A simplified procedure for semi-targeted lipidomic analysis of oxidized phosphatidylcholines induced by UVA irradiation
A simplified procedure for semi-targeted lipidomic analysis of oxidized phosphatidylcholines induced by UVA irradiation
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DOI：
10.1194/jlr.d025270
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发表时间：
2012-06-01
影响因子：
6.5
通讯作者：
Bochkov, Valery N.
中科院分区：
文献类型：
--
作者：
Gruber, Florian;Bicker, Wolfgang;Bochkov, Valery N.
氧化磷脂（OxPL）越来越多地被认为是信号传导介质，不仅是氧化应激的标志物，而且是与疾病发病机制相关的病理学的“制造商”。了解OxPL的单个分子种类的生物学作用需要了解它们在细胞和组织中的浓度动力学。在这项工作中，我们描述了一个简单的“指纹”程序的分析产生的四个最丰富的物种的多不饱和磷脂酰胆碱（OxPC）的氧化的分子种类的广谱。该方法是基于液-液萃取，然后通过反相HPLC耦合电喷雾电离MS/MS。超过500个峰对应的保留特性，极性和氧化的PC在8分钟内检测到99 m/z前体值使用一个单一的诊断产品离子从人皮肤成纤维细胞提取物。这些峰中有217个在细胞暴露于UVA照射后具有通量依赖性和统计学显著性增加，表明这些是真正的氧化或氧化片段化物质。这种靶向脂质组学分析方法可以作为表征由不同类型的氧化应激产生的OxPC的特定“特征”的简单的第一步，以便选择最具信息性的峰用于鉴定其分子结构和生物学作用。Gruber，F.，W. Bicker，O. V. Oskolkova，E. Tschachler和V.N.波奇科夫UVA照射诱导的氧化磷脂酰胆碱的半靶向脂质组学分析的简化程序。J. Lipid Res. 2012. 53：1232-1242。
Oxidized phospholipids (OxPLs) are increasingly recognized as signaling mediators that are not only markers of oxidative stress but are also "makers" of pathology relevant to disease pathogenesis. Understanding the biological role of individual molecular species of OxPLs requires the knowledge of their concentration kinetics in cells and tissues. In this work, we describe a straightforward "fingerprinting" procedure for analysis of a broad spectrum of molecular species generated by oxidation of the four most abundant species of polyunsaturated phosphatidylcholines (OxPCs). The approach is based on liquid-liquid extraction followed by reversed-phase HPLC coupled to electrospray ionization MS/MS. More than 500 peaks corresponding in retention properties to polar and oxidized PCs were detected within 8 min at 99 m/z precursor values using a single diagnostic product ion in extracts from human dermal fibroblasts. Two hundred seventeen of these peaks were fluence-dependently and statistically significantly increased upon exposure of cells to UVA irradiation, suggesting that these are genuine oxidized or oxidatively fragmented species. This method of semitargeted lipidomic analysis may serve as a simple first step for characterization of specific "signatures" of OxPCs produced by different types of oxidative stress in order to select the most informative peaks for identification of their molecular structure and biological role.-Gruber, F., W. Bicker, O. V. Oskolkova, E. Tschachler, and V. N. Bochkov. A simplified procedure for semi-targeted lipidomic analysis of oxidized phosphatidylcholines induced by UVA irradiation. J. Lipid Res. 2012. 53: 1232-1242.