外源性CO通过GC/cGMP/PKG信号通路调控脓毒症时EC/SMC互调作用的机制研究

项目名称： 外源性CO通过GC/cGMP/PKG信号通路调控脓毒症时EC/SMC互调作用的机制研究

项目编号： No.81471903

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： 孙炳伟

作者单位： 江苏大学

项目金额： 72万元

中文摘要： 脓毒症时血管内皮细胞（EC）和平滑肌细胞（SMC）微环境失稳、互调过程紊乱是导致血管功能障碍和器官功能不全的重要因素，但详细机制尚未阐明。本研究采用CLP脓毒症动物模型和EC/SMC细胞单（共）培养模型，以外源性一氧化碳（CORM）进行干预，运用基因分析、质谱分析、蛋白质水平测定和器官功能评价等手段，在分子、细胞和整体等不同层面上，研究脓毒症时EC、SMC重要结构和功能、缝隙连接蛋白Cx、微环境失稳态变化，以及EC和SMC间重要的互调因素、信号通道GC/cGMP/PKG的动态变化，并首次利用原子力显微技术和正交相位显微细胞亚表面3D特征识别技术，精确观察脓毒症时SMC收缩表型，旨在探明脓毒症时EC/SMC微环境和双向调节功能之间的相关关系，探明外源性CO通过GC/cGMP/PKG途径调节脓毒症时异常的EC/SMC微环境和相互作用的（部分）分子学基础，为发现脓毒症干预新靶位提供新的思路。

中文关键词： 脓毒症；一氧化碳；内皮细胞；血管平滑肌细胞；信号转导

英文摘要： Microenvironment and two-way adjustment disorder between endothelial cell (EC) and vascular smooth muscle cells （SMC）during sepsis are important factors leading to dysfunction of blood vessel、inadequate tissue perfusionand Organ dysfunction, the detail mechanisms, however, are not completely elucidate. This project employs CLP mice and LPS-stimulated cells as sepsis models to examine the dynamic state of EC/SMC interaction under the intervention of the exogenous CO liberated by CO-releasing molecules. The important factors will be investigated in the molecular and cellular levels through applying updated molecular biological techniques including gene analysis, mass spectrometric analysis, measurement of protein expressions, which investigate the structural and functional changes of EC/SMC, microenvironment and two-way adjustment disorder between endothelial cell and vascular smooth muscle cells ,and essential regulator of EC/SMC such as ion channels, the dynamic changes of Notch signal transduction during sepsis. Furthermore, this project also focuses on the important influences of the shear stresses induced by instabal circulating fluidin sepsis. Therefor, we are in attempt to elucidate the correlation of microenvironment and two-way adjustment between endothelial cell and vascular smooth muscle cells, to determine whether sGC/cGMP pathway and Notch signal transduction will be the novel targets of exogenous CO, in order to elucidate molecular mechnisms (even partly) responsible for the regulatory effects of CO on abnormal microenvironment and two-way adjustment of EC/SMC and signal transduction during sepsis and to provide theoretical basis and new idea for the research and treatment of sepsis.

英文关键词： Sepsis;Carbon monoxide;Endothelial cell;Vascular smooth muscle cell;Signal transduction