目的：基于中医方证代谢组学技术分析男仕口服液干预皮质酮诱导的肾阳虚证大鼠模型内源性代谢生物标记物的变化趋势，探索男仕口服液入血成分与其治疗肾阳虚证过程的相关代谢物及代谢通路的相关性，挖掘男仕口服液发挥治疗作用的体内直接作用物质及作用机理。方法：检测大鼠“下丘脑-垂体-靶器官轴”各激素水平并观察相应靶器官的组织形态学，评价男仕口服液对肾阳虚证的作用效果；利用模式识别分析各组大鼠尿液代谢谱，研究男仕口服液对肾阳虚证生物标记物影响，同时在显效基础上分析其入血成分与肾阳虚证生物标记物群的相关性，确定其发挥治疗作用的潜在药效物质基础。结果：模型大鼠血清的CRH、CORT、T3、T4、T和尿17-OHCS含量显著下降；男仕口服液可显著提升血清中CRH、CORT、T3、T4、cAMP和尿中17-OHCS水平，调节类固醇激素的生物合成、色氨酸代谢和酪氨酸代谢；PCMS相关分析发现男仕口服液显效剂量下与代谢标记物高度相关的入血成分主要为：水晶兰苷、没食子酸、鸡矢藤苷、莫诺苷葡萄糖醛酸化产物、甜菊苷C、甜菊双糖苷，这些成分是男仕口服液治疗肾阳虚证的药效关键物质基础。结论：本研究运用中医方证代谢组学技术建立了“肾阳虚证代谢标记物-男仕口服液体内直接作用物质-药效关键物质基础”的分析模式，发现水晶兰苷、没食子酸、鸡矢藤苷等成分通过改善多个代谢途径（激素合成途径、酪氨酸代谢中激素合成与释放途径等），从整体上纠正代谢轮廓的改变，体现了男仕口服液治疗肾阳虚证的多途径、多靶点、整体调节的作用特点。该研究为基于临床有效性的创新药物发现提供方法学基础。

This study aimed to explore the constituents migrating to blood and the correlation between the metabolites and metabolic pathways and find out the therapeutic basis and its mechanism behind the efficacy of Nanshi oral liquid(NSOL)by observing the changes of endogenous metabolic biomarkers closely related to kidney-yang deficiency syndrome(KYDS)induced by the administration of corticosterone.The hormone expressions in the hypothalamus-pituitary-target organs axes were detected,while the histomorphology of all target organs was observed to judge the efficacy of NSOL on the KYDS models.Then the serum and urine samples were analyzed using pattern recognition analysis for the verification of the impacts of the biomarkers related to NSOL on the KYDS models.Using the effective dose of NSOL,the correlation between the main constituents migrating to blood and the biomarkers were analyzed and finally confirmed the potential therapeutic basis.It was found that the levels of CRH,CORT,T3,T4,T in the blood and 17-OHCS in the urine significantly decreased,while the hypothalamus-pituitary-target organs axes were inhibited.The contents of CRH,CORT,T3,T4,cAMP in serum and 17-OHCS in the urine significantly increased with the administration of NSOL,and the biosynthesis of steroid hormone and the metabolisms of tryptophan and tyrosine were regulated.PCMS analysis showed that scandoside,monotropeine,gallic acid,stevioside C, steviolbioside and morroniside glucuronide conjugation were the main constituents migrating to blood which positively or negatively correlated with biomarkers under the effective dose,which were deemed as the pharmacodynamic substances of NSOL.In conclusion,technologies of Chinmedomics were adopted to successfully establish the analysis model of“KYDS markers of constituents migrating to blood under effective dose-pharmacodynamic basis of NSOL in vivo”,indicated that scandoside,monotropeine and gallic acid in NSOL improved various metabolic pathways(hormone synthesis pathways,hormone synthesis and release pathway in tyrosine metabolism,etc.)and integrally regulated the metabolic profilings.It was demonstrated that the treatment of NSOL for KYDS featuring the corresponding between“formula”and“syndrome”in traditional Chinese medicine and reflecting its multi-path and multi-target and overall regulation.This study laid a foundation for the discovery of new medications based on their efficacy in clinic.

国家自然科学基金委重点项目（81430093）：中药体内药效物质基础的系统分析方法学——中医方证代谢组学研究，负责人：王喜军；科学技术部“重大新药创制”科技重大专项（2015ZX09101043-011）：中药经典名方整合作用机制关键技术研究，负责人：王喜军；黑龙江省普通本科高等学校青年创新人才培养计划（UNPYSCT-2015118）：基于方证代谢组学的金匮肾气丸治疗肾阳虚证的药效物质基础和作用机理研究，负责人：张爱华。