DETERMINATION OF PLATELET-ACTIVATING-FACTOR BY A CHEMILUMINESCENCE METHOD AND ITS APPLICATION TO STIMULATED GUINEA-PIG NEUTROPHILS

被引：5

作者：

HASEGAWA, Y

KUNOW, E

SHINDOU, J

YUKI, H

机构：

[1] Department of Clinical Chemistry, School of Pharmaceutical Sciences, Toho University, Funabashi-shi, Chiba, 274

来源：

LIPIDS | 1991年 / 26卷 / 12期

关键词：

D O I：

10.1007/BF02536513

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A simple, rapid and sensitive chemiluminescence method has been developed to measure platelet-activating factor (PAF). Hydrogen peroxide generated from PAF, upon phospholipase D cleavage, by choline oxidase is determined as chemiluminescence by a luminol-microperoxidase system. The detection limit of PAF by this method is 5 pmol/tube. The method is reproducible with a 5.5% coefficient of variation at 10 pmol of PAF (n = 5). Lipids were extracted from guinea pig neutrophils after stimulation with cytochalasin B and N-formyl-methionyl-leucylphenylalanine, and PAF was isolated by high-performance liquid chromatography and determined by chemiluminescence measurements. The amount of PAF detected was 96.1 +/- 39.7 (mean +/- SD, n = 7) pmol/108 cells. This highly sensitive method could be useful for the determination of PAF generated under pathophysiological conditions.

引用

页码：1117 / 1121

页数：5

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