Emerging evidence indicates that inflammation in atopic dermatitis (AD) is associated with immune-mediated abnormalities in the skin. The history and severity of AD are risk factors for dust mite allergy. Bee venom (BV) is used in a complementary medicine to treat various diseases and skin disorders. Purified BV is obtained through electric stunning with a BV collector, without the harming of honeybees, followed by the removal of impurities from the collected BV and lyophilization of the final product. To evaluate the therapeutic potential of purified BV for AD, we investigated the anti-inflammatory effects of BV on house dust mite (Dermatophagoides farinae) antigen-stimulated HaCaT keratinocytes. The results showed that D. farinae induced significant increased levels of protease-activated receptor 2 (PAR2), intercellular adhesion molecule-1 (ICAM-1), and interleukin-6 (IL-6) compared to those in the normal control. However, purified BV inhibited the elevated expression of PAR2, ICAM-1 and IL-6 at the gene and protein levels. Thus, purified BV may have a therapeutic potential for the treatment and management of AD.