基于药物相互作用的香附气血药对“相使”和合的组成结构与协同增效机理研究

In the previous study of active material basis of Xiang - Fu - Si - Wu Decoction for the treatment of dysmenorrhea with syndromes of Qi stagnation and blood stasis, it was found that Qi - Blood drug pairs, which Xiangfu acted as an indispensable part and aimed at regulating Qi and blood, played a key role in revealing the composition and compatibility of this complex formula. In details, the drug pairs of Xiangfu - Danggui, Xiangfu - Chuanxiong, Xiangfu - Baishao and Xiangfu - Yanhusuo all had synergetic effect for promoting Qi circulation, relieving pain, nourishing blood and invigorating the circulation of blood. Based on drug interaction theories, the composition structure and synergistic mechanisms of Xiangfu Qi - Blood drug pairs for the mutual assistance will be further elucidated in current proposed study. Firstly, the quantitative and qualitative changes of the components in compatible drug pairs will be analyzed by modern spectroscopy tandem technology and multivariate statistical analysis. The appropriate methods for evaluating regulation of Qi and blood will be established, and the variation tendency of changed effect caused by compatibility will be further elucidated. The ADME process of main active components will be studied by Caco-2 cells, liver microsomal P450 enzymes and whole animal models. Besides, the multiple characters of the mutual assistance compatibility effect will also be analyzed. Secondly, the composition structure characters of Xiangfu Qi - Blood drug pairs for the mutual assistance compatibility will be analyzed via the equivalent line method, response surface method and so on. Finally, the synergistic mechanism of the drug pairs will be explored by metabolomics, proteomics and molecular biology approaches. The final objects of these works are to clarify the effect and structure characters of Xiangfu Qi - Blood drug pairs, and to reveal the scientific mechanisms of the mutual assistance compatibility, which will be helpful for improving effective clinical usage of traditional Chinese medicine.

在围绕治疗气滞血瘀型痛经的香附四物汤功效物质基础前期研究过程中，发现香附气血药对对揭示方剂组成及配伍规律起关键作用，香附-当归、香附-川芎、香附-白芍、香附-延胡索药对对行气止痛、养血活血有协同增效作用。据此，本项目提出基于药物相互作用的理论，开展香附气血药对“相使”和合的组成结构与协同增效机理研究，拟开展以下工作：（1）运用现代谱学串联技术，多元统计分析配伍成分量与质的变化；采用适宜的理气理血生物效应评价方法，阐明配伍效应变化趋势；采用Caco-2细胞、肝微粒体P450酶、整体动物模型研究主要效应成分的ADME过程，分析配伍功效物质多元性特点。（2）采用等效线、响应曲面法等量效关系的评价方法，分析药对配伍量效特点。（3）采用代谢组学、蛋白质组学和细胞分子生物学方法探讨药对协同增效内在机制。最终阐明香附气血药对配伍应用的结构特点，揭示其“相使”和合组成的科学内涵，为提高临床用药有效性参考。

本项目基于药物相互作用的理论，开展了香附气血药对“相使”和合的组成结构与协同增效机理研究。项目主要完成研究工作及阶段性成果如下：1. 采用现代各种谱学串联技术，对香附-当归、香附-川芎、香附-白芍、香附-延胡索药对配伍前后体外效应物质溶出规律进行分析，并建立数学模型，发现香附系列药对不同制法与不同配比的配伍化学物质变化规律有所不同，且存在优化配比，从配伍理化改变阐明了香附气血药对药物相互作用的特点。2. 采用适宜的理气理血生物效应评价方法，对香附药对配伍效应进行分析，发现香附气血药对对血虚证及机体免疫确实有改善作用，香附-当归药对的补血作用和对脾脏的修复作用较强，但香附四物汤对机体免疫细胞的恢复比各药对较好。3. 采用代谢组学、蛋白质组学和细胞分子生物学方法分析香附-当归、香附-川芎、香附-白芍、香附-延胡索药对与气血并调效应相关的生物标志物和关键作用通路，发现香附气血药对调节甘油磷脂代谢、类固醇激素生物合成和花生四烯酸代谢途径。香附-当归药对对血瘀的调节作用更明显。各药对干预蛋白涉及细胞过程比例较高。4. 采用肝脏微粒体P450酶代谢模型分析香附-当归、香附-川芎、香附-白芍、香附-延胡索药对配伍前后主要效应成分体内代谢相互作用，发现不同配伍配比的药对对肝药酶的抑制作用不同，存在最佳配伍比例。研究初步阐明了香附气血药对配伍应用的结构特点，揭示了其“相使”和合组成的科学内涵，为临床合理用药提供了参考。.研究成果在国内外学术刊物上发表论文8篇，其中SCI源期刊收录5篇；申请国家发明专利1项；研究成果获中华中医药学会科技进步一等奖。