Alkaline phosphatase prevents platelet stimulation by thromboxane-mimetics.
1. The effects of alkaline phosphatase on platelet aggregation, secretion and thromboxane B2 (TxB2) generation induced by the full dose-range of common platelet agonists were studied in human platelet-rich plasma and washed platelets. 2. Platelet aggregation and adenosine 5'-triphosphate (ATP) secretion induced by threshold and supramaximal concentrations of arachidonate and stable TxA2 and prostaglandin endoperoxide-mimetics (compounds U46619 and EP171) were abolished in the presence of alkaline phosphatase (0.5-1 u ml-1), even though the synthesis of TxB2 persisted. In contrast, platelet aggregation by PAF-acether and by supramaximal concentrations of thrombin as well as the primary wave of aggregation by adenosine diphosphate (ADP) and adrenaline were unaffected by alkaline phosphatase under conditions where the secondary wave of aggregation by ADP was blocked. 3. Alkaline phosphatase, unlike prostacyclin, failed to raise the adenosine 3':5'-cyclic monophosphate (cyclic AMP) content of the platelets. Also, the pretreatment of platelets by inorganic phosphate or by ATP plus creatine phosphate/creatine phosphokinase reversed the inhibitory effect of alkaline phosphatase. 4. Experiments performed in the guinea-pig in vivo showed that alkaline phosphatase was effective on thrombocytopenia induced by arachidonate. 5. Our results provide the first direct evidence for a specific inhibitory effect of alkaline phosphatase at a site sensitive to TxA2 and prostaglandin endoperoxides and suggest that its phosphorylation/dephosphorylation state may play an important role in modulating platelet activation. These results also suggest the presence of ecto-protein kinases on membrane platelets.
Hatmi
M
M
Unité de Pharmacologie Cellulaire, Unité Associée Institut Pasteur/INSERM No. 285, Paris, France.
Haye
B
B
Gavaret
J M
JM
Vargaftig
B B
BB
Jacquemin
C
C
D000469:Alkaline Phosphatase / Q000378:metabolism*; D001792:Blood Platelets / Q000187:drug effects*; D016084:Bronchoconstriction / Q000502:physiology; D000242:Cyclic AMP / Q000097:blood; D006801:Humans; D010766:Phosphorylation; D010975:Platelet Aggregation Inhibitors / Q000378:metabolism*; D011863:Radioimmunoassay; D013921:Thrombocytopenia / Q000097:blood; D013929:Thromboxane B2 / Q000097:blood; D013931:Thromboxanes / Q000037:antagonists & inhibitors*
D010975:Platelet Aggregation Inhibitors; D013931:Thromboxanes; D013929:Thromboxane B2; D000242:Cyclic AMP; D000469:Alkaline Phosphatase