Prohibitin 2/miR-151通路调控自噬响应对虾氧化应激损伤机制研究

氧化应激产生的活性氧诱导自噬，而自噬缓解氧化应激造成的损伤。前期工作发现，对虾血细胞中miR-151调控自噬响应环境应激，干扰Prohibitin 2后miR-151表达上调。但Prohibitin 2如何调控miR-151参与自噬尚不清楚。本项目拟探讨Prohibitin 2调控miR-151介导自噬缓解亚硝酸盐引起的氧化损伤。通过RNAi技术，明确Prohibitin 2对miR-151的调控；采用pull-down筛选与Prohibitin 2互作，并调控miR-151的候选转录因子；通过EMSA、ChIP-seq等方法，证明转录因子与Prohibitin 2互作，调控miR-151参与自噬的分子机制。揭示Prohibitin 2/转录因子/miR-151轴调控自噬缓解氧化损伤的机制，为转录因子调控miRNAs参与细胞自噬的生理学功能提供新证据，为对虾养殖业可持续发展提供新的策略。

ROS produced by oxidative stress induces autophagy, and autophagy alleviates the damage caused by oxidative stress. Previous work found that miR-151 regulates autophagy responding to environment stress in shrimp hemolymph, and the expression of miR-151 is up-regulated when Prohibitin 2 is interfered. However, how Prohibitin 2 regulating miR-151 to participate in autophagy is still unclear. This project intends to elucidate how Prohibitin 2 regulates miR-151-mediated autophagy to relieve oxidative damage caused by nitrite stress. RNA interfered will be used to clarify the regulation of Prohibitin 2 on miR-151; Pull-down will be used to screen the transcription factors that interact with Prohibitin 2 and regulate the expression of miR-151; EMSA and ChIP-seq will be used to prove that transcription factors interacting with Prohibitin 2 regulate miR-151 to participate in the regulation of autophagy. It will reveal the molecular mechanism of Prohibitin 2/transcription factor/miR-151 axis regulates autophagy to alleviate oxidative damage. This study will provide new evidence that transcription factors regulating miRNAs involve in the physiological functions of autophagy, as well as provide a new strategy for the sustainable development of shrimp aquaculture.