Atopic dermatitis (AD) is a chronic inflammatory skin disease that necessitates the development of sustainable therapeutic alternatives. Although Cannabis sativa L. is known for its valuable bioactive secondary metabolites, its potential as a functional ingredient for promoting skin health has not yet been comprehensively investigated. This study investigated the therapeutic efficacy of Cannabis sativa L. extract (CE) for managing atopic dermatitis (AD). In a DNCB-induced mouse model, dermatitis scores, mast cell infiltration, and serum IgE levels were significantly reduced by CE treatment. Additionally, the NLRP3 inflammasome and JAK1/STAT6 signaling pathways were significantly inhibited by CE treatment. In contrast, the expression of skin barrier markers, such as filaggrin and involucrin, was markedly increased by CE treatment. Collectively, these findings suggest that CE acts as a promising therapeutic candidate, offering a multi-target approach to alleviate AD and serving as a high-value functional ingredient for skin health management.