枳壳质量标志物与“陈久者良”科学内涵的研究

Fructus aurantii (FA) is a kind of common tradition Chinese medicine (TCM) with regulating-qi effect. FA is used widely in clinic, and with the saying "Fructus aurantii, the older the better". But up to now, the material basis and the potential long-storing principle are unclear. We believe that the scientific connotations exist in the parlance. During the storing, some components change in quality and/or contents, then the biological effects and quality change with it. To study the long-storing theory, we should focus on the component changes, and elucidate the material basis based on the main biological effects. Firstly, various research methods from holistic thoughts were integrated into this project such as serum pharmacochemistry, spectrum-effect relationship, spectrum-pharmacokinetics, chemometrics, network pharmacology and PK-PD model; Composition identification, pharmacodynamic evaluation and multi-component pharmacokinetics were simultaneously investigated to relate and analyze “component-pharmacodynamic-pharmacokinetic”, and the regulating-qi/ dryness material basis and mechanism were comprehensively elucidated, then the Q-markers were confirmed. Secondly, the Q-marker quality control and evaluation method was used during the full long-storing process; and the process quality evaluations were performed to multi-fractions of FA with total components and multi-indexes; and the relationship between “component-effect-quality” were revealed; then the scientific connotations of "Fructus aurantii, the older the better" were analyzed and elaborated in detail. The research ideas and methods based on Q-marker were introduced into this project, and the researches on the material basis, quality standard and the long-storing theory were related effectively. The implementation of the project can provide a research paradigm for the following TCM research fields, such as material basis, quality standard and long-storing theory.

枳壳为常用理气中药，素有“陈久者良”之说，但其物质基础不明，陈用道理不清。我们认为：“陈久者良”有其科学内涵；陈化过程中，组分可能发生了变化，生物效应和药物质量随之改变；研究陈用理论宜紧扣组分变化，立足主要功效，弄清物质基础。项目首先整合血清药物化学、谱效学、谱动学、化学计量学、网络药理学、PK-PD模型等研究成果，对枳壳进行全成分辨识、生物效应评价和多组分药动学研究，关联“组-效-动”，明确枳壳理气功效/燥性的物质基础和作用机制，确定枳壳的质量标志物；继以基于Q-marker的质量评价方法为基础，针对陈化研究的时间特点，精心设计加速试验，进行多部位、全成分、多指标的陈化全过程质量评价，解码“组分-效应-质量”，阐述“陈久者良”科学内涵。项目引入Q-marker的研究思路与方法，有机关联了物质基础、质量标准和“陈久者良”内涵的研究，能为中药物质基础、质量标准和陈用理论的研究提供参考。

陈用理论属于中医药传统理论，历代本草均有论述，至今仍为中医临床所沿用。然而，中药是否“陈久者良”尚且存疑，其科学依据更少深究。中药陈用的现代研究踟蹰不前，关键在于缺乏有效研究方法，无法应对陈药研究的时间特点。枳壳为常用理气中药，素有“陈久者良”之说，但其物质基础不明，陈用道理不清。本项目整合谱-效学、血清药物化学、质量标志物、网络药理学、分子对接技术和加速试验等技术手段，提出并验证了系统的陈化研究方法，揭示了活性成分群的动态变化规律和生物效应变化特点，综合药材性状、组分、药效/燥性等指标，全面评价了陈化过程中的质量属性，系统剖析了枳壳“陈久者良”的科学内涵。.主要内容与结果：.1. 全成分分析与多组分比较药动学研究：建立了较为系统的枳壳化学成分谱库，将化学成分鉴别，血清药物化学和药代动力学研究相结合，层层递进，互相补充，基本讲清枳壳的药效物质基础。.2. 辩证分析了枳壳的理气/燥性物质基础：引入谱-效学思路，三元关联“谱-效-燥”，发现了20个理气活性成分，21个燥性化合物。.3. 明确了枳壳的理气/燥性Q-marker：解码“组-效-动”以明确物质基础，借助网络药理学与分子对接以明晰作用机制，进行有效性验证与可测性分析以确定Q-marker。.4. 系统的中药陈药研究思路与方法：精心设计陈化实验，正常陈化、加速陈化和冰冻陈化互为补充、验证，确保了可比性和可靠性，能应对陈药研究的时间维度变化。.5. “陈久者良”科学内涵的阐释：探讨陈化过程中“组分-效应-质量”的内在关系、动态变化规律和传递规律，系统阐释枳壳“陈久者良”的科学内涵。研究发现：①枳壳的质量属性比较稳定，陈化后燥性减弱，确实“陈久者良”；②陈化过程中，活性组分发生了含量变化和转化，这是“陈久者良”的内在原因。.项目的实施为中药陈药的研究提供了系统的研究思路与方法，有助于提升枳壳的质量标准，可用于指导枳壳的包装、储存等生产过程。