Group V secretory phospholipase A2 impairs endothelial protein C receptor-dependent protein C activation and accelerates thrombosis in vivo

J Thromb Haemost. 2014 Nov;12(11):1921-7. doi: 10.1111/jth.12676. Epub 2014 Oct 12.

Abstract

Background: Endothelial protein C receptor (EPCR) must be bound to a molecule of phosphatidylcholine (PC) to be fully functional, i.e. to interact with protein C/activated protein C (APC) properly. PC can be replaced with other lipids, such as lysophosphatidylcholine or platelet-activating factor, by the action of group V secretory phospholipase A2 (sPLA2-V), an enzyme that is upregulated in a variety of inflammatory conditions. Studies in purified systems have demonstrated that the substitution of PC notably impairs EPCR function in a process called EPCR encryption.

Objectives: To analyze whether sPLA2-V was able to regulate EPCR-dependent protein C activation in vivo, and its impact on thrombosis and the hemostatic system.

Methods: Mice were transfected with sPLA2-V by hydrodynamic gene delivery. The effects on thrombosis were studied with the laser carotid artery occlusion model, and APC generation capacity was measured with ELISA. Global hemostasis was analyzed with thromboelastometry.

Results: We found that sPLA2-V overexpression in mice significantly decreased their ability to generate APC. Furthermore, a murine carotid artery laser thrombosis model revealed that higher sPLA2-V levels were directly associated with faster artery thrombosis.

Conclusions: sPLA2-V plays a thrombogenic role by impairing the ability of EPCR to promote protein C activation.

Keywords: activated protein C resistance; endothelial cell protein C receptor; group V secretory phospholipase A2; protein C; thrombosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carotid Stenosis / blood
  • Carotid Stenosis / enzymology*
  • Carotid Stenosis / genetics
  • Disease Models, Animal
  • Endothelial Protein C Receptor
  • Enzyme Activation
  • Enzyme Induction
  • Enzyme Inhibitors / pharmacology
  • Gene Transfer Techniques
  • Group V Phospholipases A2 / antagonists & inhibitors
  • Group V Phospholipases A2 / biosynthesis*
  • Group V Phospholipases A2 / genetics
  • Hemostasis
  • Humans
  • Liver / drug effects
  • Liver / enzymology*
  • Mice, Inbred ICR
  • Protein C / metabolism*
  • Receptors, Cell Surface / metabolism*
  • Signal Transduction
  • Thrombosis / blood
  • Thrombosis / enzymology*
  • Thrombosis / genetics
  • Time Factors

Substances

  • Endothelial Protein C Receptor
  • Enzyme Inhibitors
  • Procr protein, mouse
  • Protein C
  • Receptors, Cell Surface
  • Group V Phospholipases A2
  • PLA2G5 protein, human