线粒体自噬在褪黑激素缓解OTA诱导肉鸭肝脏氧化应激中的作用及机制研究

赭曲霉素A（OTA）可诱发严重的肝脏氧化应激，导致线粒体功能障碍，引发细胞自噬。褪黑激素作为自由基的清除剂，在抵御氧化应激中发挥重要作用。但褪黑激素缓解OTA诱导肝脏氧化应激的作用机制未完全阐明，线粒体自噬在其中的作用尚未揭示。本项目拟构建OTA诱导肉鸭肝脏氧化应激模型，系统采用荧光探针、流式细胞术、TEM检测等技术，通过肝脏氧化应激水平、线粒体功能及自噬相关通路指标变化，研究OTA诱导下肉鸭肝脏氧化应激损伤作用；并采用鸭肝脏原代细胞，在OTA诱导基础上通过褪黑激素调控，利用基因过表达及沉默技术，对鸭肝脏细胞线粒体自噬作用通路以及褪黑激素缓解OTA诱导氧化应激的作用机制进行体外解析；最终通过饲养对比实验，验证并揭示线粒体自噬在褪黑激素缓解OTA诱导肝脏氧化应激中的作用及机制，为阐明褪黑激素在肝脏损伤的缓解机制及针对性开展营养调控策略提供科学依据。

Ochratoxin A (OTA) could induce severe liver oxidative stress and mitochodrial dysfunction, therefore leading cell apoptosis and autophagy. As a free-redical scavenger, melatonin plays a pivotal role in resistance to oxidative stress. However, the mechanism of melatonin alleviates OTA-induced liver oxidative stress was not fully elucidated and the role of mitophagy in it was not revealed totally. This program will establish the OTA-induced liver oxidative stress model, using fluorescence probe, flow cytometry and TEM methods, according to changes of liver oxidative stress level, mitochodrial function and autophagy signal pathway, to investigate the damage of OTA-induced liver oxidative stress in ducks. Gene overexpression and RNAi methods will be used for the interpretation of liver mitophagy and mechanism of melatonin alleviates OTA-induced oxidative stress in duck liver primary cells. Feeding experiment will be used for revealing the mechanism of mitophagy in melation alleviates ochratoxin A induced liver oxidative stress. This program would illuminate the mechanism of melatonin in liver injury alleviation and provide scientific evidence for further nutritional regulation.