Inhibition of miRNA-135a-5p ameliorates TGF-β1-induced human renal fibrosis by targeting SIRT1 in diabetic nephropathy

miRNA-135a-5p upregulation has been identified in renal fibrosis in diabetic nephropathy (DN) with an incompletely known mechanism. Previous data showed that Sirtuin 1 (SIRT1) serves as a novel therapeutic target for DN and interact with the transforming growth factor-β/mothers against decapentaplegic homolog (TGF-β/Smad) signaling pathway. The aim of this study was to investigate the regulatory relationship between miR-135a-5p and SIRT1. The expression of miR-135a-5p and SIRT1 was detected using reverse transcription-quantitative PCR and western blotting. The renal fibrosis and Smad3 signaling pathway were assessed by western blotting, by analyzing protein expression of collegen1A1, α-smooth muscle actin (α-SMA), fibronectin (FN), epithelial-cadherin, Smad3 and phosphorylated Smad3 (p-Smad3). The target binding between miR-135a-5p and SIRT1 was predicted on TargetScan Human software, and confirmed by dual-luciferase reporter assay and RNA immu-noprecipitation. The results demonstrated miR-135a-5p is upregulated and SIRT1 was downregulated in the serum and renal tissue of DN patients, and TGFβ1-induced DN cell models in human HK-2 and HMCs. Knockdown of miR-135a-5p and overexpression of SIRT1 could inhibit TGFβ1-induced renal fibrosis in vitro. Moreover, SIRT1 was a downstream target for miR-135a-5p. Silencing of SIRT1 could abolish the suppressive role of miR-135a-5p knockdown in TGFβ1-induced HK-2 and HMCs. The TGFβ1 induced p-Smad3 expression in HK-2 and HMCs, which could be attenuated by miR-135a-5p knockdown via SIRT1. In conclusion, knockdown of miR-135a-5p inhibits TGFβ1-induced renal fibrosis by targeting SIRT1 and inactivating Smad3 signaling, providing a novel insight into miR-135a-5p as a potential therapeutic approach for DN.