基于脑功能网络及色氨酸代谢微环境分析逍遥散防治肝郁证的分子调控机制

As a research hotspot in recent years, the mechanismand syndrome connotation of liver stagnation syndrome have not yet been fully clear. Xiao yao San as a classic prescription for traditional chinese medicine to regulate emotional diseases, It has a unique advantage in the prevention and treatment of liver stagnation syndrome. Clinical trials have confirmed that Xiao yao San could block and reverseliver stagnation syndrome, but its regulatory mechanism and efficacy targets are still not fully elucidated. Based on our previous studies show that the incidence of liver stagnation syndromeare related to the brain functional BOLD signal disorder, accompanied by liver, brain, multi-organ tryptophan metabolism microenvironment imbalance. Thus, we hypothesis that Xiao yao San can regulate the homeostasis of brain functional network by mediating homeostasis of tryptophan metabolism in multi-systems of the liver , brain, etc. and achieve the goal of prevention and treatment of liver stagnation syndrome. The project intends to take brain functional analysis and tryptophan/ kynurenine metabolic microenvironment balance as a starting point, combined with resting state fMRI technology, metabolomics, genome, transcriptome and knockout mice platform, andinvestigate the intrinsic mechanism of liver stagnation syndrome and Xiao yao san treatment mechanism with multi-level, multi-target approach. To further clarify the pathogenesis of liver stagnation syndrome and target regulation mechanism andinvestigate the modern molecular biological mechanism and role of Xiao yao San for the treatment, we could better search for new molecular targets for the effective prevention and regulation of liver stagnation syndrome and provide new ideas for drug discoveries.

肝郁证作为近几年的研究热点，其形成机制及证侯内涵至今尚不完全清楚。逍遥散作为中医药调节情志疾病的经典方剂，在防治肝郁证领域具有独特的优势。临床试验证实逍遥散对肝郁证具有阻断和逆转作用，但其调控机制与药效靶标尚未阐明。基于我们前期的研究表明,肝郁证的发病与脑功能BOLD信号的紊乱有关，同时伴随肝、脑多器官色氨酸代谢微环境失衡。由此，我们设想逍遥散可通过介导肝脑多系统色氨酸代谢动态平衡，从而调控脑功能网络稳态，达到防治肝郁证的作用。本项目拟以脑功能分析和色氨酸/犬尿氨酸代谢微环境平衡作为切入点，结合静息态fMRI技术、代谢组学、基因组、转录组分析以及敲基因小鼠平台，多层次、多靶点分析肝郁证的内在本质以及逍遥散治疗机制。阐明肝郁证的发病基础及靶标调控机制，同时解答逍遥散治疗肝郁证的现代分子生物机制及作用基础，为肝郁证的有效预防及调控寻找新的分子靶点，并为药物开发提供新的思路。

肝郁证作为情志疾病的研究热点，其形成机制及证侯内涵尚不完全清楚。逍遥散作为中医药调节情志疾病的经典方剂，在防治肝郁证领域具有独特的优势。临床试验表明逍遥散对肝郁证具有阻断和逆转作用，但其调控机制与药效靶标尚未阐明。本课题组前期研究表明，肝郁证的发病与脑功能血氧水平依赖（BOLD）信号的紊乱有关，同时伴随肝、脑多器官色氨酸代谢微环境失衡。. 本项目以脑功能分析和犬尿氨酸/色氨酸（KYN/TRP）代谢微环境平衡作为切入点，结合静息态fMRI技术、组学分析等，多层次、多靶点分析肝郁证的内在本质以及逍遥散治疗机制。探讨了吲哚胺2，3-双加氧酶1（IDO1）干预色氨酸代谢，调节小胶质细胞功能参与调控肝郁证神经再生、可塑性及脑功能连接的作用机制并验证了逍遥散介导IDO1干预肝郁证色氨酸代谢微环境及脑功能连接的调控作用和分子机制。. 结果表明慢性温和不可预知应激（CUMS）模型小鼠海马IDO1表达水平、KYN/TRP均显著升高且伴随着小胶质细胞激活，大脑功能连接尤其是海马与边缘系统和基底神经节等脑区之间的功能连接减弱，从而导致皮质中脑边缘奖赏回路功能障碍。此外，肝郁小鼠中存在小鼠海马、杏仁核等边缘系统脑区和下丘脑、黑质基底等基底节脑区的神经元活动协同一致性的改变，以及下丘脑、海马、内嗅皮层和听觉皮层等脑区的神经元活动变化。进一步利用IDO1敲基因小鼠建立CUMS肝郁证及LPS诱导的急性炎症肝郁证模型，IDO1缺失能够改善CUMS及LPS诱导的肝郁样行为，减轻CUMS肝郁小鼠海马脑区功能连接，改善边缘系统脑区及基底节脑区的神经元活动。逍遥散通过降低肝郁证小鼠海马IDO1的表达，调节色氨酸代谢微环境，抑制小胶质细胞激活，减轻神经炎症，改善神经分化、再生及可塑性。. 本研究通过探讨逍遥散防治肝郁证发生发展的作用，明确了IDO1参与的色氨酸代谢微环境及小胶质细胞功能和神经可塑性对肝郁证调控的关键作用和分子机制，为肝郁证有效预防及调控寻找了新的分子靶点，同时为肝郁证防治药物开发提供新的思路。