高压氧介导FUNDC1去磷酸化调控小胶质细胞线粒体自噬的镇痛机制研究

Neuropathic pain is one of the thorny problems in chronic pain which demands to be solved. Hyperbaric oxygen(HBO) can alleviate neuropathic pain as a noninvasive method. But analgesic mechanism of HBO is not clear. Our previous experiments showed the analgesic effect of HBO was associated with mitophagy of microglia and FUNDC1 receptor. From point of mitophagy of microglia, we elluminate the mechanism of HBO on regulating mitophagy of microglia, production of ROS and pain threshold by contrast mitophagy of microglia, level of ROS and pain scores at different time points. At the same time we explore whether HBO regulates mitophagy through FUNDC1 receptor desphosphorylation by establishing mutant of FUNDC1 on Ser13 and Tyr18 sites. Finallly we clarify the regulatory mechanism of LC3 and FUNDC1 receptor desphosphorylation through Co-IP methods. Our study elluminate the analgesic mechanism of HBO on microglia mitophagy. It can provide new ideas for the clinical treatment in the future.

神经病理性疼痛是慢性疼痛中急需解决的问题之一。高压氧（HBO）作为一种无创的治疗手段，可有效缓解神经病理性疼痛。但HBO镇痛的分子机制不清，我们前期实验提示HBO的镇痛作用可能与小胶质细胞线粒体自噬及FUNDC1受体去磷酸化有关。本课题以小胶质细胞线粒体自噬为切入点，通过对比不同时间点下小胶质细胞线粒体自噬的相关指标与活性氧（ROS）表达水平及大鼠疼痛行为学评分之间的变化关系，明确HBO通过调控小胶质细胞线粒体自噬影响ROS产生及疼痛阈值；同时通过构建FUNDC1在Ser13与Tyr18位点的突变体，探讨FUNDC1去磷酸化在小胶质细胞线粒体自噬中的作用；最后运用免疫共沉淀的方法检测LC3与正常FUNDC1蛋白及其突变体之间的相互作用，阐明HBO影响去磷酸化的FUNDC1与LC3之间的相互作用。本项目提出并明确HBO抗神经病理性疼痛的线粒体自噬机制，为神经病理性疼痛的治疗提供新思路。

神经病理性疼痛是慢性疼痛中的棘手问题。高压氧是一种无创的可有效缓解神经病理性疼痛的治疗手段。目前，HBO镇痛的分子机制不清，本课题以小胶质细胞线粒体自噬为切入点，通过对比不同时间点下小胶质细胞线粒体自噬的相关指标与活性氧（ROS）表达水平及大鼠疼痛行为学评分之间的变化关系，明确HBO通过调控小胶质细胞线粒体自噬影响ROS生及疼痛阈值；同时探讨FUNDC1在小胶质细胞线粒体自噬中的作用及作用机制，证实DNA甲基化可能与之相关；最后通过探讨LC3与正常FUNDC1蛋白之间的相互作用，阐明HBO、FUNDC1与LC3之间的相互作用。本项目提出并明确HBO抗神经病理性疼痛的线粒体自噬机制，为神经病理性疼痛的治疗提供新思路。