基于"血清药化-药代动力学-代谢组学”的柴胡-白芍药对抗抑郁的药效物质及机制研究

Under the guidance of the theory of Traditional Chinese medicine(TCM),this paper established the method of simultaneous analysis of the multi-component in vivo to determine the direct action of the drug in the treatment of depression,in the early study of chemical composition of Bupleurum radices paeoniae alba in vitro and vivo and its therapeutic effect on depression,with the serum drug chemistry and drug metabolomics and multidimensional analysis methodology as the platform,based on the rat model of depression.Based on the metabolomics UPLC/ESI-HDMS technique, the contours of the target metabolite group were determined by using the method of principal component analysis and partial least squares discriminant analysis. The experimental data matrix was used to determine the biomarkers (Markers).In this passage,it explored and clarified the biological metabolic path,combined with advanced techniques to construct a multidimensional framework of pharmacological action,and verified the correlation between candidate Markers and direct acting substances in the body, and elucidated the effects of Pharmacodynamic substance basis and mechanism of Bupleurum radices paeoniae alba on the treatment of depression.Through the study of the relationship between the pharmacokinetics and compatibility of the main migratory components in the blood after treatment, the scientific connotation of the compatibility of Bupleurum adices paeoniae alba is established, which lays a useful theoretical foundation for the treatment of depression in Chinese medicine.

在中医药理论的指导下，在前期柴胡白芍药对体外体内化学成分研究及其治疗抑郁症药效学基础之上，以中药血清药物化学及药物代谢物组学及多维联用分析技术为核心方法学平台。基于大鼠抑郁症模型，建立药对体内多成分同时分析表征的方法，确定其治疗抑郁症的血液中体内直接作用物质；基于代谢组学UPLC/ESI-HDMS技术建立柴胡白芍药对治疗后的靶标代谢物组轮廓谱图，利用主成分分析、偏最小二乘判别分析等模式识别技术系统分析实验数据矩阵确定生物标记物（Markers）。发掘并明确其背后的生物代谢途径，结合生物信息学等先进的技术手段构建多元多维的药理作用框架全貌，验证候选Markers与体内直接作用物质的相关性，阐明柴胡白芍药对治疗抑郁症的药效物质基础和作用机制。通过药对治疗后的血液中主要移行成分药代动力学及配伍的关系研究，解析柴胡白芍药对配伍的科学内涵，为中医药治疗抑郁症奠定有益理论基础。

抑郁症是一种高发的精神类疾病，至今无有效的治疗措施。通过对抑郁症机制、中医抗抑郁理论、中药抗抑郁的研究。本研究应用代谢组学技术挖掘抑郁症血浆和尿液代谢标志物，有助于解决抑郁症尚缺客观诊断方法，有利于基础研究成果的临床推广；药物代谢动力学与代谢组学的整合研究，建立符合中药多组分、多靶点作用特征的PK/PD结合研究技术方法，在二者在分析技术方法上的相通性及研究对象上的互补性，同步揭示中药复杂体内药效物质基础与整体药效作用，并试图通过有效成分体内动力学过程与相关生物标记物群的整体量变关系，说明各成分的药效作用特点与其对整体药效作用的贡献度。应用UPLC-MS方法，及相关的文献进行对照分析柴胡-白芍药对的化学成分。鉴定出的柴胡-白芍药对化合物共40个，其中柴胡来源为9个，白芍来源为15个，未知成分16个。药对与单味药相对含量比较发现：柴胡皂苷a, c含量降低，芍药苷含量升高，说明两者配伍增加芍药苷溶出率，增强临床疗效。说明该药对配伍具有科学性。利用代谢组学技术探讨柴胡-白芍药对抗抑郁的作用机制。建立慢性不可预知温和应激抑郁（CUMS）大鼠模型，利用行为学测试评价柴胡-白芍药对的药效 ，通过UPLC-Q / TOF-MS技术，对尿液、血清进行代谢组学研究。结果显示柴胡-白芍药对能显著改善 CUMS 模型组大鼠的抑郁样行为，对 CUMS 模型大鼠代谢轮廓具有明显调节作用。其发挥抗抑郁机制涉及调节脑源性神经营养因子的表达，抑制神经毒性，调节HPA轴，增强解毒氨的能力以预防肝性脑病的发生和增强体内抗氧化能力。