MiR-30c-5p inhibits high glucose-induced EMT and renal fibrogenesis by down-regulation of JAK1 in diabetic nephropathy.

Abstract

Diabetic nephropathy (DN) is one of the most serious complications of diabetes mellitus (DM) and has become the major cause of end-stage renal failure. MicroRNAs (miRs) play key roles in many pathologic processes for initiating and progressing, including DN. Epithelial-mesenchymal transition (EMT) and renal fibrogenesis are important features of DN. However, the role of miR-30c-5p in high glucose (HG)-induced EMT and renal fibrogenesis is not clear. This study was aimed at determining the regulatory network of miR-30c-5p and JAK1 in DN. Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot assays were performed to detect expressions of miR-30c-5p, JAK1, vimentin, α-SMA, and E-cadherin. The possible binding sites between miR-30c-5p and JAK1 were predicted by TargetScan online database and verified by Luciferase report assay. The secretion of fibronectin (FN) andCollagen IV (Col IV) in the supernatant was detected by Enzyme-linked immunosorbent (ELISA) assay. MiR-30c-5p was downregulated and JAK1 was upregulated in renal fibrosis tissue and HG stimulated HK2 cells. Transfection of miR-30c-5p inhibited HG-induced EMT and renal fibrogenesis in HK2 cells, which was reversed by miR-30c-5p inhibitor. Moreover, JAK1 was confirmed as a direct target of miR-30c-5 and knockdown of JAK1 markedly inhibited HG-induced renal fibrogenesis and EMT in HK2 cells. Furthermore, overexpression of JAK1 attenuated the inhibitory effect of miR-30c-5p on HG-induced EMT and renal fibrogenesis in HK2 cells. MiR-30c-5p evidently inhibited HG-induced EMT and renal fibrogenesis by down-regulation JAK1 in DN, providing a promising therapeutic strategy for the treatment of DN.