Session: 301. Platelets and Megakaryocytes: Basic and Translational: Poster III
Hematology Disease Topics & Pathways:
Research, Bleeding and Clotting, Translational Research, Diseases, Thrombotic disorders, Technology and Procedures
To determine if the presence of EPA alters the production of 12-LOX and cyxlooxygnease-1 (COX-1) metabolites, the lipid releasate from platelets treated with EPA and stimulated with collagen was analyzed via mass spectrometry. Production of 12-HEPE, the 12-LOX metabolite of EPA, is significantly increased in the presence of EPA, while COX-1 derived metabolites of EPA remain undetected. The presence of EPA does not alter levels of the 12-LOX and COX-1 derived metabolites of arachidonic acid. To assess the ability of EPA and 12-HEPE to alter platelet activation, isolated platelets from healthy human donors were treated with EPA or 12-HEPE and stimulated with various agonists targeting different steps of the hemostatic response to vascular injury. Both EPA and 12-HEPE dose-dependently inhibit collagen and thrombin-induced platelet aggregation with 12-HEPE having increased potency compared to EPA. Furthermore, 12-HEPE is a more potent inhibitor of surface expression of platelet integrin αIIbβ3 activation and surface exposure of P-selectin analyzed via flow cytometry in comparison to EPA. Additionally, EPA fully inhibits thrombus formation in whole blood under arterial shear via Total Thrombus formation Analysis System (T-TAS), while 12-HEPE only partially inhibits thrombus formation. Similarly, only EPA attenuates platelet adhesion to collagen in whole blood under arterial shear, while 12-HEPE has no effect.
Our findings show for the first time 12-HEPE, the 12-LOX metabolite of EPA, is the most abundant metabolite produced by platelets when activated in the presence of EPA, suggesting the effects of EPA on platelets are regulated by 12-HEPE and are not by reducing the production of proplatelet arachidonic acid derived metabolites. We also show both EPA and 12-HEPE directly inhibit platelet activation, and 12-HEPE has more potent antiplatelet effects in isolated platelets. Interestingly, in whole blood, 12-HEPE has a reduced effect compared to EPA, suggesting 12-HEPE may be unstable or bound by circulating proteins in whole blood, making the local production of 12-HEPE within the platelet critical to its antiplatelet effect. These findings provide further insight into the mechanisms underlying the cardioprotective effects of EPA. A better understanding of current PUFA supplements containing EPA can inform treatment and prevention of cardiovascular diseases.
Disclosures: Holinstat: Cereno Scientific: Consultancy, Current equity holder in publicly-traded company, Patents & Royalties, Research Funding; Lexicon Pharmaceuticals: Research Funding; Veralox Therapeutics: Consultancy, Current equity holder in private company, Patents & Royalties, Research Funding.
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