基于核磁共振代謝組學(xué)的除草劑丁草胺對金魚的毒性研究
發(fā)布時間：2018-07-23 12:02
【摘要】：丁草胺,農(nóng)業(yè)中使用最廣泛的除草劑之一,在水田、河流、湖泊中的大量殘留已經(jīng)造成了嚴(yán)重的水污染。前人的研究表明丁草胺對水生植物和動物具有很高的生態(tài)毒性,然而關(guān)于丁草胺對動物造成的內(nèi)源性分子代謝變化的毒理機(jī)制的研究甚少。隨著代謝組學(xué)的檢測手段與分析方法的發(fā)展,基于核磁共振代謝組學(xué)的研究方法已經(jīng)逐漸在農(nóng)藥等環(huán)境污染物的毒理機(jī)制研究中有了廣泛的應(yīng)用。本論文首次將基于核磁共振代謝組學(xué)方法運(yùn)用到丁草胺的毒性機(jī)制研究中,并結(jié)合組織病理學(xué)檢查和生化指標(biāo)分析,全面探究了丁草胺對金魚(Carassius auratus)的四個重要器官(鰓、腦、肝、腎)的毒性作用,在內(nèi)源性小分子代謝物水平上揭示丁草胺的毒理機(jī)制。實驗設(shè)計1 mg/L和0.2 mg/L兩個丁草胺劑量組與對照組,金魚暴露10天后,取其鰓、腦、肝、腎四個組織并收集血清,分別進(jìn)行組織病理學(xué)檢查、生化指標(biāo)測試以及核磁共振1H譜測試。然后對得到的數(shù)據(jù)進(jìn)行多元統(tǒng)計分析與單變量分析,得到組織中內(nèi)源性分子代謝物變化的信息。病理組織學(xué)檢查發(fā)現(xiàn)丁草胺暴露后金魚的鰓、肝臟和腎臟都受到嚴(yán)重?fù)p害表現(xiàn)出明顯的細(xì)胞水腫甚至中度壞死。丁草胺給藥組金魚鰓中谷胱甘肽過氧化物酶(GSH-Px)的活性以及四個組織中丙二醛(MDA)含量的增加反映了染毒后金魚的抗氧化系統(tǒng)受到干擾,血清中乳酸脫氫酶(LDH)活性以及肌酐(CRE)含量增加分別表明了金魚肝功能與腎功能發(fā)生異常。對核磁共振1H譜數(shù)據(jù)進(jìn)行的正交信號校正偏最小二乘判別分析(OSC-PLS-DA)進(jìn)一步揭示了金魚體內(nèi)小分子代謝物的變化,從而解釋了一些代謝過程紊亂如氧化應(yīng)激(膽堿、磷酸膽堿、甘油磷酸膽堿、谷胱甘肽、�；撬帷⑻鸩藟A含量升高)、能量代謝紊亂(葡萄糖、麥芽糖以及6-磷酸葡萄糖含量降低,乳酸鹽、檸檬酸、琥珀酸鹽和延胡索酸鹽含量升高)、氨基酸代謝紊亂(亮氨酸、纈氨酸、異亮氨酸、酪氨酸、苯丙氨酸、精氨酸含量升高)以及神經(jīng)遞質(zhì)失衡(谷氨酸含量升高、γ-氨基丁酸含量降低)。研究結(jié)果充分表明基于1H核磁共振的代謝組學(xué)方法是研究丁草胺毒理機(jī)制以及對其風(fēng)險評估的一種有力而且實用的研究手段。
[Abstract]:Butachlor, one of the most widely used herbicides in agriculture, has caused serious water pollution in paddy fields, rivers and lakes. Previous studies have shown that butachlor is highly ecotoxic to aquatic plants and animals, but little has been done on the toxicological mechanism of endogenous molecular metabolic changes caused by butachlor. With the development of metabonomics detection and analysis methods, the method based on nuclear magnetic resonance metabonomics has been widely used in the study of toxicological mechanism of environmental pollutants such as pesticides. In this paper, based on the method of nuclear magnetic resonance metabolomics (NMR), the toxic mechanism of butachlor was studied for the first time, and combined with histopathological examination and biochemical index analysis, four important organs (Gill, brain) of butachlor on goldfish (Carassius auratus) were studied. The toxicity of liver and kidney revealed the toxicological mechanism of butachlor at the level of endogenous small molecule metabolites. The experiment designed 1 mg / L and 0.2 mg / L butachlor dose group and control group. After 10 days of exposure, goldfish were collected from four tissues of Gill, brain, liver and Kidney and collected serum for histopathological examination. Biochemical index test and 1H NMR spectrum test. Then the data were analyzed by multivariate statistical analysis and univariate analysis to get the information of the changes of endogenous molecular metabolites in tissues. Histopathological examination showed that the Gill liver and kidney of goldfish exposed to butachlor showed obvious cell edema and even moderate necrosis. The activity of glutathione peroxidase (GSH-Px) in the Gill of goldfish treated with butachlor and the increase of malondialdehyde (MDA) content in four tissues reflected the interference of antioxidant system in goldfish after exposure to butachlor. The activities of lactate dehydrogenase (LDH) and creatinine (CRE) in serum showed abnormal liver and renal function, respectively. Orthogonal signal corrected partial least squares discriminant analysis (OSC-PLS-DA) for 1H NMR data further reveals the changes of small molecular metabolites in goldfish, thus explaining some metabolic disorders such as oxidative stress (choline, choline phosphate, choline phosphate). Glycerophosphate choline, glutathione, taurine, betaine increased, energy metabolism disorder (glucose, maltose and 6-phosphate glucose content decreased, lactate, citric acid, Increased content of succinate and corydate), disorder of amino acid metabolism (leucine, valine, isoleucine, tyrosine, phenylalanine, leucine, valine, isoleucine, phenylalanine, Arginine content increased, and neurotransmitter imbalance (glutamate content increased, 緯 -aminobutyric acid content decreased). The results show that the metabonomics method based on 1H NMR is an effective and practical method to study the toxicological mechanism and risk assessment of butachlor.
