Ultraviolet B (UVB) irradiation-induced DNA damage, oxidative stress, inflammatory processes, and skin pigmentation cause pigmented spots, wrinkles, inflammation, and accelerated skin aging and cancer. Maqui berry (Aristotelia chilensis) is a natural antioxidant, anticancer, and anti-inflammatory food. We investigated the photoprotective properties of the ethyl acetate fraction of maqui berry ethanol extract (MEE) in vitro and in vivo. Spectrophotometric measurements revealed dominant extinction profile of MEE in the UVB range. MEE clearly reversed the DNA damage induced by UVB irradiation in HaCaT cells by upregulating endogenous cellular enzymatic and non-enzymatic antioxidant systems containing superoxide dismutase, catalase, and glutathione and reducing the production of nitric oxide. Moreover, MEE treatment enhanced the antioxidant ability and weakened lipid peroxidation in BALB/c mice exposed to UVB radiation. It also down-regulated interleukin (IL)-6 and tumor necrosis factor-α levels and up-regulated IL-4 levels. Moreover, MEE inhibited the UVB-triggered activation of ERK and p38 MAPK. These data suggest that MEE is an effective agent against UVB-induced photodamage.