Bao Qiankun, Liu Yajin, Song Hao, Yang Nan, Ai Ding, Zhu Yi, Zhang Xu
Tianjin Key Laboratory of Metabolic Diseases, Department of Physiology and Pathophysiology, Tianjin Medical University, Tianjin, China.
Department of Physiology and Pathophysiology, Peking University Health Science Center, Beijing, China.
Clin Exp Pharmacol Physiol. 2018 Jan;45(1):98-108. doi: 10.1111/1440-1681.12825. Epub 2017 Sep 18.
Eicosanoids are hundreds of metabolites derived from poly-unsaturated fatty acids (PUFAs), which regulate biological processes from multiple angles via a complex metabolic network. Targeted eicosanoid metabolomics is used to study the eicosanoid profile in biological samples but only for eicosanoids with available standards. To expand the coverage of eicosanoids detected, we identified the eicosanoids without available standards by estimation of the retention time and comparison of the MS/MS spectra with the reference ones which was collected in a database from literature. Scheduled multiple reaction monitoring- information dependent acquisition- enhanced product ion (sMRM-IDA-EPI) scan mode was applied in this method, which was called Spectrum Evaluation-assisted Eicosanoid Metabolomics (SEEM). By using this method, 243 eicosanoids (167 without standards) could be relatively quantified with precision over 90 percent. We applied the method to analyze the global profile of eicosanoids secreted by human umbilical vascular endothelial cells at the basal level and with n-3 PUFA treatment. 26 putative eicosanoids showed altered levels, despite no available standards. In general, n-3 PUFA treatment increased most of their own metabolites and decreased the epoxy-, hydroxyl- and keto- linoleic acid metabolites. The application of the SEEM method proved its potency of identification and quantification of eicosanoids without standards.
类二十烷酸是数百种源自多不饱和脂肪酸（PUFA）的代谢产物，它们通过复杂的代谢网络从多个角度调节生物过程。靶向类二十烷酸代谢组学用于研究生物样品中的类二十烷酸谱，但仅适用于有可用标准品的类二十烷酸。为了扩大检测到的类二十烷酸的覆盖范围，我们通过保留时间估计以及将MS/MS谱与从文献数据库中收集的参考谱进行比较，来鉴定没有可用标准品的类二十烷酸。本方法采用定时多反应监测-信息依赖采集-增强产物离子（sMRM-IDA-EPI）扫描模式，该方法称为光谱评估辅助类二十烷酸代谢组学（SEEM）。通过使用该方法，243种类二十烷酸（167种无标准品）能够以超过90%的精度进行相对定量。我们应用该方法分析了人脐静脉内皮细胞在基础水平和n-3多不饱和脂肪酸处理下分泌的类二十烷酸的整体谱。尽管没有可用标准品，但26种假定的类二十烷酸显示出水平变化。总体而言，n-3多不饱和脂肪酸处理增加了其自身的大多数代谢产物，并减少了环氧、羟基和酮基亚油酸代谢产物。SEEM方法的应用证明了其对无标准品的类二十烷酸进行鉴定和定量的能力。
