摘要
目的探讨肺动脉高压患者外周血细胞中纤溶酶原激活抑制剂-2（PAI-2）mRNA水平及可溶性鸟苷酸环化酶（sGC）激活剂对其表达的影响。方法sGC激活剂西那西呱（Cinaciguat）以100μmol／L浓度处理人肺动脉平滑肌细胞，实时定量荧光PCR和蛋白质印迹法检测PAI-2的mRNA和蛋白表达水平。收集2014年11月至2015年3月中日友好医院的8份动脉性肺动脉高压（PAH）和16份慢性血栓栓塞性肺动脉高压（CTEPH）全血样本以及2015年3月北京医院体检中心的24份健康对照全血样本，Cinaciguat与PAH组和CTEPH组及其各对照组的外周血室温孵育8h，实时定量荧光PCR检测Cinaciguat处理前后PAI-2mRNA表达水平，应用变量相关性分析明确PAI-2mRNA水平是否与肺动脉高压患者的疾病严重程度相关。结果Cinaciguat在人肺动脉平滑肌细胞中能明显上调PAI-2的mRNA和蛋白表达水平。Cinaciguat处理前，PAH组基线PAI-2mRNA表达水平显著低于PAH对照组（0．201±0．152比0．660±0．440，P=0．021），CTEPH组基线PAI-2mRNA表达水平与CTEPH对照组差异无统计学意义（0．428±0．364比0．769±0．682，P=0．152）；Cinaciguat处理后，PAH组、CTEPH组外周血白细胞中PAI-2在mRNA水平上呈增加趋势（1．352±1．127、1．203±1．008），但诱导后PAI-2mRNA水平在PAH组和PAH对照组、CTEPH组和CTEPH对照组之间差异均无统计学意义（P=0．130、0．534）。PAH组基线PAI-2mRNA水平与肺动脉收缩压显著负相关（r=-0．744，P=0．034）。结论肺动脉高压患者外周血细胞中基线PAI-2mRNA水平显著降低，sGC激活剂能上调其mRNA表达水平，PAI-2可作为PAH血管重构潜在的生物标志物。
Objective To investigate the mRNA expression level of plasminogen activator inhibitor-2 （PAI-2） in peripheral blood leucocytes and regulation by soluble guanylate cyclase （sGC） activator in pulmonary hypertension. Methods The human pulmonary arterial smooth muscle cells were treated with sGC activator Cinaeiguat. The mRNA and protein expression levels of PAI-2 were detected with Real-time PCR and Western blot. The fresh blood samples of 8 patients with pulmonary arterial hypertension （PAH） （collected at the China-Japan Friendship Hospital from November 2014 to March 2015 ） , 16 patients with chronic thromboembolic pulmonary hypertension （CTEPH） （collected at the China-Japan Friendship Hospital from November 2014 to March 2015 ） , 24 age- and gendermatched healthy controls （ collected at Beijing Hospital in March 2015 ） were treated with Cinaciguat for 8 hours. Then RNA of peripheral leukocytes was extracted and performed with reverse transcription and Real-time PCR to detect the mRNA level of PAI-2, which was compared between healthy controls and patients with pulmonary hypertension, before and after the treatment of Cinaciguat. At last, the correlation of PAI-2 mRNA level and the clinic severity of pulmonary hypertension were identified. Results The mRNA and protein expression levels of PAI-2 were promoted by Cinaciguat in human pulmonary arterial smooth muscle cells. The baseline mRNA level of PAI-2 in peripheral leukoeytes was significantly lower in PAH patients compared to the healthy controls （0. 201 ±0. 152, 0. 660 ± 0. 440, P = 0. 021 ）. There was no significant difference in the mRNA expression level of PAI-2 between the CTEPH patients and controls （ 0. 428 ±0. 364, 0. 769± 0. 682, P = 0. 152）. After Cinaeiguat treatment, the mRNA expression levels of PAI-2 were up-regulated in PAH patients and CTEPH patients （ 1. 352 ± 1. 127, 1. 203 ± 1. 008 ）, there was no significant difference in the mRNA expression level of PAI-2 among the PAH patients, CTEPH patients an
出处
《中华医学杂志》
CAS
CSCD
北大核心
2016年第16期1247-1251,共5页
National Medical Journal of China
基金
国家自然科学基金（81400036、81571384）
国家科技支撑计划（2011BA111B17）
国家重大新药创制国家科技重大专项（2015ZX09102003）
