摘 要

在生物系统中，过硫化氢的浓度差异可能会诱发许多生理疾病，例如糖尿病、阿尔兹海默症、肝硬化和唐氏综合症等。随着人们生活水平的提高，糖尿病的危害越来越大。众多研究表明2型糖尿病与血清中过硫化氢水平有关，提高血清过硫化氢水平可预防2型糖尿病的发生。因此构建可以用检测生物体内过硫化氢水平的荧光探针是很有意义的。

我们选择半花菁衍生物（具有ICT效应）作为荧光团，过硫化氢能特异性识别并断开2-氟-5-硝基苯甲酸基团，进而构建出反应型荧光探针MC-S_n。半花菁是一类结构易修饰的有机荧光母核，我们对其结构中的羟基进行修饰，引入了2-氟-5-硝基苯甲酸进而阻断了ICT效应，使得探针自身不发射荧光；而当加入过硫化氢后，其可断开2-氟-5-硝基苯甲酸并重新释放出具有ICT效应的荧光团半花菁。荧光探针MC-S_n具有光稳定性强、较低的细胞毒性和显著的颜色变化等优点。此外，荧光探针MC-S_n还可运用于对RAW 264.7细胞中内源性和外源性的过硫化物进行实时、无损、原位的检测。

关键词：荧光探针 过硫化物 半花菁 分子内电荷转移

Abstract

In the biological system, the level of hydrogen sulfide may induce many physiological diseases such as diabetes, Alzheimer's disease, cirrhosis, and Down's syndrome. There are obvious regional differences and population differences in the mortality rate of diabetes, ranging from 0 to 200 / 100,000. Data for the past 30 years show that the death rate from diabetes is higher in developed countries than in developing countries, and higher in urban areas than in rural areas. The mortality rate in developed countries is mostly between 20 and 50 per 100,000, while that in developing countries is mostly less than 10 per 100,000, that in urban areas is generally higher than 10 per 100,000, and that in rural areas is more than 10 per 100,000. The death rate of diabetes among different races is significantly different, and blacks are generally higher than whites. Our diabetes mortality rate has grown rapidly in the past 20 years. Numerous studies have shown that type 2 diabetes is related to serum hydrogen sulfide levels, and raising serum hydrogen sulfide levels can prevent the occurrence of type 2 diabetes. Therefore, it is very meaningful to construct a fluorescent probe that can be used to detect the level of hydrogen sulfide in organisms.

We chose the semi-flower cyanine derivative(with the ICT effect) as the fluorescence group, and the supersulphide can specifically identify and disconnect the 2-fluoro-5-nitrobenzoic acid group to construct the reactive fluorescence probe MC-S_n. Semi-anthocyanine is a kind of organic fluorescent mother nucleus that is easily modified. We modify the hydroxyl group in its structure and introduce 2-fluoro-5-nitrobenzoic acid to block the ICT effect, so that the probe itself does not emit fluorescence; When sulfides are added, they can disconnect 2-fluoro-5-nitrobenzoic acid and re-release fluorescent pellets with ICT effects. Fluorescent probe MC-S_n has the advantages of strong optical stability, low cytotoxicity and significant color changes. In addition, the fluorescent probe MC-S_n can also be used to detect endogenous and exogenous supersulfide in RAW 264.7 cells in real time, lossless, and in situ.

目录

摘要 I

Abstract II

第一章 绪论 1

1.1 糖尿病的研究现状....... 1

1.2 活性硫化物 1

1.2.1 活性硫化物介绍 1

1.2.2 活性硫化物的作用 1

1.2.3 常用的检测方法 2

1.3 荧光探针 3

1.3.1 荧光探针的历史研究 3

1.3.2 荧光的定义 4

1.3.3 荧光探针的介绍 4

1.3.4 荧光探针的种类 4

1.4 基于半花菁荧光团的荧光探针 4

第二章 实验材料与方法 6

2.1 实验仪器与材料 6

2.2 MC-S_n的合成路线 6

2.2.1 化合物3的合成 6

2.2.2 化合物5的合成 7

2.2.3 MC-S_n的合成 ...............................................................................................................7

2.2.4 吸收和荧光光谱...........................................................................................................7

2.2.5 双光子截面测量...........................................................................................................8

2.2.6 细胞培养和成像...........................................................................................................9

第三章 结果与讨论 10

3.1 荧光探针的体外实验 10

3.1.1 外源性H₂S_n和H₂S_n的反应动力学............................................................................10

3.1.2 MC-S_n与H₂S_n的光谱响应...........................................................................................11

3.1.3 检测MC-S_n性能..........................................................................................................13

3.1.4光稳定性和pH值影响 13

3.2 荧光探针的体内实验 15

3.2.1 细胞毒性实验 15

第四章 总结 19

4.1 总结 19

参考文献 20

致谢 23

绪论

随着我国经济的发展，人们的生活条件越来越好，糖尿病的发病率也开始逐渐上升。不仅仅在我国如此，就全世界范围来看，糖尿病仍是影响居民健康的重大疾病。目前我国每天要新增15000名糖尿病患者，因此我们必须采取措施应对糖尿病患病人数继续增加的现状。^[1]糖尿病的危害巨大，给病人的生理和心理都造成了巨大的痛苦，病人需要长期服用治疗药物，而且饮食、生活规律等方面都要特别注意，另外，糖尿病的并发症也在威胁着患者的健康，如患者脑血管出现并发症可能会导致偏瘫，眼睛出现并发症可能造成失明，心脏出现并发症可能造成患者心肌梗死，肾脏出现并发症可能会造成肾功能衰竭、尿毒症，下肢出现并发症可能造成锯腿等。

糖尿病的研究现状

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