A comprehensive analysis of membrane and morphology of erythrocytes from patients with glucose-6-phosphate dehydrogenase deficiency
A comprehensive analysis of membrane and morphology of erythrocytes from patients with glucose-6-phosphate dehydrogenase deficiency
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DOI：
10.1016/j.jsb.2015.10.015
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发表时间：
2016-06-01
影响因子：
3
通讯作者：
Jiang, Weiying
中科院分区：
文献类型：
--
作者：
Fang, Zishui;Jiang, Chengrui;Jiang, Weiying
葡萄糖-6-磷酸脱氢酶（G6 PD）缺乏症患者可由氧化应激引发急性溶血性贫血。然而，潜在的溶血机制尚不清楚。为阐明溶血机制，对溶血菌膜超微结构进行了系统的研究。采用原子力显微镜、扫描电镜、流式细胞仪和荧光显微镜等综合方法，对叔丁基过氧化氢（t-BHP）氧化G6 PD缺陷型红细胞（RBC）后的细胞膜超微结构、外化磷脂酰丝氨酸（PS）、细胞内Ca 2+浓度、细胞形态和带3蛋白分布进行了研究。结果表明，凋亡早期红细胞皱缩，膜联蛋白V（Annexin-V）与细胞膜外PS结合，膜粗糙度增加，细胞内Ca ~（2+）浓度升高，红细胞带3蛋白分布改变。当t-BHP浓度达到0.1mM时，G6 PD缺陷型红细胞膜粗糙度与对照组相比有显著性差异（p < 0.05）;当t-BHP浓度达到0.3mM时，G6 PD缺陷型红细胞外化PS与对照组相比有显著性差异（p < 0.05）。此外，红细胞的群体类型在对照组和G6 PD缺陷组之间显示出显著差异。G6 PD缺陷型红细胞在氧化应激的作用下，红细胞凋亡加重，红细胞膜粗糙度增加，带3蛋白分布异常。棘红细胞是G6 PD缺乏症患者外周血中最常见的异常红细胞形态，它可缩短红细胞的寿命。本研究的结果将有助于进一步了解G6 PD缺陷症的溶血机制。(C)2015 Elsevier Inc. All rights reserved.
Acute hemolytic anemia could be triggered by oxidative stress in the patients with glucose-6-phosphate dehydrogenase (G6PD) deficiency. However, the underlying hemolytic mechanism is unknown. To make clear the hemolytic mechanisms, a systematic study on membrane ultrastructure had been undertaken. A comprehensive method was used including atomic force microscopy, scanning electron microscopy, flow cytometer and fluorescence microscopy to analyze the membrane ultrastructure, externalized phosphatidylserine (PS), intracellular Ca2+ concentration, morphology and the distributions of band 3 protein in G6PD deficient red blood cells (RBCs) after tert-butyl-hydroperoxide (t-BHP) oxidation. The results showed that erythrocyte shrinkage, annexin-V binding to externalized PS on the membrane of early stage apoptotic cells, the increased membrane roughness and intracellular Ca2+ concentration, as well as the change of distributions of band 3 protein in RBCs. Compared with the control RBCs, as the concentration of t-BHP up to 0.1 mM, the membrane roughness of G6PD deficient RBCs showed significant difference (p < 0.05) and as the concentration of t-BHP up to 0.3 mM, externalized PS showed significant difference (p < 0.05). Furthermore, the population types of RBCs showed dramatic difference between control groups and G6PD deficient groups. Oxidative stress induced more serious erythrocyte apoptosis and resulted in increased roughness of erythrocyte membrane and abnormal distributed band 3 protein in G6PD deficient RBCs. Echinocytes are the predominant abnormal erythrocyte shape occurring in the peripheral blood from patients with G6PD deficiency, which may shorten the RBCs lifespan. The results in the present study will give an increased understanding for the hemolytic mechanism of G6PD deficiency. (C) 2015 Elsevier Inc. All rights reserved.