从miRNA调控的PMVEC通透性研究流感病毒性肺炎肺络损伤的分子机制及解毒凉血散瘀方的干预

项目名称： 从miRNA调控的PMVEC通透性研究流感病毒性肺炎肺络损伤的分子机制及解毒凉血散瘀方的干预

项目编号： No.81473520

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 吴莹

作者单位： 北京中医药大学

项目金额： 72万元

中文摘要： 流感病毒性肺炎中普遍存在肺络损伤的病变，前期研究以肺微血管内皮细胞（PMVEC）作为肺络的载体，证实ERM磷酸化是流感病毒感染PMVEC通透性升高的关键环节；miRNA是基因调控的核心成分，然而，调控ERM磷酸化、PMVEC通透性升高的miRNA分子尚未报道。本课题以原代PMVEC及流感病毒性肺炎小鼠为模型，miRNA芯片筛选流感病毒、TNF-α作用后差异表达miRNA并验证；Miranda软件及双荧光素酶报告载体预测并验证其靶基因；功能获得性或缺失性研究分析miRNA及靶基因在ERM磷酸化及PMVEC通透性升高中的作用；本课题首次从miRNA调控PMVEC通透性的角度探讨流感病毒性肺炎中肺络损伤的分子机制。并从肺络论治，观察具有解毒凉血散瘀功效的经典合方犀角地黄汤合银翘散对miRNA-靶基因相关通路的影响，探讨其改善PMVEC通透性的miRNA调控机制，为其临床应用提供依据及新的思路。

中文关键词： 流感病毒；肺炎；肺微血管内皮细胞；微小RNA；犀角地黄汤合银翘散

英文摘要： Damage of lung collaterals prevails in the pathological change of influenza virus-induced pneumonia. Using pulmonary microvascular endothelial cell (PMVEC) as the vector of lung collaterals, the previous study demonstrated that phosphorylation of ERM played an important role in PMVEC hyperpermeability induced by influenza virus. MiRNA is a key link in gene regulation, however, the miRNAs which regulate phosphorylation of ERM and PMVEC hyperpermeability, have not been reported. Therefore, using primary cultured PMVEC as in vitro model and mice with influenza virus-induced pneumonia as in vivo model, the study screens the differential expressional miRNAs induced by influenza virus and TNF-α, respectively, by using miRNA microarray chips; predicts the target genes regulated by miRNAs by using bioinformatics software, such as Miranda, and further verifies them by using dual luciferase report vectors; takes advantage of gain-of-function research or loss-of-function research to explore the effects of the differential expressional miRNAs and their target genes on ERM phosphorylation and PMVEC hyperpermeability. Based on the regulation of miRNA on PMVEC permeability, the study firstly explores the molecular mechanism of damage of lung collaterals in influenza virus-induced pneumonia. And the research observes the effect of classical combined formula Xijiao dihuang tang and Yinqiaosan, which had detoxification, cooling blood and dissipating stasis efficacy, on miRNA-target-gene and its related signaling pathway, to explore the formula's miRNA-regulating mechanism in improving PMVEC hyperpermeability and provide experimental evidence and novel idea for its clinical application.

英文关键词： influenza virus;pneumonia;pulmonary microvascular endothelial cell;micro RNA;Xijiao dihuang tang and Yinqiaosan