Objective: Intestinal fibrosis in Crohn's disease (CD) remains a clinical challenge with incompletely understood molecular mechanisms. This study systematically investigated the regulatory role and mechanism of IFN-γ in CD-associated intestinal fibrosis. Methods: Primary intestinal fibroblasts isolated from CD patients were treated with various cytokines in vitro. Amphiregulin (Areg) gene expression was assessed by RT-PCR. Based on preliminary results, fibroblasts were treated with 20 ng/mL IFN-γ for 48 h for RNA sequencing (RNA-seq). Protein-protein interaction (PPI) networks were constructed using the STRING database. Myofibroblast markers (α-SMA, Ki67) were detected by immunofluorescence to evaluate IFN-γ's effects on fibroblast activation and proliferation. Results: Key findings include: 1) Areg significantly promoted fibroblast activation, proliferation, and collagen synthesis; 2) IFN-γ markedly suppressed Areg expression and downregulated fibrosis-related genes (α-SMA, Col1a1, Col6a1); 3) IFN-γ reduced fibroblast proliferation (decreased Ki67+ cells) and activation (reduced α-SMA expression); 4) Transcriptomic analysis revealed significant enrichment of IFN-γ-regulated genes in ECM remodeling pathways, with PPI network identifying Src as a central node potentially mediating IFN-γ's anti-fibrotic effects. Conclusion: This study is the first to demonstrate that IFN-γ exerts anti-fibrotic effects by inhibiting the SRC/AREG signaling axis. We confirmed Areg as a key pro-fibrotic mediator, revealed IFN-γ's transcriptional suppression of Areg, and identified Src as a critical downstream effector of IFN-γ. These findings provide a theoretical foundation for developing novel anti-fibrotic strategies targeting the IFN-γ/SRC/AREG pathway.