甘草-附子“相杀”解毒动力学的量化表征

Chinese medicine practitioners often prescribe a combination of medicinal herbs for the treatment of diseases. The composition of the traditional Chinese medicine (TCM) formula is basically guided by the traditional compatibility，named “Seven Relation”. Even though TCM formula have demonstrated usefulness in treating patients, the compatibility guideline, especially the dose-effect relationship of Chinese formula, unfortunately, lacks the empirical interpretation of modern medicine. Therefore, the proof-of-principle studies of the compatibility on the basis of internal relationship among herbs, effects, and time in TCM formula will have scientific significance for enhancing the modern interpretation of the Laws. Studies have revealed that aconitine and glycyrrhetinic acid are the principal bioactive chemicals in Fuzi (Aconitum carmichaeli Radix) and Gancao (Glycyrrhizae Radix), respectively. Our previous studies demonstrated that aconitine and glycyrrhetinic acid possess different time-concentration relationship. Furthermore, pharmacokinetic behaviors of aconitine and glycyrrhetinic acid were both changed in combination compared with that used alone. The concentration change of biochemical component contents is a key player of the traditional compatibility laws. These lead to the hypothesis that the combination of Fuzi and Gancao guided by the inherent guidline between concentration and effect changes. To prove our hypothesis, we will model the pharmacokinetics and pharmacodynamics for TCM formula, named chromatopharmacodynamics. This mathermatical model based on Hill dose-effect relationship reflects the inherent law between TCM formula (multi-components) and their complicated effects (multi-targets). In this model, the area of fingerprint chromatogram of the medicinal herbs is dose, and Michaelis-Menten dynamics is used to associate with the herbs dose and the effects. The experimental verification will carry out using Fuzi and Gancao alone or in combination, which includes pharmacokinetic and pharmacodynamic studies of Fuzi, Gancao and Fuzi-Gancao combination. Through these studies we will obtain the data of the pharmacokinetic parameters of monomeric compounds and calculate effect coefficients, spectral effect coefficients, chromatopharmacokinetic parameters and chromatopharmacodynamic parameters by the mathematical model. This study will combine molecular biology and meta analysis techniques to build a research platform of pharmacokinetics of decreasing toxicity by compatibility. The study results will provide empirical evidence to support the mechanism of mutual restraint compatibility of Fuzi and Gancao, and lay a substantial foundation for the kinetic study methodology of TCM formula. Finally, the study will supply a new approach for quantitative research of toxic Chinese medicinals.

附子刚烈迅捷有大毒，临床常根据“相杀/相畏”理论与甘草配伍。虽然从成分结合、机体状态、汤液相态与毒效作用的关系等开展了大量研究，至今解毒的定量规律仍待阐明。药动学参数可量化反映体内过程特点，故有毒中药和/或毒性成分（组分）的药动学是方剂解毒配伍定量研究的一个切入点。课题组前期研究发现：附子甘草配伍使活性成分的吸收减少、变慢，消除加快，故认为“相杀”物质基础可能是甘草减少附子的吸收，加速其从机体消除；但没有参数可直观表征附子毒性与甘草剂量配比及吸收、消除程度的关系和规律。本项目从整合中药复方药动学和药效学的角度，结合生物学实验和数学建模技术，与复方指纹图谱定性定量分析相对接，以甘草-附子“相杀”为研究对象，用Hill量效关系和Michaelis-Menten动力学方程构建中药解毒配伍定量研究平台，测算甘草-附子“相杀”的动力学参数并加以验证，为有毒中药的配伍解毒定量研究提供新的思路。

附子刚烈迅捷有大毒，临床常根据“相杀/相畏”理论与甘草配伍。虽然从成分结合、机体状态、汤液相态与毒效作用的关系等开展了大量研究，至今解毒的定量规律仍待阐明。本项目以甘草-附子“相杀”为研究对象，从整合该药对药动学和药效学的角度出发，结合生物学实验和数学建模技术，与复方指纹图谱定性定量分析相对接，以定量表征甘草-附子“相杀”的药物-效应-时间三者的内在联系。项目组在研究时限内围绕附子甘草配伍、解毒研究概况，附子甘草体外、体内指纹图谱，中药复方药物效应动力学数学模型，以及附子甘草配伍的药效学五个方面开展了研究。主要的研究结果有：①用Hill量效关系和Michaelis-Menten动力学方程构建中药解毒配伍定量研究平台；②用网络药理学测算出附子温阳机制与影响前列腺素和白三烯代谢通路有关；③总结归纳张仲景在《伤寒论》和《金匮要略》中发挥附子不同程度的温阳作用的用法规律；④从化学、药动学和代谢组学等方面概括和评述甘草减缓有毒中药/毒性成分的作用、作用特点和机制的研究；⑤完成了附子甘草合煎液与混合液（合并液）的物质基础及其比较研究，发现附子甘草合煎液与混合液的指纹图谱总量统计矩参数有差异；⑥比较了附子单煎液、附子甘草合煎液与混合液（合并液）对离体蛙心和离体大鼠衰竭心脏的心功能的影响，发现对于衰竭离体蛙心和大鼠心脏，与附子单用和与甘草煎液简单混合比较，附子在低剂量水平配伍甘草后即可发挥明显的强心作用；⑦从超分子“印迹模板”的角度探讨建立适用于分析多成分中药的毒与效研究方法的可行性。本项目的研究为提供中药配伍定量研究新思路，运用二维特征变量曲线统计矩原理，构建了中药小复方总量统计矩PK-PD数学模型；在离体器官水平定量比较了附子单煎液、附子甘草合煎液与混合液对衰竭心脏心功能的影响，为今后进一步深入研究七情配伍，如甘草在相杀、相反中的科学内涵，探索其定量规律奠定了基础。