结核分枝杆菌MPT64抗原对巨噬细胞凋亡的抑制作用

发布时间：2018-06-08 04:16

本文选题：结核分枝杆菌 + MPT抗原　；参考：《第二军医大学学报》2013年05期

【摘要】：目的探讨结核分枝杆菌MPT64抗原对RAW264.7巨噬细胞的影响及相关机制。方法将MPT64基因在大肠杆菌中表达、纯化,获得的纯化蛋白用于后续实验。将用佛波醇酯(PMA)分化的RAW264.7巨噬细胞分为对照组、卡介菌纯蛋白衍生物(BCGPPD)诱导组、BCG-PPD+MPT64共同作用组,分别给予相应的处理。孵育16 h后,采用流式细胞术检测细胞凋亡情况,用ELISA试剂盒检测培养细胞上清的细胞因子TNF-α及IL-10水平。结果 BCG-PPD可以诱导RAW264.7巨噬细胞凋亡,BCG-PPD+MPT64组的细胞凋亡水平低于BCG-PPD组(P0.05)。细胞因子上清检测结果表明,与对照组相比,BCG-PPD作用组的TNF-α水平升高(P0.01),而IL-10水平无明显变化;与BCG-PPD组相比,BCG-PPD+MPT64共同孵育组的IL-10水平升高(P0.01),而TNF-α水平无明显变化。结论 MPT64抗原可能是一种毒力因子,能够抑制BCG-PPD诱导的巨噬细胞凋亡,该作用可能是通过提高IL-10水平来实现的。
[Abstract]:Objective to investigate the effect of Mycobacterium tuberculosis MPT64 antigen on RAW264.7 macrophages and its related mechanism. Methods MPT64 gene was expressed in Escherichia coli and purified. RAW264.7 macrophages differentiated by phorbol alcohol ester (PMA) were divided into two groups: control group, BCGPPD-induced group and BCG-PPD MPT64 group. After incubation for 16 h, apoptosis was detected by flow cytometry and the levels of cytokines TNF- 伪 and IL-10 in supernatant of cultured cells were detected by Elisa kit. Results BCG-PPD could induce apoptosis of RAW264.7 macrophages. The level of apoptosis in BCG-PPD MPT64 group was lower than that in BCG-PPD group. The results of cytokine supernatant test showed that the level of TNF- 伪 in BCG-PPD treated group was higher than that in control group, but the level of IL-10 was not changed, but the level of IL-10 in BCG-PPD MPT64 co-incubated group was higher than that in BCG-PPD MPT64 group, but the level of TNF- 伪 had no obvious change. Conclusion MPT64 antigen may be a virulence factor, which can inhibit the apoptosis of macrophages induced by BCG-PPD, which may be achieved by increasing the level of IL-10.
【作者单位】：海军医学研究所航空医学研究室;成都军区昆明总医院传染结核科;
【基金】：国家自然科学基金(31070121) 上海市自然科学基金(08ZR1405600)~~
【分类号】：R378

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