Pharmacological inhibition of the vitronectin receptor abrogates PDGF-BB-induced hepatic stellate cell migration and activation in vitro.

BACKGROUND/AIMS: Activated hepatic stellate cells (HSC) play a central role in the development of liver fibrosis. Platelet-derived growth factor (PDGF)-BB and the integrin alphavbeta3 mediate mesenchymal cell migration and proliferation. However, their contribution and interaction during fibrogenic activation of HSC remains unclear. To this aim we investigated if PDFGF-BB and alphavbeta3 interact, and how far small molecular inhibitors of alphavbeta3 modulate PDGF-BB and serum-induced migration, proliferation and fibrogenic activation of HSC.

METHODS

Rat and human HSC were subjected to migration and proliferation assays in the presence or absence of a peptide or a nonpeptide alphavbeta3 inhibitor. Activation of mitogen-activated protein kinases (ERK1/2, p38), Akt, focal adhesion kinase (FAK), paxillin and beta3 integrin was evaluated by phospho-specific Western blotting. Fibrosis related transcripts were determined by quantitative real-time PCR.

RESULTS

PDGF-BB-stimulated HSC migration which was blocked dose-dependently by the alphavbeta3 antagonists, with complete inhibition at 10(-6)M. alphavbeta3 blockage did not affect cell viability or proliferation, while it decreased phosphorylation of FAK, paxillin, beta3 integrin and p38, but not of ERK1/2 or Akt. alphavbeta3 inhibition led to downregulation of certain profibrogenic transcripts, while it upregulated fibrolytic MMP-13 mRNA.

CONCLUSIONS

Inhibition of integrin alphavbeta3 leads to abrogation of migration of HSC stimulated with PDGF-BB and to an antifibrogenic gene expression pattern.