基于代谢组学的杏香兔耳风抗宫颈炎药效物质基础及作用机制研究

Ainsliaea fragrans Champ blongs to the dominant herb in JiangXi province which has definite curative effect for anti-cervicitis. But the material basis and action mechanism of this herb were not distinct which have been the bottleneck in its further development. With the support by the National Natural Science Foundation, it had been confirmed that the total phenolic acids were the active ingredients for anti-ureaplasma urealycum and anti-cervicitis in Ainsliaea fragrans Champ. Meanwhile, the experiment result exhibited that it had the inhibitory effect of drug-resistant mutant strain and the antibacterial effect of the total phenolic acid was remarkable comparing with the monomers of dicaffeoylquinic acids which proved by the Pharmacokinetic-Pharmacodynamic models with Monte Carlo simulation. Hence, it was supposed that the curative effect resulted from the coaction of multiple ingredients and the drug targets were also pluralistic. In order to clarify the TCM (Ainsliaea fragrans Champ) metabolic pathways and the action mechanism of multiple ingredients and drug targets, the UHPLC-MS/MS metabonomics technology combined with MarkerView, MetabolitePilot, PeakView and other bioinformatics analysis method would be applied in the experiment research. It can also establish the "Metabolic fingerprinting" of biological samples (such as blood, urine, tissues and organs, etc.) from the animal models with cervicitis before and after the administration. Meanwhile, the relationships between the exogenous metabolites and endogenous biomarkers will be explored. These research efforts will provide the foundation for the secondary development of Ainsliaea fragrans Champ and expand mentality in the new drug research for anti-cervicitis.

江西特色优势药材杏香兔耳风用于宫颈炎疗效确切，但物质基础及作用机制不明确是其深入开发利用的瓶颈。在国家自然科学基金资助下，我们已证实杏香兔耳风总酚酸为抗宫颈炎有效部位，且具有抑制解脲支原体和耐药突变菌株作用，通过PK/PD 蒙特卡洛模拟，发现总酚酸抗菌效果优于各二咖啡酰奎宁酸单体，据此推测其可能为多成分和多靶点的起效机制。故本课题拟建立解脲支原体感染小鼠宫颈炎模型，运用UHPLC-MS/MS代谢组学技术，结合MarkerView、MetabolitePilot、PeakView等生物信息学分析方法，建立给药前后的生物样品（如血液、尿液、组织器官等）“代谢物指纹图谱”，探索外源性代谢产物与内源性生物标记物之间的关联，确定代谢途径和代谢通路，并通过活性验证及不同作用途径的比较分析，阐明杏香兔耳风活性物质基础及作用机制。该研究将为杏香兔耳风的二次开发奠定基础，并拓宽抗宫颈炎新药的研发思路。

本课题以江西特色优势药材杏香兔耳风为研究对象，应用代谢组学方法开展了杏香兔耳风总酚酸抗宫颈炎的物质基础及作用机制研究。通过对杏香兔耳风总酚酸的化学成分分析，共鉴定了132种化合物，为总酚酸抗宫颈炎提供了物质基础；通过大鼠体内代谢实验，结果发现了共153种化合物，其中原型为30种，代谢产物为123种；主要代谢途径为甲基化、磺酸化、葡萄糖醛酸化、还原和脱羧等，为杏香兔耳风制剂成为妇科常用药提供了科学依据.本实验应用代谢组学技术对大鼠宫颈炎模型与人类宫颈炎疾病进行了研究，分别鉴定了32种和22种差异性代谢物作为潜在标志物，且二者均发现了亚油酸、花生酸和甘油磷脂代谢差异性代谢通路，阐释了宫颈炎的发病机制。而总酚酸对宫颈炎大鼠模型中27个生物标志物具有调节作用，其中12个生物标志物与正常组相对含量相近，结果表明杏香兔耳风具有抗大鼠宫颈炎的作用，为杏香兔耳风制剂临床应用提供新的思路和理论基础。. 本课题研究期内，我们已经发表7篇学术论文，其中包括5篇SCI收录论文。通过对上述研究内容的积极开展，本课题实现了外源性代谢产物与内源性生物标记物之间的关联性，为阐明杏香兔耳风体内外多成分的活性物质基础及多靶点的起效机制，还为杏香兔耳风的二次开发奠定基础，并拓宽新药研发思路。