基于糖皮质激素代谢组学研究客观化诠释柴胡疏肝散“疏肝解郁”的作用机制

The "Relieving the Depressed Liver" law is the basic principle to relieve the liver qi stagnation types depression from the point of view of Traditional Chinese Medicine(TCM) theory, literature and clinical practice.Chaihu-Shugan-San (CHSGS), a typical prescription,shows the significant clinical effect to reduce depression, based on soothing the liver and regulating mind. However, the modern medical connotation of CHSGS in the treatment of patients with depression is unclear. Many scholars have conducted much fruitful research around the monoamine neurotransmitter hypothesis,neurotrophic factor hypothesis and MAPKs signaling pathways. Otherwise, other scholars have been interesting to investigate treatment of depression through modulating the outer periphery of Glucocorticoid (GC) level - function recovery of HPA axis and structure and function of the central nervous system. The current research have shown that CHSGS effectively relieve depression by regulating corticosterone level in CUMS rat models. Besides, the previous metabolomic studies of the applicant have shown that the closely correlation between GC level and the clinical development of depresssion. As well as, the improvement of depression closely relates to outer glucocorticoids metabolism and neurotransmitters levels.In the proposed study,we intend to further reveal the changes of the GC metabolic profiling in the liver qi stagnation types depression animal models and depression patients in clinical before and after CHSGS treatment by GC metabolomics on the basis of preliminary studies. Various mechanisms such as GC metabolic profile, neurotransmitters concentration, metabolic enzymes and transporters will be complemented and verified. It will provide the solid foundation and new ideas to clarify the mechanism of CHSGS and modern medical connotation of "Relieving the Depressed Live" law.

疏肝解郁是中医治疗肝气郁结证的基本原则，其代表方剂柴胡疏肝散临床治疗肝气瘀滞证效果显著，但其疏肝作用的现代医学内涵不明。诸多学者围绕中枢单氨神经递质假说、神经营养因子假说和MARKs信号通路等对其进行了大量卓有成效的研究，另有学者则对通过外周调节糖皮质激素（GC）水平-恢复HPA轴功能-中枢结构功能恢复而治疗抑郁症进行了饶有兴趣的探索。现有研究表明柴胡疏肝散对抑郁模型大鼠体内皮质酮的含量有调控作用。本课题组前期通过代谢组学法研究证实外周GC水平的调节与抑郁症关系密切，抑郁症的转归与人体内GC代谢和神经递质的含量密切相关。本项目拟在前期研究基础上，以柴胡疏肝散为研究对象，通过代谢组学研究方法揭示在柴胡疏肝散的干预下，肝气郁结动物模型和临床患者体液中GC代谢谱的变化，以GC和神经递质浓度、代谢酶、转运体等多种机制相互补充与验证，为阐明柴胡疏肝散的作用机制和“疏肝”现代医学内涵提供依据和新思路。

疏肝解郁是中医治疗肝气郁结的基本原则，其代表方剂柴胡疏肝散临床治疗肝气瘀滞证效果显著，但其疏肝解郁的现代医学内涵不明。我们前期研究通过代谢组学研究证实外周GC水平的调节与抑郁症关系密切，抑郁症的转归与人体内GC代谢和神经递质的含量密切相关。因此，本项目在前期研究基础上，通过代谢组学研究方法揭示在柴胡疏肝散的干预下，肝气郁结动物模型外周、中枢GC和神经递质代谢谱的变化，并以代谢酶及其上游调控等多种机制相互补充验证，为阐明柴胡疏肝散的作用机制和物质基础提供了丰富的理论依据。.本研究中，1）首先通过制备柴胡疏肝散水提物冻干粉，对柴胡疏肝散水提物冻干粉的主成分进行测定,通过建立肝郁气滞大鼠模型，检测了动物血浆及海马、前额叶皮质中多种GC和神经递质的含量，发现了GC和神经递质差异代谢物，筛选出潜在的分子标记物；2）确证了疏肝解郁方柴胡疏肝散对调控GC及神经递质代谢酶（11βHSDs、IDO、TDO等）的影响，阐明了柴胡疏肝散及其活性成分柴胡皂苷D调节GC及神经递质在体内浓度水平，发挥抗抑郁作用的分子机制和物质基础；3）发现柴胡疏肝散及活性成分柴胡皂苷D对GC及神经递质代谢酶上游信号的调节；4) 对临床抑郁症患者血浆进行代谢组学检测，分析差异代谢物质; 5）探索了柴胡疏肝散对抑郁大鼠肠道菌种的影响，发现与氨基酸代谢有关的肠道菌群发生改变。.综上，柴胡疏肝散及其有效成分柴胡皂苷D可能通过调控外周和中枢的GC、神经递质代谢酶（11βHSDs、IDO、TDO等），纠正GC、神经递质等代谢紊乱，缓解HPA轴亢进状态，从而发挥其疏肝解郁的作用。此外，柴胡疏肝散及其有效成分柴胡皂苷D还可能通过调控GC、神经递质代谢酶上游通路信号，抑制NLRP3炎症小体激活，降低炎症因子IL-1β等水平，通过缓解神经炎症发挥抗抑郁作用。.以上研究从动物整体、蛋白分子、微生物等多个层面补充与验证，阐释了中药柴胡疏肝散疏肝解郁的客观本质，为其作用机制和疏肝解郁的现代医学内涵提供了依据，将有助于推进中医疏肝解郁方的现代应用。