敲低ASAP1降低结核分枝杆菌感染小鼠RAW264.7细胞的迁移能力

ASAP1 knockdown reduces migration of RAW264.7 cells infected with Mycobacterium tuberculosis

目的观察含SH3结构域-ADP-核糖基化因子(Arf)GTP酶激活蛋白-锚蛋白重复序列和PH结构域1(ASAP1)介导小鼠RAW264.7巨噬细胞对结核分枝杆菌感染过程的影响以及ASAP1对巨噬细胞迁移能力的影响。方法建立ASAP1敲低的RAW264.7细胞系,感染结核分枝杆菌H37Ra后,利用荧光共聚焦显微镜和平板计数方法对胞内细菌的载量进行检测,采用TranswellTM法检测ASAP1敲低细胞的迁移能力。结果与对照组细胞相比,ASAP1敲低的RAW264.7细胞感染H37Ra后胞内细菌载量增多,且细胞迁移能力降低。结论敲低ASAP1降低RAW264.7细胞的迁移能力。

Objective To investigate the role of ArfGAP with SH3 domain, ankyrin repeat and PH domain 1(ASAP1) in the infection of murine macrophages RAW264.7 with Mycobacterium tuberculosis, and the function of ASAP1 in macrophage migration. Methods The expression of ASAP1 was down-regulated in RAW264.7 cells using siRNA, and then the cells were infected with H37Ra. The intracellular bacterial load was determined by fluorescence confocal microscopy and colony forming unit(CFU), and the migration ability of the cells were evaluated by TranswellTM assay. Results Compared with the control cells, the intracellular bacterial load was significantly enhanced in ASAP1-knockdown cells, whereas the migration ability of cells was reduced after knockdown of ASAP1 expression. Conclusion ASAP1 is important for RAW264.7 cell migration, and down-regulated ASAP1 expression may impair the migration ability of RAW264.7 cells.