Atopic dermatitis (AD) is a chronic, relapsing inflammatory skin disease associated with allergy and it is commonly characterized with dry skin, severe pruritus, high serum IgE levels and eosinophilia. Keratinocytes produce many cytochemokines which are involved in the pathogenesis of skin disorders. In particular, macrophage-derived chemokine (MDC/CCL22) & thymus and activation-regulated chemokine (TARC/CCL17) are Th2-type cytokines, and it has been reported that serum MDC and TARC levels are associated with AD disease activity. In present study, we investigated the anti-inflammatory activities of Prunus yedoensis Matsum barks in vitro and in vivo. In vitro, we evaluated of their inhibitory effect on the atopic dermatitis (AD)―like inflammatory markers (MDC and TARC) and Jak-STAT pathway (Jak1 and STAT1) in HaCaT human keratinocytes. In vivo, we evaluated the inhibitory effect of EtOAc fraction on the skin symptoms in AD-like murine model induced by DNCB or special diets. Among different fractions obtained from crude extract (80% EtOH), EtOAc fraction and E5 sub-fraction showed potent inhibitory activity on the mRNA and protein levels of MDC and TARC at the concentration of 50 ㎍/㎖ in HaCaT cells. In addition, E5, EtOAc sub-fraction, inhibited the activity of the STAT1 proteins associated with MDC and TARC in a dose-dependent manner. Also, the EtOAc fraction alleviated AD-like skin symptoms (skin surface hydration, epidermis thickness) in AD-like murine model induced by DNCB or special diets. These results suggest that P. yedoensis may have an anti-atopic activity by suppressing the inflammatory chemokines (MDC and TARC) and skin barrier dysfunction in vitro and in vivo.