基于代谢组技术的菊叶“清肝明目”功效物质基础及生物学机制研究

Functioning as clearing liver and improving vision and treating wind syndrome of head and vertigo, the leaves of Chrysanthemum morifolium Ramat. have been documented in ancient and modern classics of herb medicine. Focus on its chemical components and biological activities evaluation of folium Chrysanthemum, our preliminary experiments suggested that its efficacies were exerted by its phenolic acids, flavonoids and other multi-components via the inhibition of a series of pathological process featuring liver-fire. .This project would hereby establish animal models characterizing the syndrome of liver fire flaring up and metabolomics would be combined with pharmacodynamic evaluation to reveal the nature of clearing liver and improving vision of folium Chrysanthemum and the underlying mechanisms. Metabolomics approach would be adopted to discover the pattern of endogenous metabolites of the models of liver fire flaring up, deduce the possible target groups, and therefore map the target pathways and networks to characterize and understand the function of clearing liver and improving vision of folium Chrysanthemum; state-of-art statistical methods would be used to clarify the relationships between the chemical constituents of folium Chrysanthemum and metabolomics profiles, thus revealing the contribution of individual component to the efficacy and determine the material basis of its efficacy; the metabolomics results would be further combined with molecular biology techniques to disclose the biological mechanisms underlying the function of clearing liver and improving vision of folium Chrysanthemum, and to confirm its usage in medicine and in health maintenance of people, laying a solid scientific foundation for its resource effective reuse.

菊叶药用记载见于古今医药典籍，谓之具清肝明目功效，主头风，目眩，泪出。课题组对其进行的化学成分及其生物活性初步研究结果提示，菊叶“清肝明目”功效可能是其酚酸类、黄酮类等多元物质组通过抑制“肝火上炎”病理过程而实现的。据此，拟采用代谢组学技术结合药效评价，利用肝火上炎证候动物模型，以揭示菊叶清肝明目功效物质基础及其生物学机制。拟开展工作如下：（1）应用代谢组学技术，通过肝火上炎证候动物模型，研究菊叶清肝明目的生物靶标群、受影响代谢通路及代谢网络，整体表征清肝明目功效；（2）应用现代统计学方法对菊叶化学成分与其清肝明目的代谢组学进行相关分析，确定菊叶各组成成分对其功效贡献度，揭示菊叶清肝明目功效物质基础；（3）结合代谢组学结果，应用分子生物学技术揭示菊叶清肝明目功效的生物学机制，以期为菊叶药用和服务于民众健康及菊资源的有效利用提供科学依据。

作为平肝明目要药，菊花明目功效确切；《本草纲目》记载菊“茎叶根实并同”可知，菊茎叶与花具有类似功效。面向市场需求，系统评价了米菊、胎菊和朵菊中总黄酮类、总酚酸类、氨基酸类、核苷酸类、无机元素类成分、蛋白质类及挥发性成分，为制订菊花商品规格草案提供依据；系统评价药用菊不同部位，不同药用、茶用及观赏菊中黄酮类及酚酸类成分，初步揭示菊平肝明目功效物质基础，为其资源循环利用提供依据。.完成了不同类型菊花提取物及主要黄酮类、酚酸类成分抗氧化活性评价及构效关系研究，为菊资源的精细抗氧化应用奠定基础；完成了菊叶活性提取物防治肝损伤、感冒、黄斑变性研究，并申请国家发明专利，为菊叶资源化利用及产业化开发提供技术支撑。.建立了肝火上炎兔、肝炎上亢大鼠等证候动物模型；构建了基于代谢通路分析和代谢网络分析的症候动物模型及中药功效评价研究技术体系，揭示菊茎叶能够改善肝火上炎症候动物结膜充血、房水浑浊，主要通过抗炎发挥平肝明目保护作用；改善肝火上炎症候中紊乱的能量代谢、氧化应激，恢复异常的代谢向正常状态转归；下调致炎因子的表达，且乙醇提物抗炎效果优于水提物。肝炎上亢症候模型揭示了菊花组降压效果优于茎叶组，菊花水提物最佳；菊花组和茎叶乙醇组均显著降低心率；各给药组均能显著逆转高血压大鼠紊乱的芳香氨基酸代谢、三羧酸循环、无氧酵解、肠源性毒性、肠道菌群代谢等，使其向正常转归；Phenylalanine、tyrosine、acetylsalicylate、3-indolsulfate可作为评估肝阳上亢症候潜在的生物标记物，为后续研究工作奠定了基础。.初步揭示了菊茎叶水提物和乙醇提取物通过下调肝脏磷酸化NF-κB（p65）蛋白表达，增加肾脏磷酸化p38表达，主要通过NF-κB通路发挥抗炎活性，从而起平肝明目的功效。.目前已发表发表论文5篇，其中SCI收录2篇；申请国家发明专利4项；培养研究生2名。同时，含有菊茎叶挥发油的饱和芳香水正在进行菊纯露、菊花露水、菊洗手液产品的创制工作，并已经有菊茎杆样品烧制生物炭小试样品试制。