基于代谢组学的系统整合模式挖掘桂枝汤的药性配伍规律和性效关联的分子机制

This project is put forward based on the new discovery in the process of the project implementation. Herbs compatibility is an important part of herbal compatibility theory, and Guizhi Decoction is the representative of classical prescriptions. In this prescription, the strategy that co-application of sour and sweet herbs to produce Yin fluid, pungent and sweet herbs to produce Yang energy, as well as the co-application of cold and heat herbs is demonstrated. Guizhi Decoction was thus prescribed extensively in the clinical practice of TCM due to its rationality and efficency. To uncover the molecular mechanisms of herbal natures and the scientific basis of correlation between the nature and their effect of Guizhi Decoction, this project intends to introduce the system integration concept, with the different combinations of the herbs within Guizhi Decoction as as the research object. After the evaluation for natures with the mode of three-in-one, the different combinations of the herbs within Guizhi Decoction will be further studied on their main effects such as antipyretic, anti-inflammatory, analgesic, sweating, and so forth. At the same time, metabonomics, together with principal component analysis, cluster analysis, and partial least squares method will be applied to analyze changes of the endogenous metabolites and metabolic pathways, with the aim to uncover the compatibility regularity and the link between nature and effect the Guizhi Decoction, and to further enrich and improve the theory of traditional Chinese medicine and traditional Chinese medicine compatibility theory. This study will also provides new research ideas and data accumulation for understanding the scientific connotation of traditional Chinese medicine theory.

本项目是基于前期项目实施过程中的新发现提出的。药性配伍是中医配伍理论的重要组成部分，桂枝汤是经方的代表性方剂，其组方寓有酸甘化阴、辛甘化阳、寒热同用等多种基于药性理论的配伍组合，方义精深、疗效确切、应用广泛。为深入揭示桂枝汤药性配伍的科学基础和性效关联的分子机制，本项目拟引入系统整合理念，以桂枝汤方中药味不同组合模式为研究对象，在三位一体的药性评测基础上，分别对接解热、抗炎、止痛、发汗等桂枝汤最常用临床主效应，同时引入代谢组学技术，基于主成分分析、聚类分析、偏最小二乘法等化学计量学手段，以内源性代谢产物和代谢通路的变化为纽带，揭示桂枝汤方中药味之间的性效关联关系和关联特征，明确桂枝汤辛甘化阳、酸甘化阴等药性配伍组合的的科学本质，进一步丰富和完善中药药性理论和中医配伍理论，为全面解析中医药理论的科学内涵提供新的研究思路和数据积累。

本项目分别基于发汗、解热等环节对桂枝汤的药理作用以及桂枝汤配伍组方机理进行了研究，同时采用代谢组学技术方法探索了相应的分子机制。研究结果显示：桂枝汤桂枝汤全方的解热发汗作用优于方中的桂枝甘草或芍药甘草组合以及方中的单味药。桂枝汤组与对照组的差异内源性代谢物为stearic acid、L-Lactic acid、Glycerol、Palmitic acid、Linoleic acid、D-Glucose、Urea、Arachidonic acid、L-Threonine；桂枝的差异代谢物为Arachidonic acid、Palmitic acid、Linoleic acid、stearic acid、Cholesterol；白芍的差异物为L-Lactic acid、Palmitic acid、Linoleic acid、L-Serine、Oxalic acid、Urea、Arachidonic acid；甘草组的差异物为L-Lactic acid、Palmitic acid、D-Glucose、Urea、Arachidonic acid；大枣的差异代谢物为L-Lactic acid、Palmitic acid、D-Glucose、L-Serine、Oxalic acid、L-Threonine。桂枝汤发挥作用通路为亚油酸代谢、花生四烯酸代谢、淀粉和蔗糖的代谢以及甘油酯代谢；桂枝发挥作用的通路为亚油酸代谢和花生四烯酸代谢；白芍发挥作用的通路为亚油酸代谢通路、花生四烯酸代谢通路、甘氨酸，丝氨酸和苏氨酸的代谢通路和氨酰基-tRNA的生物合成；甘草发挥作用的通路为淀粉和蔗糖的代谢和花生四烯酸代谢；大枣发挥作用的通路为淀粉和蔗糖代谢及甘氨酸、丝氨酸和苏氨酸的代谢以及氨酰基-tRNA的生物合成。模块化分析发现桂枝汤发挥作用的靶点共有28个，其中26个属于细胞色素P450家族，其余两个靶点与糖酵解有关；桂枝、白芍、甘草作用的靶点属于桂枝汤细胞色素P450靶点；大枣发挥作用的靶点共有14个，其中PPT1、PPT2为棕榈酰蛋白质硫酯酶，SHMT1、SHMT2为丝氨酸羟甲基转移酶，其余靶点均与糖类代谢有关。本项目研究为桂枝汤的配伍应用的合理性和科学性提供了新的实验依据。