Effects of the dual TP receptor antagonist and thromboxane synthase inhibitor EV-077 on human endothelial and vascular smooth muscle cells

Biochem Biophys Res Commun. 2013 Nov 15;441(2):393-8. doi: 10.1016/j.bbrc.2013.10.078. Epub 2013 Oct 23.

Abstract

The prothrombotic mediator thromboxane A2 is derived from arachidonic acid metabolism through the cyclooxygenase and thromboxane synthase pathways, and transduces its effect through the thromboxane prostanoid (TP) receptor. The aim of this study was to determine the effect of the TP receptor antagonist and thromboxane synthase inhibitor EV-077 on inflammatory markers in human umbilical vein endothelial cells and on human coronary artery smooth muscle cell proliferation. To this end, mRNA levels of different proinflammatory mediators were studied by real time quantitative PCR, supernatants were analyzed by enzyme immune assay, and cell proliferation was assessed using WST-1. EV-077 significantly decreased mRNA levels of ICAM-1 and PTX3 after TNFα incubation, whereas concentrations of 6-keto PGF1α in supernatants of endothelial cells incubated with TNFα were significantly increased after EV-077 treatment. Although U46619 did not alter coronary artery smooth muscle cell proliferation, this thromboxane mimetic enhanced the proliferation induced by serum, insulin and growth factors, which was significantly inhibited by EV-077. In conclusion, EV-077 inhibited TNFα-induced endothelial inflammation and reduced the enhancement of smooth muscle cell proliferation induced by a thromboxane mimetic, supporting that the thromboxane pathway may be associated with early atherosclerosis in terms of endothelial dysfunction and vascular hypertrophy.

Keywords: Eicosanoids; Endothelial cells; Inflammation; Smooth muscle cells; Thromboxane prostanoid receptor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid / pharmacology
  • C-Reactive Protein / antagonists & inhibitors
  • C-Reactive Protein / biosynthesis
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Coronary Vessels / cytology
  • Coronary Vessels / drug effects
  • Coronary Vessels / metabolism
  • Endothelium, Vascular / drug effects*
  • Endothelium, Vascular / metabolism
  • Gene Expression / drug effects
  • Human Umbilical Vein Endothelial Cells / drug effects
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Humans
  • Intercellular Adhesion Molecule-1 / biosynthesis
  • Muscle, Smooth, Vascular / cytology
  • Muscle, Smooth, Vascular / drug effects*
  • Muscle, Smooth, Vascular / metabolism
  • Myocytes, Smooth Muscle / drug effects*
  • Myocytes, Smooth Muscle / metabolism
  • Prostaglandins F / metabolism
  • RNA, Messenger / antagonists & inhibitors
  • RNA, Messenger / biosynthesis
  • Receptors, Thromboxane / antagonists & inhibitors*
  • Serum Amyloid P-Component / antagonists & inhibitors
  • Serum Amyloid P-Component / biosynthesis
  • Thromboxane A2 / physiology
  • Thromboxane-A Synthase / antagonists & inhibitors*
  • Tumor Necrosis Factor-alpha / pharmacology

Substances

  • ICAM1 protein, human
  • Prostaglandins F
  • RNA, Messenger
  • Receptors, Thromboxane
  • Serum Amyloid P-Component
  • Tumor Necrosis Factor-alpha
  • Intercellular Adhesion Molecule-1
  • PTX3 protein
  • Thromboxane A2
  • 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid
  • C-Reactive Protein
  • Thromboxane-A Synthase
  • prostaglandin F1