Regulation of miR-496 expression by Tripterygium glycosides in response to high glucose-induced ubiquitina-tion pathway in human podocyte
Objective To investigate the effect of Tripterygium glycosides on miR-496 expression and its regulation of the ubiquitination pathway in human podocyte induced by high glucose.Methods cells were divided into the following groups:NC group,HG group,TWP-L group,TWP-M group,TWP-H group,miR-496 group,miR-NC group,and TWP-H+anti-miR-496 group.Cell proliferation was assessed using a CCK-8 assay.Cell apopto-sis was measured by flow cytometry.Protein expression of p53,COP1,caspase-3,nephrin,and podocin in cells was detected using Western blotting.The expression of miR-496 in cells was determined by qRT-PCR.Results Compared with the NC group,the HG group showed significantly decreased cell viability and expression of miR-496(0.23±0.04),COP1(0.43±0.05),nephrin(0.24±0.03),and podocin(0.31±0.04)proteins,as well as increased apoptosis rate(23.94±2.46),and expression of p53(1.26±0.15)and caspase-3(0.61±0.08)proteins(P＜0.05).Compared with the HG group,the TWP-L,TWP-M,and TWP-H groups showed significantly increased cell viabili-ty,expression of miR-496 and COP1,nephrin,and podocin proteins,as well as decreased apoptosis rate and ex-pression of p53 and caspase-3 proteins(P＜0.05).Compared with the miR-NC group,the miR-496 group showed sig-nificantly increased cell viability,expression of miR-496(0.87±0.10)and COP1(0.80±0.09),nephrin(0.58±0.06),and podocin(0.75±0.08)proteins,as well as decreased apoptosis rate(10.61±1.17),and expression of p53(0.32±0.04)and caspase-3(0.25±0.03)proteins(P＜0.05).Compared with the TWP-H group,the TWP-H+ant-miR-496 group showed significantly decreased cell viability,expression of miR-496(0.18±0.02)and COP1(0.46±0.06),nephrin(0.31±0.04),and podocin(0.34±0.04)proteins,as well as increased apoptosis rate(20.36±2.18),and expression of p53(1.19±0.13)and caspase-3(0.57±0.06)proteins(P＜0.05).Conclusion Tripterygi-um glycosides can inhibit the activation of the ubiquitin-proteasome pathway induced by high glucose in human podo-cytes,thereby suppressing podocyte apoptosis.The mechanism of action may be associated with the upregulation of miR-496 expression.
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