TFEB介导的自噬流修复在皮瓣缺血再灌注损伤中的作用及调控机制研究

缺血再灌注(I/R)损伤是移植皮瓣坏死的主要原因，但缺乏有效的治疗手段。前期发现， I/R损伤皮瓣早期发生自噬流阻滞。为明确该现象的病理机制，本研究将在皮瓣I/R损伤中，验证活性氧簇（ROS）通过破坏溶酶体功能，导致自噬流的阻滞。而自噬流的障碍能造成细胞损害。转录因子TFEB的调控能通过降低ROS水平、增强溶酶体及自噬体活性，促进自噬流修复。前期发现，TFEB的活性在皮瓣I/R损伤中显著增强。本研究将探究通过调控TFEB活性促进自噬流修复，以治疗皮瓣I/R损伤。再考证I/R损伤皮瓣中TFEB受PARP1-LKB1-AMPK-mTOR信号通路的调控。最后，在体外人体细胞模型中，证明PARP1-LKB1-AMPK-mTOR通路对TFEB的调控及TFEB介导的自噬激活对细胞的保护作用。本项目旨在为皮瓣I/R损伤阐明自噬流阻滞的新机制；并提出通过TFEB调控修复自噬流，治疗皮瓣I/R损伤的新策略。

Necrosis of the transplanted skin flap is mainly due to ischemia-reperfusion (I/R) injury. However, there is a lack of effective treatment. Previously we demonstrated that autophagy flux was impaired at the early stage of I/R injury in skin flap. To further understand the mechanism of this phenomenon, we will verify whether reactive oxygen species (ROS) can contribute to lysosome functional impairment and further the blockade of autophagy flux. It was reported that the impairment of autophagy flux can result in cell damage. It was found that transcriptional factor TFEB regulation can reduce the level of ROS and promote the activities of lysosome and autophagosome, consequently, repair autophagy flux. Besides, our preliminary data showed that the activity of TFEB was significantly enhanced in I/R injury of skin flap. Therefore, we will regulate TFEB activity to promote the repair of autophagy flux, and further alleviate I/R injury of skin flap. Furthermore, we will explore whether the activity of TFEB can be regulated by the PARP1-LKB1-AMPK-mTOR signaling pathway. Finally, we will demonstrate the protection of TFEB induced autophagy and the PARP1-LKB1-AMPK-mTOR signaling pathway for TFEB activity regulation. This study is to elucidate a new mechanism of autophagy flux blockade in I/R injury of skin flap， and to provide a strategy of taking the regulation of TFEB activity to repair autophagy flux as a target to alleviate I/R injury of skin flap.