基于"活性部位-血清成分-代谢成分”关联分析的壮药玉叶金花抗炎药效物质研究

Inflammation affects the human health in many aspects. Most of Chinese herbal medicines, originating from nature and bearing the effect of clearing heat and detoxification, have anti-inflammatory activity. Research on it should be tendency of development of these anti-inflammatory medicines. Mussaenda pubescens Ait.f., a habitually used medicine among Zhuang tribe in China has various functions, such as clearing heat, detoxification, cooling blood, as well as clearing summer heat. The applicant has discovered its remarkable anti-inflammatory activity in the early stagee, however, the anti-inflammatory constituents haven’t been clarified yet. This project aims at studying the basics of the anti-inflammatory constituents and intends to sift the anti-inflammatory activity part through pharmacology experiment model, based on Chinese traditional medicine metabonomics technology. And the chemical components in the active parts of the Mussaenda pubescens Ait. f., together with the metabolic components in the blood serum and urine through dose toxicity in mice are to be analyzed adopting the method of UPLC-QTOFMS. And also the spectrums of active part-components of blood serum-metabolic components are to be characterized, through combining methods of chemometrics of PCA, PLS-DA, VIP, etc., so as to determine the metabolic marker of the anti-inflammatory activity in Mussaenda pubescens Ait.f. The correlative compounds group will be traced reversedly through the changing rule. The chemical structure is to be separated , purified and analyzed with methods, such as Sephadex LH-20, Prep-HPLC, etc. The extracorporeal inflammation cell model is to be established to sift the anti-inflammatory activity and the anti-inflammatory constituents and mechanism of functions of the Mussaenda pubescens are to be elucidated so as to provide the scientific basis to the exploration of the national medicine in Ling nan.

炎症在多个方面影响着人类健康。清热解毒类中草药多具抗炎活性，源于天然，是抗炎药物开发的重要方向。玉叶金花为壮族习用药材，具清热解毒、凉血解暑等功效，申请人前期研究发现其抗炎活性显著，但药效物质基础尚未能阐明。本项目以探究玉叶金花抗炎物质基础为目标，以中药代谢组学技术为依托，拟通过动物药理实验模型筛选玉叶金花抗炎活性部位，采用超高压液相色谱-四级杆飞行时间质谱技术分析活性部位及给药后血清和尿液中的代谢成分；结合主成分分析、正交偏最小二乘判别分析、变量重要性投影等化学计量学方法，对“活性部位-血清成分-代谢成分”的图谱进行表征，确定玉叶金花抗炎活性的代谢标识物；通过代谢标识物的变化规律，逆向追踪其关联化合物群，采用凝胶色谱、制备液相等技术进行分离纯化并解析其化学结构；建立体外炎症细胞模型进行抗炎活性筛选，阐明壮药玉叶金花抗炎药效物质及其作用机制；为岭南民族药物的发掘提供科学依据。

玉叶金花为岭南习用药材，具清热解毒、凉血解暑等功效，但其药效物质基础尚未能阐明。所以，本课题组拟阐明玉叶金花的主要抗炎活性物质基础及相关的作用机制。经过三年研究，本课题组开展了以下几个方面的工作：对玉叶金花化学成分进行了系统研究，分离鉴定了40多种化合物。对玉叶金花水提物不同萃取部位的抗炎活性进行了研究，确立了抗炎活性部位。对环烯醚萜成分玉叶金花苷酸甲酯开展了抗炎活性及含量测定研究，研究表明玉叶金花苷酸甲酯的中、高剂量（0.10，0.15g/Kg）能显著减轻二甲苯致炎小鼠的耳肿胀度(与空白对照组比较，P＜0.05)，且高剂量的作用优于阿司匹林，其含量高达1%，是玉叶金花的重要的抗炎物质基础。开展了抗炎活性物质玉叶金花苷酸甲酯体内代谢研究，研究表明大鼠静脉注射MU后，药代动力学过程符合二室模型。分布半衰期（t1/2α）为9.892min，说明MU在大鼠体内分布较快。消除半衰期（t1/2β）为：55.384min，远大于t1/2α，说明MU在大鼠体内主要以消除过程为主，代谢速度中等。建立了体外炎症细胞模型对玉叶金花苷酸甲酯开展了抗炎机制研究。研究表明玉叶金花苷酸甲酯可以降低LPS诱导的RAW264.7细胞NO的分泌量，从而抑制巨噬细胞释放IL-1、TNF-α等促炎性细胞因子，从而抑制炎症反应。.该项目研究对玉叶金花的进一步开发提供了科学依据。发表了论文5篇，一篇英文期刊论文正在审稿中，申请专利1项，培养硕士研究生1名，培养本科毕业实习生8名。