Roflumilast-induced Local Vascular Injury Is Associated with a Coordinated Proteome and Microparticle Change in the Systemic Circulation in Pigs

Toxicol Pathol. 2015 Jun;43(4):569-80. doi: 10.1177/0192623314551971. Epub 2014 Oct 13.


Drug-induced vascular injury (DIVI) is commonly associated with phosphodiesterase (PDE) inhibitors. Despite histological characterization, qualified biomarkers for DIVI detection are lacking. We investigated whether a single administration of roflumilast (PDE-IV inhibitor) induces vascular damage and identified novel surrogate biomarkers of acute vascular injury. Pigs received postoperative 250, 375, or 500 μg of roflumilast or placebo/control. After 1.5 hr, coronary reactivity was determined by catheter-based administration of acetylcholine and sodium nitroprusside (SNP) in the coronary sinus. Immunohistochemical analysis of vessel integrity (von Willebrand factor [vWF]) and fibrin(ogen) deposition was performed in the coronary artery and aorta. Peripheral blood was collected for differential proteomics and microparticles analysis. Circulating interleukin (IL)-6 was analyzed. Roflumilast-treated animals displayed higher vasodilation to acetylcholine and SNP versus controls (p < .05). Roflumilast-treated animals showed a dose-dependent (p < .05) decrease in vessel integrity and dose-dependent increase in fibrin deposition forming a continuous layer at roflumilast-500 μg. Peripheral blood of roflumilast-500-μg-treated animals showed increased levels of total and endothelial-derived microparticles and exhibited a coordinated change in proteins kininogen-1, endothelin-1, gelsolin, apolipoprotein A-I, and apolipoprotein-J associated with vascular injury (p < .05 vs. controls). IL-6 remained unaltered. Roflumilast-induced vascular injury can be detected by novel markers in peripheral blood. Validation of these surrogate markers in human samples seems required.

Keywords: Roflumilast; biomarkers; drug-induced vascular injury; proteomics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aminopyridines / toxicity*
  • Animals
  • Benzamides / toxicity*
  • Biomarkers / blood
  • Blood Proteins / analysis
  • Blood Proteins / metabolism
  • Cell-Derived Microparticles / drug effects*
  • Cell-Derived Microparticles / metabolism
  • Cyclopropanes / toxicity
  • Female
  • Interleukin-6 / blood
  • Phosphodiesterase 4 Inhibitors / toxicity
  • Proteome / analysis
  • Proteome / drug effects*
  • Proteome / metabolism
  • Proteomics / methods
  • Swine
  • Vascular System Injuries / blood*
  • Vascular System Injuries / chemically induced*


  • Aminopyridines
  • Benzamides
  • Biomarkers
  • Blood Proteins
  • Cyclopropanes
  • Interleukin-6
  • Phosphodiesterase 4 Inhibitors
  • Proteome
  • Roflumilast