Thromboxane Synthase Inhibition Potentiates Washed Platelet Activation by Endogenous and Exogenous Arachidonic Acid

2015 ◽  
pp. 465-470
Author(s):  
H. Patscheke
Keyword(s):  
Arachidonic Acid ◽  
Platelet Activation ◽  
Thromboxane Synthase ◽  
Washed Platelet ◽  
Exogenous Arachidonic Acid

Studies on topological distribution of arachidonic acid replacement in platelet phospholipids and on enzymes involved in the phospholipid effect accompanying platelet activation

1981 ◽  
Vol 11 (6-7) ◽  
pp. 538-540 ◽  
Author(s):  
H. Chap ◽  
G. Mauco ◽  
B. Perret ◽  
M. Plantavid ◽  
F. Laffont ◽  
...  
Keyword(s):  
Arachidonic Acid ◽  
Platelet Activation

scholarly journals Fibronectin modulates formation of PF4/heparin complexes and is a potential factor for reducing risk of developing HIT

Blood ◽  
2019 ◽  
Vol 133 (9) ◽  
pp. 978-989 ◽  
Author(s):  
Krystin Krauel ◽  
Patricia Preuße ◽  
Theodore E. Warkentin ◽  
Catja Trabhardt ◽  
Sven Brandt ◽  
...  
Keyword(s):  
Platelet Activation ◽  
Platelet Rich Plasma ◽  
Enzyme Linked Immunosorbent Assay ◽  
Heparin Binding ◽  
Plasma Fibronectin ◽  
Platelet Factor ◽  
Washed Platelet ◽  
Plasma Factor ◽  
Heparin Complexes ◽  
Patient Groups

Abstract Heparin-induced thrombocytopenia (HIT) is caused by platelet-activating anti–platelet factor 4 (PF4)/heparin antibodies. Platelet activation assays that use “washed” platelets are more sensitive for detecting HIT antibodies than platelet-rich plasma (PRP)–based assays. Moreover, heparin-exposed patients vary considerably with respect to the risk of PF4/heparin immunization and, among antibody-positive patients, the risk of subsequent “breakthrough” of clinical HIT with manifestation of thrombocytopenia. We used washed platelets and PRP, standard laboratory HIT tests, and physicochemical methods to identify a plasma factor interfering with PF4/heparin complexes and anti-PF4/heparin antibody–platelet interaction, thus explaining differences in functional assays. To investigate a modulating risk for PF4/heparin immunization and breakthrough of HIT, we also tested 89 plasmas from 2 serosurveillance trials. Fibronectin levels were measured in 4 patient groups exhibiting different degrees of heparin-dependent immunization and expression of HIT. The heat-labile plasma protein, fibronectin, inhibited PF4 binding to platelets in a dose-dependent fashion, particularly in washed (vs PRP) systems. Fibronectin also inhibited PF4/heparin binding to platelets, anti-PF4/heparin antibody binding to PF4/heparin complexes, and anti-PF4/heparin antibody–induced platelet activation as a result of PF4/heparin complex disruption. In addition, plasma fibronectin levels increased progressively among the following 4 patient groups: enzyme-linked immunosorbent assay (ELISA)+/serotonin-release assay (SRA)+/HIT+ < ELISA+/SRA+/HIT− ∼ ELISA+/SRA−/HIT− < ELISA−/SRA−/HIT−. Altogether, these findings suggest that fibronectin interferes with PF4/heparin complex formation and anti-PF4/heparin antibody–induced platelet activation. Reduced fibronectin levels in washed platelet assays help to explain the greater sensitivity of washed platelet (vs PRP) assays for HIT. More importantly, lower plasma fibronectin levels could represent a risk factor for PF4/heparin immunization and clinical breakthrough of HIT.

THE REDUCED CAPACITY OF THROMBOXANE FORMATION IN THROMBIN-PREACTIVATED PLATELETS IS NOT CAUSED BY DEPLETION OF THEIR ARACHIDONIC ACID POOL

1987 ◽  
Author(s):  
R E Scharf ◽  
M Stockschläder ◽  
H J Reimers ◽  
W Schneider
Keyword(s):  
Arachidonic Acid ◽  
Human Platelets ◽  
Thrombin Receptor ◽  
Precursor Pool ◽  
Acid Pool ◽  
Exogenous Arachidonic Acid ◽  
High Concentrations ◽  
Short Time ◽  
Thromboxane Formation

