發(fā)布時(shí)間：2018-05-05 05:19

  本文選題：芳香烴受體 + 納米顆粒��； 參考：《山東大學(xué)》2017年碩士論文

【摘要】：納米材料由于其特異的理化性質(zhì)已經(jīng)被廣泛應(yīng)用于諸如航空航天、汽車工業(yè)、環(huán)保建材、體育用品、電子器件以及日常消費(fèi)品等領(lǐng)域,而且在生物醫(yī)藥領(lǐng)域如診斷、生物成像、生物傳感器和藥物運(yùn)輸?shù)纫簿哂袕V泛的應(yīng)用前景。因此人們暴露在納米環(huán)境中的機(jī)會(huì)也越來越頻繁。相對(duì)于從日常產(chǎn)品的接觸中暴露納米顆粒,應(yīng)用于醫(yī)藥領(lǐng)域的納米顆粒由于可以通過攝入和注射等方式直接進(jìn)入人體,因此人們更加關(guān)注納米顆粒在這一領(lǐng)域額應(yīng)用的安全性。芳香烴受體(Aryl hydrocarbon receptor,AhR)作為調(diào)控外源毒性化合物代謝基因的關(guān)鍵核受體,其下游被研究最多也是最經(jīng)典的靶基因是細(xì)胞色素P4501A1(CYP1A1)。近期的研究發(fā)現(xiàn)AhR在正常的生理?xiàng)l件下依然發(fā)揮著重要的作用,比如細(xì)胞增殖和分化,免疫平衡的調(diào)控等對(duì)AhR及其相關(guān)信號(hào)通路的干擾對(duì)細(xì)胞和生物體正常的生理功能可能產(chǎn)生不利影響。以往對(duì)AhR干擾物的研究往往聚焦于小分子化合物,忽略了一系列新穎的人工合成材料如納米顆粒也可能干擾AhR信號(hào)通路。已有的研究表明納米顆�？赡軙�(huì)干擾CYP1A1等AhR下游基因的表達(dá)活性�？紤]到納米顆粒在環(huán)境中的廣泛分布以及對(duì)人類健康的高暴露風(fēng)險(xiǎn),AhR信號(hào)通路是否能被納米顆粒干擾是個(gè)急需解決的問題。我們首先選擇以穩(wěn)轉(zhuǎn)熒光素酶報(bào)告基因的小鼠肝癌細(xì)胞CBG2.8D為實(shí)驗(yàn)?zāi)Ｐ?評(píng)價(jià)了幾種常見納米顆粒包括碳納米管(MWCNTs-COOH)、氧化石墨烯(GO-PEG)、納米二氧化硅顆粒(nSiO_2)、納米二氧化鈦顆粒(nTiO_2)、納米金顆粒(AuNPs)以及納米氧化鋅顆粒(nZnO)對(duì)AhR信號(hào)通路的干擾能力。實(shí)驗(yàn)結(jié)果表明上述納米顆粒除nTiO_2外,都具有不同程度干擾AhR-Luciferase通路的能力。為了進(jìn)一步闡明了納米顆粒與AhR信號(hào)通路之間相互作用的內(nèi)在分子機(jī)制,隨后我們選擇了 MWCNTs-COOH為模型,利用實(shí)時(shí)熒光定量PCR和蛋白質(zhì)免疫印跡等技術(shù),在基因和蛋白水平驗(yàn)證了 MWCNTs-COOH對(duì)AhR信號(hào)通路的干擾作用機(jī)制。結(jié)果表明MWCNTs-COOH能夠在基因和蛋白水平抑制CYP1A1的表達(dá),同時(shí)能夠改變AhR的基因表達(dá)水平和其總蛋白含量。但這種現(xiàn)象不是由于ROS所引起。隨后的激酶抑制劑實(shí)驗(yàn)表明MWCNTs-COOH對(duì)AhR-CYP1A1信號(hào)通路的抑制至少部分是由于胞外調(diào)節(jié)蛋白激酶(Extracellular signal-regulated kinase,ERK)介導(dǎo)。而內(nèi)吞抑制劑實(shí)驗(yàn)同時(shí)也表明,對(duì)AhR-CYP1A1信號(hào)通路的抑制部分需要細(xì)胞對(duì)MWCNTs-COOH的攝入。本研究不僅對(duì)闡明納米顆粒與AhR之間相互作用及其內(nèi)在機(jī)制具有理論意義,而且能夠完善納米顆粒的風(fēng)險(xiǎn)評(píng)估,對(duì)納米顆粒的安全性設(shè)計(jì)提供理論依據(jù)。
[Abstract]:Nanomaterials have been widely used in such fields as aerospace, automobile industry, environmental protection building materials, sporting goods, electronic devices and consumer goods due to their specific physical and chemical properties, as well as in biomedical fields such as diagnostics, biomedical imaging, etc. Biosensors and drug transportation also have a wide range of applications. As a result, opportunities for exposure to nanoscale environments are becoming more and more frequent. Compared with the exposure of nanoparticles from the daily products, the nanoparticles used in the field of medicine can enter the human body directly through ingestion and injection, so people pay more attention to the safety of nanoparticles in this field. Aryl hydrocarbon receptor AhRis is a key nuclear receptor for regulating the metabolic genes of exogenous toxic compounds. The most studied and classic