Effect of untreated and photobleached bovine RPE melanosomes on the photoinduced peroxidation of lipids

Abstract

Melanin is usually considered a photoprotective pigment and antioxidant agent, but it is unclear how melanosomes protect pigmented cells against oxidative stress induced by light and whether aging modulates its photoprotective function, particularly in long-lived post-mitotic cells such as the retinal pigment epithelium (RPE). To address these issues, we analyzed the effects of untreated and experimentally photobleached melanosomes from cow RPE on the peroxidation of liposomal lipids induced by irradiation with intense visible light or by a rose Bengal photosensitized reaction. Photobleached melanosomes were used as a model of photo-aged pigment granules, and the progress of lipid peroxidation was monitored by electron spin resonance (EPR) oximetry and the iodometric determination of lipid hydroperoxides. We observed that while untreated melanosomes inhibited the rose Bengal induced peroxidation of lipids only moderately, partially photobleached melanosomes had very little effect on this process. Untreated melanosomes also inhibited peroxidation of liposomal lipids induced by intense visible light; however, the inhibitory effect markedly changed with the irradiation time. On the other hand, partially photobleached pigment granules accelerated the photoinduced peroxidation of lipids. The observed effects illustrate the limited efficiency of melanin within granules to scavenge and quench reactive oxygen species randomly generated by photosensitized reactions. The photosensitizing ability of photobleached melanosomes may arise from their capacity to photogenerate hydrogen peroxide. Collectively, our data indicate that natural melanin is only a moderately efficient photoprotective pigment, which upon photoaging may lose its antioxidant efficiency and even become a photosensitizer.