DIRECT AND CONTINUOUS MEASUREMENT OF HYDROPEROXIDE-INDUCED OXIDATIVE STRESS ON THE MEMBRANE OF INTACT ERYTHROCYTES

被引：39

作者：

VANDENBERG, JJM

KUYPERS, FA

LUBIN, BH

ROELOFSEN, B

DENKAMP, JAFO

机构：

[1] Children's Hospital Oakland Research Institute, Oakland, CA 94609

[2] University of Utrecht, C.B.L.E., 3584 CH Utrecht

来源：

FREE RADICAL BIOLOGY AND MEDICINE | 1991年 / 11卷 / 03期

关键词：

ERYTHROCYTE; FREE RADICALS; MEMBRANE; PARINARIC ACID; FLUORESCENCE; HYDROPEROXIDE; HEMOGLOBIN; PEROXIDATION;

D O I：

10.1016/0891-5849(91)90121-I

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：

Having minimized spectroscopic interference by hemoglobin (Hb), peroxidation processes in intact erythrocytes could be monitored in a continuous assay using the fluorescent polyunsaturated fatty acid, parinaric acid (PnA), as a peroxidation probe. Control experiments to establish the character of the method are described in detail. As a practical application, comparative studies were performed to monitor the response of normal and sickle Hb-containing human erythrocytes to oxidative stress in the PnA assay. After 10 min of incubation with 200-mu-M cumene hydroperoxide (cumOOH), peroxidation of PnA was found to be enhanced in erythrocytes from sickle cell disease patients (SS: 48 +/- 9% (n = 6) of initial amount had been peroxidized) compared to healthy controls (AA: 30 +/- 4% (n = 9)). Pna peroxidation in erythrocytes from sickle cell trait individuals (AS: 30 +/- 3% (n = 4)) was equal to that in control cells. The increased oxidation of PnA in sickle erythrocytes was accompanied by enhanced oxidation of Hb (metHb and hemichrome formation), indicating that sickle Hb mediates enhanced cumOOH-derived radical generation. It is concluded that PnA can be a useful tool in studying membrane peroxidation processes in intact normal and pathological erythrocytes.

引用

页码：255 / 261

页数：7

共 21 条

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