title: Effect of the hydrogen sulfide donor GYY4137 on platelet activation and microvascular thrombus formation in mice.
abstract: 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.
authors.affiliation: Institute for Experimental Surgery, University of Rostock , Rostock , Germany .
authors.forename: Eberhard
authors.initials: E
authors.lastname: Grambow
authors.forename: Fabian
authors.initials: F
authors.lastname: Mueller-Graf
authors.forename: Evgenya
authors.initials: E
authors.lastname: Delyagina
authors.forename: Marcus
authors.initials: M
authors.lastname: Frank
authors.forename: Angela
authors.initials: A
authors.lastname: Kuhla
authors.forename: Brigitte
authors.initials: B
authors.lastname: Vollmar
mesh_terms: D000135:Acid Phosphatase / Q000097:blood; D000818:Animals; D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D006801:Humans; D006862:Hydrogen Sulfide / Q000737:chemistry / Q000494:pharmacology; D007527:Isoenzymes / Q000097:blood; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008854:Microscopy, Electron; D009025:Morpholines / Q000737:chemistry / Q000494:pharmacology*; D009946:Organothiophosphorus Compounds / Q000737:chemistry / Q000494:pharmacology*; D019007:P-Selectin / Q000097:blood; D015539:Platelet Activation / Q000187:drug effects*; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000378:metabolism; D000071681:Tartrate-Resistant Acid Phosphatase; D013927:Thrombosis / Q000097:blood / Q000188:drug therapy*
chemical_list: C529376:GYY 4137; D007527:Isoenzymes; D009025:Morpholines; D009946:Organothiophosphorus Compounds; D019007:P-Selectin; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D000135:Acid Phosphatase; C000605511:Acp5 protein, mouse; D000071681:Tartrate-Resistant Acid Phosphatase; D006862:Hydrogen Sulfide