miRNA-206调控IUGR相关成年期肺动脉平滑肌细胞功能失调的研究

IUGR was related with numerous diseases in adults which were found by clinical studies and epidemiological survey. Epigenetics in fetal origin of diseases is a complex process including microRNA interference. However, the underlying mechanism of this hypothesis is still unknown. Our preliminary study has shown that rats with intrauterine growth retardation (IUGR) had increased sensitivity with hypoxia. We also found the expression of miRNA-206 in pulmonary artery smooth muscle of these rats was higher then control group. We hypothesized miRNA-206 regulates the PASMCs dysfunction in adults following IUGR via multiple target pathway. In the present study, ①we will investigate the regulation mechanism of miRNA-206 in pathway of IUGR related PAH in adults through multiple techniques in vivo and in vitro. ②In vitro experiments will be conducted to confirm this epigenetic mechanism in this project. ③In vivo experiments will also be conducted to confirm whether miRNA target treatment would infect this multiple effect in IUGR, which could have an important clinical significance.

大量临床研究及流行病学调查发现，宫内生长迟缓（IUGR）与成年期慢性疾病密切相关。有证据表明表观调控机制参与其中，包括小RNA干扰，但其中的机制尚未完全阐明。 我们前期研究发现，IUGR大鼠成年期肺动脉平滑肌细胞（PASMCs）对缺氧敏感性增高；组织芯片筛选还发现，肺动脉平滑肌组织中miRNA-206表达明显增高。为验证miRNA-206通过调控缺氧相关信号通路介导IUGR大鼠成年期缺氧性肺动脉高压，本课题拟运用分子生物学检测、RNAi、原代细胞培养、活体miRNA干预等手段进行体内外实验：①发现miRNA-206调控机制参与IUGR引起成年期CH-PHT发生的作用；②初步探明miRNA-206通过多靶点调控上述作用的表观调控效应；③明确在体靶向miRNA治疗能阻止或逆转IUGR对基因表达的表观调控效应，而最终降低成年期缺氧性肺动脉高压的发生率，为胎儿来源的成年期肺动脉高压防治提供新思路。

本研究在前期研究基础上探讨了表观调控机制在IUGR致成年期肺动脉高压及肺血管重塑中的作用，我们验证了IUGR与低氧性肺动脉高压的关系，利用microRNA表达谱芯片及PCR技术对miR-206在PASMCs中的表达进行分析，筛选miR-206相关的信号通路进行分析，Luciferase发现并验证电压依赖性钾通道Kv1.5在这一表观遗传学调控机制中介导的作用；在上述实验的基础上，在体及离体验证RNA干扰后肺动脉平滑肌细胞中Kv1.5的表达改变及细胞增殖能力的变化，评价其与慢性低氧性肺动脉平滑肌重塑的关系。