腹腔注射Mcl-1信号通路阻断剂AG490、PD98059、LY294002,同时设立对应的对照组。应用ELISA筛选不同阻断剂处理小鼠的最佳剂量,采用TUNEL试剂盒检测阻断剂应用后小鼠腹腔巨噬细胞凋亡率,透射电镜观察阻断剂应用后宿主巨噬细胞的形态学变化,HE染色分析应用阻断剂后小鼠肝、肺、脾、肾脏器的病理损伤情况。结果阻断剂处理H37Rv感染组小鼠巨噬细胞凋亡率与空白对照组均有不同程度的增高,其中PD98059处理组显著高于其他处理组(P<0.05);阻断剂PD98059处理组小鼠巨噬细胞感染H37Rv数量明显减少,且出现核固缩和凋亡。H37Rv感染小鼠肝、肺、脾、肾脏器出现严重病理损伤,而阻断剂应用后的病理损害程度明显减轻(P<0.05)。结论应用阻断剂下调Mcl-1的表达可促进结核杆菌感染小鼠腹腔巨噬细胞的凋亡,减少巨噬细胞内潜伏H37Rv数量,减轻结核感染对小鼠脏器的病理损伤。作用效果以MAPK信号通路阻断剂PD98059强于JAK/STAT和PI3K信号通路阻断剂AG490和PD98059。Objective To use different blockers to down-regulate the expression of the anti-apoptotic Mcl-1 signaling pathway and to investigate the effect of down-regulating the expression of Mcl-1 on the apoptosis and inflammatory response of Mycobacterium tuberculosis infecting murine peritoneal macrophages. Methods A bacterial suspension of M.tuberculosis was prepared using the standard strain H37 Rv.BALB/c mice were infected with M.tuberculosis and then the Mcl-1 signaling pathway blockers AG490,PD98059,and LY294002 were injected intraperitoneally on day 1,3,5,and 7.A corresponding control group was created for each blocker.ELISA was used to screen for the optimal dose of each blocker.A TUNEL kit was used to detect the rate of apoptosis of murine peritoneal macrophages.After addition of a blocker,morphological changes in host macrophages were observed using transmission electron microscopy.HE staining was used to analyze pathological damage to the liver,lungs,spleen,and kidneys of mice after a blocker was administered.Results The rate of apoptosis of macrophages in mice infected with the H37 Rv strain increased with different concentrations of a blocker in comparison to the blank control group.The rate of macrophage apoptosis was significantly higher in mice treated with PD98059 than that in other groups（P〈0.05）.In addition,the number of H37 Rv cells that infected murine macrophages decreased significantly（the number decreased to 26% after treatment）.Nuclear condensation and macrophage apoptosis was evident after mice were treated with the blocker PD98059.H37 Rv infection caused severe pathological damage to the liver,lungs,spleen,and kidneys of mice,but pathological damage was significantly alleviated by treatment with a blocker.This was especially true for mice treated with the blocker PD98059（P〈0.05）. ConclusionMcl-1 signaling pathway inhibitors promoted apoptosis of peritoneal macrophages in mice infected with an M.tuberculosis strain,they reduced the number of latent M.tuberculosisin macrophages,and

关 键 词：结核 Mcl-1信号通路 腹腔巨噬细胞 细胞凋亡 

分 类 号：R378.911[医药卫生—病原生物学]