Objective: To study the effects of tetrahydroxy stilbene-2-O-β-D-glucoside (TSG) on stress-induced premature senescence of human skin fibroblasts (HSF) exposed to ultraviolet radiation B (UVB) and its possible mechanism. Methods: HSFs were repeatedly exposed to UVB at a subcytotoxic level. TSG treatment (0.02, 0.10 and 0.50 mmol/L) was given immediately after each irradiation. The HSFs were divided into six groups:blank control group, model group, UVB+0.02 mmol/L TSG group, UVB+0.10 mmol/L TSG group, UVB+0.50 mmol/L TSG and TSG group (0.50 mmol/L). Cell counting kit-8 (CCK-8) was used to evaluate the proliferative activity of cells; senescence-associated-β-galactosidase (SA-β-gal) staining was performed to estimate the degree of premature senescence in cells; TBA method and WST-1 method were used to detect intracellular malondialdehyde (MDA) and superoxide dismutase (SOD) activities; and ELISA was applied to quantify the secretion level of matrix metalloproteinase1 (MMP-1) in cultured supernatant. Results: Compared with the blank control group, the proliferative activity and SOD level in the model group decreased (P < 0.05), while the percentage of SA-β-gal-positive cells, MDA and MMP-1 levels increased (P < 0.05 or P < 0.01). Compared with the model group, the proliferative activity and SOD level increased in UVB+TSG groups (all P < 0.05), and the percentage of SA-β-gal-positive cells, MDA and MMP-1 levels decreased (P < 0.05 or P < 0.01). Conclusion: TSG can inhibit UVB-induced premature senescence of HSF, which may be related to the inhibition of oxidative stress and high expression of MMP-1.