基于药物代谢和网络药理学的中药密蒙花药效物质基础研究

发布时间：2018-12-31 15:34

【摘要】：由于中药成分复杂多样,中药药效物质基础和作用机理研究已经成为一个巨大的挑战。众所周知,中药汤剂一般采用口服给药的方式。口服给药后,中药多成分一般要经历胃肠道屏障,才能进入血液循环系统。在一系列的过程中,并不是所有的中药成分均能够进入血液循环系统而产生药效。有些中药成分能够以原型的形式吸收入血,有些中药成分可能会被肠道或肝脏的酶代谢,从而产生新的代谢物。因此,最终进入到血液循环的成分是极其复杂的。近些年来,由于分析技术手段的发展,液质联用技术由于其快速、灵敏的特点,已经成为在复杂生物样本中分析中药多成分及其代谢物的重要技术手段。然而,光分析清楚入血成分是远远不够的,还需要构建成分与药效之间的联系。近些年来,随着生物信息学的发展,网络药理学已经成为研究中药作用机理的重要技术手段。因此,本文以密蒙花为研究载体,尝试用"化学成分-序贯代谢-网络靶标"的研究模式来阐明密蒙花治疗眼结膜炎的药效物质基础和作用机理。研究主要分为以下三大部分,具体结果如下:1.利用UHPLC-LTQ-Orbitrap技术分析密蒙花提取物的化学成分研究结果表明,从密蒙花提取液中一共鉴定了 20个主要化学成分,包括11个苯乙醇苷类化合物和9个黄酮类化合物。2.密蒙花序贯代谢研究(1)苯乙醇苷类代谢行为规律研究我们选择密蒙花中两个代表性苯乙醇苷类单体红景天苷和异毛蕊花苷来阐明苯乙醇苷类成分的代谢行为规律。红景天苷序贯代谢研究结果:一共从红景天苷血液样本中检测和鉴定了四个代谢物,其主要代谢途径为水解,葡萄糖醛酸化和硫酸化。另外,红景天苷在消化液中比较稳定,其主要被肠道菌群和肝代谢,并且有一小部分被肠壁代谢。异毛蕊花苷体内代谢研究结果:一共从异毛蕊花苷尿液样本中检测到了 10个代谢产物,其主要代谢途径为水解,去羟基化,葡萄糖醛酸化,硫酸化,甲基化。总之,苯乙醇苷类的主要代谢途径为水解,去羟基化,葡萄糖醛酸化,硫酸化,甲基化等。该研究结果可以为密蒙花提取液中其他类似结构化合物代谢产物的推测提供参考。(2)黄酮类代谢行为规律研究我们选择密蒙花中两个代表性黄酮类单体蒙花苷和木犀草素来阐明黄酮类成分的代谢行为规律。蒙花苷体内代谢的研究结果:一共在尿液和粪便样品中鉴别了4个代谢物。蒙花苷的主要代谢途径为水解,葡萄糖醛酸化,去甲基化以及羟基化。木犀草素体内代谢的研究结果:一共在尿液和粪便样品中鉴别了6个代谢物。木犀草素主要代谢途径为甲基化和葡萄糖醛酸化。总之,黄酮类化合物的主要代谢途径为水解,葡萄糖醛酸化,去甲基化以及羟基化,甲基化等。该研究结果可以为密蒙花提取液中其他类似结构化合物代谢产物的推测提供参考。(3)密蒙花尿液代谢成分分析我们通过大鼠高剂量灌胃给药的方式,采集尿液样本分析,用来表征密蒙花经体内过程后可能存在的代谢物。研究结果:一共在尿液样本中检测到34个主要成分,包括5个原型成分和29个代谢产物。(4)密蒙花序贯代谢研究密蒙花序贯代谢研究主要分为以下几部分:Ⅰ.胃肠道稳定性研究;Ⅱ.肠道菌群代谢研究Ⅲ.肠壁代谢研究ⅣV.肝代谢研究研究结果表明:肠道菌群和肝脏是代谢密蒙花的主要的地方,肠壁酶会对其中一小部分成分代谢。通过序贯代谢实验,我们可以清晰的看到口服给药后,各成分在不同代谢位点的变化。3.密蒙花网络药理学研究运用Medchem Studio对密蒙花所含化学成分及体内代谢成分进行靶标预测,同时收集已知与结膜炎相关的疾病靶标数据,基于PPI蛋白相互作用数据库对密蒙花成分靶标与己知与疾病相关的靶标进行网络构建,根据节点的拓扑结构特征值筛选关键靶标,并对其进行通路富集分析。研究结果表明咖啡酸的甲基化代谢物有可能是治疗眼结膜炎的关键药效物质基础;密蒙花主要通过炎症-免疫抑制功能模块达到抗炎的效果,调控炎症-免疫通路上的相关炎症因子抑制炎症反应,从而对眼结膜炎及充血等症状起到治疗作用。该部分研究结果仍需要进一步的动物实验更加全面地验证和阐释密蒙花治疗结膜炎的分子机制。结论:本文研究结果很好的阐释了密蒙花的治疗眼结膜炎的药效物质基础和药理作用机制。同时本文的"化学成分-序贯代谢-网络靶标"的研究模式也可以为其他中药研究提供参考。
[Abstract]:Due to the complex and various components of the traditional Chinese medicine, the material base and the mechanism of action of the traditional Chinese medicine have become a great challenge. It is well known that the traditional Chinese medicine decoction is generally used in the form of oral administration. After oral administration, the multi-component of the traditional Chinese medicine is generally subject to the gastrointestinal barrier before entering the blood circulation system. In a series of processes, not all of the traditional Chinese medicine components can enter the blood circulation system to produce the medicine effect. Some of the traditional Chinese medicine components can be absorbed into the blood in the form of a prototype, and some of the traditional Chinese medicine components may be metabolized by the enzymes of the intestinal tract or the liver, thereby producing new metabolites. Therefore, the components that eventually enter the blood circulation are extremely complex. In recent years, because of its rapid and sensitive characteristics, the technology of liquid-quality combination has become an important technical means to analyze the multi-component and its metabolites in complex biological samples due to its rapid and sensitive characteristics. However, it is not enough for the light analysis to be clear that the blood component is far from enough, and it is necessary to construct the relationship between the composition