downstream target gene is cytochrome P4501A1CY CYP1A1. Recent studies have found that AhR still plays an important role in normal physiological conditions, such as cell proliferation and differentiation. The regulation of immune balance may have adverse effects on the normal physiological function of cells and organisms due to the interference of AhR and its related signaling pathways. Previous studies on AhR interferences have focused on small molecular compounds, ignoring a series of novel synthetic materials such as nanoparticles that may interfere with the AhR signal pathway. Previous studies have shown that nanoparticles may interfere with the expression activity of AhR downstream genes such as CYP1A1. Considering the widespread distribution of nanoparticles in the environment and the high exposure risk to human health, whether the AhR signaling pathway can be interfered by nanoparticles is an urgent problem to be solved. We first selected mouse hepatoma cell line CBG2.8D with stable luciferase reporter gene as the experimental model. The interference ability of several common nanoparticles, including MWCNTs-COOHU, graphene oxide GO-PEGN, nano-silica particles, nSiO2O _ 2, TIO _ 2 particles, au _ 2O _ 2, au _ (NPs) and ZnO _ 2O _ 2, to the AhR signal pathway was evaluated. The experimental results show that all the nanoparticles except nTiO_2 have the ability to interfere with the AhR-Luciferase pathway to varying degrees. In order to further elucidate the intrinsic molecular mechanism of interaction between nanoparticles and AhR signaling pathway, we then selected MWCNTs-COOH as a model, using real-time fluorescent quantitative PCR and Western blot techniques. The interference mechanism of MWCNTs-COOH on AhR signaling pathway was verified at the gene and protein levels. The results showed that MWCNTs-COOH could inhibit the expression of CYP1A1 at the gene and protein levels, and change the gene expression level and the total protein content of AhR at the same time. But this phenomenon is not caused by ROS. Subsequent kinase inhibitor experiments showed that the inhibition of AhR-CYP1A1 signaling pathway by MWCNTs-COOH was at least partly mediated by extracellular signal-regulated kinase (ERK). The endocytosis inhibitor experiment also showed that the inhibition of AhR-CYP1A1 signaling pathway required cell uptake of MWCNTs-COOH. This study not only has theoretical significance to clarify the interaction between nanoparticles and AhR and its intrinsic mechanism, but also can improve the risk assessment of nanoparticles and provide theoretical basis for the safety design of nanoparticles.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R114;TB383.1

