Objective To study the effect of New Wumei Pills(NWP)on naïve CD4+T cells in patients with ulcerative colitis(UC).Methods The medicinal serum containing NWP was prepared using SD rats,and a differentiation model of naïve CD4+T cells in vitro was established.The optimal concentration of medicinal serum containing NWP for intervening in naïve CD4+T cells was screened using the CCK-8 method.Flow cytometry was used to detect the purity of naïve CD4+T cells and the impact of the medicinal serum on the differentiation model.The expression levels of interleukin(IL)-17 and IL-10 cytokines in the cell culture supernatants were detected by ELISA,the mRNA expression of Glut1 and glycolysis-related enzymes(HK2,PKM2,LDHA)was analyzed by RT-qPCR.Results In this study,naïve CD4+T cells with a purity of 98.2％were successfully extracted,and an in vitro differentiation model of CD4+T cells was established based on these cells.The optimal concentrations of the medicinal serum containing NWP for intervening in naïve CD4+T cells,as screened by the CCK-8 method,were 2.5％,5％,and 10％.Compared with the model group,the differentiation of naïve CD4+T cells towards Th17 cells was significantly decreased,while the differentiation towards Treg cells was significantly increased after intervention with NWP.Mechanistically,compared with the model group,the gene expression levels of Glut1 and glycolysis-related enzymes significantly were decreased after intervention with the medicinal serum containing NWP.Conclusion NWP may regulate the differentiation of naïve CD4+T cells by inhibiting their glycolytic pathway,thereby restoring the Th17/Treg immune balance and achieving the purpose of treating UC.
关键词
溃疡性结肠炎/新乌梅丸/naïve/CD4+T细胞/糖酵解/免疫机制
Key words
ulcerative colitis/New Wumei Pills/naïve CD4+T cells/glycolysis/mechanisms of immunity
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