Uncovering the mechanism of arecoline's effect on oral submucous fibrosis: integrating transcriptomics and <i>in vitro</i> and <i>in vivo</i> experiments

<h4>Objective</h4>This study aimed to elucidate the potential targets and molecular mechanisms underlying arecoline-induced oral submucous fibrosis through integrated transcriptomic profiling and experimental validation.<h4>Methods</h4>Transcriptomic sequencing was first employed to identify key pathways and targets influenced by arecoline in rat oral mucosa and whole blood. Subsequently, <i>in vitro</i> experiments using human primary oral mucosal fibroblasts (hOMFs) were conducted to validate the molecular mechanisms.<h4>Results</h4><i>In vivo</i> 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. <i>In vitro</i> validation further confirmed that arecoline dose-dependently upregulated α-SMA and Col1a1 expression, enhanced fibroblast proliferation, and activated Hippo pathway effectors (YAP/TAZ).<h4>Conclusion</h4>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.