多效中药黄芪的定向药效化学成分研究

The project is based on the cognition of traditional Chinese medicine (TCM) that the component has different effects in different formulas. The multi-effect Astragali Radix is employed, which is used broadly in clinic and contains invigoration and opposite functions such as eliminating pathogenic factors, to illuminate the orientation material basis in a given formula, obtain the quality indicators associated with efficacy and provide the research basis for classifying and evaluating effectiveness of the Astragali Radix resources ultimately. It is proposed that the oriented effective components can be identified when TCM is placed in the different formulas such as huang-qi-jian-zhong-tang (tonifying Qi) and fang-ji-huang-qi-tang (reducing swelling) as the representative formulas to treat the chronic atrophic gastritis and nephrotic syndrome (NS), respectively, under different conditions (physiology or pathology). Through metabonomics, Chinese materia medica fingerprint, serum pharmacochemistry, quantitative composition activity relationship (QCAR) and separation of constituents by column chromatography, the oriented effective components, the chemical compositions, ratio relation and the trajectory of variation in vivo of Astragali Radix will be identified, and the target component group for quality control will be confirmed to evaluate the different resources of Astragali Radix. In conclusion, the pharmacological material basis of the oriented function of the Astragali Radix, as well as the scientific implications of oriented effective components exerted by muti-effect Astragali Radix on the molecular level will be elucidated in this study. The research finding will also provide a novel strategy for the identification of oriented effective components and validity evaluation of TCM resources in which the multi-effect medicine is prevalent.

本项目依据中医对单味中药在不同复方中发挥不同功效的认识，以应用广且集补益与祛邪两种相反功能于一体的多效中药黄芪为对象，阐明其在具体复方中发挥定向药效的化学物质基础，解决质控指标与药效关联问题，旨在为准确分类评价黄芪资源的有效性提供研究基础。项目选择历史悠久、临床疗效确切的张仲景经方黄芪建中汤（补气）和防已黄芪汤（消肿）为代表方剂，综合采用代谢组学、指纹图谱、中药血清化学、组效关系分析及柱色层族组成分离法等先进技术，结合传统药效指标，辨识黄芪在不同复方、不同微环境（包括生理和病理状态）中发挥不同功效的定向化学成分群，明确其化学组成、配比关系、体内代谢轨迹，为建立黄芪资源评价体系提供质控目标成分群。研究结果将明确黄芪在不同方剂中显示不同功能的化学物质基础，从分子水平上阐释黄芪发挥定向功效的科学内涵，还将为普遍存在的一药多效中药定向功效成分辨识及资源的分类评价提供研究新策略。

本项目依据中医对多效中药在不同复方中发挥不同功效的认识，选择历史悠久、临床疗效确切的经方黄芪建中汤和防已黄芪汤为代表方剂，辨识黄芪发挥不同定向药效的化学成分。首先，基于UPLC-HRMS/MS技术分别对黄芪建中汤71种化学组成和防己黄芪汤78种成分进行了快速识别，归属黄芪的分别有20和18种。同时对复方近20种成分进行了定量分析。其次，成功构建了慢性萎缩性胃炎大鼠和肾病综合证大鼠模型用于药效评价和代谢组学研究，结果均显示复方药效最佳，单味药黄芪次之，而缺黄芪的阴性方药效较差，确证了黄芪对全方的贡献。再次，整合组效相关、血清药化等多种手段构建了黄芪定向药效成分辨识体系，成功辨识了黄芪在黄芪建中汤中发挥“益气建中”药效主要为3-Hydroxy-9,10-dimethoxyptercarpan、7,2'-Dihydroxy-3,4'-dimethoxy isoflavan、Isoastragaloside I和多糖；而在防已黄芪汤发挥“利水消肿”的药效成分主要是黄芪皂苷 I、黄芪皂苷 II、黄芪皂苷 III、黄芪甲苷Kumatakenin和Dihydroxy-dimethoxy isoflavone。可见，黄芪中黄酮、皂苷以及多糖类物质均能发挥“补益”和“驱邪”作用，但“补益”功效黄酮类贡献大，而“驱邪”药效成分群皂苷类占比较高。最后，采用网络药理学、代谢组学、转录组学以及分子生物学技术阐明了黄芪在不同复方中针对不同病证发挥药效的分子机制，有趣的是黄芪无论发挥“补”还是“泄”的药效都涉及能量代谢调控，但具体机制不尽相同，值得进一步深入探讨。此外，项目还成功构建了脂质组学平台以及新的中药成分表征策略，并成功应用于本项目肾病病理机制及黄芪单味药化学成分的系统研究。本项目共发表学术论文35篇，其中研究性论文30篇，包括 SCI 论文17篇，其中SCI二区4篇；综述性文章9篇；申请专利10 件，授权2件；培养博士研究生1名，硕士研究生10名；参加会议16人次。总之，本项目明确了黄芪在不同方剂中针对不同病症的化学物质基础，能够一定程度上指导临床医生和企业合理选择原料药，也为普遍存在的一药多效中药定向功效成分辨识及资源的分类评价提供研究策略。