Effect of the hydrogen sulfide donor GYY4137 on platelet activation and microvascular thrombus formation in mice

Platelets. 2014;25(3):166-74. doi: 10.3109/09537104.2013.786823. Epub 2013 Apr 15.


This study evaluates the effect of the H2S donor GYY4137 (GYY) on adhesion molecule expression, protein S-sulfhydration and morphology of platelets in vitro and on kinetics of microvascular thrombus formation in vivo. Using flowcytometry, untreated resting, TRAP-activated, or TRAP-activated and GYY-exposed human platelets were studied for expression of P-selectin, GPIb and GPIIb/IIIa as well as for fibrinogen binding. By means of electron microscopy, platelet morphology and intracellular granule numbers were assessed. Platelet shape change was studied using immunohistochemistry for P-selectin, NSF and F-actin by SR-SIM. Biotin switch assay served for the analysis of platelet protein S-sulfhydration by GYY. Using the FeCl3 and the light/dye model in dorsal skinfold chamber-equipped mice, the effect of GYY and its vehicle DMSO was studied on venular thrombus formation and tail-vein bleeding time. Soluble (s)P-selectin plasma concentrations were measured in GYY- or DMSO-treated animals. Exposure to GYY increased the S-sulfhydration of platelet proteins. GYY reduced dose-dependently the TRAP-induced adhesion molecule expression and attenuated the morphological signs of TRAP-associated platelet activation. In mice, GYY caused a significant prolongation of venular thrombus formation and tail-vein bleeding time. Application of an anti-P-selectin antibody in DMSO-exposed animals prolonged thrombosis formation comparably as GYY did. GYY reversed the TRAP-induced distribution of P-selectin at the plasma membrane of platelets. This indicates reduced exocytosis and shedding of P-selectin, which is supported by significantly lower sP-selectin concentrations in GYY- vs. DMSO-treated mice. H2S acts anti-thrombotic and seems to regulate thrombogenesis by interference with platelet activation and adhesion molecule-mediated aggregation.

MeSH terms

  • Acid Phosphatase / blood
  • Animals
  • Blood Platelets / drug effects*
  • Blood Platelets / metabolism
  • Blood Platelets / physiology
  • Blood Platelets / ultrastructure
  • Humans
  • Hydrogen Sulfide / chemistry
  • Hydrogen Sulfide / pharmacology
  • Isoenzymes / blood
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Electron
  • Morpholines / chemistry
  • Morpholines / pharmacology*
  • Organothiophosphorus Compounds / chemistry
  • Organothiophosphorus Compounds / pharmacology*
  • P-Selectin / blood
  • Platelet Activation / drug effects*
  • Platelet Glycoprotein GPIIb-IIIa Complex / metabolism
  • Tartrate-Resistant Acid Phosphatase
  • Thrombosis / blood
  • Thrombosis / drug therapy*


  • GYY 4137
  • Isoenzymes
  • Morpholines
  • Organothiophosphorus Compounds
  • P-Selectin
  • Platelet Glycoprotein GPIIb-IIIa Complex
  • Acid Phosphatase
  • Acp5 protein, mouse
  • Tartrate-Resistant Acid Phosphatase
  • Hydrogen Sulfide