月华丸(胶囊)对耐多药结核菌感染自噬相关蛋白LC3的影响

Effect of Yuehua pills( capsules) on the autophagy-related protein microtubule-associated protein 1 light chain 3 in multidrug-resistant Mycobacterium tuberculosis infection

目的:观察月华丸(胶囊)对结核分枝杆菌感染巨噬细胞自噬的影响,探寻其抗结核分枝杆菌的药效药理作用及其机制。方法:以月华丸(胶囊)含药血清干预感染结核分枝杆菌小鼠的单核巨噬细胞(RAW264. 7),实验共分为5组:阴性对照组、10%月华丸(胶囊)含药血清组、自噬抑制剂3-甲基腺嘌呤(3-MA)+月华丸(胶囊)含药血清组、自噬诱导剂雷帕霉素(Rep)组,另设未被感染的细胞为正常组。采用免疫荧光染色法检测LC3。结果:阴性对照组细胞胞浆中未见LC3蛋白颗粒。与阴性对照组比较,3-MA+月华丸(胶囊)含药血清组细胞胞浆中可见少量LC3蛋白颗粒,10%月华丸(胶囊)含药血清组及Rep组细胞胞浆LC3蛋白颗粒均明显增多,3-MA+月华丸(胶囊)含药血清组多于10%月华丸(胶囊)含药血清组。结论:月华丸(胶囊)含药血清能诱导被耐多药结核菌感染巨噬细胞自噬。

Objective:To investigate the effect of Yuehua pills(capsules)on the autophagy of macrophages in Mycobacterium tuberculosis infection,as well as the pharmacological action of Yuehua pills(capsules)against Mycobacterium tuberculosis and related mechanism.Methods:RAW264.7 mononuclear macrophages from mice with Mycobacterium tuberculosis infection were treated with serum containing Yuehua pills(capsules).The experiment was divided into negative control group,serum containing 10%Yuehua pills(capsules)group,autophagy inhibitor 3-methyladenine(3-MA)+serum containing Yuehua pills(capsules)group,and autophagy inducer rapamycin group,and the macrophages without infection were selected as normal group.Immunofluorescent staining was used to measure the level of microtubule-associated protein 1 light chain 3(LC3).Results:LC3 protein particles were not observed in the cytoplasm in the negative control group.Compared with the negative control group,the 3-MA+serum containing Yuehua pills(capsules)group had a small number of LC3 protein particles in the cytoplasm,and the serum containing 10%Yuehua pills(capsules)group and the rapamycin group had a significant increase in LC3 protein particles in the cytoplasm;the 3-MA+serum containing Yuehua pills(capsules)group had a higher number of LC3 protein particles than the serum containing 10%Yuehua pills(capsules)group.Conclusion:Yuehua pills(capsules)can induce the autophagy of macrophages with Mycobacterium tuberculosis infection.