Introduction: Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by epidermal barrier dysfunction. Ferroptosis, a form of programmed cell death driven by lipid peroxidation, has been implicated in various diseases. However, the role of ferroptosis in AD pathogenesis remains unclear. This study aimed to elucidate the mechanism of ferroptosis in AD keratinocytes. Methods: Eighteen C57BL/6J mice were randomized into control, AD, and AD+Ferrostatin-1 (Fer- 1) groups. An AD model was induced via house dust mite (HDM) sensitization. Fer-1 was intraperitoneally administered to the intervention group. Skin histopathology was evaluated using HE staining. Levels of Fe2+, reactive oxygen species (ROS), and inflammatory cytokines were measured. Transmission electron microscopy and Western Blot were employed to assess mitochondrial morphology and ferroptosis-related protein expression. Results: HDM exposure significantly elevated Fe2+, ROS, and malondialdehyde levels, while decreasing the Oxidized/ reduced glutathione (GSH/GSSG) ratio in keratinocytes. The AD group exhibited downregulated GPX4 and SLC7A11, alongside upregulated ACSL4 and PTGS2, indicating ferroptosis activation. Fer-1 treatment mitigated lipid peroxidation, reversed aberrant expression of ferroptosis markers, and alleviated AD-like skin lesions and inflammation. Conclusions: Ferroptosis occurs in AD keratinocytes. Fer-1 ameliorates AD symptoms by inhibiting ferroptosis and regulating inflammatory responses, suggesting ferroptosis as a potential therapeutic target for AD.