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目的观察百合固金汤对巨噬细胞内自噬功能的调节作用,以探寻其可能的抗结核分枝杆菌作用机制。方法以百合固金汤含药血清干预感染结核分枝杆菌的RAW264.7巨噬细胞,0、6、12、24、36 h后RT-QPCR法检测干预前后巨噬细胞atg5、atg7、atg8、atg12表达变化;12、24、36 h后Western blot法检测干预前后巨噬细胞内自噬相关蛋白LC3-Ⅱ的表达差异;12、24、36 h后QPCR法检测用药前后胞内结核分枝杆菌DNA含量。结果药物干预24 h后,用药组巨噬细胞atg5、atg7、atg8、atg12表达量出现不同程度增加(P<0.05),自噬特异性蛋白LC3-Ⅱ表达量亦显著增高(P<0.05);药物干预36 h后,在自噬特异性蛋白LC3-Ⅱ表达量显著增高(P<0.05)的同时检测到胞内结核分枝杆菌DNA含量明显减少(P<0.05)。结论百合固金汤可能通过激活细胞自噬现象发挥抗结核的作用。
Objective To observe the regulatory effect of Lily Ginkgo Decoction on the autophagy in macrophages in order to explore its possible mechanism of action against Mycobacterium tuberculosis. Methods The serum of Lilium gingjing decoction was used to infect RAW264.7 macrophages of Mycobacterium tuberculosis. The expression of atg5, atg7, atg8, atg12 and c-myc in macrophages were detected by RT-QPCR at 0, 6, 12, The changes of expression of LC3-Ⅱ in macrophages before and after intervention were detected by Western blot at 12, 24 and 36 h. At 12, 24 and 36 h, the intracellular M. tuberculosis DNA content. Results The expression of atg5, atg7, atg8 and atg12 in macrophages increased with different degrees of increase (P <0.05) and the expression of LC3 - Ⅱ (P <0.05). After 36 h of drug intervention, the DNA content of M. tuberculosis was significantly decreased (P <0.05) while the expression of LC3-Ⅱ was significantly increased (P <0.05). Conclusion Lily Gingseng decoction may play an anti-TB role by activating autophagy.