亚致死剂量氯吡嘧磺隆胁迫对斑马鱼典型肝脏代谢物的影响

为获知水环境中亚致死剂量的氯吡嘧磺隆对斑马鱼肝脏的影响,将斑马鱼暴露在亚致死剂量的氯吡嘧磺隆药液中8 d,采用超高效液相色谱与四级杆-轨道离子阱质谱串联对斑马鱼的肝脏代谢物进行分析,采用SIEVE软件将图谱信息转换为含有保留时间、质荷比和相对峰面积的二维数据阵.同时,结合mz Cloud、Human Metabolome Database(HMDB)数据库共鉴定出33种代谢物,包括氨基酸、糖、有机酸等物质.最后,采用SIMCA软件对正离子和负离子模式下的检测结果进行主成分、偏最小二乘法和正交校正的偏最小二乘法分析.主成分和偏最小二乘法结果表明,对照组与处理组在2种检测模式下均存在差异代谢物.为了寻找造成对照组与处理组之间差异的代谢物,对代谢物进行S-Plot分析,结果表明,肌酸酐、牛磺酸、多巴胺和5b-胆甾醇硫酸酯组间差异最大(p0.01).4种代谢产物可作为斑马鱼在氯吡嘧磺隆亚致死剂量胁迫下肝脏代谢异常的潜在标记物.

Changes in metabolites in the liver of zebrafish under sublethal halosulfuron-methyl stress

We applied a sub-lethal dose of halosulfuron-methyl stress to zebrafish in order to study the changes in metabolites in the liver.Ultra-high performance liquid chromatography coupled with Q Exactive hybrid quadrupole-orbitrap mass spectrometry (UPLC-MS/MS) was applied to analyze the effects of the sub-lethal dose of halosulfuron-methyl on metabolites in the liver of zebrafish.Each chromatogram obtained from UPLC-MS/MS analysis was pre-processed using SIEVE software to generate discrete peak lists containing m/z, retention time, and relative peak area.Using the mzCloud and Human Metabolome Database, 33 metabolites were identified including amino acids, sugars, and organic acids.The pre-processed UPLC-MS/MS data were exported to SIMICA-P 13.0 for analysis by using multivariate statistical methods.Three multivariate analysis methods were used to determine the optimal separation of clusters: principal component analysis, partial least squares-discriminant analysis, and orthogonal projection to latent structure-discriminant analysis.The results revealed significant differences(p<0.01) between the control and treatment groups.A representative S-plot showed the variables responsible for the separation.Creatinine, taurine, dopamine, and 5b-cyprinol sulfate appeared to be the most affecting variables(p<0.01).These four metabolites were identified as potential indicators of halosulfuron-methyl-induced liver abnormalities.