先天性心脏病继发性肺动脉高压患者血浆中miRNA的研究

目的 应用微小RNA （microRNA,miRNA）芯片技术研究先天性心脏病合并重度肺动脉高压（pulmonary arterial hypertension,PAH）患者和不合并PAH患者血浆中miRNA表达谱的差异,并初步预测差异表达的miRNA调控的靶基因.方法 收集室间隔缺损（ventricular septal defect,VSD）合并重度PAH（PAH组）和不合并PAH患者（对照组）的血浆.分别提取总RNA,然后采用miRNA芯片进行miRNA表达谱差异分析,并对结果进行实时定量聚合酶链反应（polymerase chain reaction,PCR）验证.运用Targetscan、Pictar、Miranda软件预测可能调控的靶基因.结果 miRNA芯片结果提示：与对照组相比,左向右分流先天性心脏病继发重度PAH患者血浆中表达上调的miRNA有50个,表达下调的miRNA有36个.实时定量PCR验证miR-98与芯片结果一致,表达上调.靶基因预测软件显示内皮素（ET-1）为hsa-miR-98的重要靶基因.结论 miRNA在先天性心脏病继发重度PAH患者血浆中存在差异性表达,miRNA可能与PAH的发生、发展密切相关,血浆miR-98有可能成为先天性心脏病合并重度PAH的新的分子生物学标志物.

Study of plasma microRNA in patients with pulmonary arterial hypertension associated with congenital heart disease

Objectives To investigate the expression pattern of plasma microRNA （miRNA） in patients with severe pulmonary arterial hypertension （PAH） associated with congenital heart disease （CHD） by miRNA array,and to predict the target genes regulating miRNA differential expressions.Methods Plasmas were collected from patients with severe PAH due to ventricular septal defect （VSD） （PAH group） and patients without PAH （NPH group）.Total RNA was extracted from plasma collected and the different expression of miRNA was detected by miRNA array and verified using quantitative real-time polymerase chain reaction （qRT-PCR）.The target regulating genes were predicted by software such as Miranda,Targetscan and Pictar.Results The miRNA array results showed that 86 miRNAs were differentially expressed in PAH group,among them,50 were up-regulated and 36 were down-regulated.The qRT-PCR results of miR98 were in agreement with the miRNA array results.The target regulating gene of has-miR-98 was endothelin-1 （ET-1）predicted by software such as Miranda,Targetscan and Pictar.Conclusions There are differential expressions of plasma miRNAs in patients with PAH associated with CHD.These differentially expressed miRNAs may be involved in the development of PAH associated with CHD.It is probable that miR-98 is a new biomarker of CHD combined severe PAH.