本文選題：毒死蜱 + 斑馬魚胚胎�。� 參考：《中國農(nóng)業(yè)科學(xué)院》2015年碩士論文

【摘要】：毒死蜱是一種典型的有機(jī)磷殺蟲劑,廣泛應(yīng)用于農(nóng)業(yè)生產(chǎn)。隨著污染事件的增多,毒死蜱復(fù)雜的生物學(xué)效應(yīng)逐漸引起大家的關(guān)注。本研究主要以斑馬魚胚胎和幼魚作為研究對(duì)象,從表型、分子水平尤其是組學(xué)水平上探究毒死蜱的環(huán)境生物學(xué)效應(yīng)及其作用機(jī)理。以下是主要的研究方法和結(jié)果:1.配制高、低兩種濃度梯度的毒死蜱進(jìn)行急性毒性實(shí)驗(yàn),其中低濃度梯度為0、0.10、0.25、0.50、0.75和1.00 ppm;高濃度梯度為0、1.00、2.00、3.00和4.00 ppm,每個(gè)濃度設(shè)置3個(gè)平行組。結(jié)果顯示,處理48 h后低濃度梯度處理組胚胎孵化快于對(duì)照組,并且胚胎的孵化率與處理濃度成正相關(guān),且未出現(xiàn)明顯畸形。處理60 h后,各組胚胎幾乎全部孵化,而且胚胎存活狀況良好;在高濃度毒死蜱梯度處理組中,胚胎在48 h已經(jīng)基本全部孵化,并且出現(xiàn)了體軸彎曲、心包囊腫等畸形現(xiàn)象,并且從處理開始陸續(xù)出現(xiàn)胚胎死亡,60 h時(shí)2 ppm處理組胚胎接近半致死。2.為了探究低濃度毒死蜱的內(nèi)分泌干擾效應(yīng),使用流式細(xì)胞儀分析了其對(duì)人子宮內(nèi)膜癌細(xì)胞HEC-1B生長周期的影響。結(jié)果發(fā)現(xiàn)毒死蜱對(duì)HEC-1B具有促增殖作用,并且同等濃度下其作用不如雌二醇,具有類似雌激素的效應(yīng)。毒死蜱能夠明顯影響斑馬魚胚胎雌激素相關(guān)基因VTG和ERα、細(xì)胞增殖相關(guān)基因c-myc、cyclin D1、c-fos和c-jun的表達(dá)水平。吖啶橙染色結(jié)果表明0.75和1.00 ppm毒死蜱處理組中,可以觀察到凋亡細(xì)胞,而細(xì)胞凋亡相關(guān)基因Bcl-2和Bax的表達(dá)水平出現(xiàn)紊亂。考慮到毒死蜱的神經(jīng)毒性,本研究從基因的轉(zhuǎn)錄水平探究其神經(jīng)發(fā)育毒性。3.本研究采用轉(zhuǎn)錄組學(xué)和蛋白質(zhì)組學(xué)對(duì)其進(jìn)行生物信息學(xué)分析。其中,轉(zhuǎn)錄組學(xué)采用RNA-seq技術(shù),蛋白質(zhì)組學(xué)采用iTRAQ技術(shù)。使用非致死低濃度毒死蜱(0.05 ppm)連續(xù)處理斑馬魚胚胎9天后進(jìn)行組學(xué)分析。轉(zhuǎn)錄組數(shù)據(jù)分析差異基因和通路,蛋白質(zhì)組數(shù)據(jù)分析差異蛋白和通路,并且進(jìn)行轉(zhuǎn)錄組和蛋白質(zhì)組聯(lián)合分析。本研究結(jié)果表明:低濃度的毒死蜱具有內(nèi)分泌干擾效應(yīng),能提高癌細(xì)胞的增殖系數(shù)。其加速斑馬魚胚胎孵化脫膜的原因,可能是促進(jìn)細(xì)胞的生長和分裂,抑制細(xì)胞的凋亡。而高濃度的毒死蜱對(duì)斑馬魚主要表現(xiàn)為神經(jīng)毒性,能夠造成胚胎的畸形和死亡,并且顯著影響神經(jīng)系統(tǒng)相關(guān)基因的表達(dá)量。毒死蜱導(dǎo)致斑馬魚幼魚的代謝、轉(zhuǎn)錄的相關(guān)基因和蛋白表達(dá)上調(diào),從而影響其發(fā)育進(jìn)程;導(dǎo)致免疫系統(tǒng)、環(huán)境適應(yīng)性相關(guān)基因和蛋白表達(dá)下調(diào),可能造成免疫力降低,最終導(dǎo)致斑馬魚幼魚死亡。另外,毒死蜱處理產(chǎn)生差異表達(dá)異常顯著的基因和蛋白可以作為潛在生物標(biāo)記進(jìn)行下一步研究。
[Abstract]:Chlorpyrifos is a typical organophosphorus insecticide, widely used in agricultural production. With the increase of pollution events, the complex biological effects of chlorpyrifos have attracted more and more attention. In this study, the environmental biological effects of chlorpyrifos and its mechanism were studied from phenotypic and molecular level, especially at the group level, with zebrafish embryos and young fish as the research objects. Here are the main research methods and results: 1. The acute toxicity test of chlorpyrifos with high and low concentration gradient was carried out. The low concentration gradient was 0.100.250.50,0.50,0.75 and 1.00 ppm.The high concentration gradient was 00.000.002.003.00 and 4.00 ppms. each concentration was divided into 3 parallel groups. The results showed that the embryo hatched faster in the low concentration gradient treatment group than that in the control group after 48 h treatment, and the hatching rate of the embryos was positively correlated with the treatment concentration, and there was no obvious abnormality. After 60 h treatment, almost all the embryos hatched, and the embryo survived well. In the high concentration chlorpyrifos gradient treatment group, the embryos had almost all hatched at 48 h, and some abnormal phenomena such as body axis bending and pericardial cyst appeared. And the embryo death occurred at 60 h after treatment. The embryo of 2 ppm treatment group was close to half lethal. 2. In order to investigate the endocrine disrupting effect of chlorpyrifos at low concentration, the effects of chlorpyrifos on the HEC-1B growth cycle of human endometrial cancer cells were analyzed by flow cytometry. The results showed that chlorpyrifos could promote the proliferation of HEC-1B, and at the same concentration, the effect of chlorpyrifos was inferior to that of estradiol, which was similar to that of estrogen. Chlorpyrifos could significantly affect the expression levels of estrogen related genes VTG and ER 偽, cell proliferation related genes c-myc cyclin D1 c-fos and c-jun in zebrafish embryos. The results of acridine orange staining showed that apoptotic cells could be observed in chlorpyrifos treated with 0. 75 and 1. 00 ppm, while the expression of apoptosis-related genes Bcl-2 and Bax were disturbed. Considering the neurotoxicity of chlorpyrifos, we investigated the neurodevelopmental toxicity of chlorpyrifos from the transcriptional level. In this study, transcriptome and proteomics were used for bioinformatics analysis. Among them, RNA-seq technique was used in transcriptome and iTRAQ technique was used in proteomics. The zebrafish embryos were treated with non-lethal low concentration chlorpyrifos (0.05 ppm) for 9 days. Transcriptional data were used to analyze differential genes and pathways, proteome data were used to analyze differential proteins and pathways, and transcriptome and proteome were combined. The results showed that low concentration of chlorpyrifos had endocrine disrupting effect and could improve the proliferative coefficient of cancer cells. The reason of accelerating the hatching and demembering of zebrafish embryos may be to promote cell growth and division and inhibit cell apoptosis. The high concentration of chlorpyrifos mainly showed neurotoxicity to zebrafish, which caused embryo deformity and death, and significantly affected the expression of genes related to the nervous system. Chlorpyrifos lead to the metabolism of juvenile zebrafish and up-regulate the expression of transcription related genes and proteins, which affect the development of zebrafish, and lead to the down-regulation of the immune system and the expression of environment-adaptive genes and proteins, which may lead to the decrease of immunity. The result is the death of juvenile zebrafish. In addition, Chlorpyrifos treatment produces abnormal differentially expressed genes and proteins that can be used as potential biomarkers for further research.
【學(xué)位授予單位】：中國農(nóng)業(yè)科學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：X174

