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Excessive ultraviolet (UV) radiation exposure results in skin chronic photodamage via stimulating reactive oxygen species (ROS) and inflammation. Anthocyanins are a group of flavonoids frequently found in edible plants. Cyanidin-3-O-glucoside (C3G) as a typical anthocyanin shows effective anti-oxidative and anti-inflammatory properties. This study aims to investigate whether the topical application of C3G moisturizing gel on mice can protect the skin from UVB-induced chronic photodamage. The results of in vitro experiment showed that the active ingredient C3G can penetrate the mice skin. The dorsal of Kunming mice were treated with C3G moisturizing gel (100, 200, 300 μmol/L) after UVB exposure. The animal experiment demonstrated that C3G can reduce chronic photodamage caused by UVB. C3G could effectively ameliorate the UVB-induced epidermal barrier dysfunction including an increase in the skin hydration and decrease in the transepidermal water loss, and have statistically significance. Besides, our results also indicated that C3G inhibited UVB-induced epidermal hyperplasia, the destruction of collagen fibers, ROS levels, and the expression of COX-2 and IL-6. In brief, these results indicate that C3G can reduce UVB-induced chronic photodamage by inhibiting oxidative stress and inflammation.