Disruption of intermolecular disulfide bonds in PDGF-BB dimers by N-acetyl-L-cysteine does not prevent PDGF signaling in cultured hepatic stellate cells

Biochem Biophys Res Commun. 2005 Dec 30;338(4):1711-8. doi: 10.1016/j.bbrc.2005.10.139. Epub 2005 Nov 2.

Abstract

Oxidative stress is important in the pathogenesis of liver fibrosis through its induction of hepatic stellate cell (HSC) proliferation and enhancement of collagen synthesis. Reactive oxygen species have been found to be essential second messengers in the signaling of both major fibrotic growth factors, platelet-derived growth factor (PDGF) and transforming growth factor-beta (TGF-beta), in cultured HSC and liver fibrosis. The non-toxic aminothiol N-acetyl-L-cysteine (NAC) inhibits cellular activation and attenuates experimental fibrosis in liver. Prior reports show that NAC is capable of reducing the effects of TGF-beta in biological systems, in cultured endothelial cells, and HSC through its direct reducing activity upon TGF-beta molecules. We here analyzed the effects of NAC on PDGF integrity, receptor binding, and downstream signaling in culture-activated HSC. We found that NAC dose-dependently induces disintegration of PDGF in vitro. However, even high doses (>20mM) were not sufficient to prevent the phosphorylation of the PDGF receptor type beta, extracellular signal-regulated kinase, or protein kinase B (PKB/Akt). Therefore, we conclude that the PDGF monomer is still active. The described antifibrotic effects are therefore mainly attributable to the structural impairment of TGF-beta signaling components reported previously.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylcysteine / chemistry
  • Acetylcysteine / pharmacology*
  • Animals
  • Becaplermin
  • Disulfides / chemistry*
  • Liver / cytology
  • Male
  • Platelet-Derived Growth Factor / chemistry
  • Platelet-Derived Growth Factor / drug effects
  • Platelet-Derived Growth Factor / physiology*
  • Proto-Oncogene Proteins c-sis
  • Rats
  • Rats, Sprague-Dawley
  • Receptors, Platelet-Derived Growth Factor / drug effects
  • Receptors, Platelet-Derived Growth Factor / physiology
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / drug effects
  • Signal Transduction / drug effects
  • Signal Transduction / physiology*
  • Transforming Growth Factor beta / physiology

Substances

  • Disulfides
  • Platelet-Derived Growth Factor
  • Proto-Oncogene Proteins c-sis
  • Recombinant Proteins
  • Transforming Growth Factor beta
  • Becaplermin
  • Receptors, Platelet-Derived Growth Factor
  • Acetylcysteine