Open AccessMiR‐193‐3p attenuates the vascular remodeling in pulmonary arterial hypertension by targeting PAK4
Author(s) -
Wu Zhenhua,
Geng Jie,
Qi Yujuan,
Li Jian,
Bai Yaobang,
Guo Zhigang
Publication year - 2020
Publication title -
pulmonary circulation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.791
H-Index - 40
ISSN - 2045-8940
DOI - 10.1177/2045894020974919
Subject(s) - medicine , pulmonary artery , protein kinase b , pulmonary hypertension , hypoxia (environmental) , in vivo , western blot , proliferating cell nuclear antigen , apoptosis , lung , pathology , biology , chemistry , immunohistochemistry , biochemistry , microbiology and biotechnology , organic chemistry , oxygen , gene
Pulmonary arterial hypertension (PAH) is a progressive pulmonary vascular disease associated with dysfunction of pulmonary artery endothelial cells and pulmonary artery smooth muscle cells (PASMCs). To explore the potential mechanism of miR‐193‐3p in pulmonary arterial hypertension, human PASMCs and rats were respectively stimulated by hypoxia and monocrotaline to establish PAH model in vivo and in vitro. The expressions of miR‐193‐3p and p21‐activated protein kinase 4 (PAK4) in the lung samples of PAH patients and paired healthy samples from the healthy subjects in PHA cells and rats were detected by quantitative reverse transcriptase‐PCR. Morphological changes in lung tissues were determined using hematoxylin and eosin staining. Right ventricular systolic pressure (RVSP) and ratio of right ventricle to left ventricle plus septum (RV/LV p S) were measured. The binding relationship between miR‐193‐3p and PAK4 was analyzed by TargetScan and verified by luciferase reporter assay. Cell viability, apoptosis, and migration were detected by 3‐(4, 5‐Dimethylthiazol‐2‐ yl)‐2,5‐diphenyltetrazolium bromide (MTT) flow cytometry, and wound‐healing assays, respectively. The protein expressions of PAK4, proliferating cell nuclear antigen (PCNA), P21, p‐AKT, and AKT in vivo or in vitro were determined by Western blot. In this study, we found that in pulmonary arterial hypertension, miR‐193‐3p expression was downregulated and PAK4 expression was up‐regulated. MiR‐193‐3p directly targeted PAK4 and negatively regulated its expression. Hypoxia condition promoted cell proliferation, migration, and inhibited apoptosis accompanied with increased expressions of PCNA and p‐AKT/AKT and decreased expression of P21 in PASMCs. MiR‐193‐3p overexpression attenuated the effects of hypoxia on PASMCs via downregulating PAK4. Monocrotaline 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 monocrotaline‐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 downregulating PAK4.