Microrna-142-3p Inhibits High-Glucose-Induced Endothelial-To-Mesenchymal Transition Through Targeting Tgf-Beta 1/Smad Pathway In Primary Human Aortic Endothelial Cells

Myocardial fibrosis is an important pathological feature of diabetic cardiomyopathy (DCM) and endothelial-to-mesenchymal transition (EndMT) is an essential process for myocardial fibrosis. Recent studies have demonstrated an association between miRs and DCM. Therefore, the aim of this study is to investigate the role and the mechanism of miRNAs in the process of EndMT. We simulated the conditions occurring in EndMT by application of high glucose in primary human aortic endothelial cells (HAECs). Firstly, we compared the expression profiles of miRNAs in HAECs with or without HG treatment using microarray. Then, after addition of miR-142-3p mimics, the expression levels of EndMT markers were assessed by qRT-PCR and Western Blot. Moreover, bioinformatics analysis and luciferase assay were used to confirm the direct regulation of miR-142-3p to TGF-beta 1. Furthermore, the role of TGF-beta 1 in the inhibitory effect of miR-142-3p on EndMT was evaluated. In addition, the expressions of TGF-beta 1/Smad signaling signatures were measured by Western Blot. MiR-142-3p screened by miRNA microarray was significantly down-regulated in HAECs under HG stimulation in a dose and time dependent manner. Subsequently, we found that overexpression of miR-142-3p could inhibit HG-induced EndMT, as evidenced by decreased alpha-SMA and vimentin expression, and increased CD31 and VE-cadherin expression. Of note, transforming growth factor beta 1 (TGF-beta 1), one of the molecular mediators implicated in the progression of EndMT, was confirmed to be downstream target gene of miR-142-3p in HAECs. Moreover, TGF-beta 1 overexpression remarkably abolished the inhibitory effects of miR-142-3p overexpression on HG induced EndMT. Finally, miR-142-3p also mediated its anti-EndMT action by inactivation of TGF-beta 1/Smad pathway, as demonstrated by downregulation of TGF-beta 1, phospho-Smad2 and phospho-Smad2. Our findings demonstrated that miR-142-3p could attenuate HG-induced EndMT in HAECs, the mechanism of which may be at least partly through blocking TGF-beta 1/Smad signaling pathway. This might provide a potential therapeutic target for DCM in future.