本文關(guān)鍵詞： 熒光探針 近紅外 巰基化合物 活細(xì)胞 成像　出處：《蘭州大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

【摘要】：細(xì)胞內(nèi)的小分子巰基化合物如半胱氨酸(Cys),同型半胱氨酸(Hcy),谷胱甘肽(GSH),在生物體內(nèi)有著重要的生理功能。它們的含量水平是衡量生物體健康狀況的重要標(biāo)準(zhǔn)。熒光探針具有高靈敏度,高選擇性,操作便捷等優(yōu)點(diǎn),被廣泛應(yīng)用于檢測(cè)各種離子和分子。因此,設(shè)計(jì)合成能夠選擇性檢測(cè)細(xì)胞內(nèi)的小分子巰基化合物的熒光探針具有十分重要的科學(xué)意義。本文設(shè)計(jì)合成了兩個(gè)檢測(cè)小分子巰基化合物的探針,并將其應(yīng)用于活細(xì)胞成像。本文分為三章。第一章主要闡述了熒光的基礎(chǔ)知識(shí),熒光探針的主要機(jī)理,研究背景及小分子巰基化合物探針的研究進(jìn)展。在第二章中,設(shè)計(jì)并合成了一個(gè)基于光誘導(dǎo)電子轉(zhuǎn)移(PET)機(jī)理的選擇性識(shí)別巰基化合物的熒光探針1。我們?cè)敿?xì)研究了探針1和不同的分析物作用后的紫外吸收和熒光發(fā)射性質(zhì)。探針1在和巰基化合物作用后有一個(gè)很大的斯托克斯位移(~200 nm)并且可以在DMSO/PBS(9/1,v/v,10 mM,pH 3.5)中選擇性檢測(cè)Cys而不受GSH,Hcy,CN?和其他氨基酸的干擾,檢測(cè)限為0.124μM。而在pH 5.5時(shí),探針1對(duì)GSH,Hcy和Cys都響應(yīng),檢測(cè)限分別為0.151μM,0.128μM和0.040μM。探針1的細(xì)胞毒性很低,被成功地應(yīng)用于活細(xì)胞中巰基化合物的成像。在第三章中,合成了一個(gè)氯乙�；妮翢晒馑亟t外(NIR)熒光探針2(NFC)。它可以選擇性地識(shí)別Cys和Hcy而不受GSH和其它氨基酸(AAs)的影響。NFC對(duì)Cys和Hcy的檢測(cè)限分別為0.30和0.42μM。萘熒光素(NF)作為熒光團(tuán),其熒光強(qiáng)度由分子內(nèi)電荷轉(zhuǎn)移(ICT)過程控制。該探針合成簡(jiǎn)單,自身熒光很弱,但和Cys或Hcy反應(yīng)后,熒光明顯增強(qiáng)。我們證明了熒光的關(guān)開過程是由于NFC與Cys和Hcy先發(fā)生了親和取代反應(yīng),緊接著進(jìn)行了分子內(nèi)的環(huán)化生成了NF。該探針的毒性也比較低,被成功地應(yīng)用于檢測(cè)人血漿中的Cys和Hcy的總含量以及活細(xì)胞成像。
[Abstract]:Small molecule thiol compounds such as cysteine within the cell (Cys), homocysteine (Hcy), glutathione (GSH), have important physiological functions in organisms. Their content is an important standard to measure the organism's health. Fluorescent probe with high sensitivity, high selectivity, convenient operation and other advantages, is widely used in the detection of various ions and molecules. Therefore, the design and synthesis of which has very important scientific significance of small molecule thiol fluorescent probes for selective detection of cells. Two detection probe mercapto compounds were designed and synthesized, and its application in live cell imaging. This paper is divided into three chapters. The first chapter mainly introduces the basic knowledge of the main mechanism of fluorescence, fluorescence probe, probe the research progress of sulfhydryl compounds and small molecular background. In the second chapter, the design and synthesis based on a Photoinduced electron transfer (PET) fluorescence probe for selective recognition of sulfhydryl compounds mechanism 1. we studied in detail the probe and fluorescence emission properties of UV and 1 different analytes after absorption. In 1 probe and thiols. After a large Stokes shift (~200 nm) and the DMSO/PBS (9/1 10, v/v, mM, pH 3.5) in the selective detection of Cys but not by GSH, Hcy, CN and other amino acids? Interference, the detection limit is 0.124 M. and 5.5 in pH, 1 for Hcy and GSH probe, Cys response, detection limits were 0.151 M, 0.128 M cell toxicity and the 0.040 M. probe 1 is very low, imaging has been successfully applied to the sulfhydryl compounds in living cells. In the third chapter, the synthesis of a naphthalene fluorescein chloroacetylated near infrared fluorescent probe (NIR) 2 (NFC). It can selectively recognize Cys and Hcy but not by GSH and other the effects of amino acids (AAs). The NFC of Cys and the detection limit of Hcy were 0.30 and 0.42 M. naphthalene (NF) as fluorescein fluorophore, the fluorescence intensity of the intramolecular charge transfer (ICT) process control. The probe has simple synthesis, its fluorescence is very weak, but Cys or Hcy after the reaction, the fluorescence enhanced. We show that fluorescence the closing process is due to NFC and Cys and Hcy happened first nucleophilic substitution reaction, followed by intramolecular cyclization of the probe generated NF. toxicity is relatively low, the total content was successfully applied to detect Cys and Hcy in human plasma and live cell imaging.

【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：O657.3

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本文編號(hào)：1522980