Uncovering the mechanism of arecoline’s effect on oral submucous fibrosis: integrating transcriptomics and in vitro and in vivo experiments

Objective: This study aimed to elucidate the potential targets and molecular mechanisms underlying arecoline-induced oral submucous fibrosis through integrated transcriptomic profiling and experimental validation. Methods: Transcriptomic sequencing was first employed to identify key pathways and targets influenced by arecoline in rat oral mucosa and whole blood. Subsequently, in vitro experiments using human primary oral mucosal fibroblasts (hOMFs) were conducted to validate the molecular mechanisms. Results: In vivo experiments demonstrated that chronic topical application of arecoline significantly reduced oral opening distance and induced histopathological features of oral submucous fibrosis (OSF), including epithelial atrophy, collagen deposition, and elevated TGF-β expression. Transcriptomic analysis revealed significant enrichment of pathways associated with fibrosis, including PPAR signaling, AMPK signaling, p53 signaling, and Hippo signaling pathways. In vitro validation further confirmed that arecoline dose-dependently upregulated α-SMA and Col1a1 expression, enhanced fibroblast proliferation, and activated Hippo pathway effectors (YAP/TAZ). Conclusion: These findings highlight the Hippo signaling pathway as a critical mediator of arecoline-induced OSF, providing novel insights for therapeutic targeting and mechanistic exploration in OSF management.