基于LC-MS技术的代谢组学方法研究吡虫啉对工蜂代谢的影响

试验利用液相色谱-质谱联用技术（liquid chromatograph mass spectrometer，LC-MS）对出房的1日龄工蜂进行代谢组学分析，以明确亚致死剂量的吡虫啉对蜜蜂生长发育期代谢的影响。采用蜜蜂幼虫人工室内饲养技术，选取意大利蜜蜂（Apis mellifera ligustica）为研究对象，用含0.00001μg/μL亚致死剂量吡虫啉的日粮饲喂1日龄幼虫直至排便。记录蜜蜂的羽化率、初生重、出房时间；测量蜜蜂血淋巴的超氧化物歧化酶（super oxide dismutase，SOD）、过氧化氢酶（catalase，CAT）、过氧化物酶（peroxidase，POD）活力；采用LC-MS方法检测亚致死剂量浓度吡虫啉处理蜜蜂羽化出房时的代谢物差异，对检测数据进行模式识别分析，筛选差异代谢物。结果表明，亚致死剂量的吡虫啉引起蜜蜂出房时间显著延长（P<0.05）；使蜜蜂血淋巴的SOD和POD活性显著升高（P<0.05）；采用主成分分析（principal component analysis，PCA）和偏最小二乘法判别方法（supervised partial leastsquares-discriminant analysis，PLS-DA）分析代谢组数据，结果显示试验组和对照组明显分离。试验共鉴定出18个蜜蜂差异代谢物。本结果可对进一步研究吡虫啉胁迫蜜蜂代谢物的代谢规律、阐明蜜蜂解毒机制提供理论依据。

Metabolomic Analysis Imidacloprid Effect of Honeybee Worker Based on Liquid Chromatograph Mass Spectrometer

To identify the impact of sub-lethal dose of imidacloprid on honeybee metabolism during growth stage, the metabonomics analysis was conducted for one-day old worker bee based on liquid chromatography-mass spectrometry (LC-MS).One-day old Apis mellifera ligustica were fed with 0.00001 μg/μL sub-lethal dose of imidacloprid until defecation by the artificial indoor feeding technique.The eclosion rate, body weight at birth, out of the hive time, the activities of SOD, CAT and POD in bee hemolymph were analyzed respectively, and synchronously, differential metabolites were screened out through pattern recognition analysis by means of LC-MS.The results indicated that the out of hive time was significantly prolonged (P<0.05) and the activities of SOD and POD in bee hemolymph were significantly increased (P<0.05) with the treatment of sub-lethal dose of imidacloprid.Principal component analysis (PCA) and supervised partial leastsquares-discriminant analysis (PLS-DA) showed that the experimental group and the control group were significantly separated, the 18 different metabolites were identified.The results could provide theoretical guidance for the further research on the metabolic rules of bee metabolin under imidacloprid stress and elucidation of the bee detoxification mechanism.