摘要
目的:探讨Mcl-1信号通路阻断剂在结核分枝杆菌H37Rv感染小鼠模型中对Mcl-1表达、巨噬细胞凋亡情况及结核分枝杆菌的影响。方法:小鼠腹腔注射H37Rv菌悬液,建立感染小鼠模型,针对Mcl-1的信号通路选用JAK/STAT信号通路阻断剂AG490、MAPK信号通路阻断剂PD98059和PI3K信号通路阻断剂LY294002用腹腔注射方式作用于各组感染小鼠模型,分为H37Rv感染组、AG490处理组、PD98059处理组、LY294002处理组和对照组。通过细胞抗酸染色观察结核分枝杆菌H37Rv感染小鼠腹腔巨噬细胞的动物模型是否建立成功;通过免疫细胞化学检测结核分枝杆菌H37Rv感染巨噬细胞的Mcl-1表达情况,使用流式细胞技术检测各组巨噬细胞的凋亡率,采用结核分枝杆菌菌落计数来判断巨噬细胞凋亡对结核分枝杆菌的清除效果。结果:细胞抗酸染色结果可见感染的巨噬细胞内散在排列的红色短小抗酸结核分枝杆菌。免疫细胞化学结果显示H37Rv感染组、AG490处理组和LY294002处理组中的Mcl-1蛋白为强阳性表达,PD98059处理组中Mcl-1蛋白为弱阳性表达,对照组Mcl-1蛋白为阴性表达。流式细胞术检测发现H37Rv感染组巨噬细胞凋亡率较对照组高,PD98059处理组的凋亡率显著高于各组,差异显著(P<0.05)。结核分枝杆菌菌落计数结果显示PD98059处理组对H37Rv菌株抑菌作用最明显。结论:Mcl-1信号通路阻断剂通过抑制JAK/STAT、MAPK和PI3K信号通路增加结核分枝杆菌H37Rv感染巨噬细胞的凋亡率,抑制结核分枝杆菌生长;其中,MAPK信号通路干扰Mcl-1的作用最明显,感染的巨噬细胞凋亡率最高,抑菌作用最强。
AIM： To explore the effects of Mcl-1 signal pathway blockers on Mcl-1 expression, macrophage apoptosis and Mycobacterium tuberculosis in the model of mice infected with Mycobacterium tuberculosis H37Rv. METH- ODS： A mouse infection model was established by intraperitoneal injection of H37Rv suspension. The signaling pathway blockers AG490, PD98059 and LY294002 for JAK/STAT, MAPK and PI3K, respectively, were intraperitoneally injected into the mice infected with H37Rv. Cell acid-fast staining was used to observe whether the mouse oeritoneal macroohazesinfected with H37Rv were successfully established, lmmunocytochemical method was employed to detect Mcl-1 expression in the mouse peritoneal macrophages infected with H37Rv. The apoptotic rate in each group was measured by flow cytomer- ty. The scavenging capacity of apoptotic macrophages against H37Rv was determined by Mycobacterium tuberculosis colony counting. RESULTS ： The result of cell acid-fast staining revealed the existence of dispersive arrangement of red short anti- acid Mycobacterium tuberculosis within infected macrophages. The result of cell immunocytochemistry showed strongly posi- tive expression of Mcl-1 protein in H37Rv intection group, AG490 treatment group and LY294002 treatment group, weakly positive expression of Mcl-1 protein in PD98059 treatment group, and negative expression of Mcl-I protein in control group. The result of flow cytometry found that the macrophage apoptotic rate in H37Rv infection group was higher than that in con- trol group, while that in PD98059 treatment group was high than that in other groups with statistically significant differences （ P 〈 0.05 ）. The result of Mycobacterium tuberculosis colony counting showed that PD98059 treatment had the most signifi- cant inhibitmy effect on H37Rv strain. CONCLUSION： Mcl-1 signaling pathway blockers increase the apoptotic rate of macrophages infected with Mycobacterium tuberculosis H37Rv and inhibit the growth of Mycobacterium tuberculosis by inhibi- ting the signaling
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2015年第11期2059-2064,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81260241)
