本文選題：銅離子 + 氧化應(yīng)激��； 參考：《華中農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：銅(Cu,copper)是人體必需的微量元素,作為輔基參與合成多種生物酶。人體內(nèi)銅代謝紊亂會(huì)引發(fā)多種疾病。斑馬魚(Danio rerio)是一種重要的模式水生生物,在生態(tài)毒理學(xué)、發(fā)育生物學(xué)、分子生物學(xué)等領(lǐng)域應(yīng)用廣泛,諸如評(píng)價(jià)環(huán)境脅迫因子、構(gòu)建疾病病理模型。本論文研究了銅離子暴露引起的斑馬魚造血系統(tǒng)和相關(guān)調(diào)控機(jī)制的功能基因表達(dá)、生理代謝調(diào)節(jié)的變化。旨在探討銅離子對(duì)胚胎發(fā)育早期及成體期造血系統(tǒng)的相關(guān)影響及其機(jī)制,為促進(jìn)銅離子機(jī)體代謝、銅離子代謝相關(guān)的血液類疾病、腫瘤等癌細(xì)胞增生類疾病的預(yù)防研究提供新方法。主要結(jié)果如下:1.銅離子對(duì)斑馬魚造血系統(tǒng)發(fā)育和再生的影響通過分析外源銅離子對(duì)斑馬魚胚胎造血發(fā)生的影響,發(fā)現(xiàn)銅處理胚胎紅系細(xì)胞(RBC)的基因表達(dá)增強(qiáng)、造血干細(xì)胞(HSC)標(biāo)記基因及其相關(guān)的調(diào)控信號(hào)Notch和Wnt信號(hào)通路因子表達(dá)減弱。銅處理成魚的外周和腎臟血流式細(xì)胞分析結(jié)果顯示,處理成魚的腎臟血、外周血的細(xì)胞分化程度均明顯增強(qiáng)。結(jié)果顯示,外源銅離子破壞了魚類胚胎造血系統(tǒng)發(fā)育及其成體造血細(xì)胞的再生及平衡。2.銅離子激發(fā)斑馬魚的氧化應(yīng)激通過基因表達(dá)譜分析、轉(zhuǎn)錄水平驗(yàn)證、活性氧(ROS)含量檢測(cè),發(fā)現(xiàn)銅處理胚胎氧化應(yīng)激過程的基因表達(dá)增強(qiáng)、活性氧含量增加,氧結(jié)合和氧轉(zhuǎn)運(yùn)基因的表達(dá)增強(qiáng)。此外,預(yù)先添加抗氧化劑可以高效恢復(fù)銅處理胚胎中氧化還原類反應(yīng)的基因表達(dá)、恢復(fù)活性氧含量。3.銅調(diào)控斑馬魚造血系統(tǒng)發(fā)育和再生的作用機(jī)制抗氧化劑還可以恢復(fù)紅系細(xì)胞的基因表達(dá)、血紅蛋白的合成量,協(xié)助機(jī)體完成過量銅離子的代謝、恢復(fù)胚胎內(nèi)的銅離子含量。全基因組甲基化分析發(fā)現(xiàn),造血干細(xì)胞關(guān)鍵基因在啟動(dòng)子預(yù)測(cè)區(qū)的甲基化修飾增強(qiáng)。外源銅離子是通過激活氧化應(yīng)激基因、促進(jìn)活性氧分子的合成,增強(qiáng)造血系統(tǒng)的紅系分化、擾亂銅代謝平衡的。添加外源抗氧化劑,不僅可以恢復(fù)造血系統(tǒng)的紅系分化,而且可恢復(fù)機(jī)體銅代謝平衡。銅可能通過氧化應(yīng)激反應(yīng)和甲基化修飾DNA分子,調(diào)控造血系統(tǒng)的分化和再生。
[Abstract]:Copper (copper) is a necessary trace element in human body, which is involved in the synthesis of a variety of biological enzymes as an auxiliary group. Disorders of copper metabolism in the human body can lead to many diseases. Zebrafish (Danio rerio) is an important model aquatic organism, which is widely used in ecotoxicology, developmental biology, molecular biology and so on. In this paper, the changes of functional gene expression and physiological metabolic regulation in zebrafish hematopoietic system and related regulatory mechanisms induced by copper ion exposure were studied. The aim of this study was to investigate the effects of copper ion on hematopoietic system in early embryonic and adult stages and its mechanism. In order to promote the metabolism of copper ions, the related blood diseases related to copper ion metabolism were studied. A new method is provided for the prevention of cancer cell proliferation diseases such as tumor. The main results are as follows: 1. Effects of copper ions on the development and regeneration of hematopoietic system of zebrafish by analyzing the effects of exogenous copper ions on embryonic hematopoiesis of zebrafish, it was found that the gene expression of erythroid cells (RBC) of zebrafish embryos treated with copper was enhanced. Hematopoietic stem cells (HSC)-labeled genes and their associated regulatory signaling Notch and Wnt signal pathway factors were down-regulated. The results of peripheral and renal blood flow cytometry analysis showed that the differentiation degree of peripheral blood cells of adult fish treated with copper was significantly increased. The results showed that exogenous copper ions destroyed the development of hematopoietic system of fish embryo and the regeneration and balance of adult hematopoietic cells. Oxidative stress induced by copper ions in zebrafish was detected by gene expression profiling, transcription level verification and reactive oxygen species (Ros) content detection. It was found that the expression of genes increased and the content of reactive oxygen species increased in the process of oxidative stress induced by copper. The expression of oxygen binding and oxygen transport genes was enhanced. In addition, pretreatment with antioxidants could effectively restore the gene expression of redox reaction and the content of reactive oxygen species (Ros) in copper treated embryos. The mechanism of copper regulating the development and regeneration of hematopoietic system in zebrafish, antioxidants can also restore gene expression and hemoglobin synthesis of erythroid cells, assist the body to complete the metabolism of excessive copper ions, and recover the copper ion content in embryos. Genomic methylation analysis showed that the methylation modification of hematopoietic stem cell key genes in the promoter predictive region was enhanced. Exogenous copper ions promote the synthesis of reactive oxygen species by activating oxidative stress genes, enhance the erythroid differentiation of hematopoietic system, and disturb the balance of copper metabolism. The addition of exogenous antioxidants can not only restore the erythroid differentiation of hematopoietic system, but also restore the balance of copper metabolism. Copper may regulate the differentiation and regeneration of hematopoietic system by oxidative stress and methylation of DNA molecules.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：X503.225

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本文編號(hào)：2099416