Differential participation of phospholipase A2 isoforms during iron-induced retinal toxicity. Implications for age-related macular degeneration

Abstract

Both elevated iron concentrations and the resulting oxidative stress condition are common signs in retinas of patients with age-related macular degeneration (AMD). The role of phospholipase A2 (PLA2) during iron-induced retinal toxicity was investigated. To this end, isolated retinas were exposed to increasing Fe2+ concentrations (25, 200 or 800 μM) or to the vehicle, and lipid peroxidation levels, mitochondrial function, and the activities of cytosolic PLA2 (cPLA2) and calcium-independent PLA2 (iPLA2) were studied. Incubation with Fe2+ led to a time- and concentration-dependent increase in retinal lipid peroxidation levels whereas retinal cell viability was only affected after 60 min of oxidative injury. A differential release of arachidonic acid (AA) and palmitic acid (PAL) catalyzed by cPLA2 and iPLA 2 activities, respectively, was also observed in microsomal and cytosolic fractions obtained from retinas incubated with iron. AA release diminished as the association of cyclooxigenase-2 increased in microsomes from retinas exposed to iron. Retinal lipid peroxidation and cell viability were also analyzed in the presence of cPLA2 inhibitor, arachidonoyl trifluoromethyl ketone (ATK), and in the presence of iPLA2 inhibitor, bromoenol lactone (BEL). ATK decreased lipid peroxidation levels and also ERK1/2 activation without affecting cell viability. BEL showed the opposite effect on lipid peroxidation. Our results demonstrate that iPLA2 and cPLA2 are differentially regulated and that they selectively participate in retinal signaling in an experimental model resembling AMD.