细胞内氯离子通道蛋白1（CLIC1）被认为是氧化应激中的传感器和效应器，在细胞氧化等因素刺激后其结构发生改变并插入脂膜中充当氯离子选择性通道，导致细胞内氯离子浓度增加。为了明确CLIC1在内皮细胞损伤与炎症中的作用及丹参酮ⅡA保护内皮损伤的机制，我们拟通过动物实验观察动脉粥样硬化动物中CLIC1的表达、膜定位及内皮细胞功能变化，并利用siRNA及转基因技术，建立CLIC1低表达及高表达的HUVEC细胞系，经过氧化氢诱导后用丹参酮ⅡA进行处理，检测细胞中CLIC1的表达及膜定位、细胞内氯离子浓度和iNOS、ET-1、ICAM-1、VCAM-1的表达及炎性因子TNF-α、IL6的分泌变化，明确CLIC1参与血管内皮细胞损伤与炎症的作用，并从CLIC1调控角度深入探讨丹参酮ⅡA抗动脉粥样硬化的机制。课题的实施可望为动脉粥样硬化的防治提供一种新思路，为抗动脉粥样硬化药物的筛选提供新靶点。

Chloride intracellular channel protein 1 (CLIC1) is considered to be oxidative stress sensors and effectors , and its structure is changed after oxidation stimulating factor and inserted into the cell membrane lipid acts as chloride ion selective channels , which result in the increasing intracellular chloride ionic concentration. In order to clarify the role of CLIC1 in the damage and inflammation of endothelial cell and the protect mechanism of Tanshinone ⅡA, we intend to observe the expression and location of CLIC1 and the change of endothelial cell function. In addition, the HUVEC lines in which CLIC1 express decreased or increased were established through RNA interference technology and transgenic technology. These cells were induced with hydrogen peroxide and treated with TanshinoneⅡA. CLIC1 expression and membrane localization, concentration of intracellular chloride, expression of iNOS, ET-1, ICAM-1, VCAM-1, the secretion of IL6 and TNF-α were detected. This project illuminate the function of CLIC1 in oxidative damage of endothelial cells, and uncovered the mechanism of anti- atherosclerosis of Tanshinone ⅡA from the perspective of CLIC1. The result of this project can provide a new target for anti- atherosclerosis research.

内皮氧化损伤和炎症反应是动脉粥样硬化（AS）发病机理中的关键启动步骤。本研究探究了细胞内氯离子通道1在血管内皮细胞氧化损伤和炎症中的作用及丹参酮ⅡA的调控机制。 研究结果表明： H2O2诱导HUVEC细胞氧化损伤后ROS、MDA水平显著升高，SOD酶活力下降，炎性因子（TNF-α、IL-6）和粘附分子（ICAM-1、VCAM-1）含量明显增加，而CLIC1抑制剂IAA94预处理或CLIC1基因敲除后，细胞内氧化水平和炎症反应明显下降，提示CLIC1在加剧内皮细胞氧化损伤和炎症反应中发挥了重要的作用。 动物实验与细胞实验表明，丹参酮ⅡA具有良好的抗动脉粥样硬化作用。CLIC1基因敲除内皮细胞(CLIC1-/-)中，H2O2处理后氧化损伤不明显，而且不管是否用丹参酮ⅡA处理，发现ROS、MDA水平始终保持低水平，SOD酶活力处于高水平，提示CLIC1在丹参酮ⅡA的抗氧化过程中发挥了极为重要的作用。H2O2处理后，CLIC1-/-细胞内炎性因子和粘附分子增加，丹参酮ⅡA处理后，TNF-α、ICAM-1、VCAM-1表达降低，提示CLIC1参与丹参酮ⅡA抗炎过程，却并不是其唯一机制。 本研究明确了CLIC1在H2O2诱导的内皮细胞损伤与炎症中的作用，并从CLIC1调控角度阐明丹参酮ⅡA抗AS的机制，为AS的防治提供一种新思路和药物作用新靶点。