MiR-193-3p attenuates the vascular remodeling in pulmonary arterial hypertension by targeting PAK4
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...
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Series: | Pulmonary Circulation |
Online Access: | https://doi.org/10.1177/2045894020974919 |
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doaj-b4fc5931cb2f41fa857154a6891203772020-12-12T02:03:21ZengSAGE PublishingPulmonary Circulation2045-89402020-12-011010.1177/2045894020974919MiR-193-3p attenuates the vascular remodeling in pulmonary arterial hypertension by targeting PAK4Zhenhua WuJie GengYujuan QiJian LiYaobang BaiZhigang GuoPulmonary 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.https://doi.org/10.1177/2045894020974919 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Zhenhua Wu Jie Geng Yujuan Qi Jian Li Yaobang Bai Zhigang Guo |
spellingShingle |
Zhenhua Wu Jie Geng Yujuan Qi Jian Li Yaobang Bai Zhigang Guo MiR-193-3p attenuates the vascular remodeling in pulmonary arterial hypertension by targeting PAK4 Pulmonary Circulation |
author_facet |
Zhenhua Wu Jie Geng Yujuan Qi Jian Li Yaobang Bai Zhigang Guo |
author_sort |
Zhenhua Wu |
title |
MiR-193-3p attenuates the vascular remodeling in pulmonary arterial hypertension by targeting PAK4 |
title_short |
MiR-193-3p attenuates the vascular remodeling in pulmonary arterial hypertension by targeting PAK4 |
title_full |
MiR-193-3p attenuates the vascular remodeling in pulmonary arterial hypertension by targeting PAK4 |
title_fullStr |
MiR-193-3p attenuates the vascular remodeling in pulmonary arterial hypertension by targeting PAK4 |
title_full_unstemmed |
MiR-193-3p attenuates the vascular remodeling in pulmonary arterial hypertension by targeting PAK4 |
title_sort |
mir-193-3p attenuates the vascular remodeling in pulmonary arterial hypertension by targeting pak4 |
publisher |
SAGE Publishing |
series |
Pulmonary Circulation |
issn |
2045-8940 |
publishDate |
2020-12-01 |
description |
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. |
url |
https://doi.org/10.1177/2045894020974919 |
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