Thromboxane (TX) synthesis of washed human platelets pretreated with high concentrations of thrombin (0.5-2.0 U/ml) for 20 sec is significantly reduced upon further thrombin stimulation. Compared to controls (tyrode-pretreated platelets), thrombin-preactivated platelets recover normal TX synthesis following exposure to exogenous arachidonic acid (AA) indicating that short-time thrombin treatment does not inactivate platelet cyclooxygenase or TX synthetase (Blood 63: 858, 1984). To evaluate whether the reduced TX synthesis upon -the second thrombin exposure is due to depletion of their AA precursor pool, thrombin-pretreated platelets and tyrode-pretreated platelets (5×108/ml) were resuspended in autologous ACD plasma and incubated at 37°C with 0.2 μCi 14C-AA (20 μM) for 60 to 90 min in the presence of PGE1 (10 μM). Mean platelet uptake of 14C-AA (disappearance of radioactivity from the supernatant) was 12+3 nmoles AA/109 platelets and did not differ significantly between thrombin-pretreated platelets and controls. Thrombin-pretreated platelets released 10% or 4.5% of their 14c-activity upon further exposure to thrombin (2 U/ml) or collagen (8 μg/ml), respectively. The release from control platelets (15% with thrombin, 6.5% with collagen) did not differ from that of thrombin-pretreated platelets. However, even after incubation in ACD plasma, thrombin-pretreated platelets continued to form significantly less TXB2 (5.0±1.6 nmoles/109 platelets) than controls (9.7±2.2 nmoles/109 platelets, p< 0.05). These data indicate that the reduced capacity of thrombin-pretreated platelets is due neither to a depletion of the endogenous AA pool nor to an inactivation of cyclooxygenase or TX synthetase. The reduced TX synthesis capacity may be caused by a modification, destruction or desensitization of the platelet thrombin receptor as a consequence of the preceding thrombin stimulation.

Abstract 492: Arachidonic Acid Induces Activation of Platelet PF4 and Par-1 mRNA, Which Is Attenuated by Aspirin

2012 ◽  
Vol 32 (suppl_1) ◽  
Author(s):  
Liang Hu ◽  
Michael A Nardi ◽  
Michael Merolla ◽  
Yajaira Suarez ◽  
Jeffrey Berger
Keyword(s):  
Arachidonic Acid ◽  
Platelet Activation ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Thiazole Orange ◽  
Platelet Activity ◽  
Protein Levels ◽  
Reticulated Platelets ◽  
Mrna And Protein Expression ◽  
Dose Dependent

Arachidonic acid (AA) is converted to thromboxane A2 via the cyclooxygenase pathway; however its exact mechanism of platelet activation is uncertain. Inhibition of this pathway via aspirin highlights the importance of this pathway in decreasing thrombotic events. In the present study, we investigate the effect of AA on platelet activity indicators (leukocyte- and monocyte-platelet aggregation [LPA, MPA] and reticulated platelets [RP]), as well as the expression (mRNA and protein) of platelet markers PF4 and Par-1, previously well established platelet transcripts with quantitative determinations. To this end, whole blood was incubated with AA (150mM) for 30 min at room temperature in the absence or presence of aspirin (1mM) prior to addition of antibodies for platelet activity indicators, and isolating platelets for mRNA and protein expression. LPA and MPA were significantly increased after AA stimulation in a dose dependent manner, and were inhibited by aspirin treatment. AA significantly increased PF4 and Par-1 protein level as determined by flow cytometry and western blot assays. Pretreatment with aspirin also attenuated this increase in protein levels. Surprisingly, AA stimulation significantly increased thiazole orange staining (a measure of nucleic acids), another marker of increased platelet activity. Importantly, these results suggest that AA-mediated platelet activation produced an overall increase in platelet total RNA content. To confirm these findings, we analyzed the mRNA expression of PF4 and Par-1 by quantitative real time PCR from platelets treated with AA. Interestingly, AA significantly up-regulated the platelet mRNA transcripts of PF4 and Par-1 by 40% to 60%, and pretreatment with aspirin completely attenuated this effect supporting the specificity of the AA effect on platelet RNA. Altogether, these data suggest that platelet mRNA is affected by AA stimulation, which is attenuated by pretreatment with aspirin. However, the mechanisms responsible for the increased mRNA levels and expression of PF4 and Par-1 (processing of pre-RNA to mRNA) require further investigation. Importantly, our findings provide novel insight regarding platelet activation and a better understanding of mediators in the processes of thrombosis and hemostasis.

Sign in / Sign up

Export Citation Format

Share Document