A comprehensive analysis of membrane and morphology of erythrocytes from patients with glucose-6-phosphate dehydrogenase deficiency

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• 作者：Zishui Fang ; Chengrui Jiang ; Jia Tang ; Ming He ; Xiaoying Lin ; Xiaodan Chen ; Luhao Han ; Zhiqiang Zhang ; Yi Feng ; Yibin Guo ; Hongyi Li ; Weiying Jiang ; ^{jiangwy@mail.sysu.edu.cn" class="auth_mail" title="E-mail the corresponding author}
• 关键词：G6PD ; glucose-6-phosphate dehydrogenase ; RBCs ; red blood cells ; AFM ; atomic force microscopy ; SEM ; scanning electron microscopy ; Ra ; arithmetic average roughness ; Rq ; root-mean-square roughness ; PS ; phosphatidylserine ; NADPH ; nicotinamide adenine dinucleotide phosphate ; t-BHP ; tert-butyl-hydroperoxide
• 刊名：Journal of Structural Biology
• 出版年：2016
• 出版时间：June 2016
• 年：2016
• 卷：194
• 期：3
• 页码：235-243
• 全文大小：2779 K

文摘

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.