基于类方体内成分分析的方剂配伍对中药成分选择性吸收影响的研究

Categorized formulae, a sereial of foumulae derived from the basic formula with the development of the syndrom, embody the essence of the principle of treatment with syndrome differentiation and personalized medicine in trational Chinese medicine science, and simultaneously are the further applicaiion of the theory of formula compatiblity. Therefore, studies on the change law of the absorbed compounds and metabolites from the core ingredient medicine in the process of the derivation of categorized formulae is of great significance to clarify the compatibility law of formula and guide the modern clinical medication. Ermiao Wan is a formula treating the syndrome of damp-heat invading lower energizer, and with the change of the syndrome Sanmiao Wan and Simiao Wan are composed on the base of Ermiao Wan, respectively. In the project, the aborbed compounds and metabolites from Cortex Phellodendri with the heat-clearing and damp-drying efficacy which is the core ingredient medicine of the categorized formulae will be researched on the base of the theory of serum pharmaco-chemistry of traditional Chinese medicine. The three formulae are adminitrated orally to rat, reapectively. The compounds in blood of rat are analyzed by utilizing the technology of LC-MS/MS, and then the same compounds and the different compounds are summarized to analyze the change of the number of the compounds in blood. The same compounds are further selected as monitor indexes to research pharmacokinetics of the three formulae. By comparing the pharmacokinetic parameters such as AUC, tmax, ka et al, the absoption characteristics of the same compounds in blood will be shown. Finally, by integrating the change of the number of the compounds in blood and pharmacokinetic parameters, the effects of compatibility conditions of Ermiao Wan categorized formulae on the selective absoption of the constituents in Cortex Phellodendri will be clarified. The research result will comment the internal change law of the in vivo compounds of the core ingredient medicine in the derivation process of formula with the change of syndrom, and further contribute to develop the theory of compatibility law of traditional Chinese medical formula-based modern personalized medicine.

方剂随证候衍变产生的一组类方体现了中医辩证施治和个体化用药的精髓，是方剂配伍理论的深入应用。因此，分析类方衍变过程中核心药物被吸收进入体内的成分及其量的的变化，对于深入阐明方剂配伍规律和指导现代临床用药具有重要意义。本课题以主治湿热下注证的二妙丸及衍生方三妙丸、四妙丸为一组类方作为研究对象，应用中药血清药物化学理论和液质联用技术，分析这三个方剂分别口服给予大鼠后，来源于其清热燥湿主要中药黄柏的血中移行成分，比较黄柏被选择性吸收成分数目的异同；进而以共有体内成分为监控指标研究三个方剂的多成分药代动力学，比较AUC、tmax、ka等药动参数的变化，揭示共有体内成分吸收特征的不同，最后综合二妙丸类方配伍对黄柏成分选择性吸收的数目及其吸收特征的影响，诠释方剂随证候不同而衍变成系列类方的过程中存在核心药物体内直接作用物质的内在变化规律，进一步为形成基于中医方剂配伍规律的现代个体化给药理论奠定基础。

类方的功能主治集中体现了其核心中药功效发挥的方向，这其中必然存在着该核心中药有效成分随着这一组方剂配伍而发生变化。基于中药血清药物化学理论，类方中核心中药有效成分的变化主要应体现在这一组方剂对核心中药有效成分选择性吸收及体内表达水平的影响。本课题以关黄柏为核心中药，分析其化学成分在二妙丸、三妙丸、四妙丸这一组类方配伍中的体内吸收和表达程度，从类方的共性角度揭示二妙丸类方配伍对关黄柏成分体内选择性吸收及量的变化的影响，阐明其配伍的科学内涵。.首先，建立关黄柏化学成分的UPLC-MS/MS分析方法，鉴定或表征了关黄柏61个成分，包括生物碱类成分、柠檬苦素类成分、有机酸类成分、脂肪酸类成分、黄酮类成分等，并对二妙丸类方化学成分进行了分析，表征了分别引入苍术、牛膝、薏苡仁的化学成分。在此基础上，进一步对关黄柏及二妙丸类方分别口服给予大鼠后的血中移行成分进行了分析，关黄柏单独给药后共筛选出31个血中移行成分。二妙丸类方血中移行成分分析发现，在二妙丸类方配伍环境下检测不到γ-hydroxybutenolide deniatives Ⅱ、Rutaevin，而均能够检测到关黄柏新的代谢物M15，体现了二妙丸类方配伍对关黄柏成分的选择性吸收。.关黄柏及二妙丸类方配伍环境下20个血中移行成分的体内经时变化过程分析发现，二妙丸类方配伍能够选择性吸收木兰碱进入体内，而抑制小檗碱、巴马汀、黄柏碱等传统认为是黄柏有效成分的物质吸收进入体内，提示木兰碱与关黄柏清热燥湿功效的高度相关性。另外，二妙丸类方配伍能够减少Ⅰ相代谢产物的生成，同时增加Ⅱ相代谢产物的产生，从而促进小檗碱、黄柏碱等原形入血成分的排泄，进而减少其体内表达水平，可能是方剂配伍影响关黄柏药效发挥方向的体内代谢机制。.本课题结果表明，中药饮片只是原料，方剂才是药物；中药含有的只是化学成分或活性成分，而在发挥其某一种功效的一组方剂中将表达不同的体内直接作用物质。本课题共发表文章8篇，其中SCI收录期刊文章7篇，国内核心期刊文章1篇。