毛冬青临床主治血栓性疾病，疗效确切。前期研究发现其抗血栓活性与体内过程密切相关且具有多组分、多途径协同作用，但具体机制尚不清楚。本项目采用体内过程和整体观相结合的新思路，拟先对毛冬青化学物质组进行体内过程研究，获取其进入体内的原型、代谢物及药动学特征，经体内外物质组关联网络分析确定其潜在体内药效组分；继之针对潜在药效组分选择性敲除/敲入组分，建立血瘀证大鼠模型，考察组分缺失前后药动学和药效学差异，并用代谢组学技术分析其干预内源性代谢组应答特征的改变，将获取数据与组分进行关联分析，从药动学、药效学和代谢组学多角度揭示毛冬青多组分抗血栓的协同整合特征和规律；最后综合解读上述数据，通过对比目标成分缺失前后整体活性差异来确定各有效成分对整体药效的贡献度，解码能够表征毛冬青抗血栓的药效成分群，通过代谢网络分析发现毛冬青潜在的抗血栓作用途径和机制。本研究有望为中药药效物质基础及作用机制研究提供新思路。

Ilex pubescens is an effective traditional Chinese medicine in clinic to treat thromboembolic diseases. In our previous study, it was found that the anti-thrombotic activity of Ilex pubescens was closely associated with its ADME process, and we also found that multiple components of Ilex pubescens act synergistically. However, the mechanisms underlying these effects remain unclear. This study will explore the mechanism from a new perspective: integration of physiological disposition and holistic approach. This project will conduct as follows: (1) investigate the disposition of the components of Ilex pubescens in vivo to get the pharmacokinetic profiles of its prototype and metabolites, and then analyze the relationship of the in vitro and in vivo chemomics to confirm its potential active constituents; (2) prepare the Ilex pubescens extract samples based on potential active constituents knock-out/knock-in and compare the differences of their pharmacokinetics, pharmacodynamics and changes of endogenous metabolites in blood-stasis rat model; then correlate and analyze the data to reveal the characteristics of the synergistic anti-thrombotic activity of Ilex pubescens from the perspective of pharmacokinetics, pharmacodynamics and metabonomics; (3) comprehensively analyze the above data and compare the differences of the integrated effects before and after knocking out targeted components to figure out the exact contribution degree of each effective constituents, and elucidate the intrinsic relation and underlying mechanism by metabolic network analysis. The implementation of this project will provide a new insight for the innovation of discovering bioactive constituents and studying integrative mechanism.