法医学杂志

法医学杂志 ›› 2017, Vol. 33 ›› Issue (3): 247-251.DOI: 10.3969/j.issn.1004-5619.2017.03.007

• 论著 • 上一篇    下一篇

基于HPLC-TOF-MS代谢组学方法的溴鼠灵毒性作用评价

严  慧，卓先义，沈保华，向  平，沈  敏   

  1. （司法部司法鉴定科学技术研究所 上海市法医学重点实验室 上海市司法鉴定专业技术服务平台，上海 200063）
  • 发布日期:2017-06-25 出版日期:2017-06-28
  • 通讯作者: 沈敏，女，研究员，博士研究生导师，主要从事法医毒物学研究；E-mail：shenm@ssfjd.cn
  • 作者简介:严慧（1984—），女，博士，副研究员，主要从事法医毒物学研究；E-mail：yanh@ssfjd.cn
  • 基金资助:

    “十三五”国家重点研发计划资助项目（2016YFC080 0704）；国家自然科学基金青年科学基金资助项目（81302614）；上海市自然科学基金资助项目（13ZR1443000）；中央级公益性科研院所资助项目（GY2013G-7）；上海市法医学重点实验室资助项目（17DZ2273200）；上海市司法鉴定专业技术服务平台资助项目（16DZ2290900）

Evaluation of Brodifacoum-induced Toxicity by Metabonomics Approach Based on HPLC-TOF-MS

YAN HUI, ZHUO XIAN-YI, SHEN BAO-HUA, XIANG PING, SHEN MIN   

  1. （Shanghai Key Laboratory of Forensic Medicine, Shanghai Forensic Service Platform, Institute of Forensic Science, Ministry of Justice, PRC, Shanghai 200063, China）
  • Online:2017-06-25 Published:2017-06-28

被引次数

1

摘要： 目的 分析溴鼠灵中毒大鼠尿液的代谢特征，揭示溴鼠灵干预对大鼠毒性作用的分子机制。 方法 通过构建大鼠溴鼠灵中毒模型，采用高效液相色谱-飞行时间质谱（high performance liquid chromatography-time of flight mass spectrometry，HPLC-TOF-MS）获取大鼠尿液代谢轮廓，并用正交偏最小二乘法-判别分析（orthogonal partial least squares-discrimination analysis，OPLS-DA）进行多变量统计分析，找出与溴鼠灵毒性作用密切相关的差异代谢物。 结果 OPLS-DA得分图显示给药前后不同时间的大鼠尿液样本代谢物轨迹在各时间段内相似度较好，呈现各自聚类现象。比较溴鼠灵给药前后大鼠尿液样本，筛选出22个与溴鼠灵毒性相关的差异代谢物。 结论 溴鼠灵主要通过干扰大鼠体内的三羧酸循环、糖酵解、鞘脂代谢和色氨酸代谢等代谢通路发挥毒性作用，且溴鼠灵毒性作用具有累积效应。基于尿液HPLC-TOF-MS代谢组学方法可为溴鼠灵毒性作用的分子机制研究提供新思路。

关键词: 法医毒理学, 代谢组学, 高效液相色谱-飞行时间质谱, 溴鼠灵, 大鼠

Abstract: Objective To analyse the metabolic changes in urine of rats with brodifacoum intoxication, and to reveal the molecular mechanism of brodifacoum-induced toxicity on rats. Methods By establishing a brodifacoum poisoning rats model, the urine metabolic profiling data of rats were acquired using high performance liquid chromatography-time of flight mass spectrometry （HPLC-TOF-MS）. The orthogonal partial least squares analysis-discrimination analysis （OPLS-DA） was applied for the multivariate statistics and the discovery of differential metabolites closely related to toxicity of brodifacoum. Results OPLS-DA score plot showed that the urinary metabolic at different time points before and after drug administration had good similarity within time period and presented clustering phenomenon. Comparing the urine samples of rats before drug administration with which after drug administration, twenty-two metabolites related to brodifacoum-induced toxicity were selected. Conclusion The toxic effect of brodifacoum worked by disturbing the metabolic pathways in rats such as tricarboxylic cycle, glycolysis, sphingolipid metabolism and tryptophan metabolism, and the toxicity of brodifacoum is characterized of accumulation effect. The metabonomic method based on urine HPLC-TOF-MS can provide a novel insight into the study on molecular mechanism of brodifacoum-induced toxicity.

Key words: forensic toxicology, metabolomics, high performance liquid chromatography-time of flight mass spectrometry, brodifacoum, rats

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