【學(xué)位授予單位】：南京理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：X592;X171.5
本文編號：2139351
[Abstract]:Butachlor, one of the most widely used herbicides in agriculture, has caused serious water pollution in paddy fields, rivers and lakes. Previous studies have shown that butachlor is highly ecotoxic to aquatic plants and animals, but little has been done on the toxicological mechanism of endogenous molecular metabolic changes caused by butachlor. With the development of metabonomics detection and analysis methods, the method based on nuclear magnetic resonance metabonomics has been widely used in the study of toxicological mechanism of environmental pollutants such as pesticides. In this paper, based on the method of nuclear magnetic resonance metabolomics (NMR), the toxic mechanism of butachlor was studied for the first time, and combined with histopathological examination and biochemical index analysis, four important organs (Gill, brain) of butachlor on goldfish (Carassius auratus) were studied. The toxicity of liver and kidney revealed the toxicological mechanism of butachlor at the level of endogenous small molecule metabolites. The experiment designed 1 mg / L and 0.2 mg / L butachlor dose group and control group. After 10 days of exposure, goldfish were collected from four tissues of Gill, brain, liver and Kidney and collected serum for histopathological examination. Biochemical index test and 1H NMR spectrum test. Then the data were analyzed by multivariate statistical analysis and univariate analysis to get the information of the changes of endogenous molecular metabolites in tissues. Histopathological examination showed that the Gill liver and kidney of goldfish exposed to butachlor showed obvious cell edema and even moderate necrosis. The activity of glutathione peroxidase (GSH-Px) in the Gill of goldfish treated with butachlor and the increase of malondialdehyde (MDA) content in four tissues reflected the interference of antioxidant system in goldfish after exposure to butachlor. The activities of lactate dehydrogenase (LDH) and creatinine (CRE) in serum showed abnormal liver and renal function, respectively. Orthogonal signal corrected partial least squares discriminant analysis (OSC-PLS-DA) for 1H NMR data further reveals the changes of small molecular metabolites in goldfish, thus explaining some metabolic disorders such as oxidative stress (choline, choline phosphate, choline phosphate). Glycerophosphate choline, glutathione, taurine, betaine increased, energy metabolism disorder (glucose, maltose and 6-phosphate glucose content decreased, lactate, citric acid, Increased content of succinate and corydate), disorder of amino acid metabolism (leucine, valine, isoleucine, tyrosine, phenylalanine, leucine, valine, isoleucine, phenylalanine, Arginine content increased, and neurotransmitter imbalance (glutamate content increased, 緯 -aminobutyric acid content decreased). The results show that the metabonomics method based on 1H NMR is an effective and practical method to study the toxicological mechanism and risk assessment of butachlor.
【學(xué)位授予單位】：南京理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：X592;X171.5
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
1 馬立平;統(tǒng)計數(shù)據(jù)標(biāo)準(zhǔn)化──無量綱化方法──現(xiàn)代統(tǒng)計分析方法的學(xué)與用(三)[J];北京統(tǒng)計;2000年03期
2 范立民;馬曉燕;胡庚東;陳家長;;除草劑丁草胺對兩種魚的急性毒性研究[J];浙江海洋學(xué)院學(xué)報(自然科學(xué)版);2005年04期
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