L M Bova1, M H Sweeney, J F Jamie, R J Truscott. 1. Australian Cataract Research Foundation, Department of Chemistry, University of Wollongong, Wollongong, NSW, Australia.
Abstract
PURPOSE: Age-dependent human lens coloration may be explained by the binding of UV filters to crystallins. It has been proposed that glutathione may compete for reaction with UV filter degradation products and therefore protect crystallins from modification. To understand this process, UV filters were quantified together with oxidized and reduced glutathione in human lenses of varying age. METHODS: Lens tissues were homogenized in ethanol to extract the UV filters. Metabolites were quantified by HPLC and correlations between them in the nuclear and cortical regions of the lens were examined. RESULTS: The concentrations of the UV filters 3-hydroxykynurenine, kynurenine, and 3-hydroxykynurenine glucoside decreased linearly with age, with slightly lower levels in the nucleus than the cortex. 4-(2-Amino-3-hydroxyphenyl)-4-oxobutanoic acid glucoside was found in higher levels in the nucleus than the cortex and decreased slowly in both regions with age. Glutathionyl-3-hydroxykynurenine glucoside was present in higher concentrations in the nucleus, barely detectable in young lenses, but increased significantly after age 50. Reduced glutathione levels were lower in the nucleus and decreased in both regions with age, yet oxidized glutathione increased in the nucleus but remained constant in the cortex. CONCLUSIONS: Results are consistent with a predominantly nuclear origin for both 4-(2-amino-3-hydroxyphenyl)-4-oxobutanoic acid glucoside and glutathionyl-3-hydroxykynurenine glucoside. This is in accord with their proposed mechanism of formation, which involves an initial deamination of 3-hydroxykynurenine glucoside. This process is more pronounced in older lenses, possibly because of the barrier to diffusion. The barrier may also explain the increase in nuclear oxidized glutathione that is observed with age.
PURPOSE: Age-dependent human lens coloration may be explained by the binding of UV filters to crystallins. It has been proposed that glutathione may compete for reaction with UV filter degradation products and therefore protect crystallins from modification. To understand this process, UV filters were quantified together with oxidized and reduced glutathione in human lenses of varying age. METHODS: Lens tissues were homogenized in ethanol to extract the UV filters. Metabolites were quantified by HPLC and correlations between them in the nuclear and cortical regions of the lens were examined. RESULTS: The concentrations of the UV filters 3-hydroxykynurenine, kynurenine, and 3-hydroxykynurenine glucoside decreased linearly with age, with slightly lower levels in the nucleus than the cortex. 4-(2-Amino-3-hydroxyphenyl)-4-oxobutanoic acid glucoside was found in higher levels in the nucleus than the cortex and decreased slowly in both regions with age. Glutathionyl-3-hydroxykynurenine glucoside was present in higher concentrations in the nucleus, barely detectable in young lenses, but increased significantly after age 50. Reduced glutathione levels were lower in the nucleus and decreased in both regions with age, yet oxidized glutathione increased in the nucleus but remained constant in the cortex. CONCLUSIONS: Results are consistent with a predominantly nuclear origin for both 4-(2-amino-3-hydroxyphenyl)-4-oxobutanoic acid glucoside and glutathionyl-3-hydroxykynurenine glucoside. This is in accord with their proposed mechanism of formation, which involves an initial deamination of 3-hydroxykynurenine glucoside. This process is more pronounced in older lenses, possibly because of the barrier to diffusion. The barrier may also explain the increase in nuclear oxidized glutathione that is observed with age.
Authors: Ram H Nagaraj; Smitha Padmanabha; Maneesh Mailankot; Magdalena Staniszewska; Liew Jun Mun; Marcus A Glomb; Mikhail D Linetsky Journal: Biochim Biophys Acta Date: 2009-12-22
Authors: Olga A Snytnikova; Lyudmila V Kopylova; Elena I Chernyak; Sergey V Morozov; Nataliya G Kolosova; Yuri P Tsentalovich Journal: Mol Vis Date: 2009-12-16 Impact factor: 2.367