基于BODIPY染料的甲醛荧光探针研究

发布时间：2018-04-21 01:34

本文选题：甲醛 + 荧光探针　；参考：《曲阜师范大学》2017年硕士论文

【摘要】：甲醛是一种高活性的羰基化合物,作为代谢物的中间体普遍存在于人的各个器官组织中。在赖氨酸特异性脱甲基酶1(LSD1)和氨基脲敏感性胺氧化酶(SSAO)的催化下,氨基酸和外源性物质代谢过程中能够生成甲醛。甲醛能够与单胺、酞胺类物质发生相互作用,形成稳定的亚甲基桥;也与一些DNA和蛋白质进行交联,从而对其功能和结构造成影响。研究表明内源性甲醛的浓度失衡会造成一系列的疾病,如神经系统疾病、心脑血管疾病和癌症等。因此,检测生物体中的甲醛具有重要的生物医学意义。在各种生物检测技术中,荧光探针具有选择性好,可以实时原位检测,灵敏度高,对样品损伤小等优点,受到了广泛关注。我们报道了一种用于检测甲醛的BODIPY“关型”荧光探针BoN。探针利用氨基作为响应基团,在多种活性羰基物种存在的条件下,该探针对甲醛具有高选择性。我们还验证了LSD1对甲醛的产生起着重要的作用。探针成功的应用于活细胞外源性以及内源性的甲醛检测,同时还应用于小鼠及其海马体切片的荧光成像。实验中,检测了小鼠各个器官组织(心、肝、脾、肺、肾、脑)匀浆中甲醛的浓度。论文共分为四部分,内容如下:1、对荧光成像技术、BODIPY荧光团和探针的设计原理进行简单的介绍。对已发表的用于甲醛检测的荧光探针的设计原理、响应机制以及生物应用进行综述,开发简单灵敏的荧光探针对于研究生命活动具有重要的生物学和医学意义。2、介绍新型的BODIPY荧光探针BoN的合成路线和具体操作步骤,并介绍探针检测甲醛的机理。3、利用新型荧光探针BoN进行体外甲醛的检测,使用紫外分光光度计和荧光分光光度计对探针的光谱进行测试,同时对探针的选择性进行了分析。4、将荧光探针BoN用于细胞、组织以及活体内的甲醛的检测,使用荧光分光光度计、激光共聚焦显微镜和小动物成像系统对探针的性能进行评估,检测细胞、组织以及活体内的甲醛浓度,并验证LSD1酶对于甲醛的生成产生了重要作用。
[Abstract]:Formaldehyde is a highly active carbonyl compound. As an intermediate of metabolites, formaldehyde widely exists in human organs and tissues. Under the catalysis of Lysine-specific demethylase (LSD1) and semicarbazone sensitive amine oxidase (SSAO), formaldehyde can be produced during the metabolism of amino acids and exogenous substances. Formaldehyde can interact with monoamines and phthalamines to form a stable methylene bridge, and is also cross-linked with some DNA and proteins, thus affecting its function and structure. Studies have shown that the imbalance of endogenous formaldehyde concentration can lead to a series of diseases, such as nervous system diseases, cardiovascular and cerebrovascular diseases and cancer. Therefore, the detection of formaldehyde in organisms has an important biomedical significance. Among all kinds of biological detection techniques, fluorescent probe has many advantages, such as good selectivity, real-time in situ detection, high sensitivity and little damage to samples. We report a BODIPY "off type" fluorescence probe BoN for the determination of formaldehyde. The probe has high selectivity to formaldehyde in the presence of many active carbonyl species using amino groups as the responsive group. We also verify that LSD1 plays an important role in the production of formaldehyde. The probe was successfully applied to the detection of exogenous and endogenous formaldehyde in living cells, and also to the fluorescence imaging of mouse and hippocampal slices. The concentrations of formaldehyde in the homogenate of each organ (heart, liver, spleen, lung, kidney and brain) of mice were measured. The thesis is divided into four parts, which are as follows: 1. The design principle of the fluorescent cluster and probe is briefly introduced. The design principle, response mechanism and biological application of fluorescent probes for formaldehyde detection were reviewed. The development of simple and sensitive fluorescent probes is of great biological and medical significance for the study of life activities. The synthetic route and specific operation steps of novel BODIPY fluorescent probe BoN are introduced. The mechanism of detection of formaldehyde with probe. 3, the detection of formaldehyde in vitro with a new fluorescent probe BoN, and the spectrum of the probe with ultraviolet spectrophotometer and fluorescence spectrophotometer are also introduced. At the same time, the selectivity of the probe was analyzed. The fluorescence probe BoN was used for the detection of formaldehyde in cells, tissues and in vivo, and the performance of the probe was evaluated by fluorescence spectrophotometer, laser confocal microscope and small animal imaging system. Formaldehyde concentrations in cells, tissues and in vivo were measured, and LSD1 enzymes were demonstrated to play an important role in formaldehyde production.
【学位授予单位】：曲阜师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R446;O657.3

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