Stimulation of Phospholipid Scrambling of the Erythrocyte Membrane by 9-Cis-Retinoic Acid
Stimulation of Phospholipid Scrambling of the Erythrocyte Membrane by 9-Cis-Retinoic Acid
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DOI：
10.1159/000457014
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发表时间：
2017-01-01
影响因子：
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通讯作者：
Lang, Florian
中科院分区：
文献类型：
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作者：
Abed, Majed;Alzoubi, Kousi;Lang, Florian
背景/目的：内源性类维生素A 9-顺式视黄酸先前已被证明可引发多种细胞（包括几种肿瘤细胞）的细胞凋亡，因此被建议用于治疗恶性肿瘤。与有核细胞凋亡类似，红细胞可能进入自杀性红细胞死亡或红细胞凋亡，其特征是细胞收缩和细胞膜扰乱，磷脂酰丝氨酸易位至红细胞表面。参与红细胞死亡的细胞机制包括胞质 Ca2+ 活性 ([ Ca2+](i)) 的增加和神经酰胺的形成。本研究探讨了 9-顺式视黄酸是否会诱导红细胞凋亡，以及该作用是否涉及 Ca2+ 和/或神经酰胺。方法：采用流式细胞术评估前向散射的红细胞体积、膜联蛋白 V 结合导致的细胞表面磷脂酰丝氨酸暴露、Fluo3 荧光的 [Ca2+] i 以及使用特异性抗体的神经酰胺丰度。根据上清液中的血红蛋白浓度对溶血进行定量。结果：人红细胞暴露于 9-顺式视黄酸 (>= 0.5 μg/ml) 48 小时显着增加了膜联蛋白-V 结合细胞的百分比，并显着降低了前向散射。暴露于 9-顺-视黄酸 (= 0.5 μg/ml) 显着增加 Fluo3 荧光，并且 9-顺-视黄酸对膜联蛋白-V 结合的影响因细胞外 Ca2+ 的去除而显着减弱。暴露于9-顺式视黄酸（1μg/ml）进一步显着增加了红细胞表面的神经酰胺丰度并显着增加了溶血。结论：9-顺式视黄酸会引发红细胞细胞膜的磷脂扰乱，至少部分影响 Ca2+ 和神经酰胺的下游。 (C) 2017 作者由 S. Karger AG 出版，巴塞尔
Background/Aims: The endogenous retinoid 9-cis-retinoic acid has previously been shown to trigger apoptosis in a wide variety of cells including several tumor cells and has thus been suggested for the treatment of malignancy. Similar to apoptosis of nucleated cells, erythrocytes may enter suicidal erythrocyte death or eryptosis, which is characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. Cellular mechanisms participating in the accomplishment of eryptosis include increase of cytosolic Ca2+ activity ([ Ca2+](i)) and formation of ceramide. The present study explored, whether 9-cis-retinoic acid induces eryptosis and whether the effect involves Ca2+ and/or ceramide. Methods: Flow cytometry was employed to estimate erythrocyte volume from forward scatter, phosphatidylserine exposure at the cell surface from annexin-V-binding, [ Ca2+] i from Fluo3-fluorescence, and ceramide abundance utilizing specific antibodies. Hemolysis was quantified from hemoglobin concentration in the supernatant. Results: A 48 hours exposure of human erythrocytes to 9-cis-retinoic acid (>= 0.5 mu g/ml) significantly increased the percentage of annexin-V-binding cells and significantly decreased forward scatter. Exposure to 9-cis-retinoic acid (= 0.5 mu g/ml) significantly increased Fluo3-fluorescence, and the effect of 9-cis-retinoic acid on annexin-V-binding was significantly blunted by removal of extracellular Ca2+. Exposure to 9-cis-retinoic acid (1 mu g/ml) further significantly increased the ceramide abundance at the erythrocyte surface and significantly increased hemolysis. Conclusions: 9-cis-retinoic acid triggers phospholipid scrambling of the erythrocyte cell membrane, an effect at least in part downstream of Ca2+ and ceramide. (C) 2017 The Author(s) Published by S. Karger AG, Basel