叶黄素调控视网膜色素上皮胆固醇稳态抗年龄相关性黄斑病变的机制研究

年龄相关性黄斑病变（ARM）发生与视网膜色素上皮（RPE）胆固醇稳态失衡相关，但目前尚无系统研究。前期实验发现叶黄素可减轻RPE及视网膜损伤，但叶黄素调控RPE胆固醇稳态抗ARM的机制不清。本项目拟建立ARM模型并给予叶黄素干预：1）用脂质组学、OCT等，检测脂质异位沉积和脂毒性产物，明确RPE胆固醇代谢紊乱对ARM发生的影响及叶黄素的保护作用；2）阐明叶黄素抗ARM的RPE胆固醇稳态机制：①用CoIP和ChIP等，检测胆固醇代谢动力调控蛋白表达及SREBP/SCAP/Insig复合物的形成，探讨叶黄素维持RPE胆固醇代谢动力稳态的分子靶点；②用RNAi和甲基化CpG结合域蛋白2-染色质免疫共沉淀等，检测吞噬相关蛋白表达及胆固醇代谢酶甲基化水平改变，揭示RPE胆固醇生物转化通路间的关系及叶黄素发挥作用的分子机制。本研究将为调控RPE胆固醇稳态提供分子作用靶点，为优化ARM防治策略提供依据。

Age-related maculopathy (ARM) is associated with cholesterol homeostasis imbalance in retinal pigment epithelium (RPE); however, there is not yet systematic study about link of RPE cholesterol homeostasis imbalance and ARM. Our previous studies indicated that lutein could ameliorate RPE and retinal damage, but the mechanism of lutein regulating RPE cholesterol homeostasis against ARM was unclear. This project will establish ARM models which lutein intervention will be given. 1) Lipid heterotopic deposition and lipotoxicity will be detected by lipidomics and optical coherence tomography, etc. to investigate the impact of RPE cholesterol metabolic imbalance on ARM and the protective effect of lutein. 2) The mechanism of lutein regulating RPE cholesterol homeostasis against ARM will be clarified:①The expression of cholesterol metabolic dynamic regulatory protein and the formation of SREBP/SCAP/Insig complex will be detected by using co-immunoprecipitation and chromatin immunoprecipitation assay to explore the molecular targets of lutein for maintaining RPE cholesterol homeostasis; ② The expression of phagocytic related proteins and the methylation level of cholesterol metabolic enzymes will be detected by RNAi and methyl-CpG binding domain protein 2 ChIP to reveal the relationship between RPE cholesterol biotransformation pathways and the molecular mechanism of lutein. This study will provide molecular targets on regulating RPE cholesterol homeostasis and provide experimental basis for optimizing ARM prevention and control strategies.