摘要
为探讨miRNA-467b在实验性自身免疫性脑脊髓膜炎(experimental autoimmune encephalomyelitis,EAE)中对炎性T细胞浸润的影响,我们首先采用real-time PCR及FACS方法检测EAE小鼠脾脏单个核细胞中CXCR3、CCR5、CCR2、CCR6的mRNA和蛋白表达水平,并在MOG反应性T细胞中过表达miRNA-467b,FACS方法检测趋化因子受体CXCR3、CCR6的表达变化,并进一步通过Boyden小室(transwell实验)的方法,体外观察miRNA-467b对细胞迁移的影响。结果发现,EAE小鼠脾脏中CCR5、CCR2、CCR6的转录及蛋白表达水平显著上升,在MOG特异性的T细胞中过表达miRNA-467b后,可降低Th1表面趋化因子受体的表达,减少Th1向transwell板下室中的迁移,提示miRNA-467b可以通过影响细胞的迁移进而调节EAE的疾病进程。
We aimed to investigate the role of miRNA-467b in inflammatory T cell migration in EAE. Firstly, the expressions of CXCR3, CCR5, CCR2 and CCR6 in EAE splenocytes were measured by qRT-PCR and FACS. Secondly, synthetic miRNA- 467b mimics or normal control（NC） were transfected into MOGas-ss-specific T cells by liposome method, and the levels of CX- CR3, CCRS, CCR2 and CCR6 were detected by FACS. Finally, to further explore the effect of miRNA-467b on the migration of MOG3s 5-specific T cell, cell migration assay was detected by Boyden chemotaxis chamber. The results showed that：l. The ex- pressions of CXCR3, CCR5, CCR2 and CCR6 in EAE splenocytes were higher than those in control mice（P〈0.05）. 2. miR- NA-467b overexpression could reduce the levels of chemokine receptors in Thl cells（P〈0.05）. 3. Cell migration assay revealed that miRNA-467b can effectively reduce the migration of MOG3s_ss-specific T cells（P〈0. 05）. In conclusion, miRNA-467b could decrease the levels of chemokine receptors in Thl cells and reduce the migration of inflammatory T cells, which indicates that miRNA-467b may play an important role in EAE disease process.
出处
《现代免疫学》
CSCD
北大核心
2017年第4期294-300,共7页
Current Immunology
基金
国家自然科学基金(81373208)
