Anti-fibrotic action of pirfenidone in Dupuytren's disease-derived fibroblasts

BMC Musculoskelet Disord. 2016 Nov 11;17(1):469. doi: 10.1186/s12891-016-1326-y.

Abstract

Background: Dupuytren's disease (DD) is a complex fibro-proliferative disorder of the hand that is often progressive and eventually can cause contractures of the affected fingers. Transforming growth factor beta (TGF-β1) has been implicated as a key stimulator of myofibroblast activity and fascial contraction in DD. Pirfenidone (PFD) is an active small molecule shown to inhibit TGF-β1-mediated action in other fibrotic disorders. This study investigates the efficacy of PFD in vitro in inhibiting TGF-β1-mediated cellular functions leading to Dupuytren's fibrosis.

Methods: Fibroblasts harvested from (DD) and carpal tunnel (CT)- tissues were treated with or without TGF-β1 and/or PFD and were subjected to cell migration, cell proliferation and cell contraction assays. ELISA; western blots and real time RT-PCR assays were performed to determine the levels of fibronectin; p-Smad2/Smad3; alpha-smooth muscle actin (α-SMA), α2 chain of type I collagen and α1 chain of type III collagen respectively.

Results: Our results show that PFD effectively inhibits TGF-β1-induced cell migration, proliferation and cell contractile properties of both CT- and DD-derived fibroblasts. TGF-β1-induced α-SMA mRNA and protein levels were inhibited at the higher concentration of PFD (800 μg/ml). Interestingly, TGF-β1 induction of type I and type III collagens and fibronectin was inhibited by PFD in both CT- and DD- derived fibroblasts, but the effect was more prominent in DD cells. PFD down-regulated TGF-β1-induced phosphorylation of Smad2/Smad3, a key factor in the TGF-β1 signaling pathway.

Conclusion: Taken together these results suggest the PFD can potentially prevent TGF-β1-induced fibroblast to myofibroblast transformation and inhibit ECM production mainly Type I- and Type III- collagen and fibronectin in DD-derived fibroblasts. Further in-vivo studies with PFD may lead to a novel therapeutic application in preventing the progression or recurrence of Dupuytren's disease.

Keywords: Alpha-SMA; Carpal tunnel; Cell contraction; Cell migration; Collagen; Dupuytren’s contracture; Palmar fascia fibrosis; Smad2/Smad3.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Inflammatory Agents, Non-Steroidal / pharmacology
  • Anti-Inflammatory Agents, Non-Steroidal / therapeutic use*
  • Cell Movement / drug effects
  • Cell Proliferation / drug effects
  • Collagen Type I / metabolism
  • Collagen Type III / metabolism
  • Drug Evaluation, Preclinical
  • Dupuytren Contracture / drug therapy*
  • Fibroblasts / drug effects*
  • Fibroblasts / metabolism
  • Fibronectins / metabolism
  • Humans
  • Phosphorylation / drug effects
  • Primary Cell Culture
  • Pyridones / pharmacology
  • Pyridones / therapeutic use*
  • Smad2 Protein / metabolism
  • Smad3 Protein / metabolism
  • Transforming Growth Factor beta1 / metabolism

Substances

  • Anti-Inflammatory Agents, Non-Steroidal
  • Collagen Type I
  • Collagen Type III
  • Fibronectins
  • Pyridones
  • SMAD2 protein, human
  • SMAD3 protein, human
  • Smad2 Protein
  • Smad3 Protein
  • TGFB1 protein, human
  • Transforming Growth Factor beta1
  • pirfenidone