Indirect actions of thapsigargin on human platelets: activation of eicosanoid biosynthesis and cellular signaling pathways.
Thapsigargin, which acts by inhibition of a Ca(++)-ATPase on the dense tubule system in platelets, is a pharmacological tool to study the effects of increases in intracellular Ca++. Secondary consequences of thapsigargin treatment in platelets include extensive thromboxane B2 formation (493 +/- 106 ng/10(8) platelets) and [3H]5-hydroxytryptamine secretion (80.7 +/- 8.0%). Inhibition of cyclooxygenase by ibuprofen prevents thromboxane B2 formation (0.1 +/- 0.04 ng/10(8) platelets) and dense tubule secretion (6.5 +/- 3.8%). Aggregation in response to thapsigargin is rapid and maximal, but the rate and extent of aggregation are lowered by ibuprofen or aspirin. Mobilization of intracellular Ca++ is also significantly attenuated when eicosanoid formation is prevented, indicating the dependence of thapsigargin actions on endogenous lipid mediator formation. These studies also support the idea that formation of endogenous thromboxane A2/prostaglandin H2 is self-amplifying; thromboxane receptor antagonists inhibit endogenous thromboxane B2 formation, indicating that Ca(++)-dependent activation of phospholipase A2 is only partially responsible for eicosanoid production. Our data indicate the importance of distinguishing secondary effects of thapsigargin, especially because it may influence eicosanoid formation.
Malcolm
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KC
Department of Pharmacology, University of Colorado Health Sciences Center, Denver.
Fitzpatrick
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FA
D001792:Blood Platelets / Q000187:drug effects*; D002118:Calcium / Q000378:metabolism; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors*; D015777:Eicosanoids / Q000096:biosynthesis*; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D015539:Platelet Activation / Q000187:drug effects; D010974:Platelet Aggregation / Q000187:drug effects; D011450:Prostaglandin Endoperoxides, Synthetic / Q000378:metabolism; D044262:Prostaglandin H2; D011463:Prostaglandins H / Q000378:metabolism; D012701:Serotonin / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D013729:Terpenes / Q000494:pharmacology*; D019284:Thapsigargin; D013929:Thromboxane B2 / Q000096:biosynthesis
D015777:Eicosanoids; D007211:Indoles; D011450:Prostaglandin Endoperoxides, Synthetic; D011463:Prostaglandins H; D013729:Terpenes; C052517:L 655240; D012701:Serotonin; D044262:Prostaglandin H2; D013929:Thromboxane B2; D019284:Thapsigargin; D000252:Calcium-Transporting ATPases; D002118:Calcium