BACKGROUND/OBJECTIVE: Ferroptosis is an iron-dependent form of programmed cell death characterized by lipid peroxidation products (LPOs). A chemotherapeutic drug, 5-fluorouracil (5-FU), can induce epithelial mucositis and favor drug synergism with erastin in ferroptosis. METHODS: HaCaT cells were induced by 5-FU and erastin, treated with different TS doses, and their viability was then determined. Levels of cellular reactive oxygen species (ROS), LPOs, labile iron pool (LIP), glutathione (GSH), glutathione peroxidase 4 (GPX-4) activity, as well as IL-6, IL-1β, and TNF-α levels, and their wound healing properties were assessed. RESULTS: TS per se (at up to 25 µg/mL) was not toxic to HaCaT cells but was unable to restore the viability of 5-FU-induced cells up to the baseline levels. The compound significantly diminished increases in cellular ROS, LPOs, and LIP, while restoring GSH content and GPX-4 activity. Additionally, it suppressed the cytokine production of 5-FU-induced cells in a concentration-dependent manner. Moreover, TS exerted wound-healing effects against skin injuries and 5-FU damage significantly and dose dependently. CONCLUSIONS: The insights of this work have identified biochemical mechanisms using tea saponin extract to protect against 5-FU-induced keratinocyte ferroptosis and inflammation. This study highlights the promising adjunctive potential of tea saponin in the mitigation and management of chemotherapy-induced mucositis.