先天性心脏病引起的肺动脉高压肺血管重构的分子机制研究

Pulmonary arterial hypertension (PAH) is a common complication of congenital heart disease (CHD), particularly in patients with left-to-right (systemic-to-pulmonary) shunts. Persistent exposure of the pulmonary vasculature to increased blood flow and pressure may result in vascular remodelling and dysfunction, resulting in a progressive rise in pulmonary vascular resistance and increased pressures in the right heart. The development of PAH in patients with CHD is associated with increased mortality and high morbidity. Vascular remodeling plays a key role in the pathogenesis of PHD-CHD. The molecular pathways triggering the remodeling process are poorly understood. In order to prepare for this project, we successfully establised a novel method to create a rat model of highkinetic unilateral pulmonary hypertension surgically induced by combined surgery. In the former experiments, we obtained 6 differentially expressed proteins in patients with PAH-CHD compared with patients with CHD but without PAH by screening using 2 antibody arrays. The 6 differentially expressed proteins might be involved in pulmonar vascular remodeling by analyses on them according to literatures and bioinformayics. This project is planning to screen the key factors involved in pulmonar vascular remodeling in CHD-PAH using 6 antibody arrays which can detect 240 important vasoactive proteins. Through verification on the obtained potential key factors in a rat model of highkinetic unilateral pulmonary hypertension and studies on the key roles of the obtained potential key factors in proliferation, migration, apoptosis and extracellular matrix metabolism in cultured pulmonar arterial cells, we aimed to clarify the molecular mechanisms the key factors invole in the pathogenesis of CHD-PAH, and provide potential targets for the treatment of CHD-PAH.

先天性心脏病(先心病)是小儿常见的心血管疾病之一，而其体-肺分流导致的肺动脉高压是先心病最常见的严重并发症，是一种肺循环血流量增多引起肺小动脉收缩和进行性重构的致死性疾病。肺血管重构是先心病肺动脉高压发病的始动环节，但其分子机制不清。为了实施本课题，我们建立了首创的分流型肺动脉高压动物模型。我们在前期工作中应用2个定量抗体芯片在先心病肺动脉高压患儿筛选获得了6个差异表达显著的蛋白质，文献检索和生物信息学分析提示这些差异蛋白质可能参与肺血管重构过程。本课题拟应用能够检测240个重要血管活性蛋白的5种抗体芯片，在先心病肺动脉高压患儿筛查可能参与先心病肺动脉高压肺血管重构的关键因子，通过在动物模型水平进行验证，并在细胞水平研究这些关键因子在肺血管细胞增殖、迁移、凋亡和细胞外基质构成中所起的关键作用，阐明它们参与先心病肺动脉高压肺血管重构的分子机制，并为先心病肺动脉高压的治疗提供提供新的参考靶点。

先天性心脏病(先心病)是小儿常见的心血管疾病之一，而其体-肺分流导致的肺动脉高压是先心病最常见的严重并发症，是一种肺循环血流量增多引起肺小动脉收缩和进行性重构的致死性疾病。肺血管重构是先心病肺动脉高压发病的始动环节，但其分子机制不清。本课题应用能够检测240个重要血管活性蛋白的5种抗体芯片，在先心病肺动脉高压患儿筛查可能参与先心病肺动脉高压肺血管重构的关键因子，对经过筛查获得的差异蛋白质，经过生物信息学分析和在我们首创的分流型肺动脉高压动物模型进一步应用RT-PCR、Western、ELISA和免疫组化进行验证，最终获得4个参与肺动脉高压肺血管重构的差异表达蛋白质，既，骨桥蛋白（OPN），分泌型簇蛋白（sCLU）、神经母细胞瘤形成抑制因子1（NBL1 或DAN）和脂钙蛋白-2（Lcn2）。在细胞模型的肺动脉高压动物模型中，研究阐明了OPN，CLU、NBL1（DAN）和Lcn2在先心病肺动脉高压肺血管重构中参与信号转导、增殖和凋亡的分子机制，为先心病肺动脉高压的诊断、预警和指导治疗提供了参考。