芳香开窍药治疗脑卒中的药效物质发现及中枢-外周互动整合作用机制研究

Multi-components, multi-pathways, and synergistic effects are widely recognized as the characteristics of Traditional Chinese medicines (TCMs). It is almost impossible to find a single component in a TCM that can represent the holistic efficacy of the complex mixture. Guided by the basic theory of “Bioactive Equivalent Combinatorial Components (BECCs)”, we aim to discover the BECCs of aromatic resuscitation TCMs with clear clinical efficacy on stroke that is a serious disease with high morbidity in China. The research objectives in the project include musk-borneol pair. The following researches will be conducted. With various cell and animal models, the action pathway and therapeutic time window of TCMs for stroke treatment will be explored. Based on the chemical components of herbal prescription and herb pairs, the BECCs that represent the holistic efficacy of original TCM could be found using gold indicators for efficacy, bioinformatics-based tools, stepwise screening, and bioactivity confirmation. According to the metabolism of BECCs, the “multi-pathways and synergistic effects” of BECCs will be clarified from the perspective of central protection and peripheral regulation. The common characteristics and regularities of aromatic resuscitation TCMs could be refined. This project aims at providing novel thoughts and methods for the research of multi-component pharmacodynamic substances and crosstalk mechanisms of TCMs. Moreover, the scientific concept of BECCs is helpful to promote the theory innovation of TCM research.

中药复方具有多成分、多途径、协同整合的作用特点，单一成分往往难以代表原方药效。本项目拟以我国发病率高的脑卒中为切入点，以芳香开窍代表药对“麝香-冰片”等为研究对象，开展如下研究：通过多种细胞、整体动物模型评价，明确芳香开窍经方及药对抗脑卒中的优势作用环节和时间；在阐明复方及药对化学组成的基础上，应用脑卒中金指标模型和生物信息学手段，逐级筛选，正反确证，最终发现能够表征原复方/药对“开窍”药效的物质基础（等效成分群）；根据开窍“等效成分群”的体内代谢特点，从中枢保护和外周调控两方面，阐明“等效成分群”多途径协同作用机制，凝炼芳香开窍药“开窍”的共性特点与规律。本项目旨在为中药复方多成分药效物质和交互作用机制研究提供新的思路与方法，实现中药理论创新。

本研究聚焦于芳香开窍药麝香、冰片、郁金以及开窍经方安宫牛黄丸抗缺血性脑卒中的药效物质发现与“中枢-外周”整合作用机制展开研究。① 建立了“化学成分-体内代谢产物-内源性生物标志物”一体化研究策略，系统解析了芳香开窍药化学物质基础、安宫牛黄丸的体内外化学成分及内源性生物标志物的变化。② 应用小鼠大脑中动脉闭塞（MCAO）模型和小鼠光化学栓塞法（PT）模型开展了芳香开窍药及安宫牛黄丸抗缺血性脑卒中的药效学研究；建立斑马鱼脑缺血模型，筛选了抗缺血性脑卒中的活性成分，并通过网络药理学分析和斑马鱼脑缺血模型评价，结合MCAO整体动物模型验证，发现了安宫牛黄丸抗缺血性脑卒中的等效成分群。③ 在药效物质研究的基础上，根据芳香开窍药成分群的代谢组学特征，从中枢保护角度阐明了麝香-冰片药对中麝香酮-右旋龙脑通过调控Ca2+/Erk1/2通路和琥珀酸/Hif-1α通路抑制ROS介导的炎性反应，进而影响CREB/ claudin 5级联反应协同保护脑微血管内皮功能的作用机制；以及β-榄香烯通过抑制TLR4/MYD88/NF-κB通路阻断炎性反应，保护神经元抗缺血性脑损伤的作用路径。从外周调控角度阐释了等效成分群中姜黄素等成分抑制柠檬酸转运介导的衣康酸蓄积，通过代谢调控阻断外周巨噬细胞炎性活化减轻缺血性脑损伤的作用机制；以及汉黄芩素阻断外周脾脏单核/巨噬细胞中TREM1通路的激活，抑制线粒体DNA释放介导的炎症级联反应，减少脑梗塞部位炎症型巨噬细胞的活化与募集，进而减轻缺血性脑损伤的作用机制。相关研究内容是2022年教育部科技进步一等奖、2022年度中医药国际贡献奖—科技进步奖一等奖的组成部分。已发表论文13篇，申请发明专利1项。研究成果用于安宫牛黄丸等中成药药效物质与质量标准提升研究。