15-Hydroxyeicosatetraenoic acid-mediated potentiation of thrombin-induced platelet functions occurs via enhanced production of phosphoinositide-derived second messengers--sn-1,2-diacylglycerol and inositol-1,4,5-trisphosphate.
We investigated whether biologically relevant concentrations of the mono-hydroxyeicosatetraenoic acids (mono-HETEs) modulate platelet functions. We report that 15-HETE, an eicosanoid produced by endothelial cells, granulocytes, and lymphocytes, potentiated platelet aggregation, nucleotide release, and elevation in intracellular calcium levels induced by a threshold concentration of thrombin (0.025 U/mL). Significant potentiation effects on these responses were observed at concentrations between 1 and 100 nmol/L. 15-HETE at these concentrations enhanced thrombin-induced platelet aggregation by 32% to 57%, nucleotide release by 40% to 65%, and elevation of intracellular calcium by 31% to 52% (P < .05 to .01). Both 12-HETE and 5-HETE, the structural isomers of 15-HETE, also potentiated thrombin-induced platelet aggregation and nucleotide release. While 12-HETE showed a small but significant effect at 100 pmol/L, 5-HETE had effects similar to those of 15-HETE at micromolar concentrations. To understand the mechanism of the HETE modulation of platelet functions, we studied the effect of 10 and 100 nmol/L 15-HETE on the production of sn-1,2-diacylglycerol (DAG) and inositol-1,4,5-trisphosphate (1,4,5-IP3). 15-HETE enhanced thrombin-induced production of DAG and 1,4,5-IP3 in a time- and concentration-dependent manner. 15-HETE also potentiated agonist-induced phosphorylation of the 47-Kd platelet protein. These studies demonstrate an important modulatory role for 15-HETE on platelet functions. Since this eicosanoid is elevated in pathologic states associated with platelet hyperfunction, including diabetes mellitus and atherosclerosis, an elucidation of its mechanism(s) of action appears relevant to our understanding of the genesis of atherothrombotic vascular disease.
Setty
B N
BN
Division of Hematology-Oncology, St Christopher's Hospital for Children, Temple University School of Medicine, Philadelphia, PA 19134-1095.
Werner
M H
MH
Hannun
Y A
YA
Stuart
M J
MJ
D000255:Adenosine Triphosphate / Q000097:blood; D000328:Adult; D001792:Blood Platelets / Q000187:drug effects / Q000502:physiology*; D004075:Diglycerides / Q000097:blood*; D004357:Drug Synergism; D015777:Eicosanoids / Q000096:biosynthesis / Q000097:blood; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000494:pharmacology*; D066298:In Vitro Techniques; D015544:Inositol 1,4,5-Trisphosphate / Q000097:blood*; D007700:Kinetics; D010716:Phosphatidylinositols / Q000097:blood*; D010750:Phosphoproteins / Q000097:blood / Q000302:isolation & purification; D010766:Phosphorylation; D010974:Platelet Aggregation / Q000187:drug effects*; D011869:Radioligand Assay; D015290:Second Messenger Systems / Q000187:drug effects*; D013917:Thrombin / Q000494:pharmacology*
D004075:Diglycerides; D015777:Eicosanoids; D006893:Hydroxyeicosatetraenoic Acids; D010716:Phosphatidylinositols; D010750:Phosphoproteins; C025984:15-hydroxy-5,8,11,13-eicosatetraenoic acid; D015544:Inositol 1,4,5-Trisphosphate; D000255:Adenosine Triphosphate; D013917:Thrombin