基于苯并吡喃腈荧光团检测偶氮还原酶的荧光探针的设计、合成和生物成像

发布时间：2018-10-12 15:56

【摘要】：荧光探针及荧光生物成像技术近年来取得了迅猛发展,该技术可以对活体细胞和动物体内的微小物种进行荧光成像检测。偶氮还原酶是生命体内一种重要的酶,肿瘤中的偶氮还原酶水平与缺氧程度密切相关,对偶氮还原酶水平的检测可用于评价肿瘤的缺氧程度(肿瘤的缺氧程度是判断其为良性或恶性的重要指标),因而,对偶氮还原酶水平的检测具有重要意义。利用荧光探针及荧光生物成像技术对活体细胞及活体组织中的偶氮还原酶进行检测,在生物医学及临床诊断学中具有重要的意义。本文设计合成了一种检测偶氮还原酶的光物理性能优良的荧光探针,并将该探针成功应用于活体细胞、秀丽线虫和实验小鼠肿瘤中的偶氮还原酶荧光成像研究。主要工作内容如下:一、我设计合成了3种苯并吡喃腈荧光团,研究了这些荧光团的光物理性质,发现荧光团C3在DMF PBS缓冲液(0.02 M,pH 7.4)(V/V=5:5)中的最大发射波长位于675 nm,达到了近红外荧光发射区(650-900 nm)。二、我基于荧光团C3设计合成了2种新型的检测偶氮还原酶的荧光探针S1和S2,研究了这2种荧光探针对偶氮还原酶的荧光识别和荧光成像性能,发现探针S1和S2均对偶氮还原酶表现出由红色荧光到绿色荧光发射的光谱变化。我们选用光物理性能更为优良的探针S1为主要研究对象,对探针S1与偶氮还原酶的作用机理及探针S1在生物荧光成像(细胞、秀丽线虫和小鼠肿瘤组织)中的应用进行了研究,发现探针S1对偶氮还原酶具有良好的光谱响应和生物相容性,可成功检测活体细胞和活体组织中的偶氮还原酶。
[Abstract]:In recent years, fluorescent probes and fluorescent imaging techniques have been developed rapidly, which can be used to detect small species in living cells and animals. Azoreductase is an important enzyme in human body. The level of azoreductase in tumor is closely related to the degree of hypoxia. The determination of azoreductase level can be used to evaluate the degree of anoxia of tumor (the degree of anoxia of tumor is an important index to judge whether it is benign or malignant), so it is important to detect the level of azoreductase. The detection of azoreductase in living cells and tissues by fluorescence probe and fluorescence biometric imaging is of great significance in biomedicine and clinical diagnosis. A photophysical fluorescent probe for the detection of azoreductase was designed and synthesized. The probe was successfully applied to the fluorescence imaging of azoreductase in living cells, nematodes and mice. The main work is as follows: 1. I have designed and synthesized three kinds of benzopyranonitrile fluorescent groups and studied their photophysical properties. It was found that the maximum emission wavelength of fluorescent group C3 in DMF PBS buffer solution (0.02 MN pH 7.4) (V/V=5:5) was at 675 nm, and reached the near infrared fluorescence emission region (650-900 nm). Secondly, I designed and synthesized two new fluorescent probes S1 and S2 for detecting azo reductase based on fluorescent group C3, and studied the fluorescence recognition and imaging properties of these two fluorescent probes for azoreductase. It was found that both S _ 1 and S _ 2 showed spectral changes from red fluorescence to green fluorescence emission for azo reductase. The mechanism of probe S1 and azoreductase and the application of probe S1 in biological fluorescence imaging (cell, nematode and mouse tumor tissue) were studied. It was found that the probe S1 had good spectral response and biocompatibility to azo reductase and could be successfully used to detect azo reductase in living cells and tissues.
【学位授予单位】：云南师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.3

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