EFFECT OF HYDROGEN-PEROXIDE EXPOSURE ON NORMAL HUMAN-ERYTHROCYTE DEFORMABILITY, MORPHOLOGY, SURFACE CHARACTERISTICS, AND SPECTRIN-HEMOGLOBIN CROSS-LINKING
EFFECT OF HYDROGEN-PEROXIDE EXPOSURE ON NORMAL HUMAN-ERYTHROCYTE DEFORMABILITY, MORPHOLOGY, SURFACE CHARACTERISTICS, AND SPECTRIN-HEMOGLOBIN CROSS-LINKING
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DOI：
10.1172/jci112196
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发表时间：
1985-01-01
影响因子：
15.9
通讯作者：
MOHANDAS, N
中科院分区：
文献类型：
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作者：
SNYDER, LM;FORTIER, NL;MOHANDAS, N
为了进一步确定在人红细胞膜中形成血影蛋白-血红蛋白交联的条件并检查其对膜功能的可能影响，我们将正常人红细胞在叠氮磷酸盐缓冲液（pH 7.4）中在浓度为45至180 μ M的H2 O2中孵育长达3小时。观察到的化学变化表明高铁血红蛋白形成发生得早且浓度低（45 μ M）。以棘细胞形成增加为特征的形态学变化以剂量依赖性方式发生。此外，降低细胞变形能力与增加膜刚度相称的被发现。最后，在细胞识别的单核细胞吞噬作用和粘附在体外，以及降低磷脂酰胆碱可及性蜂毒磷脂酶A2，被发现在H2 O2处理的红细胞与对照组相比，增加。后两种变化都与血影蛋白-血红蛋白交联的程度密切相关。除了这些蛋白质介导的相互作用，脂质过氧化反应也发生后，过氧化氢暴露，所示的荧光氨基丙烯衍生物的生成。添加抗氧化剂，丁基羟基甲苯，减少了荧光衍生物，但没有阻止对膜功能的影响。这表明，脂质过氧化，虽然存在，是不必要的膜的变化发现。相反，血影蛋白-血红蛋白聚集和膜功能的改变是完全防止事先暴露的红细胞一氧化碳。
To further define the conditions for forming spectrin-hemoglobin cross-linking in human erythrocyte membranes and to examine its possible effects on membrane function, we incubated normal human erythrocytes for up to 3 h in concentrations of H2O2, varying from 45 to 180 .mu.M, in an azide phosphate buffer, pH 7.4. The chemical changes observed indicated that methemoglobin formation occurred early and at a low concentration (45 .mu.M). Morphologic changes characterized by increased echinocyte formation occurred in a dose-dependent fashion. In addition, decreased cell deformability commensurate with increased membrane rigidity was found. Finally, an increase in cell recognition as determined by monocyte phagocytosis and adherence in vitro, as well as decreased phosphatidylcholine accessibility to bee venom phospholipase A2, was found in H2O2-treated erythrocytes compared with controls. Both of these latter changes were closely correlated with the extent of spectrin-hemoglobin cross-linking. In addition to these protein-mediated interactions, lipid peroxidation also occurred after H2O2 exposure, as shown by generation of fluorescent amino propene derivatives. The addition of the antioxidant, butylated hydroxytoluene, decreased the fluorescent derivatives, but did not prevent the effects on membrane function. This suggests that lipid peroxidation, though present, was not necessary for the membrane changes found. In contrast, spectrin-hemoglobin aggregation and the alterations in membrane function were completely prevented by prior exposure of the erythrocytes to carbon monoxide.