Mechanism of tRF-013354 mediating adriamycin-induced podocyte injury
Objective:To investigate the role and mechanism of tRNA-derived fragment-013354(tRF-013354)in adriamycin(ADR)-induced podocyte injury.Methods:The podocyte injury model was induced by ADR.Podocytes were respectively transfected with tRF-013354 mimic and tRF-013354 NC-mimic,and the transfection efficiency was verified by real-time PCR(RT-PCR).CCK-8 kit was used to detect the survival rate of podocytes.Immunofluorescence was employed to detect the localization and expression levels of podocyte injury markers(nephrin and podocin).The protein expression levels of nephrin,podocin and Wnt/β-catenin signaling pathway-related proteins(non-phosphorylated β-catenin and Wnt1)were detected by Western blot,and the expression of tRF-013354 was measured by RT-PCR.After blocking the Wnt/β-catenin signaling pathway with the Wnt signaling antagonist Dickkopf-1(DKK1),the protein expression levels of non-phosphorylated β-catenin,nephrin and podocin,and the expression of tRF-013354 were respectively detected with Western blot and RT-PCR.Results:After 24 h intervention with 1 ug/ml ADR,adriamycin-induced injury model was successfully constructed,Wnt/β-catenin signaling pathway was activated,and tRF-013354 was significantly down-regulated(P＜0.05).Overexpression of tRF-013354 in podocytes could reduce the injury of podocytes.When DDK1 was used to block Wnt/β-catenin signaling pathway,ADR+DKK1 group showed less injury of podocytes and up-regulated expression of tRF-013354 than ADR group(P＜0.05).Conclusion:tRF-013354 may act as a downstream effector of Wnt/β-catenin signaling pathway to attenuate ADR-induced podocyte injury.
万方数据
维普
