阻断小鼠Mcl-1表达信号通路对BCG感染的影响

目的 探讨下调Mcl-1表达信号通路对BCG感染的小鼠模型的影响。方法 首先制备好BCG菌悬液感染BALB/c 小鼠,再分别用调控Mcl-1表达的不同信号通路阻断剂腹腔注射感染小鼠模型,并同时设立对应的对照组,于处理后的1 d、3 d、5 d、7 d处死小鼠并收集腹腔巨噬细胞。应用TUNEL检测不同阻断剂在不同时间点处理后小鼠巨噬细胞凋亡率,细胞免疫化学分析不同阻断剂处理下Mcl-1的表达,HE染色和脏器指数分析不同阻断剂处理对小鼠脏器病变的影响。结果 与对照组比,信号通路阻断剂处理后,BCG感染组巨噬细胞凋亡率均有不同程度的增高,其中PD98059处理组巨噬细胞凋亡率最高(F=40.621, P<0.001)。而且阻断剂PD98059处理后,巨噬细胞内BCG的菌落数量明显减少(t=3.392, P<0.001),小鼠肝、肺、脾、肾的病变程度明显减轻(F=59.24, F=811.134, F=34.091, F=9.543, P<0.05),脏器病理损伤也有所改善。结论 应用阻断剂PD98059阻断Mcl-1表达信号通路可有效控制结核病的潜伏感染和持续感染。

Influences of blocking the signals pathway of Mcl-1 expression on mouse infected with BCG

In the study, we aimed to investigate the impact of inhibiting the signals pathway of Mcl-1 expression on mouse model infected with BCG. Firstly, bacteria suspension of BCG was prepared to infect BALB/c mice, and then different signaling pathways inhibitors treatment was used to establish a mouse model, a corresponding control group at the same time was set up. The mice were executed and the mouse peritoneal macrophages at the treatment of 1 d, 3 d, 5 d, 7 d were collected. The apoptosis of the macrophages treated with BCG at different time points after different inhibitors-treated was analyzed by TUNEL technique, cell immunochemical analyzed the expression of Mcl-1 after different inhibitors-treated, HE staining analyzed the pathological injury of organs in mice after inhibitors-treated, and the degree of pathological changes was analyzed by the viscera index. Result showed that the apoptosis rate of macrophages was increased at different degree in inhibitors-treated BCG infection group compare with the control group, and PD98059 treatment group was highest (F=40.621, P<0.001).Moreover, the number of BCG in macrophages was decreased significantly(t=3.392, P<0.001), the pathological injury of liver, lung, spleen, kidney organs in mice were relieved and organs lesions were decreased significantly (F=59.24, F=811.134, F=34.091, F=9.543, P<0.05). In summary, the application of blocker PD98059 blocking Mcl-1 signaling pathway can effectively control the latent infection and persistent infection of tuberculosis. This study provides more theoretical basis for the introduction of Mcl-1 intervention to prevention and control of tuberculosis infection.