MiR‐193‐3p attenuates the vascular remodeling in pulmonary arterial hypertension by targeting PAK4

Abstract

Pulmonary arterial hypertension (PAH) is a progressive pulmonary vascular disease associated with dysfunction of pulmonary artery endothelial cells (PAEC) and pulmonary artery smooth muscle cells (PASMC). To explore the potential mechanism of miR-193-3p in PAH, human PASMCs and rats were respectively stimulated by hypoxia and monocrotaline (MCT) to establish PAH model in vivo and in vitro. The expressions of miR-193-3p and PAK4 in the lung samples of PAH patients and paired healthy samples from the healthy subjects, in PHA cells and rats were detected by qRT-PCR. The pathological features of lung tissues were determined using HE staining. The binding relationship between miR-193-3p and PAK4 was analyzed by TargetScan and verified by luciferase reporter assay. Cell viability, apoptosis, migration were detected by MTT, flow cytometry and wound healing assays, respectively. The protein expressions of PAK4, PCNA, P21, p-AKT and AKT in vivo or in vitro were determined by Western blot. In this study, we found that in PAH, miR-193-3p expression was downregulated and PAK4 expression was up-regulated. MiR-193-3p directly targeted PAK4 and negatively regulated its expression. Hypoxia induction promoted cell proliferation, migration and apoptosis resistance accompanied with increased the expressions of PCNA and p-AKT/AKT and decreased that of P21 in PASMCs. MCT treatment increased p-AKT/AKT and decreased P21 expression, and caused pulmonary vascular remodeling in the model rats. MiR-193-3p overexpression attenuated pulmonary vascular remodeling, decreased p-AKT/AKT and increased P21 levels via downregulating PAK4 in MCT-induced rats. The results in this study demonstrated that upregulation of miR-193-3p reduced cell proliferation, migration, and apoptosis of PAH in vitro and pulmonary vascular remodeling in PAH in vivo through downregualting PAK4.

Astragaloside IV restrains pyroptosis and fibrotic development of pulmonary artery smooth muscle cells to ameliorate pulmonary artery hypertension through the PHD2/HIF1α signaling pathway

“…Finally, the sections were observed under a microscope CKX53 (Olympus, Tokyo, Japan). Pulmonary vessel area and wall thickness were measured as previous described [ 36 ]: Pulmonary arterioles around 100 μm were selected for measurement, and 10 cross-sectional pulmonary arteries were randomly selected from each slice. Wall thickness (%) = (external diameter - internal diameter)/ external diameter×100%, and vessel area (%) = (total area - internal area) / total area×100%.…”