Abstract: Objective To investigate the role of ubiquitin-specific protease 25 (USP25) in the development of allergic rhinitis (AR) induced by epithelial-derived thymic stromal lymphopoietin (TSLP) and potential mechanism. Methods Nasal mucosal tissues were collected from 40 patients undergoing nasal septum or turbinate surgery. The patients were divided into two groups (n=20): a control group and an AR group. The expression of USP25 in nasal mucosa was detected by quantitative reverse transcription PCR and Western blot. The localization in AR nasal mucosa was assessed by immunohistochemistry. Furthermore, 60 C57BL/6J mice were randomly assigned to four groups: control, AR, AR + negative control for overexpression (oe-NC), and AR + USP25 overexpression (oe-USP25). Except for those in the control group, the remaining mice were sensitized with ovalbumin (OVA) to establish an AR model. The AR+oe-NC and AR+oe-USP25 groups received either a control plasmid (pcDNA) or a USP25 overexpression plasmid (pcDNA-USP25) complexed with liposomes. After treatment, AR symptoms were evaluated. Histopathological changes in nasal mucosa were observed using hematoxylin and eosin (H&E) staining, and Th1/Th2 cytokine levels in nasal lavage fluid were measured by ELISA. Then, the human nasal epithelial RPMI2650 cells were divided into six groups: control, house dust mite (HDM), HDM + pcDNA, HDM + pcDNA-USP25, HDM + pcDNA-USP25 + siRNA negative control (siRNA-NC), and HDM + pcDNA-USP25 + siRNA targeting transforming growth factor β-activated kinase 1 (TAK1). Corresponding siRNA and pcDNA transfections were performed. The expression of USP25, TSLP, and TAK1 in mouse nasal mucosa and RPMI2650 cells was detected by Western blot. Results USP25 expression was elevated in the nasal mucosa of AR patients and mainly localized in the epithelial layer. USP25 overexpression alleviated AR symptoms in mice, reduced nasal mucosal damage, restored Th1/Th2 balance, downregulated TAK1 expression, and upregulated TSLP expression. In vitro experiments indicated that after USP25 overexpression, knockdown of TAK1 reversed the USP25-induced suppression of TSLP expression in HDM-stimulated cells. Conclusions USP25 may alleviate Th2-type inflammation in AR by inducing TRAF3 expression and inhibiting TSLP signaling in nasal epithelial cells.