and the drug effect. In recent years, with the development of bioinformatics, network pharmacology has become an important technical means to study the mechanism of traditional Chinese medicine. Therefore, this paper attempts to elucidate the material base and mechanism of the treatment of eye conjunctivitis by using the "chemical composition-sequence-through-metabolic-network target" 's research mode as the research carrier. The research is mainly divided into the following three parts, and the specific results are as follows: 1. The results of the study on the chemical constituents of the extract of Mimonflower by UHPLC-LtQ-Orbitrap show that 20 major chemical components, including 11 benzhydrins and 9 flavonoids, were identified. In this paper, the metabolic behavior of the benzene-ethanol-like components was studied by the study of the law of the metabolic behavior of the phenethyl alcohol (1). The results showed that four metabolites were detected and identified from the blood samples of Rhodiola rosea, and the main metabolic pathways were hydrolysis, glucuronidation and sulfation. In addition, the rhodiola root is more stable in the digestive juice, and is mainly metabolized by the intestinal flora and the liver, and a small part is metabolized by the intestinal wall. The results showed that 10 metabolites were detected in the urine samples of the pistil flower. The main metabolic pathways were hydrolysis, dehydroxylation, glucuronidation, sulfation, and methylation. In conclusion, the main metabolic pathway of the phenethyl alcohol is hydrolysis, dehydroxylation, glucuronidation, sulfation, methylation and so on. The results of this study can be used to provide reference for the estimation of the metabolites of other similar structural compounds in the extract. (2) Studies on the law of the metabolic behavior of the flavonoids, we selected two representative flavonoids, Monflower and luteolin, to elucidate the law of the metabolic behavior of the flavonoids. A total of 4 metabolites were identified in the urine and stool samples as a result of the study of the metabolism in the body of the Mongolian flower. The main metabolic pathway of the Mongolian flowers is hydrolysis, glucuronidation, demethylation and hydroxylation. The results of the study of the in vivo metabolism of luteolin: a total of 6 metabolites were identified in urine and stool samples. The main metabolic pathway of luteolin is methylation and glucuronidation. In general, the main metabolic pathway of flavonoids is hydrolysis, glucuronidation, demethylation and hydroxylation, methylation, etc. The results of this study can be used to provide reference for the estimation of the metabolites of other similar structural compounds in the extract. (3) The analysis of the urine metabolites in the miconflower was carried out by means of the high dose of the rat, and the urine samples were collected and used to characterize the metabolites that might be present after the in-vivo process. Results: A total of 34 major components were detected in the urine sample, including 5 prototype components and 29 metabolites. (4) The study on the cross-metabolism of the dense-masked flower is divided into the following parts: 鈪，

本文编号：2396776