利用代谢组学技术研究橘属中药物质基础及其体内作用

Because of the high edible and medicinal value, Citrus plants become a hot research object all over the world. Nine Chinese medicines from Citrus species are listed in the Chinese Pharmacopoeia, which belong to qi-regulation drug in TCM. However, the efficacy during them is different. The efficacy based on the chemical substances is important to ensure their safety and effectiveness in clinical use. Metabonomics is a well-established field in systems biology. Plants metabonomics could be applied in research of chemical substances in plants..Hypothesis: The chemical substances in different Citrus Chinese medicines are varied, which can lead to diversify responses of endogenous metabolites. Even, the metabolic pathway will be disturbed due to the irritation. The relationship between metabolism pathway and chemical substances could be used to indicate different effective mechanism...In the present study, nine Chinese medicines of Citrus species were investigated using LC-MSn complemented by NMR technique. Furthermore, a multivariate statistical method was applied to distinguish different Citrus Chinese medicines according to their chemical substances. Meanwhile, the endogenous metabolites disturbances after administrated with nine Citrus Chinese medicines are revealed by NMR-based metabonomics. The data between the two metabonomics will be fitted based on decision tree analysis and support vector machine analysis. The established two-dimension metabonomics strategy will be assist in quality assessment of Citrus Chinese medicines.

橘属植物极高的食用及药用价值，使其成为国内外研究的热点。中国药典收录的橘属中药达九种，同为理气药，但其功效各有偏重。本课题拟从品种、药用部位及生长阶段不同的橘属中药物质基础及其体内作用切入，借助代谢组学技术在定性定量描述中药成分代谢物，客观反映机体内源性生物代谢物的优势，初步探讨橘属中药功效差异机理。.假说：基于代谢组学技术研究橘属中药物质基础及其体内作用，对于揭示其功效差异机理具有实际意义。.以九种橘属中药为研究对象，采用LC-MSn辅以NMR分析及检测手段，结合多变量统计学方法，开展基于化学表征差异鉴别橘属中药的植物代谢组学研究；利用NMR方法，开展不同橘属中药引起大鼠生物代谢物差异性应答的体内代谢组学研究，借助生物信息学方法进行代谢通路分析；利用多元回归方法关联分析两组代谢组学数据，构建橘属中药物质基础及其体内作用的新型研究模式，探索橘属中药功效差异机理，为临床合理用药提供依据。

本项目收集了九种橘属中药，分别建立其活性提取部位的质量控制方法及提取工艺（发表文章并申请专利）。采用LC-MSn辅以NMR分析及检测手段，结合多变量统计学方法，开展基于化学表征差异鉴别橘属中药的植物代谢组学研究，分别建立初生及次生代谢产物化学分类学方法，可有效区分亲缘相近，但品种、药用部位及生长阶段不同的橘属中药（发表文章并申请专利）。通过分析不同橘属中药引起大鼠内源性代谢物的差异性应答，发现药效物质基础的差异分别引起体内不同代谢通路的激活，借助网络药理学方法阐释代谢通路与其调节蛋白及功效差异的相关性（发表文章）。基于以上结果，筛选出橘属中生物活性较强的四味中药，建立了体内ADMET分析，进一步阐释体内活性成分及其代谢轮廓（发表文章）。筛选出橘属中药活性提取部位分别表现出多样性生物活性，其中，枳实、枳壳分别通过抑制MAPK信号通路和激活PPAR-AKT信号通路对LPS诱导的炎症反应发挥不同的抗炎作用，枳实中活性提取部位对药源性肝损伤具有保护作用等（发表文章）。通过以上研究，我们建立了橘属中药体内体外量-效关联性研究模式，对于系统阐释中药药效物质基础具有借鉴意义，并为临床合理用药提供依据。