Comparative analysis of short-term and long-term LL-37-induced rosacea-like mouse models:Histopathological features and inflammatory immune responses

Yiling Wu; Chuanxi Zhang; Hui Jin; Ruiping Zheng; Tian Li; Fuyu Jin; Yaqian Li; Xuemin Gao; Hong Xu; Zhongqiu Wei; Jie Yang

doi:10.1002/ame2.70020

Abstract：Background: It is well recognized that developing new animal models, refining the existing mouse models, and thoroughly characterizing their features are essential for gaining a deeper understanding of rosacea pathogenesis and for advancing therapeutic strategies in this direction. Accordingly, we aimed to characterize the pathological features of a long-term LL-37-induced mouse model of rosacea and to compare the disease manifestations and pathophysiological characteristics between short-term and long-term LL-37-induced models. A key focus was to investigate differential gene expression and the underlying mechanisms of immune system dysregulation in these models. Methods: We comparatively assessed skin lesion manifestations, the extent of inflammatory infiltration, sebaceous gland alterations, fibrosis, and angiogenesis in both models. Assessments were performed using photographic documentation, hematoxylin-eosin (HE) staining, Van Gieson's (VG) staining, immunohistochemistry, and Western blotting. Furthermore, we employed RNA sequencing to analyze differential gene expression in mouse skin. The RNA sequencing data were validated using immunofluorescence staining and Western blotting, with a specific focus on gene variations and mechanisms related to immune system dysregulation. Results: Mice subjected to long-term LL-37 induction developed rosacea-like pathological features, including angiogenesis, thickened skin tissue, and sebaceous gland hypertrophy. In the short-term LL-37-induced model, immune dysregulation primarily involved the innate immune response. However, long-term LL-37 induction resulted in significant activation of both innate and adaptive immune responses. Conclusion: The long-term LL-37-induced mouse model offers a valuable animal model for the detailed investigation of the pathological mechanisms driving moderate-to-severe rosacea with prolonged disease duration. Importantly, this model provides a significant experimental foundation for exploring the potential role of immune system dysregulation in rosacea pathogenesis.