基于核磁共振氢谱的氟咯草酮90d染毒大鼠尿液特征代谢物研究  

^1HNMR-based study on characteristic metabolites in urine of mice following 90d of flurochloridone exposure

作　　者：薛黎明[1] 金玉娥[1] 张素慧[2] 周志俊[2] 卢大胜[1] 汪国权[1] 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)

机构地区：[1]上海市疾病预防控制中心化学品毒性检定所,上海200336 [2]复旦大学公共卫生学院,上海200032

出　　处：《环境与职业医学》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)。

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[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 p

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 

分 类 号：R114[医药卫生—卫生毒理学]