Based on the preparation of strawberry leaf water extract (SLWE), the composition, antioxidative activities and non-enzymatic glycosylation (NEG) inhibition rate of SLWE was explored. In addition, the effects of SLWE on the survival rate, hyaluronic acid (HA), collagen I (COL I) and hydroxyproline (HYP), matrix metalloproteinase 1 (MMP 1) and matrix metalloproteinase 9 (MMP 9) of normal growing human skin fibroblasts (HFF-1) were evaluated, in order to estimate the effect in delaying skin aging of SLWE. Through building UVB causes HFF-1 damage protection model, the effects of SLWE on reactive oxygen species (ROS), malondialdehyde (MDA), MMP-1 and superoxide dismutase (SOD) in the protection group were measured to explore the protective mechanism of SLWE on UVB-induced HFF-1 damage. The results showed that SLWE was rich in Vc, quercetin, chlorogenic acid, gallic acid and catechin, among which quercetin and epicatechin were 12.86 mg/L and 10.13 mg/L respectively. SLWE had good free radical scavenging ability, which was in order, O2-? > DPPH > ABTS > ferrous chelation > ?OH. The NEG inhibition rate of 1% (volume fraction) SLWE was 87.17%. More than 3% (volume fraction) SLWE could improve contents of HA, COL I and HYP (p < 0.05) in HFF-1, and 6% (volume fraction) SLWE could decrease the content of MMP 1 (p < 0.01), suggesting that SLWE was helpful to anti-aging through lifting the skin moisture, increasing the collagen content and inhibiting collagen degradation. In UVB causes HFF-1 damage protection model, SLWE could effectively ROS generated by UVB stimulation, 6% (volume fraction) SLWE could significantly reduce MDA, inhibit MMP-1 expression, and improve the activity of SOD, indicating that SLWE could help alleviate cell damage caused by oxidative stress.