NK1R对小胶质细胞迁移能力影响的实验研究

Effects of NK1R on the migration of BV2 microglial cells

目的探讨神经激肽1受体(NK1R)对小胶质细胞迁移能力的影响:方法以NK1R激动剂P物质(SP)和抑制剂CP-99994处理BV2细胞,采用MTT法检测BV2细胞的增殖能力,采用划痕实验、Transwell迁移实验检测BV2细胞的迁移能力,蛋白免疫印迹法检测核内NF-κB p65亚基的表达水平。以NF-κB特异性抑制剂预处理BV2细胞,检测SP对BV2细胞迁移能力的影响。结果与对照组比较,SP组BV2细胞的增殖能力、迁移能力和核内p65亚基显著上调,差异有统计学意义(P<0.0l),然而这种增强效应能被NK1R抑制剂CP-99994所消除。此外,CP-99994显著削弱了SP引起核内p65亚基的升高,并且抑制SP诱导的BV2细胞迁移。结论NK1R的活化能促进BV2细胞的迁移,其机制可能与NF-κB信号通路的活化有关。

Objective To investigate the effects of neurokinin 1 receptor (NK1R) on the migration of BV2 microglia cells. Methods BV2 cells were treated with NK1R agonist substance P (SP) and inhibitor CP-99994. Then the migration of BV2 cells was detected by the scratch test and Transwell assay. The expression of NF-κB p65 subunit was measured by Western blot. BV2 cells were also pretreated with NF-κB antagonist, PDTC, and its effects on BV2 cell migration were determined. Results Compared with the control, SP treatment significantly enhanced the proliferation and migration abilities of BV2 cells, and dramatically increased the expression of NF-κB p65 in the nucleus P<0.01. However, such increases were reversed by NK1R inhibitor CP-99994. Furthermore, CP-99994 remarkably inhibited the increased expression of p65 in the nucleus caused by SP treatment, and suppressed the migration of BV2 cells induced by SP. Conclusions Activated NK1R can stimulate the migration of BV2 cells which may be associated with the activation of NF-κB signaling pathway.