Objective:To observe the role of microRNA-92a(miRNA-92a or miR-92a)in the differentiation of CD4+T cells in the central nervous system of mice with experimental autoimmune encephalomyelitis(EAE);to study the process of miR-92a regulating cell differentiation through the glycolytic pathway,and to investigate the molecular mechanism by which miR-92a affects the pathological process of EAE targeting mTOR as a downstream key molecule.Methods:After EAE models were successfully constructed on C57BL/6J or miR-92a-/-mice,spinal cord CD4+T cells were isolated and cultured in vitro,the proportions of Th1,Th2,Th17 and Treg cells were measured by flow cytometry,the level of glycolysis was measured using Seahorse,and the level of related gene changes was measured using RT-qPCR;naive CD4+T cells cultured in vitro were induced to differentiate into Th1 or Treg cells,on the basis of which miR-92a levels were regulated and combined with glycolytic agonists or inhibitors to detect cell differentiation;mTOR and p-Akt expression changes in CD4+T cells of C57BL/6J or miR-92a-/-mice were detected using Western Blot,and glycolysis levels in CD4+T cell and cell differentiation were measured after overexpression of mTOR with plasmid transfection.Results:miR-92a could lead to the destruction of CD4+T cell differentiation balance,the ratio of Th1 and Th17 cells was increased,and that of Th2 and Treg cells decreased after EAE;miR-92a regulated CD4+T cell differentiation by promoting glycolysis;Treg cell differentiation was promoted after inhibiting glycolysis;miR-92a could increase the level of glycolysis by activating the mTOR signaling pathway,thus affecting cell differentiation.Conclusion:miR-92a promotes T cell glycolysis and disrupts the balance of CD4+T cell differentiation by activating mTOR signaling pathway,which is involved in the pathological process of MS and EAE.
关键词
多发性硬化/哺乳动物雷帕霉素靶蛋白/神经炎症/糖酵解/miRNA-92a/T细胞/小鼠
Key words
multiple sclerosis(MS)/mammalian target of rapamycin(mTOR)/neuroinflammation/glycolysis/miRNA-92a(miR-92a)/T cell/mouse
维普
万方数据