疏肝解郁方中核心药对柴胡-白芍散收相合协同抗抑郁的配伍机制研究

Radix Bupleuri - Radix Paeoniae Alba (Chaihu-Baishao) herb pair are basic herbs of many famous shuganjieyu prescriptions such as Xiaoyaosan, Chaihu-shugan-san, Sinisan. Radix Bupleuri, Xinsan, Shuganjieyu, and Radix Paeoniae Alba, Suanshou, Yangxuerougan, they are central herbs of shuganjieyu and tiaochangqingzhi prescriptions. However, modern scientific connotation of importance of Chaihu- Baishao in prescription and composition rules of Chaihu-Baishao herb pairs are unclear. Our previous study results also show that Radix Bupleuri and Radix Paeoniae Alba play main role in antidepressant effect of Xiaoyaosan. On the basis of previous study, the project combind metabonomics pharmacokinetics, and network technology to study overall impact of components both to each other in vitro and in vivo，influence of metabolic characteristics of the active ingredients each other and the differences of significant metabolites and target regulated by herb pair and single herb in depression animal model rats, which clarify the chemical and pharmacological mechanism of compatibility. On this basis, we select Xiaoyaosan as a representative of shuganjieyu prescription, compare the chemical components and mechanism of herb pair and prescription, combined with the results of the drug compatibility mechanism to clarify the antidepressant mechanism of the herb pair in the Xiaoyaosan. The completion of this project, not only can reveal Chaihu-Baishao compatibility mechanism in antidepressant effect, but also can provide scientific basis for clarifying composition rules of shuganjieyu prescription based on Chaihu–Baishao herb pair, at the same time, which can provide ideas and methods to confirm the scientificity of Clinical medicine of Traditional Chinese Medicine.

柴胡-白芍系多个疏肝解郁名方如逍遥散、柴胡疏肝散、四逆散等的基础药物。柴胡辛散，疏肝解郁；白芍酸收，养血柔肝，二者散收相合，构成疏肝解郁、调畅情志类方剂的核心药对，现代研究表明此药对抗抑郁作用确切。然而，该药对配伍抗抑郁及其在疏肝解郁方中核心地位的现代科学内涵尚不清楚。本项目在课题组前期发现柴胡、白芍对逍遥散抗抑郁作用贡献较大的基础上，拟整合代谢组学、药代动力学和网络药理学技术对柴胡、白芍配伍前后体外、内化学成分、药代行为及作用机制进行系统研究，以阐明二者配伍的协同增效机制。在此基础上，以逍遥散为疏肝解郁方代表，将药对回归于复方，验证柴胡-白芍药对在复方抗抑郁作用的核心地位，并阐明该药对在逍遥散抗抑郁中的机制。本项目的完成，不仅可揭示柴胡-白芍散收药对协同抗抑郁的配伍机制，而且可为以柴胡-白芍药对为基础药物的疏肝方配伍规律的阐释提供科学依据，同时，为证实中医临床用药的科学性提供研究思路。

柴胡辛散，疏肝解郁；白芍酸收，养血柔肝，二者散收相合，构成多个疏肝解郁、调畅情志类方剂的核心药对。因此，阐明该药对在疏肝解郁方中的核心地位及二者配伍机制的现代科学内涵十分重要。本项目首先从药对对复方化学成分贡献和对抑郁模型动物内源性代谢物调节两个角度阐明该药对在疏肝解郁代表方逍遥散中的核心地位；其次，整合代谢组学和质谱背景扣除技术研究药对配伍前后体内外化学成分变化，发现柴胡、白芍配伍能显著促进柴胡中皂苷类原型成分在体内的代谢转化，促进白芍中主要原型成分在体内的吸收；同时，基于多成分药代动力学研究发现，配伍能提高柴胡中5种成分及白芍中6种成分在体内的生物利用度。此外，基于PK-PD相关性研究发现，柴胡及白芍配伍后生物利用度显著提高的成分与体内抗炎及神经保护代谢物呈显著正相关，提示柴胡-白芍配伍能增强神经保护能力及抗炎活性；其次，通过动物行为学、代谢组学发现，同等剂量下柴胡-白芍药对的抗抑郁作用显著优于柴胡、白芍单用药组。并应用网络药理学、代谢组学、分子生物学技术进一步阐明了药对发挥协同抗抑郁作用的机制与协同抑制前列腺素-过氧化物合成酶1、2的活性及调节抑郁大鼠花生四烯酸代谢紊乱有关，并发现柴胡、白芍配伍可通过抑制黄嘌呤脱氢酶转换为黄嘌呤氧化酶，进而抑制NLRP3炎症小体的激活，从而发挥抗抑郁作用。最后，运用多种协同效应计算模型，研究柴胡-白芍抗抑郁代表性成分（柴胡皂苷A、芍药内酯苷）配伍协同抗抑郁作用。并通过代谢组学及分子生物学技术，明确柴胡皂苷A可通过调节嘌呤代谢紊乱并进一步降低ROS水平、芍药内酯苷可通过调节谷氨酸-谷氨酰胺代谢紊乱、且两者可共同抑制 NLRP3 蛋白表达进而抑制由ROS所导致的炎症反应来发挥协同增效作用。综上，本项目整合代谢组学、药代动力学、网络药理学和分子生物学技术从体内外化学、体内过程、药理机制多角度明确了柴胡-白芍药对协同抗抑郁的配伍机制，也为证实中医临床用药的科学性提供研究思路。