Oxidative-induced apoptosis to an immortalized ganglion cell line is caspase independent but involves the activation of poly (ADP-ribose)polymerase and apoptosis-inducing factor
Oxidative-induced apoptosis to an immortalized ganglion cell line is caspase independent but involves the activation of poly (ADP-ribose)polymerase and apoptosis-inducing factor
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DOI：
10.1016/j.brainres.2007.10.073
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发表时间：
2008-01-10
期刊：
影响因子：
2.9
通讯作者：
Osborne, Neville N.
中科院分区：
文献类型：
--
作者：
Li, Guang-Yu;Osborne, Neville N.
本工作的目的是表征氧化诱导死亡的分子基础，这一过程与青光眼等眼部疾病有关，在RGC-5细胞中，一种永生化的视网膜神经节细胞（RGC）系。通过用过氧化氢处理RGC-5细胞诱导氧化应激，并与已知的光损伤（1000 lx，400 - 760 nm）的作用进行比较。过氧化氢以剂量依赖性方式导致RGC-5细胞活力丧失。细胞活力的丧失是通过细胞凋亡，其特征在于DNA的破坏（TUNEL方法）、膜磷脂酰丝氨酸的存在（APOPercentage（TM）方法）、PARP-1和AIF的活化。氧化应激引起的ROS刺激达到最高水平之前，最佳的凋亡。过氧化氢诱导的细胞凋亡并不导致caspase-3的激活，也不受caspase抑制剂Z-VAD-favor的影响。然而，PARP-1抑制剂NU-1025抵消了过氧化氢和光的影响。提供的证据表明，这两种形式的氧化应激导致AIF被裂解的产品位于胞质隔室。光诱导的细胞凋亡的线粒体解偶联剂M3778的存在下，但增强钴的存在下衰减。与此相反，过氧化氢诱导的细胞凋亡不受M3778的影响，但衰减钴。结果表明，光引起的氧化应激依赖于功能线粒体，过氧化氢或光引起的细胞凋亡的分子机制相似但不完全相同。(c)2007年由Elsevier B. V.出版。
The aim of the present work was to characterize the molecular basis of oxidative-induced death, a process that has been implicated in eye diseases like glaucoma, in RGC-5 cells, an immortalized retinal ganglion cell (RGC) line. Oxidative stress was induced by treatment of RGC-5 cells with hydrogen peroxide and compared to a known effect of a light insult (1000 lx, 400 - 760 nm). Hydrogen peroxide causes a loss of viability of RGC-5 cells in a dose-dependent manner. Loss of cell viability was by apoptosis characterized by breakdown of DNA (TUNEL method), presence of membrane phosphatidylserine (APOPercentage (TM) method), activation of PARP-1 and AIF. Oxidative stress caused a stimulation of ROS which reached maximum levels before optimum apoptosis. Hydrogen-peroxide-induced apoptosis did not result in an activation of caspase-3 and was unaffected by the caspase inhibitor Z-VAD-fmk. However, the PARP-1 inhibitor NU-1025 counteracted the effects of hydrogen peroxide and light. Evidence is provided to show that both forms of oxidative stress caused AIF to be cleaved with the product located to the cytosolic compartment. Light-induced apoptosis was attenuated by the presence of the mitochondrial uncoupler M3778 but potentiated by the presence of cobalt. In contrast, hydrogen-peroxide-induced apoptosis was unaffected by M3778 but attenuated by cobalt. The results show that oxidative stress caused by light is dependent an functional mitochondria and that the molecular mechanisms of apoptosis caused by hydrogen peroxide or light are similar but not identical. (c) 2007 Published by Elsevier B.V.