Ascorbic acid 2-O-alpha-glucoside, a stable form of ascorbic acid, rescues human keratinocyte cell line, SCC, from cytotoxicity of ultraviolet light B.

Abstract

Environmental exposure to ultraviolet light B (UVB, wave lengths 290-320 nm) of the solar spectrum causes major damage, including an inflammatory response, in skin. In the present study, we estimated the ability of a stable derivative of ascorbic acid, ascorbic acid 2-O-alpha-glucoside (AA-2G), to reduce UVB damage, using the human keratinocyte cell line, SCC, established from squamous cell carcinoma. By pre- (9 h) and post-cultivation with AA-2G, a significant preventive effect on the decrease in the absolute number of surviving cells by exposure to UVB (typical dose, 20 mJ/cm2) was measurable by a neutral red-uptake assay. The release of lactate dehydrogenase from the cell membrane damaged by UVB was inhibited by AA-2G. In agarose gel electrophoresis, relatively high molecular weight DNA fragments were detected in irradiated cells after 6 h post-irradiation, suggesting that the mechanism of cell death was necrosis. Quantitative analysis of DNA content by flow cytometry indicated that AA-2G suppressed both an increase in debris with degraded nuclei and a decrease in cells in G1 and S phases, but not in the G2/M phase, by UVB exposure. These data suggest that AA-2G shows a photoprotective effect against UVB-induced damage in human epithelial cells.