Page 78 - 南京医科大学自然版

南京医科大学学报（自然科学版） 第45卷第11期
·1608 · Journal of Nanjing Medical University（Natural Sciences） 2025年11月

·基础研究·

IFN⁃γ通过抑制SRC/Areg信号轴减轻克罗恩病肠纤维化

林俊杰，孙君健，王 璐，王 舒，张红杰，赵小静 *
南京医科大学第一附属医院消化内科，江苏 南京 210029

［摘 要］ 目的：系统探讨干扰素⁃γ（interferon⁃gamma，IFN⁃γ）在克罗恩病（Crohn’s disease，CD）肠纤维化中的调控作用及机
制。方法：提取分离CD患者肠成纤维细胞，体外予以不同的细胞因子［20 ng/mL干扰素⁃γ（interferon⁃gamma，IFN⁃γ）、50 ng/mL 白
介素（interleukin，IL）⁃17A、10 ng/mL IL⁃1β、100 ng/mL IL⁃33、200 ng/mL IL⁃36α、20 ng/mL 肿瘤坏死因子（tumor necrosis factor，
TNF）⁃α］处理 48 h，RT⁃PCR检测人肠成纤维细胞中双调蛋白（amphiregulin，Areg）基因表达水平。20 ng/mL IFN⁃γ处理人肠成纤
维细胞48 h后，进行转录组测序（RNA⁃seq），结合STRING数据库构建蛋白质互作（protein⁃protein interaction，PPI）网络。免疫荧光
检测 α平滑肌肌动蛋白（α⁃smooth muscle actin，α⁃SMA）、抗原Kiel 67（antigen Kiel 67，Ki67）等肌成纤维细胞标志物，评估IFN⁃γ
对人肠道成纤维细胞活化、增殖的影响。结果：Areg显著促进肠成纤维细胞活化、增殖及胶原合成；IFN⁃γ可显著抑制肠成纤维
细胞Areg表达，并下调α⁃SMA、COL1A1和COL6A1等纤维化相关基因，同时IFN⁃γ抑制成纤维细胞增殖与活化能力。RNA⁃seq分
析发现IFN⁃γ调控的差异基因显著富集于细胞外基质（extracellular matrix，ECM）重构通路，蛋白互作（protein⁃protein interaction，
PPI）网络鉴定出肉瘤原癌基因（sarcoma proto⁃oncogene，SRC）为核心节点，提示其可能介导IFN⁃γ的抗纤维化作用。结论：Areg
是促CD肠纤维化关键介质，IFN⁃γ通过转录调控抑制Areg表达，并发现SRC可能是IFN⁃γ下游的关键效应分子。IFN⁃γ可能通过
抑制SRC/Areg信号轴来发挥其抗纤维化作用。这些发现为开发靶向IFN⁃γ/SRC/Areg通路的抗纤维化策略提供了理论依据。
［关键词］ IFN⁃γ；双调蛋白；肠成纤维细胞；克罗恩病；肠纤维化
［中图分类号］ R574.62 ［文献标志码］ A ［文章编号］ 1007⁃4368（2025）11⁃1608⁃09
doi：10.7655/NYDXBNSN250631

IFN ⁃ γ alleviates intestinal fibrosis in Crohn’s disease through inhibiting the SRC/Areg
signaling axis

*
LIN Junjie，SUN Junjian，WANG Lu，WANG Shu，ZHANG Hongjie，ZHAO Xiaojing
Department of Gastroenterology，the First Affiliated Hospital of Nanjing Medical University，Nanjing 210029，China

［Abstract］ Objective：This study systematically investigates the regulatory role and mechanism of interferon⁃gamma（IFN⁃γ）in
Crohn’s disease（CD）⁃associated intestinal fibrosis. Methods：Intestinal fibroblasts were isolated from CD patients and treated with
various cytokines［IFN⁃γ，interleukin（IL）⁃17A，IL⁃1β，IL⁃33，IL⁃36α，tumor necrosis factor（TNF）⁃α］in vitro. The mRNA expression
level of amphiregulin（Areg）in human intestinal fibroblasts was detected using RT⁃PCR. Following treatment with 20 ng/mL IFN⁃γ for
48 h，transcriptome sequencing（RNA⁃seq）was performed，and a protein⁃protein interaction（PPI）network was constructed using the
STRING database. Immunofluorescence was employed to detect myofibroblast markers such as alpha⁃smooth muscle actin（α⁃SMA）
and antigen Kiel 67（Ki67），evaluating the effects of IFN⁃γ on the activation and proliferation of human intestinal fibroblasts. Results：
Areg significantly promoted the activation，proliferation，and collagen synthesis of intestinal fibroblasts，whereas IFN ⁃ γ markedly
suppressed Areg expression in intestinal fibroblasts and downregulated fibrosis ⁃ related genes including α ⁃ SMA，COL1A1，and
COL6A1，while also inhibiting fibroblast proliferation and activation. RNA⁃seq analysis revealed that differentially expressed genes
regulated by IFN⁃γ were significantly enriched in the extracellular matrix（ECM）remodeling pathway. PPI network analysis identified
SRC as a core node，suggesting its potential role in mediating the anti⁃fibrotic effects of IFN⁃γ. Conclusion：Areg is a key mediator
promoting intestinal fibrosis in CD. IFN⁃γ inhibits Areg expression through transcriptional regulation，and it is found that SRC may be
a key effector molecule downstream of IFN⁃γ. IFN⁃γ may exert its anti⁃fibrotic effect by inhibiting the SRC/Areg signaling axis. These
findings provide a theorectial basis for the development of anti⁃fibrotic strategies targeting the IFN⁃γ/SRC/Areg pathway.
［Key words］ IFN⁃γ；amphiregulin；intestinal fibroblast；Crohn’s disease；intestinal fibrosis
［J Nanjing Med Univ，2025，45（11）：1608⁃1615，1625］
［基金项目］ 国家自然科学基金（82200582，82370535）；中国博士后科学基金（2024M761212）
通信作者（Corresponding author），E⁃mail：zhaoxj91718@163.com（ORCID：0000⁃0001⁃5156⁃3864）
∗