Phloretin suppresses thrombin-mediated leukocyte-platelet-endothelial interactions

Mol Nutr Food Res. 2014 Apr;58(4):698-708. doi: 10.1002/mnfr.201300267. Epub 2013 Dec 16.

Abstract

Scope: Thrombin playing a pivotal role in coagulation cascade may influence the onset and progression of atherosclerosis as a pro-inflammatory mediator. This study investigated whether phloretin found in apple tree leaves, severed a linkage between thrombosis and atherosclerosis by thrombin.

Methods and results: Human endothelial cells were pre-treated with 1-20 μM phloretin and stimulated with 10 U/mL thrombin. Phloretin attenuated adhesion of THP-1 monocytes and platelets to thrombin-inflamed endothelial cells with concurrent inhibition of protease-activated receptor (PAR-1) induction. The thrombin induction of endothelial CD40, endothelial integrin β3 and P-selectin, and monocytic CD40L was dampened by phloretin. Additionally, phloretin inhibited monocyte secretion of MCP-1, IL-6 and IL-8 responsible for pro-inflammatory activity of thrombin inducing endothelial CD40. The monocyte COX-2 induction and PGE2 secretion due to thrombin were down-regulated by phloretin, deterring endothelial CD40 expression. Thrombin promoted production of PAI-1 and tissue factor in monocytes was attenuated by phloretin through blocking PAR-1 and CD40. Thrombin up-regulated the induction of endothelial connective tissue growth factor independent of PAR-1 activation, which was reversed by phloretin.

Conclusion: Phloretin disturbed tethering and stable adhesion of monocytes and platelets onto endothelium during increased thrombosis by thrombin. Phloretin would be a potent agent preventing thrombosis and atherosclerosis.

Keywords: CD40; Phloretin; Platelet; Protease-activated receptor-1; Thrombin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blood Platelets / drug effects*
  • CD40 Antigens / metabolism
  • Cells, Cultured
  • Cyclooxygenase 2 / metabolism
  • Cytokines / metabolism
  • Dinoprostone / metabolism
  • Endothelial Cells / drug effects
  • Endothelial Cells / metabolism
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / drug effects*
  • Humans
  • Inflammation Mediators / metabolism
  • Leukocytes / drug effects
  • Phloretin / pharmacology*
  • Plasminogen Activator Inhibitor 1 / metabolism
  • Platelet Adhesiveness / drug effects
  • Receptor, PAR-1 / metabolism
  • Thrombin / metabolism*
  • Thrombin / pharmacology

Substances

  • CD40 Antigens
  • Cytokines
  • Inflammation Mediators
  • Plasminogen Activator Inhibitor 1
  • Receptor, PAR-1
  • SERPINE1 protein, human
  • Cyclooxygenase 2
  • PTGS2 protein, human
  • Thrombin
  • Dinoprostone
  • Phloretin