基于核磁共振代谢组学的除草剂丁草胺对金鱼的毒性研究
发布时间：2018-07-23 12:02
【摘要】：丁草胺,农业中使用最广泛的除草剂之一,在水田、河流、湖泊中的大量残留已经造成了严重的水污染。前人的研究表明丁草胺对水生植物和动物具有很高的生态毒性,然而关于丁草胺对动物造成的内源性分子代谢变化的毒理机制的研究甚少。随着代谢组学的检测手段与分析方法的发展,基于核磁共振代谢组学的研究方法已经逐渐在农药等环境污染物的毒理机制研究中有了广泛的应用。本论文首次将基于核磁共振代谢组学方法运用到丁草胺的毒性机制研究中,并结合组织病理学检查和生化指标分析,全面探究了丁草胺对金鱼(Carassius auratus)的四个重要器官(鳃、脑、肝、肾)的毒性作用,在内源性小分子代谢物水平上揭示丁草胺的毒理机制。实验设计1 mg/L和0.2 mg/L两个丁草胺剂量组与对照组,金鱼暴露10天后,取其鳃、脑、肝、肾四个组织并收集血清,分别进行组织病理学检查、生化指标测试以及核磁共振1H谱测试。然后对得到的数据进行多元统计分析与单变量分析,得到组织中内源性分子代谢物变化的信息。病理组织学检查发现丁草胺暴露后金鱼的鳃、肝脏和肾脏都受到严重损害表现出明显的细胞水肿甚至中度坏死。丁草胺给药组金鱼鳃中谷胱甘肽过氧化物酶(GSH-Px)的活性以及四个组织中丙二醛(MDA)含量的增加反映了染毒后金鱼的抗氧化系统受到干扰,血清中乳酸脱氢酶(LDH)活性以及肌酐(CRE)含量增加分别表明了金鱼肝功能与肾功能发生异常。对核磁共振1H谱数据进行的正交信号校正偏最小二乘判别分析(OSC-PLS-DA)进一步揭示了金鱼体内小分子代谢物的变化,从而解释了一些代谢过程紊乱如氧化应激(胆碱、磷酸胆碱、甘油磷酸胆碱、谷胱甘肽、牛磺酸、甜菜碱含量升高)、能量代谢紊乱(葡萄糖、麦芽糖以及6-磷酸葡萄糖含量降低,乳酸盐、柠檬酸、琥珀酸盐和延胡索酸盐含量升高)、氨基酸代谢紊乱(亮氨酸、缬氨酸、异亮氨酸、酪氨酸、苯丙氨酸、精氨酸含量升高)以及神经递质失衡(谷氨酸含量升高、γ-氨基丁酸含量降低)。研究结果充分表明基于1H核磁共振的代谢组学方法是研究丁草胺毒理机制以及对其风险评估的一种有力而且实用的研究手段。
[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, 纬 -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.
【学位授予单位】：南京理工大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：X592;X171.5
本文编号：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, 纬 -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.
【学位授予单位】：南京理工大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：X592;X171.5
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相关期刊论文 前2条
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2 范立民;马晓燕;胡庚东;陈家长;;除草剂丁草胺对两种鱼的急性毒性研究[J];浙江海洋学院学报(自然科学版);2005年04期
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