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Transforming growth factor β signalling and matrix metalloproteinases in the mucosa overlying Crohn’s disease strictures
  1. A Di Sabatino1,
  2. C L Jackson2,
  3. K M Pickard2,
  4. M Buckley2,
  5. L Rovedatti1,
  6. N A B Leakey3,
  7. L Picariello4,
  8. P Cazzola1,
  9. G Monteleone5,
  10. F Tonelli4,
  11. G R Corazza1,
  12. T T MacDonald3,
  13. S L Pender2
  1. 1
    First Department of Medicine, Fondazione IRCCS Policlinico S. Matteo, Centro per lo Studio e la Cura delle Malattie Infiammatorie Croniche Intestinali, University of Pavia, Pavia, Italy
  2. 2
    Division of Infection, Inflammation and Repair, University of Southampton, School of Medicine, Southampton, UK
  3. 3
    Centre for Infectious Disease, Institute of Cell and Molecular Science, Barts and the London School of Medicine and Dentistry, London, UK
  4. 4
    Department of Clinical Physiopathology, University of Florence, Florence, Italy
  5. 5
    Dipartimento di Medicina Interna e Centro di Eccellenza per lo Studio delle Malattie Complesse e Multifattoriali, Università Tor Vergata, Rome, Italy
  1. Professor T T MacDonald, Centre for Infectious Disease, Institute of Cell and Molecular Science, Barts and the London School of Medicine and Dentistry, Whitechapel, London E1 2AT, UK; t.t.macdonald{at}qmul.ac.uk

Abstract

Background and Aims: In addition to its crucial role in dampening tissue-damaging immune responses in the gut, transforming growth factor β (TGFβ) is a potent profibrogenic agent inducing collagen synthesis and regulating the balance between matrix-degrading matrix metalloproteinases (MMPs) and their inhibitors (TIMPs). TGFβ signalling was investigated by analysis of Smad proteins and MMPs/TIMPs in the mucosa overlying strictures in patients with Crohn’s disease (CD).

Methods: Specimens were collected from macroscopically normal mucosa overlying strictured and non-strictured gut of patients with fibrostenosing CD. Isolated myofibroblasts were cultured with anti-TGFβ blocking antibody or TGFβ1. TGFβ transcripts were analysed by quantitative reverse transcription-PCR (RT-PCR). Smad proteins and MMPs were determined by immunoblotting. MMP-12 activity was measured by a real-time MMP-12 activity assay. An in vitro wound-healing scratch assay was used to assess myofibroblast migration.

Results: TGFβ transcripts, phosphorylated Smad2–Smad3 (pSmad2–3) and TIMP-1 proteins were higher in mucosa overlying strictures than in mucosa overlying non-strictured areas. In contrast, mucosa overlying strictured gut had lower expression of Smad7, MMP-12 and MMP-3. Myofibroblasts from mucosa overlying strictured gut showed higher TGFβ transcripts, a greater pSmad2–3 response to TGFβ, increased TIMP-1, lower Smad7, increased collagen production and reduced migration ability compared with myofibroblasts from mucosa overlying non-strictured gut. TGFβ blockade increased myofibroblast MMP-12 production and migration, more obviously in myofibroblasts isolated from mucosa overlying non-strictured compared with strictured gut.

Conclusions: Changes in TGF-β signalling and MMP production were identified in the mucosa overlying strictures in CD which may give a window into the process of fibrosis.

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Footnotes

  • Competing interests: None.

  • Ethics approval: Appropriate local Ethics Committee approval was obtained for this study

  • Patient consent: Informed consent was obtained in all cases.

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