Department of Pulmonary and Critical Care Medicine, First Affiliated Hospital of Soochow University, Suzhou 215006, China.
Department of Pulmonary and Critical Care Medicine, First Affiliated Hospital of Soochow University, Suzhou 215006, China.
Microvasc Res. 2020 Jul;130:103988. doi: 10.1016/j.mvr.2020.103988. Epub 2020 Feb 10.
Resveratrol has shown benefit for pulmonary hypertension improvement. Our previous reports showed NR4A3/cyclin D1 pathway promoted pulmonary arterial smooth muscle cells (PASMCs) proliferation. This study tried to explore the mechanism underlying this process, focusing on the role of resveratrol in regulation of miRNA and NR4A3.
Rats were injected with monocrotaline (MCT) to establish pulmonary hypertension (PH) models. Resveratrol was used to prevent pulmonary vascular remodeling. Primary rat PASMCs were cultured in vitro and stimulated by platelet-derived growth factor (PDGF) with or without resveratrol. Cells proliferation and expression of miR-638 as well as NR4A3 were evaluated.
MCT resulted in significant pulmonary vascular remodeling and down-regulation of miR-638, which could be suppressed by resveratrol. Moreover, PDGF-induced PASMC proliferation and miR-638 down-regulation were both significantly prevented by resveratrol treatment in vitro. MiR-638 mimics markedly inhibited PASMC proliferation and percentage of PCNA-positive cells in vitro. But anti-miR-638 could markedly promote cells proliferation and percentage of PCNA-positive cells. The luciferase reporter assay showed that NR4A3 was a direct target of miR-638. The loss-of-function and gain-of-function experiments indicated that NR4A3 promoted proliferation via cyclin D1 pathway.
Our data indicated that resveratrol prevented MCT-induced pulmonary vascular remodeling via miR-638 regulating NR4A3/cyclin D1 pathway.
白藜芦醇已显示出对肺动脉高压改善的益处。我们之前的报告表明，NR4A3/细胞周期蛋白 D1 通路促进肺动脉平滑肌细胞（PASMC）增殖。本研究试图探讨这一过程的机制，重点研究白藜芦醇在调节 miRNA 和 NR4A3 中的作用。
用野百合碱（MCT）注射大鼠建立肺动脉高压（PH）模型。用白藜芦醇预防肺血管重构。原代大鼠 PASMC 体外培养，用血小板衍生生长因子（PDGF）刺激，或用白藜芦醇预处理。评估细胞增殖和 miR-638 以及 NR4A3 的表达。
MCT 导致明显的肺血管重构和 miR-638 下调，白藜芦醇可抑制其发生。此外，白藜芦醇在体外还可显著抑制 PDGF 诱导的 PASMC 增殖和 miR-638 下调。miR-638 模拟物显著抑制 PASMC 增殖和 PCNA 阳性细胞的百分比。但抗 miR-638 可显著促进细胞增殖和 PCNA 阳性细胞的百分比。荧光素酶报告基因检测表明，NR4A3 是 miR-638 的直接靶标。功能丧失和功能获得实验表明，NR4A3 通过细胞周期蛋白 D1 通路促进增殖。
我们的数据表明，白藜芦醇通过 miR-638 调节 NR4A3/细胞周期蛋白 D1 通路预防 MCT 诱导的肺血管重构。
