四逆散调节慢性应激动物胆汁酸代谢的抗抑郁机制研究

Depression is a common mental disorder with high morbidity in modern society, however, its pathological mechanisms are elusive and clinical antidepressants are relatively insufficient. According to the understanding of depression in the theory of traditional Chinese medicine, the method of Shu-Gan-Jie-Yu is commonly used to treat depression in clinic. The applicant found liver-brain inflammation axis was activated in chronic unpredictable mild stress (CUMS) depressive animals in previous work, yet the underlying mechanisms mediating liver and brain inflammation were little unknown. This project proposes a hypothesis based on prior work and literature research, that the altered bile acid metabolism may be key pathological events mediating the liver-brain inflammation axis activation in depressive animals and Shu-Gan-Jie-Yu classic formula Si-Ni-San has anti-depressant effects through regulating bile acid metabolism. This project will focus on analyzing and defining the altered serum bile acid profile in CUMS mice, then explore the molecular mechanisms of specific altered bile acids mediating central neuroinflamamtion. It will also mainly study the regulatory effects of Si-Ni-San on bile acid metabolism in CUMS animals, and demonstrate the anti-depressant pharmacological mechanisms of Si-Ni-San by regulating bile acid metabolism thus alleviating liver-brain inflammation axis activation. The findings of this project will provide basic research evidence of liver-smoothing traditional Chinese medicine such as Si-Ni-San for clinical prevention and treatment of depression, and provide new strategies and targets for developing new antidepressants.

抑郁症是现代社会高发疾病，其病理机制尚不明确且临床药物相对匮乏。中医依据对抑郁症的辨证，临床实践多用疏肝解郁法治疗抑郁症。申请人前期工作也发现慢性不可预计温和应激（CUMS）抑郁症模型动物肝-脑炎性轴异常活化的病理特征，但对介导肝-脑炎性联动的机制还有待解读。本项目基于前期工作基础与文献调研提出假说：胆汁酸代谢异常可能作为关键环节介导抑郁症模型动物肝-脑炎性轴异常活化；疏肝解郁经典方四逆散则可通过调节胆汁酸代谢而发挥抗抑郁作用。本项目拟在分析以明确CUMS小鼠血胆汁酸成分变化的基础上，探究特定胆汁酸成分改变引起中枢神经炎症的分子机制；重点研究四逆散对CUMS动物胆汁酸代谢的调控作用，阐明其通过调节胆汁酸代谢进而抑制肝-脑炎性轴活化的抗抑郁药理机制。本项目成果为四逆散等疏肝类中药临床防治抑郁症提供基础研究证据支撑的同时，也为开发新型抗抑郁药物提供新策略和新靶标。

抑郁症是现代社会高发疾病，其病理机制尚不明确且临床治疗药物相对匮乏。疏肝解郁经典方四逆散在临床上可有效治疗抑郁症但亟需基础研究推动。本研究基于前期工作基础，提出四逆散通过调节胆汁酸代谢发挥抗抑郁作用的工作假说。首先，对四逆散水提物成分进行定性分析，以四逆散水提物实测成分开展其抗抑郁作用的网络药理学研究为先导。其次，开展四逆散成分定量分析及入血成分定性分析工作。四逆散水提物成分定性分析共计鉴定119个化合物，主要含黄酮类和三萜皂苷类化合物。本研究建立了一种同时检测四逆散中没食子酸、芍药内酯苷、芍药苷、橙皮苷、槲皮素、甘草酸、柠檬苦素和川陈皮素8种成分的高效液相色谱（HPLC）定量分析方法，其中芍药苷含量最高。四逆散入血成分主要包括没食子酸、柠檬苦素、橙皮苷、柚皮苷、芍药苷和芍药内酯苷等，进一步明确了四逆散抗抑郁药效物质基础。最后，按计划完成动物模型的实证研究，建立血浆胆汁酸靶标代谢组学检测方法，证实慢性不可预计温和应激（CUMS）大鼠血浆胆汁酸代谢异常，如鹅脱氧胆酸（CDCA）、甘氨鹅脱氧胆酸（GCDCA）、甘氨脱氧胆酸（GDCA）、甘氨胆酸（GCA）含量显著增加。CUMS大鼠还呈现以肝脏和前额叶皮层核转录因子-κB（NF-κB）/NOD样受体蛋白3（NLRP3）炎症小体激活为主的炎症反应。本研究重点聚焦胆汁酸代谢异常所引起的肝-脑炎性轴活化，以阐明四逆散调节胆汁酸代谢发挥抗抑郁作用的分子机制和作用特色，为四逆散及其有效成分临床防治抑郁症提供一定实验依据。