Itch is the most bothersome symptom of atopic dermatitis and is often exacerbated by stress. Allopregnanolone (ALLO), one of the neurosteroids in the brain, is shown to increase rapidly following acute stress, serving as a homeostatic response to stress. We have previously demonstrated that an intracisternal injection of ALLO markedly increases itch-related scratching behavior in atopic dermatitis model mice. The present study examines whether acute stress exacerbates itching in those mice by comparing scratching behavior under three different conditions: 1) conventional conditions (5 mice in the housing cage), 2) confinement stress (under isolated conditions in a small invisible chamber), and 3) confinement stress + forced swim stress (FSS) (under isolated conditions in the same chamber immediately following FSS). Scratching was significantly increased under confinement stress conditions compared to conventional conditions. FSS further exacerbated scratching behavior. Our results show that acute stress exacerbates itch-related scratching in atopic dermatitis mice. This exacerbation of scratching may involve ALLO transiently increased in the brain as a response to stress.