Laboratory of Food and Biodynamics, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan.
Biochem Biophys Res Commun. 2010 Mar 19;393(4):631-6. doi: 10.1016/j.bbrc.2010.02.043. Epub 2010 Feb 12.
Lipid peroxidation products have a high reactivity against the primary amino groups of biomolecules such as aminophospholipids, proteins, and DNA. Until now, many papers have reported about the modification of biomolecules derived from lipid peroxides. Our group has also reported that aminophospholipids, such as phosphatidylethanolamine (PE), can be modified by lipid peroxidation including 13-hydroperoxyoctadecadienoic acid (13-HPODE). The aim of this study was to examine the oxidative stress in vivo by detecting the formation of N-(hexanoyl)phosphatidylethanolamine (HEPE) and N-(hexanoyl)phosphatidylserine (HEPS), a novel hexanoyl adduct, using a liquid chromatography/tandem mass spectrometry (LC/MS/MS) and a monoclonal antibody. Consequently, we observed that the formation of HEPE and HEPS occurred in the red blood cell (RBC) ghosts modified by 13-HPODE and the oxidative stress model induced by carbon tetrachloride (CCl(4)) using LC/MS/MS monitoring hexanoyl ethanolamine (HEEA), a head group of HEPE, and hexanoyl serine (HESE) as a part of HEPS. Furthermore, we obtained a novel type of monoclonal antibody against HEPE. This antibody could recognize HEPE in the liver of rats with oxidative stress in vivo.
脂质过氧化物产物与生物分子的伯氨基具有高反应性，如氨基磷脂、蛋白质和 DNA。到目前为止，许多论文都报道了脂质过氧化物衍生的生物分子的修饰。我们的小组还报告说，氨基磷脂，如磷脂酰乙醇胺（PE），可以通过包括 13-羟基十八碳二烯酸（13-HPODE）在内的脂质过氧化作用修饰。本研究的目的是通过使用液相色谱/串联质谱（LC/MS/MS）和单克隆抗体检测新型己酰化加合物 N-（己酰基）磷脂酰乙醇胺（HEPE）和 N-（己酰基）磷脂酰丝氨酸（HEPS）的形成，来检测体内氧化应激。结果表明，在 13-HPODE 修饰的红细胞（RBC）血影和四氯化碳（CCl4）诱导的氧化应激模型中观察到 HEPE 和 HEPS 的形成，使用 LC/MS/MS 监测 HEPE 的头基己酰乙醇胺（HEEA）和 HEPS 的一部分己酰丝氨酸（HESE）。此外，我们获得了针对 HEPE 的新型单克隆抗体。该抗体可识别体内氧化应激大鼠肝脏中的 HEPE。
