LncRNA TCONS_00107023/miR-15a-5p/cxcl10通路在肺动脉高压复合炎症疾病中机制的研究

Pulmonary arterial hypertension (PAH) is a progressive pulmonary vascular disease, which is characterized by a marked increase in pulmonary arterial pressure and right ventricular hypertrophy. It was observed that severe PAH once got an acute inflammation, the cardiac function would deteriorate rapidly or even lead to death. As inflammation obviously aggravated damage of PAH, the underlying mechanisms is needed to be studied. In our preliminary experiment, we found the express of a new lncRNA-lncRNA TCONS_00107023 and cxcl10mRNA was significantly increased.On the basis of the online biological tools and the progress of the study, we assumed that miR-15a-5p is a key miRNA in the known partial pathway.So we investigate the role of lncRNA TCONS_00107023/miR-15a-5p/cxcl10 pathway in PAH with acute inflammation.The study intends to regulate the pathway to observe the biological function through morphology, QPCR, WB, immunohistochemistry and hemodynamics in vivo and in vitro .

肺动脉高压是一种非常常见的心血管疾病，主要是由于肺血管形态学和肺循环血液动力学改变而导致的严重的病理现象，临床上表现为明显增高的肺动脉压力和右心室肥厚。在围术期管理中严重肺动脉高压患者一旦出现肺部感染，心功能将急剧恶化甚至危及生命，研究这种现象的具体机制将有助于改善肺动脉高压的围术期治疗。前期研究通过高通量测序筛选出并且验证了一种尚未研究过的lncRNA-lncRNA TCONS_00107023，其表达显著升高，此外cxcl10mRNA表达量亦显著升高，通过在线生物学工具预测，结合国内外研究进展，发现miRNA-15a-5p可能也参与调控，我们拟研究lncRNA TCONS_00107023/miR-15a-5p/cxcl10这一通路在该疾病中的具体作用机制。本项目拟从细胞和动物实验，调控通路各节点表达强度后通过形态学、QPCR、WB、免疫组化、血流动力学观察这一通路的生物学作用。

肺动脉高压是临床常见心血管疾患，围术期死亡率高，在肺动脉高压的基础上，感染或炎症的发生会进一步诱发右心功能的急剧恶化。长期以来，肺动脉高压合并右心衰竭一直是围术期治疗非常棘手的难题。我们前期高通量测序结果显示，脂多糖注射后肺动脉高压大鼠右室心肌lncRNA XLOC_034025(也称tcon_00052110)表达水平升高。因此本研究研究目的为进一步探索该新发现LncRNA对肥厚性心肌细胞中丙酮酸激酶2/丙酮酸激酶1表达比例的影响；方法：采用qRT-PCR方法检测LncRNA XLOC_034025在H9C2肥厚细胞中的表达。利用RACE和FISH在心肌细胞中验证和定位lncRNA XLOC_034025。采用Western blotting、qRT-PCR、免疫荧光、RNA pulldown等方法，论证lncRNA XLOC_034025通过与PTBP1蛋白相互作用调控PKM2/PKM1表达的分子机制。结果:LncRNA XLOC_034025在肥厚型H9C2细胞中高过表达。LncRNA XLOC_034025的敲低导致PTBP1的不稳定性，导致PKM2 / PKM1的比例增加。PKM2的增加和PKM1的减少减轻了线粒体氧化应激和钙负荷。结论:LncRNA XLOC_034025通过与PTBP1蛋白相互作用调控PKM2/PKM1的表达比例。