Cysteine-rich 61 (Cyr61) upregulated in pulmonary arterial hypertension promotes the proliferation of pulmonary artery smooth muscle cells
Cysteine-rich 61 (Cyr61) upregulated in pulmonary arterial hypertension promotes the proliferation of pulmonary artery smooth muscle cells
复制标题
肺动脉高压中富含半胱氨酸 61 (Cyr61) 上调促进肺动脉平滑肌细胞增殖
DOI：
10.7150/ijms.19282
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发表时间：
2017-01-01
影响因子：
3.6
通讯作者：
Zhang, Zhuoli
中科院分区：
文献类型：
--
作者：
Gao, Lan;Fan, Yong;Zhang, Zhuoli
背景资料：本研究旨在探讨富含半胱氨酸61（Cyr 61）在肺动脉高压（PAH）患者及野百合碱（MCT）诱导的PAH大鼠肺动脉平滑肌细胞（PASMCs）中的表达，并进一步探讨Cyr 61对PASMCs增殖的影响及其可能机制。方法和结果：从20例特发性PAH患者、20例结缔组织病（CTD）相关PAH患者、29例年龄、性别和疾病匹配的CTD无PAH患者和28例健康对照者中采集血浆样本。ELISA法检测血浆Cyr 61水平。单次皮下注射MCT（60 mg·kg-1）建立MCT诱导的PAH（MCT-PAH）大鼠模型。取大鼠肺组织和肺动脉，分离培养PASMCs进行体外实验。采用免疫组化、western blot和实时荧光定量聚合酶链反应检测肺组织、肺动脉和PASMCs中Cyr 61的表达。外源性重组Cyr 61蛋白刺激PAH大鼠的PASMC，并用小干扰RNA敲低。采用Cell Counting Kit-8法检测细胞增殖情况，Western blot法检测p-AKT和AKT的表达。结果表明，PAH患者血浆Cyr 61水平显著高于CTD非PAH患者和健康对照组，且以CTD合并PAH患者为著。与野生型大鼠相比，PAH大鼠肺组织、肺动脉和PASMCs中Cyr 61的表达明显增强。外源性重组Cyr 61蛋白可促进PASMCs增殖，并呈剂量依赖性。特异性siRNA抑制PASMCs中Cyr 61的表达后，细胞增殖受到抑制，p-AKT表达下降。结论：PAH患者血浆Cyr 61浓度明显升高。Cyr 61可通过AKT途径促进PASMCs增殖，提示Cyr 61可能参与PAH的发病过程。
Background: We aimed to evaluate the expression of cysteine rich 61 (Cyr61) in patients with pulmonary arterial hypertension (PAH) as well as monocrotaline (MCT) induced PAH rat, and further investigate the effects and potential mechanisms of Cyr61 on the proliferation of pulmonary arterial smooth muscle cells (PASMCs). Methods and Results: Plasma samples were collected from 20 patients with idiopathic PAH, 20 connective tissue disease (CTD) associated PAH, 29 age-, gender- and disease matched CTD without PAH patients, and 28 healthy controls. ELISA was used to detect the level of Cyr61 in plasma. MCT-induced PAH (MCT-PAH) rat model was established by a single subcutaneous injection of MCT (60mg·kg-1). Lung tissues and pulmonary arteries of rats were collected, while the PASMCs were dissected and cultivated for in vitro experiments. Expression of Cyr61 in the lung tissues, pulmonary arteries and PASMCs were tested by immunohistochemical staining, western blot and quantitative real-time polymerase chain reaction. PASMCs from PAH rats were stimulated by exogenous recombinant Cyr61 protein and knocked down by small interfering RNA. Cell Counting Kit-8 assay was used to identify cell proliferation and the expression of p-AKT and AKT were analysed by western blot. The results showed plasma level of Cyr61 in PAH patients, especially CTD-PAH patients, were significant higher than that of CTD without PAH patients and healthy controls. Compared with wild rats, Cyr61 was overexpressed in the lung tissue, pulmonary arterial and PASMCs in PAH rats. Exogenous recombinant Cyr61 protein promoted the proliferation of PASMCs in a dose-dependent manner. While the expression of Cyr61 in PASMCs was inhibited by specific siRNA, cell proliferation was restrained and the expression of p-AKT declined. Conclusion: Plasma Cyr61 concentration in PAH patients was highly increased. Cyr61 could promote PASMCs proliferation via AKT pathway, indicating that Cyr61 may play a role in the pathogenesis of PAH.