T16 modulated extracellular matrix remodeling in fibroblasts via paracrine activation of TGF-β1 through M2 macrophage polarization

Mol Biol Rep. 2025 Jun 26;52(1):642. doi: 10.1007/s11033-025-10733-7.

Abstract

Background: T16 has demonstrated significant potential for regulating extracellular matrix (ECM) remodeling by modulating immune and fibroblast functions. This study investigated how T16 influences fibroblast-mediated extracellular matrix (ECM) remodeling through macrophage polarization.

Methods: Uterosacral ligaments were collected from 15 patients with pelvic organ prolapse (POP) and 6 patients without pelvic organ prolapse (POP). Fibroblasts isolated from these tissues were cultured and co-cultured with THP-1-derived macrophages in the following experimental groups: control, T16 + fibroblasts, M1 macrophages + fibroblasts, and T16 + M1 macrophages + fibroblasts. Cell proliferation was assessed using the CCK-8 assay, while protein and mRNA expression levels were analyzed by western blotting and qRT-PCR, respectively.

Results: Our findings revealed that T16 promoted macrophage polarization toward the anti-inflammatory M2 phenotype, which, in turn, exerted paracrine effects on fibroblasts. Specifically, M2 macrophages induced by T16 significantly enhance the activation of transforming growth factor beta-1 (TGF-β1), a key regulator of extracellular matrix (ECM) synthesis and remodeling. This paracrine signaling pathway facilitates fibroblast-mediated ECM stabilization, characterized by increased collagen synthesis and extracellular matrix turnover.

Conclusion: These results highlight the therapeutic potential of T16 in conditions involving extracellular matrix (ECM) dysfunction, such as pelvic organ prolapse (POP), by targeting the crosstalk between immune cells and fibroblasts. Further research is required to explore its applications in regenerative medicine and tissue engineering.

Keywords: Extracellular matrix (ECM) remodeling; Fibroblasts; Immune modulation; M2 macrophages; Macrophage polarization; Pelvic organ prolapse (POP); T16; TGF-β1; Uterosacral ligaments.

MeSH terms

  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Coculture Techniques
  • Extracellular Matrix* / drug effects
  • Extracellular Matrix* / metabolism
  • Female
  • Fibroblasts* / drug effects
  • Fibroblasts* / metabolism
  • Humans
  • Ligaments / metabolism
  • Macrophage Activation / drug effects
  • Macrophages* / drug effects
  • Macrophages* / metabolism
  • Middle Aged
  • Paracrine Communication / drug effects
  • Pelvic Organ Prolapse / metabolism
  • Pelvic Organ Prolapse / pathology
  • THP-1 Cells
  • Transforming Growth Factor beta1* / metabolism

Substances

  • Transforming Growth Factor beta1
  • TGFB1 protein, human