基于ccRNA-miRNA-lncRNA-mRNA复杂调控网络的复苏合剂（温肾潜阳法）治疗脓毒症急性肺损伤内皮功能障碍作用机制研究

Vascular endothelium injury and vascular leakage are the central links in the development of acute lung injury (ALI). In our previous study, it was confirmed that the use of Fusu agent （Warming kidney and subsiding yang methods) to treat sepsis-associated ALI can reduce the pulmonary water index in patients. Its mechanism of action is mainly a multi-target endothelial protective effect.In order to further elucidate the mechanism of its core regulation, this study is preparing to use high-throughput sequencing of whole transcriptomes. Firstly, based on the previous differential selection of endothelial cells, we constructe a post-transcriptional regulatory network based on ALI of ccRNA, miRNA, lncRNA and mRNA and Fusu agent to intervene endothelial function, and locate the key nodes of the network.Secondly, at the level of tissues and cells, we verify the function, mechanism, and targeted treatment strategy of the key nodes of the resuscitation mixture control screen; finally, we use the peripheral blood of patients as samples and endothelial-specific ncRNA in the core regulatory network as the target to study the molecular markers of endothelial injury in ALI ,and clinical validation by drug intervention.We innovatively (not previously reported) establish a regulatory network of post-transcriptional expression of endothelial dysfunction associated with sepsis-associated ALI and elucidate the core regulatory targets for multi-targeted endothelial protection of Fusu agent （Warming kidney and subsiding yang methods).It is used to enhance its scientific connotation and also lays the foundation for the study of related molecular biomarkers.

血管内皮损伤和血管渗漏是急性肺损伤(ALI)发生中心环节.课题组前期研究证实，复苏合剂(温肾潜阳法)治疗脓毒症ALI可降低患者肺水指数,作用机制主要为多靶点内皮保护作用,为了进一步阐明其核心调控机制,本研究拟借助全转录组高通量测序,首先,在前期筛选内皮细胞差异表达因素基础上,构建基于ccRNA、miRNA、lncRNA与mRNA的ALI及复苏合剂干预内皮功能转录后表达调控网络,定位该网络关键节点;其次,在组织和细胞水平验证复苏合剂调控筛得关键节点的功能,机制与靶向治疗策略;最终,以患者外周血为样本,以核心调控网络中具有内皮特异性ncRNA为目标,行ALI内皮损伤分子生物学标志物研究,并经药物干预以临床验证。创新性(未见报道)建立脓毒症ALI内皮功能损伤相关转录后表达调控网络,阐明复苏合剂(温肾潜阳法)多靶点内皮保护作用的核心调控靶点,提升其科学内涵，同时也为相关分子生物学标志物研究奠定基础

肺毛细血管渗漏是急性肺损伤（ALI）发生的中心环节，本课题在前期研究基础上，从整体水平、组织水平、细胞水平、分子水平多层次构建了复苏合剂（温肾潜阳法）干预ALI内皮功能的转录调控网络。本项目：（1）体内实验研究表明复苏合剂可改善脓毒症急性肺损伤大鼠肺组织病理损伤、降低大鼠血清TNF-α、IL-1β水平，mRNA/lncRNA表达谱及circRNA-miRNA-mRNA表达谱差异显示：复苏合剂通过多个lncRNA/circRNA作为miRNA的海绵吸附体发挥作用，并与NF-κB、RAGE等信号通路密切相关；（2）细胞层面研究显示复苏合剂可以降低LPS处理所致的IL-1β和TNF-a升高，并从ncRNA层面验证了复苏合剂可降低miR-146a-5p mRNA的表达；（3）临床研究表明复苏合剂能降低ARDS患者28天病死率和90天病死率，改善PaO2/FiO2，减少呼吸机使用时间和ICU住院时间，并与NLR信号通路等密切相关。本研究建立了脓毒症ALI内皮功能损伤及复苏合剂干预的mRNA/lncRNA表达谱及circRNA-miRNA-mRNA表达谱，阐明了复苏合剂多靶点内皮保护作用的核心调控靶点，为解决急性肺损伤肺毛细血管渗漏病理机制难题提供了临床有效治疗方法与药物靶点，对提高脓毒症所致急性肺损伤临床疗效、降低死亡率具有重要现实意义。