Simvastatin impairs smad-3 phosphorylation and modulates transforming growth factor beta1-mediated activation of intestinal fibroblasts

Br J Surg. 2009 May;96(5):541-51. doi: 10.1002/bjs.6577.

Abstract

Background: Transforming growth factor (TGF) beta1, acting through the smad pathway, is critical to fibroblast-mediated intestinal fibrosis. Simvastatin exhibits antifibrotic properties. This study assessed the effects of simvastatin on TGF-beta1-mediated intestinal fibroblast activation.

Methods: Human intestinal fibroblasts were activated with TGF-beta1 with or without simvastatin or the cholesterol pathway intermediates farnesyl pyrophosphate (FPP) and geranylgeranyl pyrophosphate (GGPP). Collagen-Ialpha2 expression was assessed by reverse transcriptase-polymerase chain reaction. Connective tissue growth factor (CTGF) and smad phosphorylation were evaluated by western blot, and plasminogen activator inhibitor (PAI) 1 activity by enzyme-linked immunosorbent assay. Fibroblast filamentous (F)-actin accumulation was assessed by confocal microscopy and contraction by a fibroblast-populated collagen lattice (FPCL) model.

Results: TGF-beta1 treatment of fibroblasts induced smad-2/3 phosphorylation, CTGF and collagen-Ialpha2 production, F-actin bundling, FPCL contraction and PAI-1 activation. Pretreatment with simvastatin inhibited the induction of CTGF and collagen-Ialpha2, PAI-1 activation, F-actin bundling and FPCL contraction. The inhibitory effect of simvastatin on PAI-1 activation was reversed by GGPP and FPP. Simvastatin pretreatment inhibited TGF-beta1-mediated phosphorylation of smad-3.

Conclusion: Simvastatin abrogates TGF-beta1-mediated intestinal fibroblast activation by inhibition of smad-3 phosphorylation. These findings offer a mechanism for the antifibrotic effects of simvastatin and a therapeutic entry point in the treatment of intestinal fibrosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism
  • Blotting, Western
  • Cell Communication
  • Cells, Cultured
  • Colon / cytology*
  • Connective Tissue Growth Factor / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Fibroblasts / drug effects*
  • Fibroblasts / physiology
  • Humans
  • Hydroxymethylglutaryl-CoA Reductase Inhibitors / pharmacology*
  • Microscopy, Confocal
  • Phosphorylation / drug effects
  • Plasminogen Activator Inhibitor 1 / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Simvastatin / pharmacology*
  • Smad3 Protein / metabolism*
  • Transforming Growth Factor beta1 / drug effects*
  • Transforming Growth Factor beta1 / physiology

Substances

  • Actins
  • Hydroxymethylglutaryl-CoA Reductase Inhibitors
  • Plasminogen Activator Inhibitor 1
  • Smad3 Protein
  • Transforming Growth Factor beta1
  • Connective Tissue Growth Factor
  • Simvastatin