Interleukin-4 changes the transcriptome, ECM-associated components and function of mare endometrial fibroblast: Insights from healthy and fibrotic cells

Fibrosis remains incompletely understood, particularly in terms of how immune mediators shape stromal programs. We used a spontaneous large‑animal model—endometrosis (equine endometrial fibrosis) to define how interleukin‑4 (IL‑4) reprograms fibroblasts from healthy and fibrotic endometrium. Primary fibroblasts were exposed to IL‑4 (10 ng/mL) for 48 or 96 h. At 48 h, bulk transcriptomes revealed 1307 differentially expressed genes (DEGs; 648 up, 659 down) and 1271 DEGs (645 up, 626 down) in fibroblasts derived from endometria without or with endometrosis, respectively. Enrichment analyses implicated cellular metabolism, extracellular matrix (ECM) organization and remodeling, and signaling pathways commonly linked to fibrogenesis. IL‑4 also affected the long non‑coding RNA (lncRNA) expression, with 143 and 135 differentially expressed lncRNAs in fibroblasts derived from healthy or fibrotic endometria, respectively; linking these lncRNAs to DEGs involved in inflammation, ECM organization, and cytokine signaling. Moreover, IL‑4 increased proliferation and viability in fibroblasts derived from healthy or fibrotic endometria, while selectively reducing migration in fibroblasts derived from endometria without fibrosis after 96 h. IL‑4 further altered mRNA expression, protein abundance, and gelatinolytic activity of matrix metalloproteinases in a manner contingent on the fibrosis status of the tissue of origin, indicating stage‑dependent control of ECM turnover. Collectively, these data identify IL‑4 as a potent modulator of fibroblast function in a spontaneous large‑animal fibrosis model, revealing fibrosis stage‑dependent responses. Graphical Abstract Interleukin-4 (IL-4) modulates the transcriptome, extracellular matrix (ECM)–associated components, and functional characteristics of mare endometrial fibroblasts derived from healthy and fibrotic uteri. IL-4 treatment induces distinct transcriptomic and proteomic changes, alters cell proliferation, migration, collagen contractility, and MMP activity, and elicits differential responses between non-fibrotic and fibrotic cells, highlighting fibrosis-dependent effects of IL-4 on endometrial remodeling. Similar content being viewed by others Data availability The raw reads have been deposited in the Sequence Read Archive (SRA) under accession number PRJNA1154451.

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The authors would like to thank Agnieszka Bacławska, Witold Krzywiec, Ewa Liszewska, and Krzysztof Witek from the Institute of Animal Reproduction and Food Research, PAS, Olsztyn, for their technical support. Graphical abstract was created using images from Servier Medical Art (https://smart.servier.com), licensed under a Creative Commons Attribution 3.0 Unported License (CC BY 3.0). Funding This study was carried out as a part of the Sonata project (2019/35/D/NZ9/02989), financed by the National Science Centre, Poland. Author information Authors and Affiliations Contributions A.W.—writing original draft, study design, methodology, formal analysis, visualization, investigation, statistical analysis; A.S—investigation; K.M.—bioinformatic analysis of NGS results, visualization; T.M.—bioinformatic analysis of NGS results, visualization; M.M.K.—formal analysis; A.S.-M.—conceptualization, study design, funding acquisition, methodology, formal analysis, editing original draft. Corresponding author Ethics declarations Ethics approval This study used mare endometrial tissue obtained post-mortem from mares at a commercial slaughterhouse, where the animals were processed solely for meat production. As the samples were collected after death from non-experimental animals, no ethical approval was required. Competing interests The authors declare no competing interests. Additional information Publisher's Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Supplementary Information Below is the link to the electronic supplementary material. ESM 1 (download DOCX ) (5.25 MB DOCX) ESM 2 (download XLSX ) (765 KB XLSX) Rights and permissions Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. About this article Cite this article Wójtowicz , A., Sadowska, A., Myszczyński, K. et al. Interleukin-4 changes the transcriptome, ECM-associated components and function of mare endometrial fibroblast: Insights from healthy and fibrotic cells. Cell Tissue Res 403, 25 (2026). https://doi.org/10.1007/s00441-026-04049-6 Received: Accepted: Published: Version of record: DOI: https://doi.org/10.1007/s00441-026-04049-6