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 鄭文;唐文浩;;土壤粒度與含水量對(duì)太陽光降解百草枯的影響[J];安徽農(nóng)業(yè)科學(xué);2007年08期

2 陳昂;張戰(zhàn)泓;劉勇;羅香文;何明遠(yuǎn);張松柏;張德詠;;3種植物源農(nóng)藥對(duì)鵪鶉和斑馬魚的急性毒性評(píng)價(jià)[J];安徽農(nóng)業(yè)科學(xué);2010年12期

3 劉鑒樟;郭聰;劉慧君;;丁硫克百威對(duì)雌性鵪鶉肝臟及生殖腺的影響[J];現(xiàn)代農(nóng)業(yè)科技;2008年19期

4 汪芳;王軍;;毒死蜱和咪草煙對(duì)土壤微生物呼吸的影響[J];環(huán)境科學(xué)與管理;2010年09期

5 陳偉國;董瑞華;張國平;;農(nóng)藥對(duì)家蠶毒性評(píng)價(jià)若干問題的探討[J];蠶桑通報(bào);2011年02期

6 梁楊;徐立;馬曉敏;溫斐斐;余茂德;吳福安;黃先智;;對(duì)桑椹肥大性菌核病菌具有抑制活性的中草藥篩選[J];蠶業(yè)科學(xué);2011年02期

