心肌脂滴比较蛋白组学研究及心肌脂滴蛋白CGI-58对心脏功能的影响

项目名称： 心肌脂滴比较蛋白组学研究及心肌脂滴蛋白CGI-58对心脏功能的影响

项目编号： No.81471082

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 张慧娜

作者单位： 中国科学院生物物理研究所

项目金额： 73万元

中文摘要： 心脏能量代谢紊乱与心功能低下密切相关，其中甘油三酯代谢障碍是主要环节。脂滴是细胞内甘油三酯代谢的主要场所，在维持细胞能量平衡中发挥关键作用。而决定甘油三酯水解酶定位及活性的脂滴蛋白CGI-58在心肌脂代谢和心脏功能中的作用还未见阐述。本项目将探索CGI-58对心肌脂代谢和心脏功能的影响及其机制。预实验结果表明正常与心功能不全鼠心肌细胞脂滴上CGI-58定位有显著差异，提示CGI-58表达降低和脂滴定位减少可能与心功能不全心肌能量代谢紊乱相关。我们将利用分子生物学和转基因手段考察增加和干扰CGI-58对心肌细胞和心功能低下动物模型的心肌中脂滴大小数量，甘油三酯含量，脂代谢及心脏功能的影响。并利用免疫共沉淀，荧光共振能量转移和激共聚焦显微镜实时观察CGI-58对心肌细胞脂代谢酶定位及活性的调节机制。本研究将为心功能不全能量代谢失衡提供理论依据，并为心功能不全的防治提供新的思路和靶标。

中文关键词： 脂滴；心肌细胞；比较蛋白组学；甘油三酯代谢；CGI-58

英文摘要： Myocardial metabolic disorder especially the aberrant metabolism in triglyceride (TG) hydrolyzation plays a pivotal role in the pathogenesis of cardiac dysfunction. Lipid droplet (LD) is the main organelle in TG storage and metabolism and plays a pivotal role in sustaining intracellular energy homeostasis. LD protein CGI-58 is the key regulator of adipose triglyceride lipase (ATGL) through determining the activity and LD location of ATGL. However, till now, the research about LDs in cardiomyocyte is less well developed, and the effect of CGI-58 on cardiomyocyte lipid metabolism and heart function is not well delineated. This project will explore the role of CGI-58 in cardiomyocyte triglycerol（TG）metabolism as well as in heart function. We have identified the different LD proteins from normal and diastolic dysfunction rat hearts. The preliminary results of proteome and immunoblotting presented that CGI-58 was significantly decreased on LD of diastolic dysfunction rat hearts, suggesting that the reduced CGI-58 may be implicated in the lipid metabolic disorder in cardiac dysfunction. Through constructing heart specific CGI-58 transgenic mouse and manipulating the expression level and location of CGI-58 by using molecular biology techniques, we will detect the variation of the amount and size of LD, the amount and metabolism of TG and fatty acid as well as heart function in vivo and in vitro under conditions of normal and cardiac dysfunction. Using co-immunoprecipitation (Co-IP), fluorescent energy resonance transfer (FERT) and confocal assay, we will investigate the regulation mechanism of CGI-58 on the activity and location of lipid metabolic enzyme ATGL. This research will provide the theory basis for the aberrant lipid metabolism in cardiac dysfunction, and may promote new directions in future research concerning the etiology and treatment of cardiac dysfunction.

英文关键词： Lipid droplet;Cardiomyocyte;Comparison proteome;TG metabolism;CGI-58