Long-term inhibition of Rho-kinase suppresses neointimal formation after stent implantation in porcine coronary arteries: involvement of multiple mechanisms

Arterioscler Thromb Vasc Biol. 2004 Jan;24(1):181-6. doi: 10.1161/01.ATV.0000105053.46994.5B. Epub 2003 Oct 30.

Abstract

Objective: We recently demonstrated that Rho-kinase, an effector of the small GTPase Rho, is substantially involved in the pathogenesis of arteriosclerosis. In this study, we examined whether Rho-kinase is also involved in in-stent restenosis and if so, what mechanism is involved.

Methods and results: Pigs underwent stent implantation in the left coronary artery with or without administration of fasudil (30 mg/kg per day orally), a specific Rho-kinase inhibitor, starting 2 days before the procedure for a duration of 4 weeks. On day 28, reductions in coronary diameter and neointimal formation associated with macrophage accumulation, collagen deposition, and transforming growth factor (TGF)-beta1 expression were noted at the stent site, and all were significantly suppressed by fasudil. On day 7, fasudil significantly increased the frequency of TUNEL-positive apoptotic cells, while it tended to reduce that of bromodeoxyuridine-positive proliferating cells in the neointima. Western blot analysis on day 7 demonstrated that phosphorylations of the ezrin/radixin/moesin family (a marker of Rho-kinase activity in vivo) and protein expression of monocyte chemoattractant protein-1and bcl-2 were upregulated at the stent site and were significantly suppressed by fasudil.

Conclusions: These results indicate that long-term inhibition of Rho-kinase suppresses in-stent neointimal formation by multiple mechanisms, including reduced vascular inflammation, enhanced apoptosis, and decreased collagen deposition.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / analogs & derivatives*
  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / pharmacology*
  • Animals
  • Apoptosis / drug effects
  • Blood Proteins / metabolism
  • Catheterization / adverse effects
  • Chemokine CCL2 / biosynthesis
  • Chemokine CCL2 / genetics
  • Collagen / metabolism
  • Coronary Restenosis / enzymology*
  • Coronary Restenosis / pathology
  • Coronary Restenosis / prevention & control
  • Coronary Stenosis / surgery
  • Coronary Stenosis / therapy
  • Cytoskeletal Proteins / metabolism
  • Drug Evaluation, Preclinical
  • Enzyme Inhibitors / pharmacology*
  • Genes, bcl-2
  • Intracellular Signaling Peptides and Proteins
  • Male
  • Membrane Proteins / metabolism
  • Microfilament Proteins / metabolism
  • Phosphoproteins / metabolism
  • Phosphorylation / drug effects
  • Protein Processing, Post-Translational / drug effects
  • Protein Serine-Threonine Kinases / antagonists & inhibitors
  • Protein Serine-Threonine Kinases / physiology*
  • Proto-Oncogene Proteins c-bcl-2 / biosynthesis
  • Single-Blind Method
  • Stents*
  • Swine
  • Tunica Intima / drug effects
  • Tunica Intima / enzymology
  • Tunica Intima / pathology*
  • rho-Associated Kinases

Substances

  • Blood Proteins
  • Chemokine CCL2
  • Cytoskeletal Proteins
  • Enzyme Inhibitors
  • Intracellular Signaling Peptides and Proteins
  • Membrane Proteins
  • Microfilament Proteins
  • Phosphoproteins
  • Proto-Oncogene Proteins c-bcl-2
  • ezrin
  • moesin
  • radixin
  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
  • Collagen
  • Protein Serine-Threonine Kinases
  • rho-Associated Kinases
  • fasudil