四氯化碳对大鼠肝微粒体脂质的共价修饰。

Molecular toxicology Pub Date : 1989-07-01
B S Kaphalia, G A Ansari
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引用次数: 0

摘要

本研究在体外厌氧条件下和体内大鼠肝微粒体14CCl4代谢过程中分离和鉴定与14C标签结合的各种脂质。通过薄层色谱法鉴定的两种主要放射性组分分别为中性脂和磷脂,分别对应脂肪酸和甘油三酯,分别对应磷脂酰胆碱(PC)和磷脂酰乙醇胺(PE)。在PC和PE馏分中发现了大约89%与磷脂相关的放射性。磷脂酶A2 (EC 3.1.1.4)水解PC和PE释放出约50%的总放射性,作为脂肪酸对应的脂质部分。用三氟化硼在甲醇中甲基化了放射性中性脂以及PC和PE水解的脂质部分。通过反相高效液相色谱(HPLC)对甲基化脂质进行分离,体外实验得到的脂质中14C标记的洗脱谱与体内实验得到的相似。油酸甲酯后立即洗脱的主要放射性组分经化学电离质谱鉴定为三氯甲基十四烯酸甲酯和三氯甲基三烯酸甲酯。质谱分析也表明油酸形成了二氯苯加合物。然而,类似的质谱检测三氯甲基化脂质在中性脂质和从体内研究中分离的磷脂中并不明显。作为非极性馏分洗脱的14c标记脂质显示出含有3个以上氯的高分子量。本文还讨论了基于高效液相色谱和质谱分析的三氯甲基化脂的二聚化和交联反应。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Covalent modification of hepatic microsomal lipids of rats by carbon tetrachloride.

The present study was undertaken to isolate and identify various lipids bound to 14C label during hepatic microsomal metabolism of 14CCl4 in vitro under anaerobic conditions and in vivo in rats. The two major radioactive fractions identified by thin-layer chromatography each for neutral lipids and phospholipids from in vitro and in vivo experiments corresponded to fatty acids and triglycerides and to phosphatidylcholine (PC) and phosphatidylethanolamine (PE), respectively. Approximately 89% of the radioactivity associated with phospholipids was found in PC and PE fractions. Hydrolysis of PC and PE with phospholipase A2 (EC 3.1.1.4) released about 50% of the total radioactivity as lipid moieties corresponding to fatty acids. The radioactive neutral lipids and the lipid moieties hydrolyzed from PC and PE were methylated with boron trifluoride in methanol. These methylated lipids were separated by reversed-phase high-performance liquid chromatography (HPLC), and the elution profiles of 14C label found for the lipids obtained from in vitro experiments were similar to those from in vivo. The major radioactive fractions eluted immediately after methyl oleate were identified as trichloromethyloctadecenoic and trichloromethyleicosatrienoic acid methyl esters by chemical ionization mass spectrometry. The mass spectral analysis of these fractions also indicated the formation of dichlorocarbene adduct of oleic acid. However, similar mass spectrometric detection of trichloromethylated lipids was not evident in neutral lipids and phospholipids isolated from in vivo studies. The 14C-labeled lipids eluted as a nonpolar fraction exhibited a high molecular weight containing more than three chlorines. Dimerization and cross-linking of trichloromethylated lipids based on HPLC and mass spectral analysis are also discussed in this paper.

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