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Relative suppression of the sodium-dependent Vitamin C transport in mouse versus human lens epithelial cells
Authors:Mark E. Obrenovich  Xingjun Fan  Makoto Satake  Simon M. Jarvis  Lixing Reneker  John R. Reddan  Vincent M. Monnier
Affiliation:(1) Department of Pathology, Case Western Reserve University, Cleveland, OH, USA;(2) Department of Biochemistry, Case Western Reserve University, Cleveland, OH, USA;(3) Research School of Biosciences, University of Westminster, London, UK;(4) Department of Ophthalmology, University of Missouri, Columbia, MO, USA;(5) Department of Biological Sciences, Oakland University, Rochester, MI, 48039, USA;(6) Institute of Pathology, Case Western Reserve University, 2085 Adelbert Road, Cleveland, OH 44106, USA;(7) Present address: Tissue Engineering Laboratory, Innovation Research Institute, Teijin Ltd., Tokyo, Japan
Abstract:Vitamin C is a major antioxidant and UV absorbent in the human lens. In the rodent lens, the levels are very low for unknown reasons. Searching for clues to explain this suppression, we investigated the comparative uptake of Vitamin C in cultured human and mouse lens epithelial cells. When compared to human HLE-B3 lens epithelial cells, 14C-ASA uptake was 4- to 10-fold impaired in confluent mouse lens 17EM15 (p < 0.0001) and 21EM15 (p < 0.001) cells, respectively. High glucose concentrations reduced the uptake by 30–50% in all cells (p < 0.005). Incubation of cells with 6-deoxy-6-fluoro-ascorbic (F-ASA), i.e. a probe specific for the sodium-dependent Vitamin C uptake (SVCT2), revealed a 10-fold uptake suppression into mouse 17EM15 relative to human HLE-B3 and JAR choriocarcinoma cells (a control), that could be overcome by overexpressing hSVCT2 using two different promoter constructs. The relative Vitamin C uptake differences suggest either low expression of SVCT2, molecular differences between the transporters themselves or their biological regulation, since a recent study has shown that exogenous feeding of ascorbic acid to rats increased only modestly lenticular uptake (Mody et al., Acta Ophthalmol Scand 83: 228–223, 2005). Elucidation of the mechanism by which SCVT2 activity is suppressed in mouse lens may help unravel a major question of evolutionary significance for night vision in nocturnal animals.
Keywords:aging  ascorbic acid  diabetes  glycation  oxidation  SVCT2
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