【摘要】：由于中藥成分復(fù)雜多樣,中藥藥效物質(zhì)基礎(chǔ)和作用機(jī)理研究已經(jīng)成為一個(gè)巨大的挑戰(zhàn)。眾所周知,中藥湯劑一般采用口服給藥的方式�？诜o藥后,中藥多成分一般要經(jīng)歷胃腸道屏障,才能進(jìn)入血液循環(huán)系統(tǒng)。在一系列的過程中,并不是所有的中藥成分均能夠進(jìn)入血液循環(huán)系統(tǒng)而產(chǎn)生藥效。有些中藥成分能夠以原型的形式吸收入血,有些中藥成分可能會(huì)被腸道或肝臟的酶代謝,從而產(chǎn)生新的代謝物。因此,最終進(jìn)入到血液循環(huán)的成分是極其復(fù)雜的。近些年來,由于分析技術(shù)手段的發(fā)展,液質(zhì)聯(lián)用技術(shù)由于其快速、靈敏的特點(diǎn),已經(jīng)成為在復(fù)雜生物樣本中分析中藥多成分及其代謝物的重要技術(shù)手段。然而,光分析清楚入血成分是遠(yuǎn)遠(yuǎn)不夠的,還需要構(gòu)建成分與藥效之間的聯(lián)系。近些年來,隨著生物信息學(xué)的發(fā)展,網(wǎng)絡(luò)藥理學(xué)已經(jīng)成為研究中藥作用機(jī)理的重要技術(shù)手段。因此,本文以密蒙花為研究載體,嘗試用"化學(xué)成分-序貫代謝-網(wǎng)絡(luò)靶標(biāo)"的研究模式來闡明密蒙花治療眼結(jié)膜炎的藥效物質(zhì)基礎(chǔ)和作用機(jī)理。研究主要分為以下三大部分,具體結(jié)果如下:1.利用UHPLC-LTQ-Orbitrap技術(shù)分析密蒙花提取物的化學(xué)成分研究結(jié)果表明,從密蒙花提取液中一共鑒定了 20個(gè)主要化學(xué)成分,包括11個(gè)苯乙醇苷類化合物和9個(gè)黃酮類化合物。2.密蒙花序貫代謝研究(1)苯乙醇苷類代謝行為規(guī)律研究我們選擇密蒙花中兩個(gè)代表性苯乙醇苷類單體紅景天苷和異毛蕊花苷來闡明苯乙醇苷類成分的代謝行為規(guī)律。紅景天苷序貫代謝研究結(jié)果:一共從紅景天苷血液樣本中檢測和鑒定了四個(gè)代謝物,其主要代謝途徑為水解,葡萄糖醛酸化和硫酸化。另外,紅景天苷在消化液中比較穩(wěn)定,其主要被腸道菌群和肝代謝,并且有一小部分被腸壁代謝。異毛蕊花苷體內(nèi)代謝研究結(jié)果:一共從異毛蕊花苷尿液樣本中檢測到了 10個(gè)代謝產(chǎn)物,其主要代謝途徑為水解,去羥基化,葡萄糖醛酸化,硫酸化,甲基化�？傊�,苯乙醇苷類的主要代謝途徑為水解,去羥基化,葡萄糖醛酸化,硫酸化,甲基化等。該研究結(jié)果可以為密蒙花提取液中其他類似結(jié)構(gòu)化合物代謝產(chǎn)物的推測提供參考。(2)黃酮類代謝行為規(guī)律研究我們選擇密蒙花中兩個(gè)代表性黃酮類單體蒙花苷和木犀草素來闡明黃酮類成分的代謝行為規(guī)律。蒙花苷體內(nèi)代謝的研究結(jié)果:一共在尿液和糞便樣品中鑒別了4個(gè)代謝物。蒙花苷的主要代謝途徑為水解,葡萄糖醛酸化,去甲基化以及羥基化。木犀草素體內(nèi)代謝的研究結(jié)果:一共在尿液和糞便樣品中鑒別了6個(gè)代謝物。木犀草素主要代謝途徑為甲基化和葡萄糖醛酸化。總之,黃酮類化合物的主要代謝途徑為水解,葡萄糖醛酸化,去甲基化以及羥基化,甲基化等。該研究結(jié)果可以為密蒙花提取液中其他類似結(jié)構(gòu)化合物代謝產(chǎn)物的推測提供參考。(3)密蒙花尿液代謝成分分析我們通過大鼠高劑量灌胃給藥的方式,采集尿液樣本分析,用來表征密蒙花經(jīng)體內(nèi)過程后可能存在的代謝物。研究結(jié)果:一共在尿液樣本中檢測到34個(gè)主要成分,包括5個(gè)原型成分和29個(gè)代謝產(chǎn)物。(4)密蒙花序貫代謝研究密蒙花序貫代謝研究主要分為以下幾部分:Ⅰ.胃腸道穩(wěn)定性研究;Ⅱ.腸道菌群代謝研究Ⅲ.腸壁代謝研究ⅣV.肝代謝研究研究結(jié)果表明:腸道菌群和肝臟是代謝密蒙花的主要的地方,腸壁酶會(huì)對(duì)其中一小部分成分代謝。通過序貫代謝實(shí)驗(yàn),我們可以清晰的看到口服給藥后,各成分在不同代謝位點(diǎn)的變化。3.密蒙花網(wǎng)絡(luò)藥理學(xué)研究運(yùn)用Medchem Studio對(duì)密蒙花所含化學(xué)成分及體內(nèi)代謝成分進(jìn)行靶標(biāo)預(yù)測,同時(shí)收集已知與結(jié)膜炎相關(guān)的疾病靶標(biāo)數(shù)據(jù),基于PPI蛋白相互作用數(shù)據(jù)庫對(duì)密蒙花成分靶標(biāo)與己知與疾病相關(guān)的靶標(biāo)進(jìn)行網(wǎng)絡(luò)構(gòu)建,根據(jù)節(jié)點(diǎn)的拓?fù)浣Y(jié)構(gòu)特征值篩選關(guān)鍵靶標(biāo),并對(duì)其進(jìn)行通路富集分析。研究結(jié)果表明咖啡酸的甲基化代謝物有可能是治療眼結(jié)膜炎的關(guān)鍵藥效物質(zhì)基礎(chǔ);密蒙花主要通過炎癥-免疫抑制功能模塊達(dá)到抗炎的效果,調(diào)控炎癥-免疫通路上的相關(guān)炎癥因子抑制炎癥反應(yīng),從而對(duì)眼結(jié)膜炎及充血等癥狀起到治療作用。該部分研究結(jié)果仍需要進(jìn)一步的動(dòng)物實(shí)驗(yàn)更加全面地驗(yàn)證和闡釋密蒙花治療結(jié)膜炎的分子機(jī)制。結(jié)論:本文研究結(jié)果很好的闡釋了密蒙花的治療眼結(jié)膜炎的藥效物質(zhì)基礎(chǔ)和藥理作用機(jī)制。同時(shí)本文的"化學(xué)成分-序貫代謝-網(wǎng)絡(luò)靶標(biāo)"的研究模式也可以為其他中藥研究提供參考。
[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，

本文編號(hào)：2396776