The Anti-Fibrotic Effects of IFN-γ on TGF-β1-Treated Endometrial Stromal Cells

Background: Endometrial fibrosis represents a distinctive pathological feature of diverse gynecological disorders, marked by the abnormal trans-differentiation of endometrial stromal cells (ESCs) and elevated deposition of extracellular matrix (ECM). Transforming growth factor-beta1 (TGF-β1) emerges as an essential driver of fibrogenesis. The present investigation sought to elucidate the regulatory pathways of Interferon-γ (IFN-γ) in driving fibrotic changes mediated by TGF-β1 within human ESCs populations.Methods: A TGF-β1-induced in vitro ESCs fibrosis model was established. ESCs were subjected to various concentrations and time durations of TGF-β1 stimulation. The fibroproteins α-smooth muscle actin (α-SMA), collagen type I alpha 1 (COL1A1), and Vimentin (VIM) were evaluated through a combination of quantitative reverse transcription-polymerase chain reaction (RT-qPCR), Western blotting, and immunofluorescent (IF) staining to assess cell fibrogenesis. IFN-γ pretreatment and intervention groups were designed to evaluate its inhibitory effects on fibrosis, with an emphasis on the TGF-β1/Smad signaling network.Results: TGF-β1 strongly induced ESCs to undergo fibrotic changes, leading to the elevation of COL1A1 and α-SMA expressions while simultaneously reducing Vimentin levels. This effect was demonstrated in both dose and timing. IFN-γ notably suppressed the production of fibrotic markers induced by TGF-β1, mitigated the reduction in Vimentin expression, and effectively dampened the phosphorylation of p-Smad3 levels.Conclusion: IFN-γ inhibits TGF-β1- mediated fibrosis in ESCs by targeting the TGF-β1/Smad signaling pathway, providing new insights into the mechanism of endometrial fibrosis and potential theoretical support for targeted therapy of related diseases.背景：子宫内膜纤维化是多种妇科疾病的特征性病理表现，以子宫内膜间质细胞（endometrial stromal cells, ESCs）异常转分化及细胞外基质（extracellular matrix, ECM）沉积增加为主要特征。转化生长因子-β1（transforming growth factor-beta1, TGF-β1）是纤维化发生的关键驱动因子。本研究旨在阐明干扰素-γ（interferon-γ, IFN-γ）在人ESCs群体中介导TGF-β1诱导的纤维化改变的调控通路。 方法：构建TGF-β1诱导的体外ESCs纤维化模型。将ESCs予以不同浓度、不同时长的TGF-β1刺激。通过定量逆转录聚合酶链反应（quantitative reverse transcription-polymerase chain reaction, RT-qPCR）、蛋白质印迹法（Western blotting）及免疫荧光染色（immunofluorescent staining, IF）联合检测成纤维标志物α-平滑肌肌动蛋白（α-smooth muscle actin, α-SMA）、Ⅰ型胶原α1链（collagen type I alpha 1, COL1A1）及波形蛋白（Vimentin, VIM）的表达水平，以评估细胞纤维化程度。设置IFN-γ预处理组与干预组，以评估其对纤维化的抑制作用，重点关注TGF-β1/Smad信号通路网络。 结果：TGF-β1可强力诱导ESCs发生纤维化改变，表现为COL1A1与α-SMA表达水平升高，同时降低Vimentin的表达水平，且该效应呈剂量与时间依赖性。IFN-γ可显著抑制TGF-β1诱导的成纤维标志物生成，缓解Vimentin表达的下调，并有效降低p-Smad3的磷酸化水平。 结论：IFN-γ通过靶向调控TGF-β1/Smad信号通路，抑制ESCs中TGF-β1介导的纤维化过程，为子宫内膜纤维化的发病机制研究提供了新视角，也为相关疾病的靶向治疗提供了潜在理论依据。