荧光探针法研究药物与生物大分子作用机理及其测定

发布时间：2018-02-03 08:41

本文关键词： 牛血清白蛋白荧光探针荧光猝灭链霉素槲皮素脱落酸　出处：《山东师范大学》2011年硕士论文　论文类型：学位论文

【摘要】：药物与生物大分子之间的结合机理和作用机理是药学科学领域中一个重要的组成部分。药物在人体中的结合部位和作用机理直接影响着药物对疾病的治疗作用。因此,在模拟人体环境条件下研究药物与生物大分子的结合部位和作用机理是十分必要的,也可以确保人们用药的安全、合理和有效。蛋白质是生物体内重要的大分子,它能与许多内源或外源性的化合物结合,具有存储和转运的功能。药物进入人体一般要经过血浆的贮存和运输,到达受体部位发生药理作用,因此研究血清白蛋白与药物的结合作用机理具有重要的理论和现实意义。目前,用于表征蛋白质-药物分子结合作用的方法很多,其中荧光探针技术因其具有灵敏度高、选择性好、简便等特点而得到广泛应用。荧光探针法就是利用加入的药物与探针分子作用使得生物大分子或探针分子的荧光发生变化来研究药物分子与生物大分子之间的作用机理,从而进一步了解药物分子在人体中的作用机理。本论文主要采用荧光探针法研究药物分子与牛血清白蛋白的结合部位和作用机理,从而进一步了解和研究药物在人体中的作用机理和发挥药效的途径。本论文共分为五章第一章综述了药物分子与血清白蛋白相互作用的研究现状及研究意义。首先简要介绍了蛋白质的结构和理化性质,然后依次对药物小分子与血清白蛋白相互作用的研究方法、研究内容和研究进展展开了评述。第二章利用荧光光谱和紫外光谱法研究了伊文思蓝(EB, Evans blue)和链霉素(SM, streptomycin)对牛血清白蛋白间的竞争反应机制。伊文思蓝能够猝灭牛血清白蛋白的荧光发生静态猝灭。然而当加入链霉素后,牛血清白蛋白的荧光恢复。这说明EB和链霉素对BSA发生了竞争反应。求得了牛血清白蛋白与链霉素的结合常数为6.52×10~6 L/moL。第三章利用荧光光谱法研究了牛血清白蛋白与脱落酸的相互作用。脱落酸与牛血清白蛋白作用使牛血清白蛋白发生荧光猝灭。根据Stern-Volmer方程和荧光寿命研究了荧光猝灭的类型及机理。其猝灭机理是静态猝灭,即脱落酸和牛血清白蛋白形成了一种稳定的复合物。利用同步荧光光谱确定了脱落酸对牛血清白蛋白的微区结构的影响。通过实验求算了脱落酸和牛血清白蛋白的结合常数和结合点数K =6.38×10~4L.mol~(-1), n =1.452。第四章利用荧光探针法筛选评价了天然抗氧化药剂槲皮素的抗氧化能力,以1, 3-二苯基异苯并呋喃(DPBF)作为荧光探针,研究了抗氧化药物槲皮素对单线态氧的清除能力,讨论了其作用机制,并实验了干扰物质的影响。第五章利用荧光猝灭法建立了测定植物中脱落酸的新方法。
[Abstract]:The binding mechanism and action mechanism between drugs and biomolecules is an important part in the field of pharmaceutical science. The binding site and action mechanism of drugs in human body directly affect the therapeutic effect of drugs on diseases. So... It is necessary to study the binding site and action mechanism of drugs and biomolecules under simulated human environment, and to ensure the safety of drug use. Reasonable and effective. Protein is an important macromolecule in organism, it can bind with many endogenous or exogenous compounds, and has the function of storage and transportation. Drugs entering the human body generally have to be stored and transported through plasma. Therefore, it is of great theoretical and practical significance to study the mechanism of binding of serum albumin to drugs. At present, there are many methods used to characterize protein-drug molecular binding, among which fluorescent probe technology has high sensitivity and good selectivity. Fluorescence probe method is to make use of the action of added drug and probe molecule to change the fluorescence of biological macromolecule or probe molecule to study the action between drug molecule and biological macromolecule. Using mechanism. In order to further understand the action mechanism of drug molecules in human body, this paper mainly uses fluorescence probe method to study the binding site and action mechanism of drug molecules with bovine serum albumin (BSA). In order to further understand and study the mechanism of action of drugs in the human body and the ways to give full play to the drug effect. This paper is divided into five chapters. In the first chapter, the research status and significance of the interaction between drug molecules and serum albumin are reviewed. Firstly, the structure and physicochemical properties of proteins are briefly introduced. Then, the research methods, contents and progress of the interaction between drug small molecules and serum albumin were reviewed in turn. In chapter 2, Evans blueand streptomycin SM were studied by fluorescence and UV spectra. The mechanism of competitive reaction between bovine serum albumin by streptomycin.Evans blue can quench the fluorescence of bovine serum albumin by static quenching, however, when streptomycin is added. The fluorescence recovery of bovine serum albumin indicates that EB and streptomycin react competitively to BSA, and the binding constant of bovine serum albumin to streptomycin is 6.52 脳 10 ~ (-6) L / mol ~ (-1). In chapter 3, the interaction between bovine serum albumin and abscisic acid was studied by fluorescence spectrometry. The fluorescence quenching of bovine serum albumin was induced by the interaction of abscisic acid and bovine serum albumin. The type and mechanism of fluorescence quenching are studied by the equation and fluorescence lifetime. The quenching mechanism is static quenching. The effect of abscisic acid on the microstructures of bovine serum albumin (BSA) was determined by synchronous fluorescence spectroscopy. The abscisic acid and bovine serum albumin (BSA) were calculated by experiments. Binding constants and binding points K of the. 6.38 脳 10 ~ (4) L. Moll (. -1). N 1.452. In chapter 4th, the antioxidant ability of quercetin was evaluated by fluorescence probe method, and 1,3-diphenyl isobenzofuran (DPBF) was used as the fluorescent probe. The scavenging ability of quercetin on singlet oxygen was studied, the mechanism of its action was discussed, and the effect of interfering substances was tested. In Chapter 5th, a new method for the determination of abscisic acid in plants was established by fluorescence quenching method.
【学位授予单位】：山东师范大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R346

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