The effects of hydrogen sulfide on platelet-leukocyte aggregation and microvascular thrombolysis.
The volatile transmitter hydrogen sulfide (H2S) is known for its various functions in vascular biology. This study evaluates the effect of the H2S-donor GYY4137 (GYY) on thrombus stability and microvascular thrombolysis. Human whole blood served for all in vitro studies and was analyzed in a resting state, after stimulation with thrombin-receptor activating peptide (TRAP) and after incubation with 10 or 30 mM GYY or its vehicle DMSO following TRAP-activation, respectively. As a marker for thrombus stability, platelet-leukocyte aggregation was assessed using flow cytometry after staining of human whole blood against CD62P and CD45, respectively. Furthermore, morphology and quantity of platelet-leukocyte aggregation were studied by means of scanning electron microscopy (scanning EM). Therefore, platelets were stained for CD62P followed by immuno gold labeling. In vivo, the dorsal skinfold chamber preparation was performed for light/dye induction of thrombi in arterioles and venules using intravital fluorescence microscopy. Thrombolysis was assessed 10 and 22 h after thrombus induction and treatment with the vehicle, GYY, or recombinant tissue plasminogen activator (rtPA). Flow cytometry revealed an increase of CD62P/CD45 positive aggregates after TRAP stimulation of human whole blood, which was significantly reduced by preincubation with 30 mM GYY. Scanning EM additionally showed a reduced platelet-leukocyte aggregation and a decreased leukocyte count within the aggregates after preincubation with GYY compared to TRAP stimulation alone. Further on, morphological signs of platelet activation were found markedly reduced upon treatment with GYY. In mice, both GYY and rtPA significantly accelerated arteriolar and venular thrombolysis compared to the vehicle control. In conclusion, GYY impairs thrombus stability by reducing platelet-leukocyte aggregation and thereby facilitates endogenous thrombolysis.
a Institute for Experimental Surgery, Rostock University Medical Center , Rostock , Germany.|b Department of General , Thoracic, Vascular and Transplantation Surgery, Rostock University Medical Center , Rostock , Germany.
Eberhard
E
Grambow
a Institute for Experimental Surgery, Rostock University Medical Center , Rostock , Germany.
Christian
C
Leppin
a Institute for Experimental Surgery, Rostock University Medical Center , Rostock , Germany.
Katja
K
Leppin
c Institute for Biostatistics and Informatics in Medicine and Aging Research, Rostock University Medical Center , Rostock , Germany.
Günther
G
Kundt
b Department of General , Thoracic, Vascular and Transplantation Surgery, Rostock University Medical Center , Rostock , Germany.
Ernst
E
Klar
d Medical Biology and Electron Microscopy Centre, Rostock University Medical Center , Rostock , Germany.
Marcus
M
Frank
a Institute for Experimental Surgery, Rostock University Medical Center , Rostock , Germany.
Brigitte
B
Vollmar
D000328:Adult; D001160:Arterioles / Q000378:metabolism; D001792:Blood Platelets / Q000378:metabolism*; D005260:Female; D006801:Humans; D006862:Hydrogen Sulfide / Q000494:pharmacology*; D017493:Leukocyte Common Antigens / Q000378:metabolism; D007962:Leukocytes / Q000378:metabolism*; D008297:Male; D009025:Morpholines / Q000494:pharmacology; D009946:Organothiophosphorus Compounds / Q000494:pharmacology; D019007:P-Selectin / Q000378:metabolism; D010974:Platelet Aggregation / Q000187:drug effects*; D000071681:Tartrate-Resistant Acid Phosphatase / Q000378:metabolism; D015912:Thrombolytic Therapy; D014699:Venules / Q000378:metabolism
C529376:GYY 4137; D009025:Morpholines; D009946:Organothiophosphorus Compounds; D019007:P-Selectin; C546937:SELP protein, human; C000605510:ACP5 protein, human; D000071681:Tartrate-Resistant Acid Phosphatase; D017493:Leukocyte Common Antigens; C516769:PTPRC protein, human; D006862:Hydrogen Sulfide
Dorsal skinfold chamber; hemostasis; microcirculation; platelet leukocyte aggregation; thrombolysis; thrombus stability