發(fā)布時(shí)間：2018-03-06 10:20

  本文選題：重金屬　切入點(diǎn)：生物發(fā)光　出處：《山東大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：作為常見的污染物,重金屬(Heavy Metal)是比重相對(duì)輕金屬更大的一類金屬,可以由工業(yè)污染、交通尾氣以及食物、化妝品中引入。由于脂溶性高、易產(chǎn)生螯合作用等特性,重金屬在自然界或人體內(nèi)存在著蓄積的現(xiàn)象,當(dāng)蓄積達(dá)到一定量時(shí),容易引發(fā)機(jī)體各器官、各系統(tǒng)退行性功能損傷,因此危害巨大。除了傳統(tǒng)的檢測手段之外,最近新發(fā)展起來的熒光小分子探針檢測具有較強(qiáng)的優(yōu)勢(shì),但這些檢測手段都存在無法應(yīng)用于活體檢測的缺點(diǎn),因此本工作主要以重金屬鈀和汞為例,探索將前沿的生物發(fā)光檢測手段應(yīng)用于重金屬的檢測中。生物發(fā)光(Bioluminescence)是一種自然界普遍存在的,依賴于生物正常生命活動(dòng)的化學(xué)發(fā)光。其本質(zhì)是酶和底物之間的相互作用,經(jīng)過一系列生物反應(yīng)產(chǎn)生化學(xué)能,并以光能的形式釋放出來。生物發(fā)光成像技術(shù)憑借其高靈敏性、高生物相容性以及可視化等特點(diǎn)已經(jīng)成為很重要的檢測手段,被廣泛應(yīng)用于各個(gè)領(lǐng)域。其中螢火蟲生物發(fā)光系統(tǒng)是研究最為完善的一類生物發(fā)光系統(tǒng)�；谝褕�(bào)道的重金屬鈀和汞的熒光小分子探針的設(shè)計(jì)機(jī)理,本論文設(shè)計(jì)合成了兩類五個(gè)生物發(fā)光探針,以開發(fā)用于重金屬鈀和汞的體內(nèi)外檢測技術(shù)。主要包括三個(gè)部分:一、前言;二、重金屬鈀的生物發(fā)光檢測及其應(yīng)用;三、重金屬汞的生物發(fā)光檢測及其應(yīng)用。第一部分介紹了重金屬的概況、其污染和毒性研究現(xiàn)狀及傳統(tǒng)檢測方法;總結(jié)目前已有的重金屬鈀和汞的熒光小分子檢測探針及其基本檢測原理;概括了生物發(fā)光的原理及螢火蟲螢光素系統(tǒng)。第二部分主要介紹了本課題研究的2個(gè)重金屬鈀生物發(fā)光探針。鈀在珠寶裝飾行業(yè)、交通行業(yè)、電子和電氣行業(yè)、醫(yī)療行業(yè)以及工業(yè)催化行業(yè)均有相應(yīng)的應(yīng)用,但鈀進(jìn)入體內(nèi)后會(huì)引起很大危害,因此本部分設(shè)計(jì)合成了兩個(gè)生物發(fā)光鈀探針。兩個(gè)鈀探針均以炔丙基醚基為水解基團(tuán),與鈀作用后釋放螢光素酶底物,從而實(shí)現(xiàn)檢測鈀的目的。該部分對(duì)探針的合成路線、體外應(yīng)用、細(xì)胞水平應(yīng)用及動(dòng)物水平的應(yīng)用進(jìn)行了探討。結(jié)果表明兩個(gè)鈀探針均可以實(shí)現(xiàn)體外對(duì)鈀的線性檢測,但CH-1的強(qiáng)度較弱,因此在動(dòng)物水平僅以CH-2為檢測探針,結(jié)果表明該探針可以實(shí)現(xiàn)對(duì)鈀在小鼠體內(nèi)的檢測。第三部分主要介紹了本課題研究的3個(gè)汞生物發(fā)光探針。汞是一種非常重要的重金屬,在各個(gè)行業(yè)領(lǐng)域中都有應(yīng)用,包括制造行業(yè)、醫(yī)藥行業(yè)、化妝品行業(yè)以及礦產(chǎn)業(yè)等。汞是一中毒性非常大的重金屬,臨床上重金屬中毒的案例不在少數(shù),而且隨著工業(yè)發(fā)展,汞的大規(guī)模污染也有發(fā)生,造成很大的危害,因此本部分設(shè)計(jì)合成了三個(gè)汞離子探針。分別以乙烯基醚基和丁炔基醚基為水解基團(tuán),在汞的作用下釋放螢光素酶底物,用以檢測汞。該部分對(duì)三個(gè)探針的合成路線,體外檢測,細(xì)胞水平應(yīng)用以及動(dòng)物水平的應(yīng)用進(jìn)行了探討。結(jié)果表明,CHG-1由于反應(yīng)溫度原因,只能實(shí)現(xiàn)體外檢測,而CHG-3檢測范圍窄,靈敏性差,因此選擇檢測范圍窄且易于反應(yīng)的CHG-2作為體內(nèi)水平檢測的探針,CHG-2體內(nèi)可以實(shí)現(xiàn)對(duì)汞中毒的檢測,而且有對(duì)汞中毒體內(nèi)蓄積狀態(tài)進(jìn)行檢測的潛在價(jià)值。
[Abstract]:As the common pollutants, heavy metals (Heavy, Metal) is a kind of light metal metal proportion is relatively larger, from industrial pollution, traffic exhaust and food, cosmetics was introduced. Due to the high fat soluble, easy to produce the effect of chelating characteristics of heavy metals in the accumulation phenomenon in nature or human memory, when the accumulation reached a certain amount of time, easy to cause the body organs function, degenerative system damage, so the damage is huge. In addition to the traditional means of detection, fluorescent molecular probe detection recently developed has a strong advantage, but these measures cannot be applied to in vivo detection there are shortcomings, so this work is mainly to heavy metal palladium and mercury as an example, to explore the frontiers of bioluminescence detection methods in the detection of heavy metal application. Bioluminescence (Bioluminescence) is a kind of universal existence in nature, depends on