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

Prosracyclin (PGI(2)) has been shown to inhibit proliferation in vascular smooth muscle cells. To clarify the underlying molecular mechanism, we investigated the vasoprotection of beraprost (a PGI(2) 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 (PPAR delta) and inducible nitric oxide synthetase (iNOS) expressions, which were associated with the antiproliferative action of beraprost according to inhibition experiments by [H-3]thymidine incorporation. Additionally, elimination of iNOS activity by PPAR delta antagonists suggested that iNOS is the downstream target of PPAR delta. Furthermore, beraprost increased both consensus PPAR delta-responsive element (PPRE)-driven luciferase activity and the binding activity of the PPAR delta to the putative PPRE in the iNOS promoter; nevertheless, it was abolished by PPAR delta 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, PPAR delta 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 PPAR delta antagonist. Taken together, the results suggest that the causal relationship between PPAR delta and iNOS contributes to the vasoprotective action of beraprost in RASMCs.