基于“组-效-动”三元定向关联的黄芪“益气建中”功效的质量标志物研究

The current quality evaluation of Astragali Radix is reliant on two evaluating indexes, astragaloside IV and calycosin-7-O-β-D-glucoside, which is difficult to evaluate its effectiveness relevant to its oriented pharmacodynamic action associated with the compatible environment in formulas under the particular pathologic conditions. In this project, a classical prescription Huangqi jianzhong tang (HQJZ) selected as the compatible environment of Astragali Radix, and the qi deficiency of spleen and stomache-induced chronic atrophic gastritis chosen as the particular pathologic conditions, were directionally applied to identify the active constituents related to its oriented efficacy of tonifying qi and strengthening stomache and spleen. Consequently, based on the oriented relevance "composition-activity-pharmacokinetics" strategy by integrating multidisciplinary technologies including pharmaceutical analysis, pharmacology, metabonomics and quantitative composition activity relationship, the relationship between the chemical composition of Astragali Radix and pharmacodynamic markers in terms of metabolic regulation was illustrated to screen the effective basis related to its efficacy of tonifying qi and strengthening stomache and spleen. Meanwhile, a pharmacokinetic research was utilized to depict their pharmacokinetics characteristics in HQJZ. At last, the quality markers of Astragali Radix related to the efficacy of tonifying qi and strengthening stomache and spleen were optimized through multivariate combination analysis of their three dimensions including the chemical composition, the regulation of pharmacodynamic markers and pharmacokinetics characteristics. On this foundation, our study will provide the scientific basis for the quality control of traditional Chinese medicine based on its oriented pharmacodynamic action.

黄芪现行的质量评价方法仅以黄芪甲苷和毛蕊异黄酮苷为质控指标，与针对特定病症和复方配伍环境中的定向药效关联不强，难以准确评价黄芪的有效性。而药效成分的阐明是解决中药质量控制问题的关键。故，本项目以辨识黄芪“益气建中”的定向药效成分群为主要研究目标，选用黄芪“益气建中”经方黄芪建中汤为“配伍环境”，以脾胃气虚所致的慢性萎缩性胃炎为“特定病症背景”，在“组-效-动”三元定向关联的研究思路指导下，综合采用药物分析、药理学、代谢组学和组效相关分析等多学科技术与方法，从代谢调控的角度探讨黄芪化学成分组成与其起效标志物群的相互关系，锁定黄芪发挥“益气建中”定向功效的药效成分群，同时应用药代动力学技术研究药效成分的药动学特征，通过多元组合分析对“化学组成”—“起效标志物群”—“药动学特征”三个维度关联分析，优选复方中黄芪的“益气建中”质量标志物，为基于定向功效的黄芪质量有效性评价提供科学依据。

性萎缩性胃炎为“特定病症背景”，辨识黄芪在复方中的定向药效成分和定向质量标志物。取得了以下结果：①建立了基于LC-MS技术的黄芪建中汤的化学指纹图谱和多指标成分定量的分析方法，其中黄芪化学成分定性了24个，定量了12个。②从样本制备（血清和血浆比较）、数据处理（静态和动态结合分析时序数据）、分析方法（NMR和LC-MS技术结合）等多方面优化了代谢组学研究方法，结果显示黄芪建中汤对慢性萎缩性胃炎（CAG）大鼠代谢紊乱的调控作用，可能是基于其对能量平衡的调节，炎症免疫系统的改善，氧化应激的调节和肠道菌群的调节。③建立了黄芪建中汤在正常大鼠和CAG大鼠体内定性定量的分析方法，为系统解析该方的体内过程奠定了基础。④结合传统药效指标、血清代谢组学、线粒体功能指标以及线粒体代谢组学多个角度初步阐释了黄芪对全方的药效贡献，且优于另一君药“饴糖”。⑤灰色关联分析、偏最小二乘回归分析和皮尔逊相关分析显示，复方中黄芪的25个化学成分（包括多糖）与实验大鼠体重、胃液pH值、胃蛋白酶活性以及血清代谢组学数据之间均存在显著相关性，为复方中黄芪发挥“益气建中”定向功效的药效成分群。⑥基于中药质量标志物筛选“五原则”，最后将芒柄花素、7,2'-二羟基-3',4'-二甲氧基异黄烷、毛蕊异黄酮、毛蕊异黄酮葡萄糖苷和黄芪甲苷选为黄芪的质量标志物。本项目共发表学术论文23篇，其中研究性论文16篇，包括SCI论文13篇，其中SCI二区2篇；综述性文章7篇；申请专利1件，授权1件；培养硕士研究生3名；参加会议5人次，发表会议论文2篇。总之，本项目构建了基于“化学组成”—“起效标志物群”—“药动学特征”三个维度关联分析的复方中君药定向功效质量标志物的优选策略，为基于定向功效的中药材质量有效性评价提供科学依据。