多成分"代谢-暴露反应-质效关联”模式下的当归补血汤药效物质辨识与质量控制研究

The quality control methods are very important and difficult for the development of traditional Chinese medicine (TCM). How to establish corresponding with the clinical effect of the quality evaluation system is an important scientific problem in the development of Chinese medicine. Our former study indicated that metabolic components may be involved in a major component in curative effect, but how to find and evaluate it remains unknown..In this paper, we will use the multi-component analysis and metabolic methods to track and screen the effective components of Danggui Buxue decoction. Through the study of the correlation between metabolism and efficacy, we will confirm the prototypes or metabolic markers of associated efficacy, and then establish a metabolic-pharmaceutical system. At the same time, we will trace the metabolic markers back to the original decoction, define the components and their proportions, and then establish a quality-pharmacological effect model. Eventually, we explore the material basis and mechanism of action of Chinese medicine, and present the comprehensive method for effective basis identification and quality evaluation by the model of “multicomponent drug metabolism-exposure and response-association in quality and curative effect”. The implementation of the project will further clarify the effective basis and mechanism of Danggui Buxue Decoction, enhance its basic research level of pharmacodynamics, and provide important enlightenment and reference for the quality evaluation of Chinese herbal compound..This project will publish five to seven papers, including at least three SCI articles and draft one key technical standard operating procedure. Also，training of three master's degree will be executed based on the grant.

中药复方的质量评价是中医药发挥疗效和相关研究的重点与难点之一。前期研究发现当归补血汤部分潜在的质量标记物或代谢标记物，其代谢规律与复方体内暴露水平和药效大小相关，但如何确定药效成分并用于质量评价亟待研究解决。本项目拟采用多成分分析与代谢方法，跟踪筛选有效组分，通过代谢与疗效相关性研究，确定关联药效的原型或代谢标志物，构建代谢-药效体系；同时追溯代谢标记物在原方中的母体，明确组分内含成分及比例，利用疗效主成分量化质控，建立质量-药效模式，进而探索中药药效物质基础及作用机理，最终建立多成分“代谢-暴露反应-质效关联”模式下的当归补血汤药效物质辨识和质量评价方法。项目实施将进一步明确当归补血汤的药效成分及作用机理，提升其药效物质基础研究水平，并为中药复方质量评价研究提供重要启示和参考。基于本项目将发表5-7篇论文（SCI3-4篇），起草关键技术标准操作规程1部，培养3名硕士。

本项目采用高效液相色谱法、液质联用技术建立当归补血汤特征图谱，定性鉴别出31种化合物，并结合多元统计分析方法对复方整体质量进行综合评价，为复方质量标准建立提供重要支撑，也为后续代谢研究提供较为全面的药效物质基础成分。以序贯代谢理论为指导，采用在体封闭肠环-腹主动脉采血技术，从肠无菌、肠道菌、肠壁酶、肝代谢四个部位探讨当归补血汤在体多成分代谢规律，最终鉴定出11个代谢成分，并遴选出毛蕊异黄酮、毛蕊异黄酮-7-O-β-D-葡萄糖苷、芒柄花素、芒柄花苷、黄芪甲苷、藁本内酯、洋川芎内酯I这7种原型成分为当归补血汤在体内吸收代谢过程中重要的代谢枢纽，可考虑作为当归补血汤的潜在质量标志物。基于补血药效方面，以血虚大鼠模型为研究对象，以外周血常规为评价指标，发现当归补血汤在12g/kg、24g/kg剂量下均具有明显的补血功效；基于补血药物代谢动力学方面，以正常和血虚大鼠为研究对象进行对比，发现当归补血汤中关键代谢成分（阿魏酸、毛蕊异黄酮、毛蕊异黄酮-7-O-β-D-葡萄糖苷、芒柄花素、芒柄花苷、藁本内酯、洋川芎内酯I）在病理状态下暴露更高，且分布的潜在靶器官为心脏；基于活血药效方面，以急性血瘀大鼠模型为研究对象，以血液流变学、凝血四项、血脂水平为评价指标，发现当归补血汤在12 g/kg剂量下具有明显的活血功效，为后续质效关联提供药效基础。本项目已经发表SCI论文1篇，在投SCI论文2篇，核心期刊论文2篇，硕士毕业论文2篇；起草关键技术标准操作规程1部，培养5名硕士研究生。