科研之友
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摘要
Purpose Under conditions of oxidative stress, cell apoptosis is triggered through the mitochondrial intrinsic pathway. Increased levels of reactive oxygen species (ROS) are linked to excess cell loss and mediate the initiation of apoptosis in a diverse range of cell types. The aims of this study were to assess intracellular Ca2+ release, ROS production, and caspase-3, and -9 activation in ARPE-19 cells during the blue light-mediated cell death, and to examine a potential protective effect of melatonin and amfenac, in the apoptotic cascade. Methods ARPE-19 cells were cultured in their medium. First, MTT tests were performed to determine the protective effects of amfenac and melatonin. Cells were then exposed to blue light irradiation in an incubator. Intracellular Ca2+ release experiments, mitochondrial membrane depolarization, apoptosis assay, glutathione (GSH), glutathione peroxidase (GSH-Px), and ROS experiments were done according to the method stated in the Materials and methods section. Results Cell death was clearly associated with increased levels of ROS production, as measured by 2',7'-dichlorofluorescein fluorescence, and associated increase in Ca2+ levels, as measured by Fura-2-AM. Blue light-induced cell death was associated with an increased level of caspase-3 and 9, suggesting mediation via the apoptotic pathway. Cell death was also associated with mitochondrial depolarization. Melatonin was shown to delay these three steps. Conclusion Melatonin, amfenac, and their combination protect ARPE-19 cells against blue light-triggered ROS accumulation and caspase-3 and -9 activation. The antiapoptotic effect of melatonin and amfenac at doses inhibiting caspase synthesis modified Ca2+ release and prevented excessive ROS production, suggesting a new therapeutic approach to age-related macular degeneration.
- 出版日期2014-6
