發(fā)布時間：2018-04-21 01:34

  本文選題：甲醛 + 熒光探針��； 參考：《曲阜師范大學》2017年碩士論文

【摘要】：甲醛是一種高活性的羰基化合物,作為代謝物的中間體普遍存在于人的各個器官組織中。在賴氨酸特異性脫甲基酶1(LSD1)和氨基脲敏感性胺氧化酶(SSAO)的催化下,氨基酸和外源性物質(zhì)代謝過程中能夠生成甲醛。甲醛能夠與單胺、酞胺類物質(zhì)發(fā)生相互作用,形成穩(wěn)定的亞甲基橋;也與一些DNA和蛋白質(zhì)進行交聯(lián),從而對其功能和結(jié)構(gòu)造成影響。研究表明內(nèi)源性甲醛的濃度失衡會造成一系列的疾病,如神經(jīng)系統(tǒng)疾病、心腦血管疾病和癌癥等。因此,檢測生物體中的甲醛具有重要的生物醫(yī)學意義。在各種生物檢測技術(shù)中,熒光探針具有選擇性好,可以實時原位檢測,靈敏度高,對樣品損傷小等優(yōu)點,受到了廣泛關(guān)注。我們報道了一種用于檢測甲醛的BODIPY“關(guān)型”熒光探針BoN。探針利用氨基作為響應(yīng)基團,在多種活性羰基物種存在的條件下,該探針對甲醛具有高選擇性。我們還驗證了LSD1對甲醛的產(chǎn)生起著重要的作用。探針成功的應(yīng)用于活細胞外源性以及內(nèi)源性的甲醛檢測,同時還應(yīng)用于小鼠及其海馬體切片的熒光成像。實驗中,檢測了小鼠各個器官組織(心、肝、脾、肺、腎、腦)勻漿中甲醛的濃度。論文共分為四部分,內(nèi)容如下:1、對熒光成像技術(shù)、BODIPY熒光團和探針的設(shè)計原理進行簡單的介紹。對已發(fā)表的用于甲醛檢測的熒光探針的設(shè)計原理、響應(yīng)機制以及生物應(yīng)用進行綜述,開發(fā)簡單靈敏的熒光探針對于研究生命活動具有重要的生物學和醫(yī)學意義。2、介紹新型的BODIPY熒光探針BoN的合成路線和具體操作步驟,并介紹探針檢測甲醛的機理。3、利用新型熒光探針BoN進行體外甲醛的檢測,使用紫外分光光度計和熒光分光光度計對探針的光譜進行測試,同時對探針的選擇性進行了分析。4、將熒光探針BoN用于細胞、組織以及活體內(nèi)的甲醛的檢測,使用熒光分光光度計、激光共聚焦顯微鏡和小動物成像系統(tǒng)對探針的性能進行評估,檢測細胞、組織以及活體內(nèi)的甲醛濃度,并驗證LSD1酶對于甲醛的生成產(chǎn)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.
【學位授予單位】：曲阜師范大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R446;O657.3

【參考文獻】

相關(guān)期刊論文 前1條

1 ;Formaldehyde stress[J];Science China(Life Sciences);2010年12期

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本文編號：1780370