Mechanism of Panax Notoginseng Saponins Regulates HIF-1α/PDHK1 Pathway-mediated Aerobic Oxidation of CD4+T Cells to Promote Treg Differentiation
OBJECTIVE To observe the regulation of aerobic oxidation mediated by HIF-1α/PDHKl pathway by PNS,and to explore its mechanism of promoting the differentiation of Naive CD4+T cells into Treg cells.METHODS Naive CD4+T cells were isolated from the spleen of C57BL/6 mice by magnetic beads and induced to differentiate into Treg cells for in vitro culture.Naive CD4+T cells were divided into PNS treatment group(5,10,20 μg·mL-1),PNS combined with HIF-1α inhibitor(PX-478)group,and the control group was set up.The proportion of Treg cells differentiation was detected by flow cytometry.The expression of HIF-1α and PDHK1 protein was detected by Western blotting.The expression of HIF-1α,PDHK1 and FOXP3 mRNA was detected by real-time fluorescence quantitative PCR.The level of IL-10 in cell culture supernatant was detected by enzyme-linked immunosorbent assay.RESULTS PNS could significantly increase the proportion of Treg cells and the secretion level of IL-10,and increase the expression of FOXP3 mRNA in cells.At the same time,the expression of HIF-1α and PDHK1 protein and mRNA was inhibited.When the cells were treated with 10 μmol·L-1 PX-478 and then treated with 10 μg·mL-1 PNS,the expression of PDHK1 and FOXP3 and the differentiation ratio of Treg cells were not significantly different from those treated with 10 μmol·L-1 PX-478 alone.CONCLUSION PNS can reduce the expression level of PDHK1 by HIF-1α to enhance the aerobic oxidation of Naive CD4+T cells and promote their differentiation into Treg cells.
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