the biological Chemical normal life activity light. Its essence is the interaction between the enzyme and the substrate, after a series of biological reaction and chemical energy, light energy is released. Bioluminescence imaging technology with its high sensitivity, high biocompatibility and visualization features detection means has become very important and widely used in various fields. The firefly bioluminescence system is a kind of biological sound emitting system as the research design. The mechanism of the fluorescence molecular probes of heavy metals have been reported palladium and mercury based on two kinds of five bioluminescent probes were designed and synthesized in this paper, in order to develop for heavy metal palladium and mercury in vivo detection technology mainly includes three parts: first, introduction; two, heavy metal palladium bioluminescence detection and its application; three, mercury bioluminescence detection and its application. The first part is the introduction of a large amount of money The general situation, the pollution and toxicity research and traditional detection methods; summarizes the current detection of fluorescent molecules for heavy metal palladium and mercury probe and its basic detection principle; summarizes the principle and firefly bioluminescence in fluorescent system. The second part mainly introduces the research of 2 metal palladium bioluminescence probe. Palladium traffic in the jewelry industry, decoration industry, electronic and electrical industry, medical industry and industrial application of catalytic industry has a corresponding, but palladium enters the body will cause great harm, so the design and synthesis of two bioluminescent probes. Two palladium palladium probe with propargyl ether base for the hydrolysis of groups with palladium prime Enzyme Fluorescence liberating effect, so as to achieve the purpose of detection PD. The part of synthetic routes of the probe in vitro application, application level and application level of animal cells is discussed. The results show that the two palladium probe can achieve linear for PD detection in vitro, but weaker CH-1, so in the animal level only using CH-2 as the detection probe, the results show that the probe can be achieved on the palladium in the detection of mice. The third part mainly introduces the research of 3 mercury biological luminescent probe. Mercury is a kind of heavy metal is very important, are widely used in various industries, including manufacturing industry, pharmaceutical industry, cosmetics industry and mining industry. Mercury is very toxic heavy metals in clinical heavy metal poisoning in a few cases, but with the development of industry, large-scale pollution of Mercury also can occur. Great harm, so the synthesis of three mercury ion probe design. This part respectively with vinyl ether and ether groups for the hydrolysis of Ding Queji, in the role of mercury release luciferase substrate for detection of mercury. 璇ラ儴鍒嗗涓変釜鎺㈤拡鐨勫悎鎴愯礬綰，

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