Anti-oxidative and anti-aging activities of 2-O-α-glucopyranosyl-L-ascorbic acid on human dermal fibroblasts

A stable ascorbic acid derivative, 2-O-α-glucopyranosyl-l-ascorbic acid (AA-2G), was evaluated and compared with ascorbic acid for its protective effect against cellular damage and senescence induced by hydrogen peroxide (H2O2). Pretreatment with AA-2G for 72 h promoted the proliferation of normal human dermal fibroblasts (NHDF) and protected against cell damage induced by H2O2. In contrast, ascorbic acid increased the proliferation and protected against cell damage, only when culture medium containing ascorbic acid was replaced every 24 h during the pretreatment period. These results suggest that the effect of AA-2G is longer-lasting compared to that of ascorbic acid. Senescence associated-β-galactosidase (SA-β-gal) activity, a classical biomarker of cellular senescence, was increased in H2O2-exposed NHDF cells, but pretreatment or posttreatment with ascorbic acid or AA-2G significantly inhibited the increase in SA-β-gal levels. AA-2G was more potent than ascorbic acid in down-regulating SA-β-gal activity. Expression of SIRT1, which has attracted attention as an anti-aging factor in recent years, was significantly decreased in H2O2-exposed NHDF cells compared to untreated cells. However, pretreatment NHDF cells with AA-2G before H2O2 exposure significantly inhibited this decrease in SIRT1 expression, whereas ascorbic acid had no effect. After H2O2 exposure, the expression levels of p53 and p21 were increased in NHDF cells and pretreatment with AA-2G inhibited this increase. Together, these results suggest that AA-2G protects dermal fibroblasts from oxidative stress and cellular senescence. These characteristics indicate that AA-2G could become a promising material for its anti-aging properties.