Purpose: Prunus persica Flos (PPF, peach flowers) extracts were reported to have anti-oxidant and anti-inflammatory activities. In this study, we investigated whether PPF extracts prevent the human skin cells from oxidative stress and matrix metalloproteinases (MMPs) induced by UVB irradiation which ultimately leads to photoaging, or not. Methods: Ethanol extract (Et) of PPF and its dichloromethane (DM), ethylacetate (EA), and n-butanol (Bt) fractions were pretreated in UVB-irradiated (75 mJ/cm 2) human keratinocytes (HaCaT) or human dermal fibroblasts (HDF). Cellular reactive oxygen species (ROS) generation was measured using DCF-DA assay, and mRNA expression and production of MMPs were measured by real-time PCR and Western blotting, respectively. Results: The treatment of Et, DM, EA, and Bt significantly inhibited UVB-induced ROS generation and MMP-1 production in HaCaT cells. The treatments of EA and Bt significantly decreased not only the mRNA expression of MMP-1, MMP-3, and MMP-9 but also the protein production of MMP-1 and MMP-3 in HDF cells, while they increased the expression of type-1 procollagen in mRNA and protein levels. Et and DM significantly inhibited only MMP-1 and MMP-3 production without an preventive effect on type-1 procollagen production in UVB-irradiated HDF cells. Conclusion: The treatments with EA and Bt fractions from PPF extract protected human skin cells from UVB-induced oxidative stress and collagen loss. These findings suggest that EA and Bt fractions from PPF extract have the potential of natural resource for a skin anti-photoaging product in the food and cosmetic industry.