MicroRNA-663 prevents monocrotaline-induced pulmonary arterial hypertension by targeting TGF-β1/smad2/3 signaling
MicroRNA-663 prevents monocrotaline-induced pulmonary arterial hypertension by targeting TGF-β1/smad2/3 signaling
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MicroRNA-663 通过靶向 TGF-β1/smad2/3 信号传导预防野百合碱诱导的肺动脉高压
DOI：
10.1016/j.yjmcc.2021.07.010
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发表时间：
2021-08-09
影响因子：
5
通讯作者：
Zhu, Ni
中科院分区：
文献类型：
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作者：
Li, Pan;Song, Jingwen;Zhu, Ni
目的：由于过度生长因子产生和肺动脉平滑肌细胞（PASMC）增殖导致的肺血管重塑是肺动脉高压（PAH）的标志性特征。最近的研究表明 miR-663 是肿瘤发生和动脉粥样硬化的有效调节剂。然而，miR-663是否参与肺血管重塑尚不清楚。方法和结果：通过定量RT-PCR，我们发现miR-663在正常人PASMCs中高表达。相比之下，PAH 患者的 miR-663 循环水平显着降低。此外，原位杂交显示PAH患者肺血管系统中miR-663的表达降低。此外，MTT和细胞划痕实验表明，miR-663模拟物的转染显着抑制血小板衍生生长因子（PDGF）诱导的PASMCs增殖和迁移，而miR-663表达的敲低则增强了这些作用。从机制上讲，双荧光素酶报告基因检测显示 miR-663 直接靶向 TGF-131 的 3'UTR。此外，蛋白质印迹和ELISA结果表明，miR-663减少PDGF诱导的TGF-131表达和分泌，进而抑制下游smad2/3磷酸化和I型胶原蛋白表达。最后，气管内滴注腺-miR-663可有效抑制野百合线（MCT）诱导的PAH大鼠模型中肺血管重塑和右心室肥厚的发展。结论：这些结果表明 miR-663 是 PAH 的潜在生物标志物。 MiR-663 通过靶向 TGF-131/smad2/3 信号传导，减少 PDGF-BB 诱导的 PASMC 增殖，并预防 MCT-PAH 中的肺血管重塑和右心室肥厚。这些发现表明 miR-663 可能是诊断和治疗 PAH 的一种有吸引力的方法。
Objective: Pulmonary vascular remodeling due to excessive growth factor production and pulmonary artery smooth muscle cells (PASMCs) proliferation is the hallmark feature of pulmonary arterial hypertension (PAH). Recent studies suggest that miR-663 is a potent modulator for tumorigenesis and atherosclerosis. However, whether miR-663 involves in pulmonary vascular remodeling is still unclear. Methods and results: By using quantitative RT-PCR, we found that miR-663 was highly expressed in normal human PASMCs. In contrast, circulating level of miR-663 dramatically reduced in PAH patients. In addition, in situ hybridization showed that expression of miR-663 was decreased in pulmonary vasculature of PAH patients. Furthermore, MTT and cell scratch-wound assay showed that transfection of miR-663 mimics significantly inhibited platelet derived growth factor (PDGF)-induced PASMCs proliferation and migration, while knockdown of miR-663 expression enhanced these effects. Mechanistically, dual-luciferase reporter assay revealed that miR-663 directly targets the 3'UTR of TGF-131. Moreover, western blots and ELISA results showed that miR-663 decreased PDGF-induced TGF-131 expression and secretion, which in turn suppressed the downstream smad2/3 phosphorylation and collagen I expression. Finally, intratracheal instillation of adeno-miR-663 efficiently inhibited the development of pulmonary vascular remodeling and right ventricular hypertrophy in monocrota-line (MCT)-induced PAH rat models. Conclusion: These results indicate that miR-663 is a potential biomarker for PAH. MiR-663 decreases PDGF-BB-induced PASMCs proliferation and prevents pulmonary vascular remodeling and right ventricular hypertrophy in MCT-PAH by targeting TGF-131/smad2/3 signaling. These findings suggest that miR-663 may represent as an attractive approach for the diagnosis and treatment for PAH.