Eberhard Matthias, Ferlinz Klaus, Alizzi Katia, Cacciato Patrizia Maria, Faggio Caterina, Föller Michael, Lang Florian
Department of Physiology, University of Tübingen, Tübingen, Germany.
Cell Physiol Biochem. 2010;26(4-5):761-6. doi: 10.1159/000322343. Epub 2010 Oct 29.
FTY720 is a potent anti-inflammatory drug known to trigger suicidal death or apoptosis of a variety of nucleated cells. Erythrocytes may similarly undergo suicidal erythrocyte death or eryptosis. Hallmarks of eryptosis include cell membrane scrambling and cell shrinkage, which are triggered by increase in cytosolic Ca(2+) concentration and ceramide. The present study explored whether FTY720 stimulates eryptosis. Cell membrane scrambling was determined from annexin V-binding, cell shrinkage from forward scatter in FACS analysis, cytosolic Ca(2+) concentration from Fluo3 fluorescence, ceramide formation from fluorescence-labeled antibody binding and hemolysis from the hemoglobin concentration in the supernatant. Within 48 hours exposure to FTY720 (10 μM) significantly increased annexin V-binding, decreased forward scatter and increased cytosolic Ca(2+) concentration but did not significantly modify ceramide formation. The effects of FTY720 were significantly blunted in the nominal absence of extracelluar Ca(2+). In conclusion, at toxic concentrations, FTY720 stimulates suicidal cell death, an effect at least partially due to stimulation of Ca(2+) entry.
FTY720是一种强效抗炎药物，已知可引发多种有核细胞的自杀性死亡或凋亡。红细胞可能同样会经历自杀性红细胞死亡或细胞凋亡。细胞凋亡的标志包括细胞膜磷脂酰丝氨酸外翻和细胞皱缩，这是由胞质Ca(2+)浓度和神经酰胺增加所触发的。本研究探讨了FTY720是否会刺激细胞凋亡。通过膜联蛋白V结合来测定细胞膜磷脂酰丝氨酸外翻，通过流式细胞术分析中的前向散射来测定细胞皱缩，通过Fluo3荧光来测定胞质Ca(2+)浓度，通过荧光标记抗体结合来测定神经酰胺形成，通过上清液中的血红蛋白浓度来测定溶血。在暴露于FTY720（10 μM）48小时内，膜联蛋白V结合显著增加，前向散射降低，胞质Ca(2+)浓度增加，但神经酰胺形成没有显著改变。在名义上没有细胞外Ca(2+)的情况下，FTY720的作用显著减弱。总之，在毒性浓度下，FTY720会刺激自杀性细胞死亡，这种作用至少部分是由于刺激了Ca(2+)内流。
