The presence of oxidized phosphatidylserine on Fas-mediated apoptotic cell surface
The presence of oxidized phosphatidylserine on Fas-mediated apoptotic cell surface
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DOI：
10.1016/j.bbalip.2005.08.011
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发表时间：
2005-10-01
影响因子：
4.8
通讯作者：
Yamada, K
中科院分区：
文献类型：
--
作者：
Matsura, T;Togawa, A;Yamada, K
越来越多的证据表明，磷脂酰丝氨酸（PS）的氧化与其跨膜迁移从内到外小叶的质膜在细胞凋亡过程中。然而，没有直接的证据氧化PS（PSox）的细胞表面上进行凋亡的存在。本研究是为了检测Fas参与Jurkat细胞后外化到细胞表面的PSox。用抗Fas抗体处理Jurkat细胞诱导caspase-3活化、染色质浓缩、PS外化、活性氧的产生、细胞内谷胱甘肽耗竭、线粒体跨膜电位的破坏和细胞色素c从线粒体的释放。为了确定外部化的PS和磷脂酰乙醇胺（PE），Jurkat细胞用抗Fas抗体处理，然后用膜不可渗透的荧光胺（一种用于可视化含有伯氨基的脂质的探针）标记。提取其总脂质并进行二维高效薄层色谱（HPTLC）。用N，N，N '，N”-四甲基-对苯二胺二盐酸盐喷洒HPTLC板以检测磷脂氢过氧化物。PSox在正常细胞内少量存在，但不在细胞表面。用抗Fas抗体处理增加了细胞内的PSox，并导致PSox出现在细胞表面。与此相反，Fas连接的细胞表面的PE没有被氧化。因此，本研究首次证明了PSox在凋亡细胞内和表面的存在。(c)2005 Elsevier B.V保留所有权利。
A growing body of evidence suggests that phosphatidylserine (PS) oxidation is linked with its transmembrane migration from the inner to the outer leaflet of the plasma membrane during apoptosis. However, there is no direct evidence for the presence of oxidized PS (PSox) on the surface of cells undergoing apoptosis. The present study was performed to detect PSox externalized to the cell surface after Fas engagement in Jurkat cells. Treatment of Jurkat cells with anti-Fas antibody induced caspase-3 activation, chromatin condensation, PS externalization, generation of reactive oxygen species, intracellular glutathione depletion, disruption of mitochondrial transmembrane potential and release of cytochrome c from mitochondria. To determine externalized PS and phosphatidylethanolamine (PE), Jurkat cells were treated with anti-Fas antibody and then labeled with membrane-impermeable fluorescamine, a probe for visualizing lipids that contain primary amino groups. Their total lipids were extracted and subjected to two-dimensional high-performance thin-layer chromatography (HPTLC). The HPTLC plate was sprayed with N,N,N',N"-tetramethyl-p-phenylenediamine dihydrochloride to detect phospholipid hydroperoxides. PSox was present in small amounts within but not on the surface of normal cells. Treatment with anti-Fas antibody increased PSox within the cells and caused PSox to appear on the cell surface. In contrast, PE on the surface of Fas-ligated cells was not oxidized. Thus, the present study demonstrates for the first time the presence of PSox both within and on the surface of apoptotic cells. (c) 2005 Elsevier B.V All rights reserved.