Background and Aims: In addition to its crucial role in dampening tissue-damaging immune responses in the gut, transforming growth factor b (TGFb) is a potent profibrogenic agent inducing collagen synthesis and regulating the balance between matrix-degrading matrix metalloproteinases (MMPs) and their inhibitors (TIMPs). TGFb 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 nonstrictured gut of patients with fibrostenosing CD. Isolated myofibroblasts were cultured with anti-TGFb blocking antibody or TGFb1. TGFb 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: TGFb transcripts, phosphorylated Smad2– Smad3 (pSmad2–3) and TIMP-1 proteins were higher in mucosa overlying strictures than in mucosa overlying nonstrictured 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 TGFb transcripts, a greater pSmad2–3 response to TGFb, increased TIMP-1, lower Smad7, increased collagen production and reduced migration ability compared with myofibroblasts from mucosa overlying non-strictured gut. TGFb 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-b signalling and MMP production were identified in the mucosa overlying strictures in CD which may give a window into the process of fibrosis.
Transforming growth factor beta signalling and matrix metalloproteinases in the mucosa overlying Crohn's disease strictures / Di Sabatino A; Jackson CL; Pickard KM; Buckley M; Rovedatti L; Leakey NA; Picariello L; Cazzola P; Monteleone G; Tonelli F; Corazza GR; Mac Donald TT; Pender SL. - In: GUT. - ISSN 0017-5749. - STAMPA. - 58:(2009), pp. 777-789.
Transforming growth factor beta signalling and matrix metalloproteinases in the mucosa overlying Crohn's disease strictures.
PICARIELLO, LUCIA;TONELLI, FRANCESCO;
2009
Abstract
Background and Aims: In addition to its crucial role in dampening tissue-damaging immune responses in the gut, transforming growth factor b (TGFb) is a potent profibrogenic agent inducing collagen synthesis and regulating the balance between matrix-degrading matrix metalloproteinases (MMPs) and their inhibitors (TIMPs). TGFb 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 nonstrictured gut of patients with fibrostenosing CD. Isolated myofibroblasts were cultured with anti-TGFb blocking antibody or TGFb1. TGFb 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: TGFb transcripts, phosphorylated Smad2– Smad3 (pSmad2–3) and TIMP-1 proteins were higher in mucosa overlying strictures than in mucosa overlying nonstrictured 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 TGFb transcripts, a greater pSmad2–3 response to TGFb, increased TIMP-1, lower Smad7, increased collagen production and reduced migration ability compared with myofibroblasts from mucosa overlying non-strictured gut. TGFb 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-b signalling and MMP production were identified in the mucosa overlying strictures in CD which may give a window into the process of fibrosis.File | Dimensione | Formato | |
---|---|---|---|
777.full.pdf
Accesso chiuso
Tipologia:
Altro
Licenza:
Tutti i diritti riservati
Dimensione
1.58 MB
Formato
Adobe PDF
|
1.58 MB | Adobe PDF | Richiedi una copia |
I documenti in FLORE sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.