桑源中药客体特征及“同源异效”的超分子作用机理研究

The mechanism research on the variety classification of traditional Chinese medicines is an important basis for the standardization of traditional Chinese medicines. At present, there is still no identification standard about structural and quantitative perspective for the variety classification of Chinese medicines, which restricts the standardization process of traditional Chinese medicines. This project is aimed at this scientific bottleneck problem. The research on the mechanism of supramolecular action of “the same efficacy from heterologous traditional Chinese medicine” of Lonicera japonica and Lonicerae flos was the base, which was that the different chemical components in the Lonicera japonica and Lonicerae flos in vivo had the similar "imprint template" to the similar target molecule and it had a similar supramolecular biological efficacy. Then the hypothesis was that “the phenomenon of ‘different efficacy from homologous traditional Chinese medicine ’was determined by the degree of difference in the 'imprint template' in vivo”. It was proposed to use Morus alba L. as a model drug to reveal the guest components and metabolites characteristics of "imprint template"、the relationship between the the guest components and metabolites characteristics of "imprint template" and efficacy、the corresponding relationship between the degree of dissimilarity of the “imprinted template” and the different efficacy. Combined with “the same efficacy from heterologous traditional Chinese medicine” research, the mechanism of supramolecular action of “different efficacy from homologous traditional Chinese medicine” would be clarified and a new method for studying the characteristics of supramolecular "imprinted templates" variety classification of traditional Chinese medicines would be established.

中药入药品种归属分类的科学机理研究是中药标准化的重要基础。目前中药入药品种归属问题从结构和量化角度仍没有鉴定标准，这制约了中药的标准化进程。本项目针对这一科学瓶颈问题，在前期金银花山银花的“异源同效”中药超分子作用机理研究的基础上，基于金银花山银花中不同的化学成分入体后作用于靶点分子的“印迹模板”相似，具有相似的超分子生物效应，提出“中药‘同源异效’现象是由其入体后的代谢产物的客体‘印迹模板’差异程度决定”的假说。拟以桑源中药为模型药物，通过桑源中药客体成分及其代谢产物的“印迹模板”特征、“印迹模板”特征与功效的关联性研究，揭示桑源中药客体成分及代谢产物的“印迹模板”相异程度与“异效”的对应关系，阐明中药“同源异效”现象的超分子作用机理，结合前期“异源同效”研究，建立中药入药品种归属的超分子“印迹模板”特征研究的新方法。

本项目基于中医药超分子“气析”理论，在前期金银花山银花“异源同效”中药超分子作用机理研究的基础上，对“同源异效”桑源中药的超分子“印迹模板”相异程度与“异效”的对应关系进行研究。运用UPLC测定桑枝、桑叶、桑白皮及桑椹4种桑源药材中主要化学成分的含量；运用段带总量统计矩法和信息熵对桑源药材HPLC指纹图谱进行分析，4种桑源药材指纹图谱AUCT、MCRTT、VCRTT、H值成对配对t检验P均小于0.05，具有显著性差异，4味桑源药材均在3-11，20-24min同时有吸收峰，说明4味桑源药材在上述相同时间段有相同或相似的化学成分，在保留时间3-4、16-18、25-27、31-32min时VCRTT值均较大，说明4味桑源药材间的成分在这些时间段内存在显著差异；建立桑源药材化学成分数据库，计算桑枝、桑叶、桑白皮、桑椹黄酮类成分0-8阶MCI平均值分别为4.57，4.59，6.41，4.24，生物碱类成分0-8阶MCI平均值分别为2.65，4.55，2.58，2.78，0-8阶物芯指数（CI）平均值分别为5.51，5.49，5.44，2.88，建立中药成分（群）MCI和CI与药效相关联的方法，从“点”（单味药各成分MCI的计算）到“面”（单味药整体CI的计算）静态定量表征药材成分（群）的结构特点和构效关系；通过色谱-质谱技术分析鉴定得到桑源药材的41种活性成分，联合网络药理学和分子对接技术比较分析桑源药材不同部位改善胰岛素抵抗(IR)物质基础；通过体外印迹实验与体内药效学评价研究比较了“异效”的程度。通过桑源中药超分子“印迹模板”特征、“印迹模板”特征与功效的关联性研究，揭示了桑源中药超分子“印迹模板”相异程度与“异效”的对应关系，建立了“构-效-印”的中药入药品种归属的超分子“印迹模板”特征研究的新方法。