In this study, we investigated the protective effects of a peptide (YGDEY, Tyr‐Gly‐Asp‐Glu‐Tyr) isolated from tilapia skin gelatin hydrolysates (TGHs), against UVB‐induced photoaging in human keratinocytes (HaCaT) cells. Results showed that YGDEY significantly decreased levels of intracellular reactive oxygen species (ROS), increased antioxidant factors (Superoxide Dismutase, SOD and Glutathione, GSH) expression and maintained balance between GSH and GSSG in HaCaT cells. Comet assay shows that YGDEY can protect DNA from oxidative damage. Furthermore, it significantly inhibited MMP‐1 (collagenase) and MMP‐9 (gelatinase) expression and increased Type I procollagen production. In addition, the molecular docking study showed that YGDEY may form active sites with MMP‐1 and MMP‐9. Moreover, Western blot analysis was utilized to measure the protein levels of UVB‐induced mitogen‐activated protein kinase (MAPK) and nuclear factor‐kappa B (NF‐κB) signaling pathways. Therefore, these results suggested that YGDEY has a therapeutic effectiveness in prevention of UVB‐induced cellular damage, and it is a candidate worthy of being developed as a potential natural antioxidant and food additive.