【相似文獻(xiàn)】

相關(guān)期刊論文 前2條

1 姜龍;程冰川;李魚;;多溴二苯醚取代特征對(duì)其芳香烴受體結(jié)合能力的效應(yīng)分析[J];化學(xué)學(xué)報(bào);2014年06期

2 周海龍;張林寶;廖春陽(yáng);韋雙雙;鄭繼平;薛欽昭;;持久性有機(jī)污染物對(duì)水生動(dòng)物芳香烴受體通道的毒理機(jī)制及其早期監(jiān)測(cè)(英文)[J];生態(tài)毒理學(xué)報(bào);2010年01期

相關(guān)會(huì)議論文 前5條

1 裴新輝;趙斌;;芳香烴受體轉(zhuǎn)錄調(diào)控的機(jī)理研究[A];中國(guó)化學(xué)會(huì)第28屆學(xué)術(shù)年會(huì)第2分會(huì)場(chǎng)摘要集[C];2012年

2 趙斌;;基于芳香烴受體通路的二惡英毒理機(jī)制研究進(jìn)展[A];第八屆全國(guó)分析毒理學(xué)大會(huì)暨中國(guó)毒理學(xué)會(huì)分析毒理專業(yè)委員會(huì)第五屆會(huì)員代表大會(huì)論文摘要集[C];2014年

3 田文靜;裴新輝;趙斌;;芳香烴受體蛋白單克隆抗體的制備及應(yīng)用[A];中國(guó)化學(xué)會(huì)第28屆學(xué)術(shù)年會(huì)第2分會(huì)場(chǎng)摘要集[C];2012年

4 藺遠(yuǎn);穆云松;張愛茜;王連生;;芳香烴受體與其配體的分子動(dòng)力學(xué)研究[A];第五屆全國(guó)環(huán)境化學(xué)大會(huì)摘要集[C];2009年

5 朱從會(huì);趙斌;;芳香烴受體在環(huán)境毒理學(xué)研究中的新認(rèn)識(shí)[A];中國(guó)毒理學(xué)會(huì)第六屆全國(guó)毒理學(xué)大會(huì)論文摘要[C];2013年

相關(guān)博士學(xué)位論文 前1條

1 王朝奎;白介素27和芳香烴受體在Vogt-小柳原田綜合征和Behcet病發(fā)病機(jī)制中作用的研究[D];重慶醫(yī)科大學(xué);2014年

相關(guān)碩士學(xué)位論文 前8條

1 桂曉玲;芳香烴受體及肝臟代謝酶在飲用水有機(jī)提取物染毒大鼠肝臟中的表達(dá)及意義[D];貴陽(yáng)醫(yī)學(xué)院;2015年

2 魏永義;常見納米顆粒干擾芳香烴受體信號(hào)通路機(jī)制的初步研究[D];山東大學(xué);2017年

3 王卓;基于芳香烴受體系統(tǒng)的痕量二惡英生物檢測(cè)方法的研究[D];天津醫(yī)科大學(xué);2007年

4 過倩萍;SUMO化修飾對(duì)芳香烴受體AhR調(diào)控機(jī)制的研究[D];大連理工大學(xué);2010年

5 李磊;益艾康膠囊對(duì)HIV感染者外周血Th22細(xì)胞及其關(guān)鍵轉(zhuǎn)錄因子芳香烴受體表達(dá)的影響[D];河南中醫(yī)學(xué)院;2015年

6 王鈺瑩;芳香烴受體在腸易激綜合征小鼠模型Th17細(xì)胞活化中的作用[D];鄭州大學(xué);2014年

7 楊尚儒;芳香烴受體AHR通過破壞DNA成環(huán)負(fù)向調(diào)控LPS誘導(dǎo)的骨橋蛋白OPN的表達(dá)[D];山東大學(xué);2014年

8 趙娜;二惡英生物檢測(cè)體系相關(guān)蛋白質(zhì)的表達(dá)[D];天津醫(yī)科大學(xué);2005年

，

本文編號(hào)：1846380