本项目以体内外二个层面为研究视角，沿着TFEB靶向作用LAL调控脂类自噬的思路，朝着系统阐述TFEB靶向作用LAL调控脂类自噬改善糖尿病心肌病脂毒性机制的目标开展工作。体外研究以心肌脂毒性细胞模型为对象，首先在细胞中验证TFEB靶向作用LAL；随后采用高表达载体和CRISPR/Cas9方法上下调LAL基因，采用免疫荧光共定位、流式细胞术和RT-PCR/Western Blot等技术检测脂类自噬、脂类堆积及细胞功能状态，证实TFEB靶向作用LAL调控脂类自噬改善心肌细胞功能。体内研究以db/db小鼠模型为对象，导入高表达LAL病毒载体，运用心脏功能学、心血流动力学、超微组织学和检测脂类自噬特异基因表达等方法，评价脂类堆积、自噬及心功能，系统阐述TFEB靶向作用LAL调控脂类自噬改善糖尿病心肌病脂毒性损害的机制，此机制具有重要的理论意义及应用价值，为深刻认识和有效治疗糖尿病心肌病奠定理论基础。

To explore the mechanism of cardiac lipotoxicity in diabetic heart, LAL will be focused on how to control lipophagy by TFEB for ameliorating cardiac lipotoxicity in the vivo and vitro of diabetic heart. In lipotoxicity model of the rat neonatal cardiomyocyte, the role of TFEB targeting on LAL has to be certificated before demonstrating that LAL attenuates cardiac lipotoxicity through TFEB in the diabetic cardiomyopathy, CRISPR/Cas9 and transfection of adeno-assocatied virus will be used to knockout and overexpress LAL gene respectively to regulate lipophagy. Co-localization under fluorescence microscope, flow cytometry and RT-PCR/Western Blot are performed to check status of lipophagy, lipotoxity and gene function in myocytes. The model of db/db mouse is used to appraise the effect of the LAL on cardiac lipotoxicity, cardiac function and lipophagy by overexpression of LAL gene. In this study, our aim is to provide a new strategy for the treatment of diabetic cardiomyopathy.