肺动脉高压（PH）的主要临床表现为肺动脉压力的进行性升高，它可分为原发性肺动脉高压和继发性肺动脉高压。前者是不明原因的肺动脉功能失调，而后者则是许多肺部疾病极具破坏性的并发症。其中，慢性低氧性肺动脉高压（CHPH）和慢性血栓栓塞性肺动脉高压（CTEPH）具有较高的死亡率和死亡率。然而，关于这两种疾病的发病机制目前尚未完全阐明。本研究通过成功建立肺动脉平滑肌细胞原代培养及低氧性肺动脉高压动物模型，探讨CHPH肺血管重构发生的分子机制及其干预措施；同时完善国人CTEPH患者生物标本及遗传数据库，建立了CTEPH患者血栓内膜剥脱术标本细胞分离与培养的方法，并研究其发病的分子机制。本课题发现慢性低氧可引起BMP4表达上调，依赖于HIF-1α的BMP4作用于BMP受体，上调ERK1/2和p38MARK的表达，从而引起TRPC1、TRPC6蛋白表达的上调，使得肺血管平滑肌细胞内SOCE上调，造成细胞增殖和血管重塑，引起肺动脉高压。西地那非则通过cGMP-PKG-PPARγ/NFAT轴下调CHPH大鼠肺动脉TRPC1、TRPC6的表达，降低肺动脉平滑肌细胞的SOCE，从而抑制低氧引起的肺动脉平滑肌细胞内钙离子失衡，抑制低氧引起的肺动脉平滑肌细胞增殖和血管重塑。而丹参酮IIA亦可通过下调CHPH大鼠肺动脉TRPC1、TRPC6的表达，降低肺动脉平滑肌细胞的SOCE，从而发挥保护作用。在CTEPH的研究中发现纤维蛋白原Aα Thr312Ala (A/G)是CTEPH发病的独立危险因素且其纤维蛋白不易溶解。在CTEPH患者与正常对照组的血浆及平滑肌细胞中，miRNA表达谱存在差异。其中let-7b在CTEPH患者血浆中表达明显下调，且let-7b通过影响其靶基因TGF-β影响了血液中ET-1的表达水平和肺血管内皮与平滑细胞的增殖和迁移能力。此外，CTEPH患者的肺血管平滑肌细胞与正常的肺动脉平滑肌细胞的增殖、凋亡、代谢等功能也存在着差异。以上研究成果为探讨了CHPH和CTEPH的病理生理学机制奠定了基础，从不同侧面和研究水平探讨导致CHPH和CTEPH发生发展的决定性因素，为CHPH和CTEPH的预防、治疗提供新的分子遗传标志和治疗靶点。 Pulmonary hypertension (PH) is characterized by progressive increases in pulmonary arterial pressure, which is categorized into primary or secondary PH. Primary PH is a disease of unknown etiology associated with the dysfunction of pulmonary arteries. Secondary PH is a devastating complication of chronic pulmonary disease. Chronic hypoxic pulmonary hypertension (CHPH) and chronic thromboembolic pulmonary hypertension (CTEPH) are members of secondary PH and contribute significantly to the high morbidity and mortality rates. However, the pathogenesis of CHPH and CTEPH remains not clearly understood. We used primary cultured pulmonary arterial smooth muscle cells (PASMCs) and animal models of CHPH to investigate the mechanisms of the remodeling of pulmonary arteries and the treatment for CHPH. Moreover, we collected the information of CTEPH patients and established biological databases including biological samples and gene function, isolated and cultured PASMCs from endarterectomized tissue of CTEPH patients to study the molecular basis of the development of CTEPH. We found that the HIF-1α dependent upregulation of BMP4 bind to its receptor induced the increase of store-operated Ca2+ entry (SOCE), canonical transient receptor potential 1 (TRPC1), TRPC6 expression by activating p38MAPK and ERK1/2 signaling pathways in PASMSc after exposed to hypoxia.. Sildenafil and sodium tanshinone IIA sulfonate treatment respectively ameliorated hypoxia-induced PASMCs proliferation and attenuated hypoxia induced enhancement of SOCE, up-regulation of TRPC1, TRPC6 expression. Furthermore, Fibrinogen Aa Thr312Ala (A/G) specifically contributes to CTEPH by increasing fibrin resistance. Differentially expressed miRNAs were detected in the plasma and PASMCs of CTEPH and control patients. Let-7b decreased in CTEPH patients and inhibited the proliferation and migration of pulmonary arterial endothelial cells (PAECs) and PASMCs through the upregulation of TGF-β and ET-1. The distinguishing features of proliferation, apoptosis and metabolism between PASMCs of CTEPH and control patients were also existed. This study identifies the molecular basis of the pathogenesis of the CHPH and CTEPH, investigates the critical factors in the development and progression of CHPH and CTEPH from different approaches, meanwhile, provides novel genetic molecules and therapeutic targets for prevention and reversion of CHPH and CTEPH.
首次提出内源性硫化氢（H2S）是心血管调节的新型气体信号分子。研究过程中，继续深入探索了H2S在动脉粥样硬化、高血压、肺动脉高压等心血管损伤性疾病发病中的调节作用及其机制，发现H2S可通过抗炎症、抗氧化及抗内质网应激作用发挥重要的心血管保护效应。在此基础上，揭示含硫氨基酸代谢途径中产生的另一种气体分子二氧化硫（SO2）在心血管系统存在生成体系，并对其心血管调节作用进行了探索，发现并提出SO2是心血管调节的又一新型气体信号分子。同时，探索了皮质抑素(cortistatin, CST)、intermedin1-53、ghrelin及obestatin等新型血管活性肽在血管钙化等心血管疾病发病中的作用机制；以MicroRNA的克隆、检测和功能研究方法及技术平台为基础，筛选并确定miR-1、miR-133、miR-499、miR-26等miRNA在心血管重塑发生过程中发挥重要调节作用。5, n=5). As the sponsor of the conference, we organized the First and Second National Congress of Gasotransmitter H2S (2007 and 2010). The principal investigator got the Chang Jiang Scholars Program and was awarded the young & middle–aged experts with outstanding contributions by Ministry of Health of China. The members of our research group was awarded the New star of Beijing Scientist and Technology and Pujiang Scholar. 3 post doctorates were trained, 24 graduates got doctor degree and 20 graduated got master degrees.