PMID: 11923269Mar 30, 2002Paper

Involvement of oxidative mechanisms in blue-light-induced damage to A2E-laden RPE

Investigative Ophthalmology & Visual Science
J R SparrowK Nakanishi

Abstract

The lipofuscin fluorophore A2E is known to be an initiator of blue-light-induced apoptosis in retinal pigment epithelial cells (RPE). The purpose of this study was to evaluate the role of oxidative mechanisms in mediating the cellular damage. Human RPE (ARPE-19) cells that had accumulated A2E were exposed to blue light in the presence and absence of oxygen, and nonviable cells were quantified. Potential suppressors (histidine, azide, 1,4-diazabicyclooctane [DABCO], and 1,3-dimethyl-2-thiourea [DMTU]) and enhancers (deuterium oxide [D(2)O] and 3-aminotriazole [3-AT]) of oxidative damage, were also screened for their ability to modulate the frequency of nonviable cells. A2E in PBS, with and without an oxygen-depleter or singlet-oxygen quencher and A2E-laden RPE, were exposed to 430-nm light and examined by reversed-phase high performance liquid chromatography (HPLC) and fast atom bombardment mass spectrometry (FAB-MS). The death of blue-light-illuminated A2E-laden RPE was blocked in oxygen-depleted media. When A2E-laden RPE were transferred to D(2)O-based media and then irradiated (480 nm), the number of nonviable cells was increased, whereas the latter was decreased in the presence of histidine, DABCO, and azide. Conversely, no ...Continue Reading

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