Molecular mechanisms of the antiproliferative effect of beraprost, a prostacyclin agonist, in murine vascular smooth muscle cells

J Cell Physiol. 2008 Feb;214(2):434-41. doi: 10.1002/jcp.21214.

Abstract

Prostacyclin (PGI2) has been shown to inhibit proliferation in vascular smooth muscle cells. To clarify the underlying molecular mechanism, we investigated the vasoprotection of beraprost (a PGI2 agonist) both in vivo and in vitro. Beraprost eliminated increases in proliferation of rat aortic smooth muscle cells (RASMCs) by 12-O-tetradecanoylphorbol 13-acetate, and enhanced the peroxisome proliferator-activated receptor-delta (PPARdelta) and inducible nitric oxide synthetase (iNOS) expressions, which were associated with the antiproliferative action of beraprost according to inhibition experiments by [(3)H]thymidine incorporation. Additionally, elimination of iNOS activity by PPARdelta antagonists suggested that iNOS is the downstream target of PPARdelta. Furthermore, beraprost increased both consensus PPARdelta-responsive element (PPRE)-driven luciferase activity and the binding activity of the PPARdelta to the putative PPRE in the iNOS promoter; nevertheless, it was abolished by PPARdelta antagonists. Deletion of PPRE (-1,349/-1,330) in the iNOS promoter region (-1,359/+2) strongly reduced promoter-driven activity, representing a novel mechanism of iNOS induction by beraprost. Consistent with this, PPARdelta and the concomitant iNOS induction by beraprost were also evident in vivo. Beraprost-mediated protection in a murine model of balloon angioplasty was significantly attenuated by 13S-HODE, a PPARdelta antagonist. Taken together, the results suggest that the causal relationship between PPARdelta and iNOS contributes to the vasoprotective action of beraprost in RASMCs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aorta, Thoracic / cytology
  • Cell Proliferation / drug effects*
  • Cell Survival / drug effects
  • Dose-Response Relationship, Drug
  • Enzyme Induction / drug effects
  • Epoprostenol / agonists*
  • Epoprostenol / analogs & derivatives*
  • Epoprostenol / pharmacology
  • Formazans / analysis
  • Formazans / metabolism
  • Gene Deletion
  • Genes, Reporter
  • Luciferases / metabolism
  • Male
  • Muscle, Smooth, Vascular / drug effects*
  • Nitric Oxide Synthase Type II / biosynthesis
  • Organ Culture Techniques
  • PPAR delta / drug effects
  • PPAR delta / metabolism
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Response Elements / drug effects
  • Response Elements / genetics
  • Tetradecanoylphorbol Acetate / pharmacology
  • Tetrazolium Salts / analysis
  • Tetrazolium Salts / metabolism
  • Thymidine / antagonists & inhibitors
  • Time Factors
  • Transfection

Substances

  • Formazans
  • PPAR delta
  • RNA, Messenger
  • Tetrazolium Salts
  • MTT formazan
  • beraprost
  • Epoprostenol
  • Luciferases
  • Nitric Oxide Synthase Type II
  • Tetradecanoylphorbol Acetate
  • Thymidine