钙激活氯通道ANO1在ox-LDL诱导单核及巨噬细胞活化中的作用研究

Atherosclerosis seriously harms human health. Vascular inflammation mediated by monocyte migration and macrophage activation is crucial in the development of the atherosclerosis. Recent study has found that ANO1, a calcium activated chloride channel, can regulate the inflammatory response in the airway epithelium. However, the relationship between ANO1 and the vascular inflammation mediated by macrophage has not been reported. Our study shows that ANO1 is expressed in the human THP-1 cells-derived macrophage and affects the secretion of inflammatory factors; Ox-LDL, a atherogenic risk factors, can significantly enhance expression of ANO1 in macrophages derived from human THP-1 cells. Therefore, we speculate that ANO1 might participate in the vascular inflammation by promoting monocytes migration and macrophages activation. Using human THP-1 cells and advanced scientific research methods (including molecular biology, patch clamp, confocal imaging and genetic interference technology, ect.) , we will explore the relationship between ANO1 and monocyte migration or macrophage activation thereby to provide a new mentality and a new therapeutic target for prevention and treatment of atherosclerosis.

动脉粥样硬化严重危害人类健康。由单核巨噬细胞迁移及活化介导的血管炎症反应在动脉粥样硬化的发生阶段至关重要。最新研究发现钙激活氯通道ANO1参与气道上皮炎症反应的调节，而与单核巨噬细胞介导的血管炎症反应之间的关系尚未见报导。我们的研究结果表明ANO1在THP-1源性巨噬细胞中表达并影响巨噬细胞炎症因子的分泌功能；ox-LDL（致动脉粥样硬化因子）能够显著提高ANO1在THP-1源性巨噬细胞中表达。我们推测：ANO1可能促进ox-LDL诱导的单核巨噬细胞迁移及活化，参与血管炎症反应。为证实该假说，我们拟以人THP-1及其诱导分化的巨噬细胞作为实验模型，采用分子生物学、膜片钳及基因干扰等技术，从细胞和分子水平来探讨ANO1与单核巨噬细胞迁移及活化所介导的炎症因子分泌功能之间的关系，通过调节ANO1功能性表达来预防单核巨噬细胞介导血管炎症的发生，从这一角度入手，为动脉粥样硬化的防治提供新的靶向。

动脉粥样硬化是一种独立的疾病，同时也是导致心脑肾等多器官损害的高危因素，严重危害人类健康。由单核细胞迁移进而转变成巨噬细胞介导的血管炎症反应在动脉粥样硬化的发生发展过程中至关重要。钙激活氯通道ANO1与炎症反应之间关系密切，我们推测：ANO1可能促进ox-LDL（致动脉粥样硬化因子）诱导的单核巨噬细胞迁移及活化，参与血管炎症反应。为证实该假说，我们以人THP-1及THP-1源性巨噬细胞作为研究对象，利用分子生物学、基因编辑、细胞免疫化学及膜片钳等技术研究了钙激活氯通道ANO1在THP-1细胞迁移及THP-1源性巨噬细胞炎症反应中的作用。结果表明：钙激活氯通道ANO1在THP-1/THP-1源性巨噬细胞膜上存在着功能性表达；ox-LDL呈浓度和时间依赖性的促进THP-1/THP-1源性巨噬细胞ANO1的蛋白表达。钙激活氯通道ANO1能够促进THP-1细胞的迁移，抑制ANO1的活性或敲减其功能性表达能够显著抑制ox-LDL诱导的THP-1细胞的迁移；钙激活氯通道ANO1能够促进THP-1源性巨噬细胞炎症因子IL-1β、CRP的蛋白表达；抑制ANO1的活性或敲减其功能性表达能够显著抑制ox-LDL诱导的炎症因子IL-1β、CRP的蛋白表达；ox-LDL可能通过促ANO1功能性表达，进而激活NF-κB信号通路从而促进THP-1源性巨噬细胞的炎症反应。从单核细胞迁移及巨噬细胞炎症因子分泌的角度来探讨ANO1的功能性表达与动脉粥样硬化血管炎症反应之间的关系及可能的信号通路，探讨ANO1在动脉粥样硬化血管炎症反应中的角色，为动脉粥样硬化的防治提供新的药物作用靶点。