Lipid metabolism of leukocytes in the unstimulated and activated states
Lipid metabolism of leukocytes in the unstimulated and activated states
复制标题
非刺激状态和激活状态下白细胞的脂代谢
DOI：
10.1007/s00216-020-02460-8
复制
发表时间：
2020-02-14
影响因子：
4.3
通讯作者：
Giera, Martin
中科院分区：
文献类型：
--
作者：
Alarcon-Barrera, Juan Carlos;von Hegedus, Johannes H.;Giera, Martin
脂质组学已成为研究细胞脂质代谢的有力技术。由于脂质组包含许多同分异构体和同量异序物质，导致不同脂质类别之间存在显著重叠，因此需要尖端的分析技术来全面分析脂质代谢。就在最近，差分迁移率光谱法（DMS）已经发展成为这样一种技术，有助于克服几个分析挑战。我们在这里开始应用DMS和Lipidyzer（TM）平台，以获得在静息和激活状态下白细胞相关脂质代谢的全面概述。首先，我们使用HL 60细胞测试平台的线性和重复性。我们获得了良好的线性大多数分析的13种脂质类（相关系数> 0.95），和良好的重复性（%CV < 15）。通过比较中性粒细胞（PMNs），单核细胞（CD14+）和淋巴细胞（CD4+）的脂质组，我们揭示了白细胞特异性脂质模式以及通过差异刺激发生的脂质组学变化。例如，在静息状态下，与CD4+和CD14+细胞相比，PMN被证明含有更高量的三酰甘油酯。另一方面，CD4+和CD14+细胞含有较高水平的磷脂和神经酰胺。刺激后，CD 4+细胞和中性粒细胞中的二酰基甘油酯、己糖神经酰胺、磷脂酰胆碱、磷酸乙醇胺和溶血磷酸乙醇胺上调，而CD 14+细胞没有显示出显着变化。通过探索显著上调的脂质类的脂肪酸含量，我们主要发现非常长和多不饱和脂肪酸的浓度增加。我们的研究结果表明Lipidyzer（TM）平台用于研究细胞脂质代谢的有用性。它的应用使我们能够探索白细胞的脂质体。
Lipidomics has emerged as a powerful technique to study cellular lipid metabolism. As the lipidome contains numerous isomeric and isobaric species resulting in a significant overlap between different lipid classes, cutting-edge analytical technology is necessary for a comprehensive analysis of lipid metabolism. Just recently, differential mobility spectrometry (DMS) has evolved as such a technology, helping to overcome several analytical challenges. We here set out to apply DMS and the Lipidyzer (TM) platform to obtain a comprehensive overview of leukocyte-related lipid metabolism in the resting and activated states. First, we tested the linearity and repeatability of the platform by using HL60 cells. We obtained good linearities for most of the thirteen analyzed lipid classes (correlation coefficient > 0.95), and good repeatability (%CV < 15). By comparing the lipidome of neutrophils (PMNs), monocytes (CD14+), and lymphocytes (CD4+), we shed light on leukocyte-specific lipid patterns as well as lipidomic changes occurring through differential stimulation. For example, at the resting state, PMNs proved to contain higher amounts of triacylglycerides compared to CD4+ and CD14+ cells. On the other hand, CD4+ and CD14+ cells contained higher levels of phospholipids and ceramides. Upon stimulation, diacylglycerides, hexosylceramides, phosphatidylcholines, phosphoethanolamines, and lysophosphoethanolamines were upregulated in CD4+ cells and PMNs, whereas CD14+ cells did not show significant changes. By exploring the fatty acid content of the significantly upregulated lipid classes, we mainly found increased concentrations of very long and polyunsaturated fatty acids. Our results indicate the usefulness of the Lipidyzer (TM) platform for studying cellular lipid metabolism. Its application allowed us to explore the lipidome of leukocytes.