通过AAV8-TBG-LOX-1特异性异位表达于肝细胞实现肝脏降解循环Ox-LDL减轻血管脂质驱动炎症反应抑制动脉粥样硬化研究

肝脏脂质代谢失衡与氧化脂质在血管壁沉积驱动炎症反应是动脉粥样硬化形成的关键，两者关系密切交织。肝脏胆固醇降解不足将促进脂质颗粒在外周循环时氧化修饰和血管壁沉积，血管壁将氧化型低密度脂蛋白（Ox-LDL）作为异物通过模式识别受体吞噬并处理，最终形成包裹脂质的斑块；其中内皮细胞LOX-1受体吞噬是启动和促进这一机制的重要环节；而作为脂质代谢核心的肝细胞因为缺乏摄取Ox-LDL的受体而无法处理氧化的脂质颗粒。本研究拟构建AAV8-TBG-LOX-1特异性肝脏转染体系，使肝细胞持续异位表达LOX-1受体，通过ApoE及LOX-1基因单敲、双敲小鼠及离体细胞探讨肝脏摄取降解Ox-LDL的效果，观察全身胆固醇代谢改变，阐明这一策略对肝脏脂质代谢和血管壁脂质沉积的分子调变机制。最终从脂质代谢平衡、脂质驱动血管炎症反应及转基因策略对机体及相关细胞脂质代谢分子途径的转变，探寻抗动脉粥样硬化的新策略和新靶点。

Liver lipid metabolism imbalance and oxidized lipid-driving inflammation in the vascular wall play a key role in the formation of atherosclerosis. And these two pathogenic progresses have a crosstalk and are closely intertwined. Low-density lipoprotein is oxidized to oxidized low-density lipoprotein (Ox-LDL) in peripheral circulation and processed by the pattern recognition receptor in the vascular wall to form lipid plaques when the liver cholesterol is insufficiently degraded. Among this process, the LOX-1 receptor in the endothelial cell associated phagocytosis pathway is an important link to initiate and promote this pathogenic mechanism. And the hepatocytes which are the core cells in the lipid processing cannot degrade oxidized lipid particles due to the lack of receptors that can take up Ox-LDL. In this study, a specific liver transfection virus system AAV8-TBG-LOX1 was constructed to enable hepatocytes to continuously express ectopic LOX-1 receptors to enhance the liver to process and degrade Ox-LDL. We will explore the ability of taking up and degrading Ox-LDL in liver via LOX-1 receptor, observe the changes of cholesterol metabolism in ApoE knockout mice and ApoE/LOX-1 double knockout mouse models, and use the isolated hepatocytes, endothelial cells, macrophages cells to elucidate the molecular modulation mechanisms of this strategy on liver lipid metabolism and vascular lipid deposition. We will comprehensively observe and analyze the changes in the distribution of cholesterol metabolism throughout the animal body, especially the promoting effect of liver bile efflux; and the changes in the formation of atherosclerosis in the aortic wall. And we will clarify the detailed molecular mechanism of this gene-level intervention on cholesterol metabolism performance. In summary, we will study the liver lipid metabolism balance, vascular lipid-driven inflammation via the AAV8-TBG-LOX-1 transgenic intervention and provide new strategies and new targets for anti-atherosclerosis.