基于代谢组学的火炭母药材抗炎作用与品质相关性研究

Polygonum chinense L. Chinese medicine , which is one of the main ingredients of herbal tea, has its exact anti-inflammatory effect both in experimental and clinical researches. However, the source of it is highly confusion, and it is difficult to separate Polygonum chinense L.from P. chinense Linn. Var. hispidum Hook.f. by appearance. As a result, there is always a combination of the two sources in the herb market that can seriously affect its quality and efficacy. According to our pilot study, it found that DNA barcodes failed to distinguish between the two variants, but the chemical composition of the two plants showed significant differences. It indicated that the two variants cannot be used in combination. Therefore, exploring the chemical indicators associated with effects can be a possible way to control the quality of Polygonum chinense L. Chinese medicine effectively.This project plans to apply a combinational strategy of metabolomics and chemical fingerprint to find a way in quality control of Polygonum chinense L. Chinese medicine. Firstly, applying metabolomics and pharmacodynamics to figure out the biomarkers and regulating effect of the two variants based on inflammatory model induced by Carrageenin. And then, establishment of chemical fingerprints of the two variants and find the different variables by means of multivariate analyses. Thirdly, comprehensive analyzing of the results of the first two steps to find chemical indicators of Polygonum chinense L. Chinese medicine associated with its effect that can be used to distinguish the two variants.

火炭母药材临床和实验研究均显示其具有确切的抗炎作用。但是其资源来源比较混乱，火炭母（Polygonum chinense L.）与其变种粗毛火炭母（P. chinense Linn. var. hispidum Hook.f.）形态难以区分，故药材市场上常混合在一起，严重影响其质量和疗效。本课题组前期研究发现DNA条形码不能用于区分火炭母及其变种，但化学研究显示二者化学成分差异大，应区别使用。因此寻找与疗效相关的化学成分指标，才有可能有效控制火炭母药材质量控制。本课题拟应用代谢组学结合化学指纹方法， 确定二者对角叉菜胶致大鼠足趾肿胀模型大鼠代谢紊乱的调控作用及其差异生物标志物，建立UPLC-Q-TOF化学成分谱并结合多元统计分析找出二者的差异化学标志物，通过种内化学成分和代谢调控作用、种间差异生物标志物和化学标志物关联度分析，为建立与疗效相关且能区分二者的质量控制指标提供科学依据。

火炭母药材是我国传统凉茶的重要组成之一，临床和实验研究均显示其具有确切的抗炎作用。但是其资源来源比较混乱，火炭母（Polygonum chinense L.）与其变种粗毛火炭母（P. chinense L. var. hispidum Hook. f.）在植物形态上难以区分，故药材市场上常混合在一起使用。鉴于目前关于粗毛火炭母的研究甚少，且本课题组前期化学研究显示火炭母和粗毛火炭母化学成分有一定的差异，二者是否可以混用尚无定论，因此本课题分别建立了火炭母和粗毛火炭母的UPLC-Q-TOF/MS成分谱，从火炭母和粗毛火炭母80%乙醇提取物中分别鉴定出60和66个化合物，结合多元统计分析发现槲皮苷，槲皮素，原花青素B-O-没食子酸酯，表儿茶素没食子酸酯，山奈酚，短叶苏木酚，短叶苏木酚甲酯为二者的差异化学标志物。通过LPS诱导的诱导的RAW264.7巨噬细胞NO生成炎症活性确定了火炭母乙酸乙酯部位为抗炎有效部位。建立慢性溃疡性结肠炎的大鼠模型，发现火炭母对慢性溃疡性结肠炎具有显著治疗效果，并通过代谢组学方法初步阐释了火炭母对慢性溃疡性结肠炎起效机制，依据UPLC-Q-TOF/MS获得的火炭母的化学成分结合中药网络药理学方法，构建了“化合物⁃核心靶点⁃通路”网络，预测发现3,3'-di-O-methylellagic acid，avicularin，batatifolin，poanthocyanidin B1，quercetin 3-O-β-D-glucuronide等25个关键活性成分，主要涉及炎症反应、免疫系统、氧化应激等多种途径，推测火炭母可能参与增强机体免疫功能、降低氧化应激损伤等起到抗炎作用，进一步确定了对火炭母与抗炎活性相关的质量标志物。通过测定粗毛火炭母药材的体外抗氧化活性，确定粗毛火炭母乙酸乙酯部位为其抗氧化活性部位，并鉴定85个化合物。应用代谢组学方法对不同采收期获得的粗毛火炭母药材进行抗氧化活性与化学成分关联度分析，发现粗毛火炭母中老鹳草素，诃子鞣酸，儿茶素没食子酸酯与粗毛火炭母抗氧化活性为正相关，可以作为粗毛火炭母抗氧化活性相关质量标志物。发表SCI论文二篇。本项目通过寻找火炭母和粗毛火炭母与疗效相关的化学成分指标物，不仅为二者的合理使用提供了科学依据，而且为火炭母和粗毛火炭母药材的质量标准的制定提供研究基础。