Sphingosine kinase 1 (Sphk1) negatively regulates platelet activation and thrombus formation
Sphingosine 1-phosphate (S1P) is a powerful regulator of platelet formation. Enzymes generating S1P include sphingosine kinase 1. The present study thus explored the role of sphingosine kinase 1 in platelet formation and function. Activation-dependent platelet integrin αIIbβ3activation and secretion of platelets lacking functional sphingosine kinase 1 ( sphk1−/−) and of wild-type platelets ( sphk1+/+) were determined utilizing flow cytometry and chronolume luciferin assay. Cytosolic Ca2+activity ([Ca2+]i) and aggregation were measured using fura-2 fluorescence and aggregometry, respectively. In vitro platelet adhesion and thrombus formation were evaluated using a flow chamber with shear rates of 1,700 s−1. Activation-dependent increase of [Ca2+]i, degranulation (release of alpha and dense granules), integrin αIIbβ3activation, and aggregation were all significantly increased in sphk1−/−platelets compared with sphk1+/+platelets. Moreover, while platelet adhesion and thrombus formation under arterial shear rates were significantly augmented in Sphk1-deficient platelets, bleeding time and blood count were unaffected in sphk1−/−mice. In conclusion, sphingosine kinase 1 is a powerful negative regulator of platelet function counteracting degranulation, aggregation, and thrombus formation.