结核分枝杆菌BCG和H37Ra对巨噬细胞凋亡的影响及其机制研究  

Effect and Its Mechanism of Mycobacterium tuberculosis BCG and H37Ra on Macrophage Apoptosis

作　　者：何玉龙 张文清[1] 何承文[1] 李雪[1] 郝秀静[1] 李敏[1] 李勇 吴月红[1,2] 

机构地区：[1]宁夏大学西部特色生物资源保护与利用教育部重点实验室,银川750021 [2]浙江理工大学生命科学学院,杭州310018

出　　处：《农业生物技术学报》2015年第5期571-578,共8页Journal of Agricultural Biotechnology

基　　金：宁夏自治区自然科学基金项目资助(No.NZ12105)

摘　　要：在结核分枝杆菌(Mycobacterium tuberculosis,MTB)与宿主巨噬细胞的相互作用中,巨噬细胞的凋亡对抵御及杀灭MTB起着重要的作用。为了探讨MTB的BCG和H37Ra两个弱毒株对巨噬细胞凋亡的影响及其相关机制,本研究比较了体外培养的BCG和H37Ra两种菌株感染小鼠(Mus musculus)巨噬细胞后对细胞凋亡的影响,并检测了肿瘤坏死因子-alpha(tumor necrosis factor-α,TNF-α)、凋亡相关蛋白半胱氨酸蛋白酶-3(Caspase-3)和B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)的表达情况。结果发现,与对照组相比,BCG和H37Ra感染后显著提高了巨噬细胞的凋亡率(P<0.05),TNF-α和Caspase-3的表达及活性也显著增加(P<0.01或P<0.05),同时极显著抑制了Bcl-2的表达(P<0.01)。但BCG感染组细胞凋亡率极显著高于H37Ra组(P<0.01),BCG感染组Caspase-3的表达和活性极显著高于H37Ra组(P<0.01),而Bcl-2和TNF-α的表达在BCG和H37Ra感染组间差异不显著(P>0.05)。结果提示,BCG和H37Ra两个弱毒株感染后对巨噬细胞凋亡的影响不同,可能与Caspase-3的表达相关,而与Bcl-2和TNF-α的表达无关。本研究为深入研究MTB毒力与巨噬细胞凋亡的相互作用机制提供了基础资料。Apoptosis plays an important role in the process of resisting and killing Mycobacterium tuberculosis （MTB） in macrophage. In order to research the effect of 2 attenuated strains BCG and H37Ra on macrophage apoptosis and its mechanisms, the present study compared the effect on apoptosis in BCG and H37Ra infected mouse （Mus musculus） macrophages in vitro, and detected the expression levels of cytokine of tumor necrosis factor- α （TNF- α）, apoptosis- related proteins Caspase- 3 and B- cell lymphoma- 2 （Bcl- 2） respectively. The results showed that in BCG and H37Ra infected macrophages, the ratio of apoptosis significantly increased （P〈0.05）, and the concentration of TNF-α, the expression and activity of Caspase-3 increased significantly （P〈0.01 or P〈0.05）, meanwhile the protein and mRNA expression of Bcl-2 inhibited significantly （P〈 0.01）, comparing with the control group. The ratio of cells apoptosis in BCG infected group was significantly higher than that in H37Ra infected group （P〈0.01）; moreover, the expression and activity of Caspase-3 of the cells in BCG infected group were significantly higher than that in H37Ra group （P〈0.01）. However, there was no significant difference in the expression of Bcl-2 and TNF-oL between in BCG and in H37Ra infected groups （P〉0.05）. These results initially confirmed that the difference of BCG and H37Ra MTB infected macrophage apoptosis was related to Caspase-3, not to the expression of Bcl-2 and TNF-α. The present study results may provide basic data for further research on the interaction between MTB and macrophages.

关 键 词：巨噬细胞 凋亡 半胱氨酸蛋白酶-3 B淋巴细胞瘤-2(Bcl-2) 肿瘤坏死因子-alpha(TNF-α) 

分 类 号：S858.23[农业科学—临床兽医学]