Blue light induced A2E oxidation in rat eyes--experimental animal model of dry AMD.

Previous studies have shown that short-wavelength blue visible light induces retinal injury and may be a risk factor for age related macular degeneration. A2E is a blue light absorbing retinal chromophore that accumulates with age. Our previous in vitro studies have determined that, although A2E itself has a low phototoxic efficiency, the oxidation products of A2E that are formed in the presence of visible light can contribute to observed retinal pigment epithelial photodamage. The purpose of this study was to investigate the effects of blue light on retinal phototoxicity and its relationship to A2E, oxidized A2E and its isomers. Sprague-Dawley albino rats were dark adapted for 24 h. Control rats remained in the dark while experimental rats were exposed to blue light (λ = 450 nm, 3.1 mW cm(-2)) for 6 h. Isolated retinas were homogenized in Folch extraction mixture and then in chloroform. The dried extracts were reconstituted and divided for determination of organic soluble compound. Esters of fatty acids were determined with GC-MS, A2E and other chromophores using HPLC, and A2E oxidation products with LC-MS. Exposure of rat eyes to blue light did not significantly change the fatty acid composition of the retina. The A2E concentration (normalized to fatty acid content) in blue light exposed animals was found to be lower than the A2E concentration in control rats. The concentrations of all-trans-retinal-ethanolamine adduct and iso-A2E a precursor and an isomer of A2E respectively, were also lower after blue-light exposure than in the retinas of rats housed in the dark. On the other hand, the amount of oxidized forms of A2E was higher in the animals exposed to blue light. We conclude that in the rat eye, blue-light exposure promotes oxidation of A2E and iso-A2E to the products that are toxic to retinal tissue. Although high concentrations of A2E may be cytotoxic to the retina, the phototoxicity associated with blue light damage to the retina is in part a result of the formation of toxic A2E oxides. This effect may partially explain the association between blue light induced retinal injury and macular degeneration.

Melanin-porphyrin interaction monitored by delayed luminescence and photoacoustics.

Even though melanin is commonly viewed as a photoprotective agent, the molecular mechanism of the melanin-related photoprotective action remains unclear. We studied the interaction of a synthetic dopa melanin with positively and negatively charged porphyrins using an array of spectroscopic techniques. Thus absorption, fluorescence, time-resolved delayed luminescence in the microsecond time range, and photoacoustic spectra at different modulation frequencies of both porphyrins and their mixtures with dopa melanin were measured in buffered aqueous solution at room temperature. It has been confirmed that dopa melanin forms a complex with the cationic porphyrin in its ground state, which significantly modifies the optical properties of the dye molecule. Although no such complex can be detected for the anionic porphyrin, some interaction between melanin and the porphyrin molecule can be detected in its excited state. Both porphyrins show delayed luminescence with a decay time of 35-38 microseconds, which is substantially reduced by melanin. Our photoacoustic measurements indicate that upon photoexcitation, melanin efficiently liberates heat. Our data also suggest that the thermal properties of melanin can be modified by porphyrins, particularly by the cationic form of the dye. The ionic complexes of dopa melanin with the positively charged porphyrin exhibit properties typical for a supermolecular system that liberates heat as a whole.