Evidence for a Role for Phospholipase C, but not Phospholipase A2, in Platelet Activation in Response to Low Concentrations of Collagen

Leslie K. Lockhart; Caroline Pampolina; Brent R. Nickolaychuk; Archibald McNicol

doi:10.1055/s-0037-1615763

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2001; 85(05): 882-889
DOI: 10.1055/s-0037-1615763

Review Article

Schattauer GmbH

Evidence for a Role for Phospholipase C, but not Phospholipase A₂, in Platelet Activation in Response to Low Concentrations of Collagen

Leslie K. Lockhart

¹Departments of Oral Biology and Pharmacology & Therapeutics, University of Manitoba, Winnipeg, Manitoba, Canada, Present address: Hill Top Research, Inc., P.O. Box 429501, Cincinnati, OH 45242, USA

,

Caroline Pampolina

¹Departments of Oral Biology and Pharmacology & Therapeutics, University of Manitoba, Winnipeg, Manitoba, Canada, Present address: Hill Top Research, Inc., P.O. Box 429501, Cincinnati, OH 45242, USA

,

Brent R. Nickolaychuk

¹Departments of Oral Biology and Pharmacology & Therapeutics, University of Manitoba, Winnipeg, Manitoba, Canada, Present address: Hill Top Research, Inc., P.O. Box 429501, Cincinnati, OH 45242, USA

,

Archibald McNicol

¹Departments of Oral Biology and Pharmacology & Therapeutics, University of Manitoba, Winnipeg, Manitoba, Canada, Present address: Hill Top Research, Inc., P.O. Box 429501, Cincinnati, OH 45242, USA

› Author Affiliations

Abstract

Summary

The release of arachidonic acid is a key component in platelet activation in response to low concentrations (1-20 g/ml) of collagen. The precise mechanism remains elusive although a variety of pathways have been implicated. In the present study the effects of inhibitors of several potentially key enzymes in these pathways have been examined. Collagen (1-10 g/ml) caused maximal platelet aggregation which was accompanied by the release of arachidonic acid, the synthesis of thromboxane A₂, and p38^MAPK phosphorylation. Preincubation with the dual cyclooxygenase/lipoxygenase inhibitor BW755C inhibited aggregation and thromboxane production, and reduced p38^MAPK phosphorylation. A phospholipase C inhibitor, U73122, blocked collagen-induced aggregation and reduced arachidonic acid release, thromboxane synthesis and p38^MAPK phosphorylation. Pretreatment with a cytosolic phospholipase A₂ inhibitor, AACOCF₃, blocked collagen-induced aggregation, reduced the levels of thromboxane formation and p38^MAPK phosphorylation but had no significant effect on arachidonic acid release. In contrast inhibition of PKC by Rö31-8220 inhibited collagen-induced aggregation, did not affect p38^MAPK phosphorylation but significantly potentiated arachidonic acid release and thromboxane formation. Collagen caused the tyrosine phosphorylation of phospholipase C 2 which was inhibited by pretreatment with U73122, unaffected by AACOCF₃ and enhanced by Rö31-8220. These results suggest that cytosolic phospholipase A₂ plays no role in the arachidonic acid release in response to collagen. In contrast, the data are consistent with phospholipase C 2 playing a role in an intricately controlled pathway, or multiple pathways, mediating the release of arachidonic acid in collagen-stimulated platelets.

Keywords

Platelets - collagen - arachidonic acid - phospholipase

Full Text

References

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