摘要
[背景]氟咯草酮(FLC)是一种吡咯烷酮类芽前施用的选择性除草剂,用于防制多种阔叶类和禾本科杂草,常施用于向日葵、马铃薯、胡萝卜、小麦和玉米等作物以防其周边杂草生长。[目的]研究农药FLC 90 d染毒大鼠尿液的代谢物特征,寻找染毒特征性代谢物,探索FLC毒理机制。[方法]48只Wistar大鼠,按体重随机分为31.25、125.0、500.0 mg·kg^-1 FLC染毒组和溶媒对照组,每组12只,雌雄各半。灌胃染毒90 d后,收集大鼠的尿液。采用600 MHz核磁共振氢谱(1HNMR)法检测,得到一维氢谱图,以相对化学位移δ为0.04区间进行分段积分。采用模式识别技术主成分分析和偏最小二乘法判别分析(PLS-DA)法分析,寻找FLC染毒特征性代谢物。[结果]四组雄性大鼠PLS-DA分析交叉验证模型合理(P <0.05),FLC染毒雄性大鼠各剂量组与溶媒对照组散点分类明确;而雌性大鼠模型不合理(P> 0.05),各组散点分类不明显。各组内部雌雄大鼠PLS-DA分析,溶媒对照组雌雄大鼠差异有统计学意义(P <0.05),各染毒组模型均不合理(P> 0.05)。单个染毒组与溶媒对照组PLS-DA模型合理(P <0.001),预测值> 0.6,受试者特征曲线下面积> 0.75,分类明确。尿液核磁图谱共鉴定出35个代谢物,FLC染毒的差异有统计学意义的代谢物有脯氨酸、肌酸/肌酐、肌醇、氧化三甲胺、牛磺酸、甘露醇、苯丙氨酸;雄性高剂量染毒组与溶媒对照组的特征代谢物共15个,其中乙酸和尿酸升高,苯丙氨酸、乳酸、牛磺酸、3-羟基丁酸、脯氨酸、氧化三甲胺、二甲胺、肌酸、肌酐、肌醇、葡萄糖、甘露醇、柠檬酸盐降低。而雌性染毒仅柠檬酸盐、肌酸、肌酐和苯丙氨酸4个代谢物降低。[结论]FLC染毒对雄性大鼠更敏感,代谢物差异变化更明显,呈现一定量效关系;同时随染毒剂量的增高,组内雌雄大鼠代谢物的差异降低。高剂量染毒雄性大鼠的15个特征代谢物主要参与糖代谢和多种氨基酸代谢,呈现一定的肝毒性和肾毒性。
[Background] Flurochloridone(FLC), a kind of selective preemergence herbicides, is mainly used to control a variety of broad-leaved and gramineous weeds, commonly applied in sunflower, potato, carrot, wheat, and corn.[Objective] This study aims to describe the urinary metabolites in rats after 90 d of FLC exposure, identify characteristic metabolites, and explore the toxicological mechanism of FLC. [Methods] Forty-eight Wistar rats were randomly divided into four groups by weight: 31.25, 125.0, and 500.0 mg·kg^-1 FLC exposure groups and a solvent control group, with 12 rats in each group, half male and half female. Urine samples were collected after 90 d of intragastric administration. After 600 MHz 1H nuclear magnetic resonance(1HNMR) spectroscopy, onedimensional hydrogen spectrum was obtained, and the spectral region was integrated into regions with chemical shifts δ over an interval of 0.04. Principal component analysis and partial least squares discriminant analysis(PLS-DA) were applied to determine the characteristic metabolites of FLC. [Results] The PLS-DA cross validation analysis results showed that the male rat model was reasonable(P < 0.05), with obvious classification of scattered points among the three dose groups and the solvent control group, while the female rat model was unreasonable(P > 0.05), with obscure classification results. The PLS-DA intragroup analysis results showed that there was a significant difference between male and female rats in the solvent control group(P < 0.05), but not in the three exposure groups(P > 0.05). The PLS-DA model between single FLC exposure group and the solvent control group was reasonable(P < 0.001), with accuracy > 0.6 and area under curve of Receiver Operating Characteristic > 0.75, providing a clear classification between groups. Thirty-five metabolites were identified by urinary nuclear magnetic resonance, and the significantly changed metabolites after FLC exposure were proline, creatine/creatinine, inositol, trimethylamine oxide, taurine, mannitol, and phenylalanine;15 characteristic metabolites were obtained between the male rats in the high-dose FLC group and the solvent control group, including increased acetate and uric, and decreased phenylalanine, lactic acid, taurine, 3-hydroxybutyric acid, proline, trimethylamine oxide, dimethylamine, methylamine, creatine, creatinine, inositol, glucose, mannitol, and citrate. However, only citrate, creatine, creatinine, and phenylalanine were significantly decreased in the female rats after FLC exposure.[Conclusion] Male rats are more sensitive to FLC as the changes of metabolites are more obvious, showing a dose-response relationship. With the increase of FLC exposure dose, the differences of metabolites are reduced between male and female rats in the same group. The 15 characteristic metabolites in male rats exposed to high dose of FLC are mainly involved in carbohydrate metabolism and amino acid metabolism, indicating hepatotoxicity and nephrotoxicity.
作者
薛黎明
金玉娥
张素慧
周志俊
卢大胜
汪国权
XUE Li-ming;JIN Yu-e;ZHANG Su-hui;ZHOU Zhi-jun;LU Da-sheng;WANG Guo-quan(Division of Chemical Toxicity and Safety Assessment,Shanghai Municipal Center for Disease Control and Prevention,Shanghai 200336,China;School of Public Health,Fudan University,Shanghai 200032,China)
出处
《环境与职业医学》
CAS
CSCD
北大核心
2020年第3期197-204,共8页
Journal of Environmental and Occupational Medicine
基金
上海市卫计委面上项目(201740068)
上海市自然科学基金项目(18ZR1432200)
国家自然科学基金项目(81872643)。
关键词
氟咯草酮
核磁共振氢谱
代谢组学
特征性代谢物
糖代谢
flurochloridone
~1H nuclear magnetic resonance
metabolomics
characteristic metabolite
glucose metabolism
