An Zhe, Wang Dan, Yang Guang, Zhang Wen-Qi, Ren Jin, Fu Jin-Ling
Department of Cardiology, China-Japan Union Hospital of Jilin University Department of Breast Surgery, The Second Hospital of Jilin University Department of Molecular Biology, College of Basic Medical Sciences, Jilin University Department of Respiratory Medicine, the Second Hospital of Jilin University Department of Ophthalmology, The Second Hospital of Jilin University, Changchun, Jilin, P.R. China.
Medicine (Baltimore). 2017 May;96(20):e6746. doi: 10.1097/MD.0000000000006746.
The purpose of this study was to elucidate the role of microRNA-130a (miR-130a) in obstructive sleep apnea hypopnea syndrome (OSAHS)-associated pulmonary hypertension (PHT) by targeting the growth arrest-specific homeobox (GAX) gene.
A total of 108 patients with OSAHS-associated PHT were recruited as the OSAHS-associated PHT group and 110 healthy individuals were randomly selected as the normal control group. Human umbilical vein endothelial cells (HUVECs) were selected and divided into the control, miR-130a mimic, mimic negative control (NC), miR-130a inhibitor, and inhibitor-NC groups. The dual luciferase reporter gene assay was used to identify the relationship between miR-130a and the GAX gene. The quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting were applied for the relative expressions of miR-130a and the mRNA and protein expressions of GAX. Serum levels of endothelin-1 (ET-1), vascular endothelial growth factor (VEGF), nitric oxide (NO), and super oxide dismutase (SOD) were detected. Cell apoptosis and angiogenic activity were analyzed by flow cytometry and cell tube formation assay.
GAX was a target gene of miR-130a. Compared with the normal control group, the relative expression of miR-130a and the serum levels of ET-1 and VEGF were increased, whereas the mRNA expression of GAX and the serum levels of NO and SOD were decreased in the OSAHS-associated PHT group. Compared with the control, mimic-NC, and inhibitor-NC groups, the relative expressions of miR-130a in the miR-130a mimic group were enhanced, whereas the expression of miR-130a in the miR-130a inhibitor group was reduced. However, the mRNA and protein expressions of GAX showed an opposite trend in the miR-130a mimic and miR-130a inhibitor groups. In comparison to the control, mimic-NC, and inhibitor-NC groups, the miR-130a mimic group had an increase of ET-1 and VEGF expressions, whereas the expressions of NO and SOD were reduced. However, the miR-130a inhibitor group exhibited an opposite trend. The apoptosis rate and tube formation number in the miR-130a mimic group were obviously increased, whereas the miR-130a inhibitor group showed an obvious decrease.
These data provided strong evidence that miR-130a may be involved in the progression of OSAHS-associated PHT by down-regulating GAX gene.
本研究旨在通过靶向生长停滞特异性同源框（GAX）基因，阐明微小RNA - 130a（miR - 130a）在阻塞性睡眠呼吸暂停低通气综合征（OSAHS）相关肺动脉高压（PHT）中的作用。
共招募108例OSAHS相关PHT患者作为OSAHS相关PHT组，并随机选取110名健康个体作为正常对照组。选取人脐静脉内皮细胞（HUVECs），分为对照组、miR - 130a模拟物组、模拟物阴性对照组（NC）、miR - 130a抑制剂组和抑制剂阴性对照组。采用双荧光素酶报告基因检测法鉴定miR - 130a与GAX基因之间的关系。应用定量实时聚合酶链反应（qRT - PCR）和蛋白质印迹法检测miR - 130a的相对表达以及GAX的mRNA和蛋白质表达。检测血清内皮素 - 1（ET - 1）、血管内皮生长因子（VEGF）、一氧化氮（NO）和超氧化物歧化酶（SOD）水平。通过流式细胞术和细胞管形成试验分析细胞凋亡和血管生成活性。
GAX是miR - 130a的靶基因。与正常对照组相比，OSAHS相关PHT组中miR - 130a的相对表达以及ET - 1和VEGF的血清水平升高，而GAX的mRNA表达以及NO和SOD的血清水平降低。与对照组、模拟物阴性对照组和抑制剂阴性对照组相比，miR - 130a模拟物组中miR - 130a的相对表达增强，而miR - 130a抑制剂组中miR - 130a的表达降低。然而，miR - 130a模拟物组和miR - 130a抑制剂组中GAX的mRNA和蛋白质表达呈现相反趋势。与对照组、模拟物阴性对照组和抑制剂阴性对照组相比，miR - 130a模拟物组中ET - 1和VEGF的表达增加，而NO和SOD的表达降低。然而，miR - 130a抑制剂组呈现相反趋势。miR - 130a模拟物组中的凋亡率和管形成数量明显增加，而miR - 130a抑制剂组则明显降低。
这些数据提供了强有力的证据，表明miR - 130a可能通过下调GAX基因参与OSAHS相关PHT的进展。
