Department of Anesthesiology, State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100037, China.
State Key Laboratory of Cardiovascular Disease, FuWai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100037, China.
J Mol Cell Cardiol. 2019 May;130:23-35. doi: 10.1016/j.yjmcc.2019.03.002. Epub 2019 Mar 8.
To analyze the effects of miR-455-3p-1 and its possible mechanisms in pulmonary arterial hypertension (PAH).
A microarray assay was used to examine the expressed genes between normal and PAH. The expressed genes in PAH was assessed by qRT-PCR. The targeted interaction between miRNAs and FGF7 was confirmed using a dual luciferase reporter assay. A CCK-8 assay and cell count were used to analyze the pulmonary artery smooth muscle cells (PASMCs) activity and proliferation level, respectively. Apoptotic PASMCs were detected by flow cytometry. In addition, the mRNA and protein expression levels of RAS/ERK signaling pathway were determined by qRT-PCR and a Western blot assay, respectively. A PAH rat model was used to identify the effects of miR-455-3p-1 in vivo.
FGF7 was upregulated in PAH. MiR-455-3p-1 was downregulated in PAH. MiR-455-3p-1 targeted FGF7. MiR-455-3p-1 decreased the expression of FGF7. Moreover, the effect of FGF7 on PASMCs was suppressed by miR-455-3p-1. MiR-455-3p-1 upregulation was associated with reduced mRNA and protein levels of core RAS/ERK signal genes, suggesting the inhibition of the RAS/ERK pathway. Furthermore, miR-455-3p-1 upregulation improved the RVSP, mPAP, ratio of RV/LV + S, CO and RV function of PAH rat model in vivo.
Our findings illustrate a role for miR-455-3p-1 in modulating FGF7-RAS/ERK signaling and suggest that an agomir of miR-455-3p-1 could inhibit the proliferation of PASMCs and mitigate PAH in vivo.
分析 miR-455-3p-1 在肺动脉高压（PAH）中的作用及其可能机制。
使用微阵列分析检测正常和 PAH 之间表达的基因。通过 qRT-PCR 评估 PAH 中的表达基因。使用双荧光素酶报告基因检测证实 miRNA 和 FGF7 之间的靶向相互作用。CCK-8 测定和细胞计数分别用于分析肺动脉平滑肌细胞（PASMC）的活性和增殖水平。通过流式细胞术检测凋亡 PASMC。此外，通过 qRT-PCR 和 Western blot 测定分别确定 RAS/ERK 信号通路的 mRNA 和蛋白表达水平。使用 PAH 大鼠模型鉴定 miR-455-3p-1 在体内的作用。
FGF7 在 PAH 中上调。miR-455-3p-1 在 PAH 中下调。miR-455-3p-1 靶向 FGF7。miR-455-3p-1 降低了 FGF7 的表达。此外，miR-455-3p-1 下调可抑制 PASMCs 中的 FGF7 作用。miR-455-3p-1 的上调与核心 RAS/ERK 信号基因的 mRNA 和蛋白水平降低有关，提示 RAS/ERK 途径的抑制。此外，miR-455-3p-1 的上调改善了体内 PAH 大鼠模型的 RVSP、mPAP、RV/LV+S 比、CO 和 RV 功能。
我们的研究结果表明，miR-455-3p-1 在调节 FGF7-RAS/ERK 信号方面发挥作用，并表明 miR-455-3p-1 的激动剂可抑制 PASMC 的增殖并减轻体内 PAH。
