Collagen synthesis in human skin fibroblasts is stimulated by a stable form of ascorbate, 2-O-alpha-D-glucopyranosyl-L-ascorbic acid.
We evaluated the effect of 2-O-alpha-D-glucopyranosyl-L-ascorbic acid (AA-2G) on collagen synthesis in cultured human skin fibroblasts and on proliferation of fibroblasts. At concentrations of 0.1-0.5 mmol/L, AA-2G effectively stimulated collagen synthesis with an effectiveness comparable to that of L-ascorbic acid. On the other hand, 6-O-alpha-D-glucopyranosyl-L-ascorbic acid showed a weak effect. The stimulation of collagen synthesis by AA-2G was attenuated by the addition of a collagen synthesis inhibitor, L-azetidine 2-carboxylic acid, in a dose-dependent manner. In addition, AA-2G-induced stimulation of collagen synthesis could be completely inhibited by the addition of castanospermine, an inhibitor of neutral alpha-glucosidase. Relatively high alpha-glucosidase activity, which would contribute to release of ascorbic acid from AA-2G, could be detected in the lysate of cultured fibroblasts. The stimulatory activity of AA-2G on collagen synthesis was observed after 5 d in culture, whereas L-ascorbic acid tended to lose its stimulatory activity. Continuous supplementation of AA-2G (0.25 mmol/L) to culture medium for 24 d enhanced the cell growth four times that of the control. These results indicate that AA-2G is gradually cleaved by the cellular alpha-glucosidase to release L-ascorbic acid, which adequately stimulates collagen synthesis and proliferation of human skin fibroblasts.
Yamamoto
I
I
Department of Immunochemistry, Faculty of Pharmaceutical Sciences, Okayama University, Japan.
Muto
N
N
Murakami
K
K
Akiyama
J
J
D000469:Alkaline Phosphatase / Q000378:metabolism; D001205:Ascorbic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D002478:Cells, Cultured; D002675:Child, Preschool; D003094:Collagen / Q000096:biosynthesis*; D004247:DNA / Q000302:isolation & purification; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D005959:Glucosidases / Q000378:metabolism; D065089:Glycoside Hydrolase Inhibitors; D006801:Humans; D007212:Indolizines / Q000494:pharmacology; D012867:Skin / Q000187:drug effects* / Q000378:metabolism; D054836:Indolizidines
D000469:Alkaline Phosphatase; D001205:Ascorbic Acid; D003094:Collagen; D004247:DNA; D005959:Glucosidases; D065089:Glycoside Hydrolase Inhibitors; D007212:Indolizines; C065378:ascorbic acid 2-O-glucoside; C037806:castanospermine; D054836:Indolizidines