从JAK2/STAT3通路研究A-FABP介导的巨噬细胞“自噬-凋亡对话”促动脉粥样硬化机制

项目名称： 从JAK2/STAT3通路研究A-FABP介导的巨噬细胞“自噬-凋亡对话”促动脉粥样硬化机制

项目编号： No.81500658

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 李卉

作者单位： 中南大学

项目金额： 17万元

中文摘要： 巨噬细胞凋亡是斑块破裂的重要因素。自噬/凋亡平衡决定了细胞命运，并影响动脉粥样硬化AS进程。A-FABP介导毒性脂质所致巨噬细胞凋亡而促AS发展，但其机制尚未阐明。本项目在我们先前证明 A-FABP增强脂毒性的作用可能与线粒体损伤及自噬有关的基础上，拟从整体、细胞以及亚细胞水平，综合运用多种技术对自噬、线粒体损伤、线粒体功能及凋亡展开全方位评价。利用ApoE-/-小鼠模型，体内论证A-FABP与AS发展的关系及对粥样斑块中巨噬细胞线粒体损伤及自噬/凋亡变化的作用。然后在巨噬细胞中使其沉默和过表达进行体外验证，并采用多种阻断策略依次抑制JAK2和自噬，明确作用的信号传导途径，及自噬对A-FABP介导的线粒体损伤及凋亡的影响。本课题首次以自噬-凋亡的交叉对话与A-FABP关系为切入点，从线粒体角度揭示了A-FABP对巨噬细胞凋亡/自噬平衡的调控及其信号传导途径，为AS发病机制提供新的理论依据。

中文关键词： 脂肪酸结合蛋白；自噬；巨噬细胞凋亡；线粒体；动脉粥样硬化

英文摘要： Macrophage apoptosis is a major cause of plaque rupture. The balance between autophagy and apoptosis determines cell fate, and influences atherosclerosis(AS) progression. A-FABP deficiency can mediate protection from lipid induced-apoptosis in macrophages and the course of AS, although the underlying mechanisms remain unknown. Our previous studies have suggested that one potential way A-FABP can modulate stress responses to toxic lipids may be through alteration in mitochondria function and autophagy. Here, a comprehensive evaluation is presented of status of autophagy, mitochondrial function and apoptosis by utilizing multiple techniques. We demonstrate in vivo the relationship between AS and A-FABP, and confirm the critical role of A-FABP in mediating macrophage mitochondrial damage and autophagy/apoptosis imbalance in the setting of dyslipidemia in AS utilizing ApoE-/- mouse model. Then in vitro A-FABP is silenced/ overexpressed in RAW264.7 macrophage cells to futher confirm its role as an obligatory intermediate for autophagy/apoptosis imbalance. Various blocking strategies are developed to inhibit JAK2 and autophagy to uncover the signaling of lipid-induced autophagy/apoptosis imbalance in macrophages, and examine the impact of autophagy on A-FABP-mediated mitochondrial injury and apoptosis. In conclusion, the study will uncover a previous unknown function for A-FABP in lipid-induced autophagy/apoptosis imbalance in macrophages, while addressing the essential role of mitochondrial function in A-FABP-mediated lipotoxic signaling.

英文关键词： fatty acid binding protein;autophagy ;macrophage apoptosis;mitochondria;atherosclerosis