7 陳偉國;魯興萌;;農(nóng)藥對(duì)家蠶的毒性和安全性評(píng)價(jià)研究[J];蠶業(yè)科學(xué);2012年02期

8 于力;劉杰;王霞;李專;楊國強(qiáng);李成;盧振蘭;李莉;;魚組合測廢水急性毒性技術(shù)的特點(diǎn)與理論基礎(chǔ)[J];環(huán)境科學(xué)與技術(shù);2012年S2期

9 高曉華;曹海鵬;侯三玲;胡鯤;楊先樂;;水產(chǎn)用聚維酮碘對(duì)異育銀鯽養(yǎng)殖的安全性評(píng)價(jià)[J];動(dòng)物學(xué)雜志;2013年02期

10 孫海燕;陳偉國;董瑞華;戴建忠;;氟鈴脲對(duì)家蠶的毒性測定[J];北方蠶業(yè);2014年01期

相關(guān)會(huì)議論文 前3條

1 陳偉國;張國平;;完善農(nóng)藥對(duì)家蠶毒性評(píng)價(jià)制度和方法的探討[A];中國蠶學(xué)會(huì)桑樹病蟲害防治學(xué)術(shù)研討會(huì)論文集[C];2008年

2 廖秀麗;劉佳霖;羅術(shù)東;吳杰;;2種熊蜂對(duì)5種設(shè)施農(nóng)業(yè)常用農(nóng)藥的敏感性研究[A];國家蜂產(chǎn)業(yè)技術(shù)體系蜜蜂育種與授粉功能研究室學(xué)術(shù)研討會(huì)暨中國養(yǎng)蜂學(xué)會(huì)蜜蜂育種專業(yè)委員會(huì)第四屆第一次會(huì)議暨中國養(yǎng)蜂學(xué)會(huì)蜜源與蜜蜂授粉專業(yè)委員會(huì)第五屆第一次會(huì)議論文匯編[C];2012年

3 劉佳霖;伍翔;廖秀麗;和紹禹;羅術(shù)東;吳杰;;4種殺蟲劑對(duì)小峰熊蜂工蜂和雄性蜂的急性經(jīng)口毒性測定[A];國家蜂產(chǎn)業(yè)技術(shù)體系蜜蜂育種與授粉功能研究室學(xué)術(shù)研討會(huì)暨中國養(yǎng)蜂學(xué)會(huì)蜜蜂育種專業(yè)委員會(huì)第四屆第一次會(huì)議暨中國養(yǎng)蜂學(xué)會(huì)蜜源與蜜蜂授粉專業(yè)委員會(huì)第五屆第一次會(huì)議論文匯編[C];2012年

相關(guān)博士學(xué)位論文 前1條

1 徐敦明;植物源殺蟲物質(zhì)鬼臼毒素的酶免疫分析及環(huán)境安全性研究[D];西北農(nóng)林科技大學(xué);2005年

相關(guān)碩士學(xué)位論文 前10條

1 謝夢(mèng)醒;環(huán)嗪酮·敵草隆在甘蔗、土壤中的殘留消解動(dòng)態(tài)及敵草隆的土壤吸附行為研究[D];安徽農(nóng)業(yè)大學(xué);2010年

2 李橋;楊梅葉抑菌活性研究[D];浙江農(nóng)林大學(xué);2010年

3 仉歡;56%磷化鋁片劑對(duì)番茄、黃瓜根結(jié)線蟲的控制效果及其土壤微生物生態(tài)效應(yīng)[D];山東農(nóng)業(yè)大學(xué);2011年

4 趙志強(qiáng);毒死蜱和丁硫克百威對(duì)香蕉根際土壤微生物的影響及其降解特征的研究[D];海南大學(xué);2011年

5 代雪芳;四聚乙醛在甘藍(lán)中的殘留動(dòng)態(tài)研究[D];中國農(nóng)業(yè)科學(xué)院;2011年

6 祁志軍;0.2%苦皮藤素乳油環(huán)境安全性評(píng)價(jià)[D];西北農(nóng)林科技大學(xué);2001年

7 崔淑華;戊唑醇及其混劑在小麥中的殘留動(dòng)態(tài)及對(duì)土壤微生物影響的研究[D];山東農(nóng)業(yè)大學(xué);2005年

8 王文軍;WTO環(huán)境下漢中綠色食品蔬菜發(fā)展戰(zhàn)略研究[D];西北農(nóng)林科技大學(xué);2004年

9 范昆;1，3-二氯丙烯對(duì)番茄根結(jié)線蟲的控制效果和環(huán)境生態(tài)效應(yīng)[D];山東農(nóng)業(yè)大學(xué);2006年

10 來有鵬;幾種農(nóng)藥對(duì)家蠶和河蝦安全性評(píng)價(jià)試驗(yàn)技術(shù)的研究[D];南京農(nóng)業(yè)大學(xué);2008年

，

本文編號(hào)：1984336