TLR2和TLR6在鸟分枝杆菌诱导巨噬细胞凋亡中的作用

发布时间：2018-11-20 06:21

【摘要】：【目的】建立鸟分枝杆菌感染巨噬细胞模型,探讨Toll样受体2(TLR2)和Toll样受体6(TLR6)在鸟分枝杆菌诱导巨噬细胞凋亡中的作用,为阐明鸟分枝杆菌致病机制提供依据。【方法】用鸟分枝杆菌感染U937细胞,于感染0h、8h、16h、24h和48h后分别提取细胞总蛋白,并通过Western blot方法检测TLR2和TLR6的表达;分别提取细胞培养上清液,利用ELISA技术检测肿瘤坏死因子(TNF-α)含量的变化情况。用鸟分枝杆菌感染阻断TLR2和TLR6后的U937细胞,通过Western blot方法分别检测其促凋亡因子BAX和抗凋亡因子BCL-2的表达量,用ELISA技术检测细胞培养上清液中TNF-α含量变化情况,用流式细胞仪检测U937细胞的凋亡变化。【结果】鸟分枝杆菌感染U937细胞可引起TLR2、TLR6和TNF-α表达上调;用鸟分枝杆菌感染阻断TLR2和TLR6后的U937细胞,可使U937细胞内促凋亡因子BAX和TNF-α表达量下降(P0.05),抗凋亡因子BCL-2表达量升高(P0.05),并可以明显降低感染鸟分枝杆菌后的细胞的凋亡率(P0.05)。【结论】TLR2和TLR6与巨噬细胞凋亡相关,巨噬细胞通过TLR2和TLR6来识别鸟分枝杆菌,而鸟分枝杆菌反过来通过促进TLR2和TLR6的表达来影响巨噬细胞相关凋亡通路,从而诱导巨噬细胞凋亡。
[Abstract]:[objective] to study the role of Toll like receptor 2 (TLR2) and Toll like receptor 6 (TLR6) in inducing macrophage apoptosis induced by Mycobacterium avium. [methods] U937 cells were infected with Mycobacterium avium. The total protein was extracted from U937 cells at 0 h, 8 h, 16 h and 48 h after infection, and the expression of TLR2 and TLR6 was detected by Western blot method. The supernatant of cell culture was extracted, and the content of tumor necrosis factor (TNF- 伪) was detected by ELISA technique. U937 cells of TLR2 and TLR6 were blocked by Mycobacterium avium infection. The expression of BAX and BCL-2 were detected by Western blot, and the content of TNF- 伪 in the supernatant of cell culture was detected by ELISA technique. The apoptosis of U937 cells was detected by flow cytometry. [results] the expression of TLR2,TLR6 and TNF- 伪 was up-regulated in U937 cells infected by Mycobacterium avium. When U937 cells were blocked by Mycobacterium avium infection, the expression of pro-apoptotic factor BAX and TNF- 伪 in U937 cells decreased (P0.05), and the expression of anti-apoptotic factor BCL-2 increased (P0.05). TLR2 and TLR6 were associated with macrophage apoptosis. Macrophages recognized Mycobacterium avium by TLR2 and TLR6, and the apoptosis rate of mycobacterium avium was significantly decreased after infection (P0.05). [conclusion] TLR2 and TLR6 are associated with macrophage apoptosis, and macrophages recognize Mycobacterium avium by TLR2 and TLR6. On the other hand, Mycobacterium avium can induce macrophage apoptosis by promoting the expression of TLR2 and TLR6.
【作者单位】：广西医科大学广西高校生物分子医学研究重点实验室生物化学与分子生物学教研室
【基金】：国家自然科学基金项目(81260245) 广西自然科学基金项目(2015GXNSFAA139077)资助
【分类号】：R378.91

【相似文献】