Fish oil fatty acids and human platelets: Dose-dependent decrease in dienoic and increase in trienoic thromboxane generation

doi:10.1016/0006-2952(96)00473-X

Biochemical Pharmacology

Volume 52, Issue 8, 25 October 1996, Pages 1211-1217

https://doi.org/10.1016/0006-2952(96)00473-X Get rights and content

Abstract

Dietary enrichment of membrane phospholipids with n-3 (fish-oil-derived) fatty acids has attracted attention as a putative therapeutic regimen for suppression of inflammatory and coagulatory events. Use of n-3 fatty-acid-enriched lipid infusions for parenteral nutrition results in micromolar concentrations of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DCHA) in the plasma-free fatty acid fraction. We investigated the influence of free EPA and DCHA on platelet thromboxane (Tx) A₂ and A₃ formation by using a recently developed high performance liquid chromatography-ELISA technique for separate quantification of the stable hydrolysis products TxB₂ and TxB₃. Washed human thrombocytes were incubated with free arachidonic acid (AA; 1 μM), A23187 (0.1 μM) or thrombin (5 U/mL) for stimulation; all regimens provoked large quantities of TxA₂ in the absence of TxA₃. Simultaneous admixture of free EPA or free DCHA to the incubation medium (concentration range, 0.01–50 μM) largely suppressed platelet TxA₂ generation in response to all stimuli used in a dose-dependent manner. The effective concentration with 50% influence of arachidonic acid was 4.2 μM, whereas the inhibitory concentration with 50% effect of EPA and DCHA were both in the same order of magnitude but differed with the nature of the agonist (0.2–7 μM). Platelet (co-)incubation with EPA, but not DCHA, provoked dose-dependent synthesis of n-3-lipid-derived thromboxane: kinetics of formation and absolute quantities of TxA₃ approximated 20% of the respective TxA₂ data upon stimulation with AA. Both EPA and DCHA dose-dependently suppressed U46619-provoked platelet aggregation. We conclude that EPA and DCHA are potent competitive inhibitors of TxA₂ generation by intact platelets, with EPA acting as poor substrate and DCHA being no substrate for the cyclooxygenase/thromboxane synthase complex. Enrichment of the plasma-free fatty acid fraction with n-3 lipids may offer a therapeutic regimen to suppress the synthesis of the potent proaggregatory and vasoconstrictory agent TxA₂.

References (36)

PC Weber et al.
The conversion of dietary eicosapentaenoic acid to prostanoids and leukotrienes in man
Prog Lipid Res
(1986)
J Dyerberg et al.
Eicosapentaenoic acid and prevention of thrombosis and atherosclerosis?
Lancet
(1978)
S Fischer et al.
The prostacyclin/thromboxane balance is favourably shifted in Greenland eskimos
Prostaglandins
(1986)
M Croset et al.
Different metabolic behavior of long-chain n-3 polyunsaturated fatty acids in human platelets.
Biochim Biophys Acta
(1988)
SF O'Keefe et al.
Trans n-3 eicosapentaenoic and docosahexaenoic acid isomers exhibit different inhibitory effects on arachidonic acid metabolism in human platelets compared to the respective cis fatty acids
J Lipid Res
(1990)
JC Bordet et al.
Modulation of prostanoid formation by various polyunsaturated fatty acids during platelet-endothelial cell interactions
Prostagl Leukotr Essent Fatty Acids
(1990)
PG Swann et al.
Enrichment of platelet phospholipids with eicosapentaenoic acid and docosahexaenoic acid inhibits thromboxane A₂/prostaglandin H₂ receptor binding and function
J Biol Chem
(1990)
K Yamazaki et al.
Changes in fatty acid composition in rat blood and organs after infusion of docosahexaenoic acid ethyl ester
Am J Clin Nutr
(1991)
T Hamazaki et al.
The infusion of trieicosapentaenoyl-glycerol into humans and the in vivo formation of prostaglandin 13 and thromboxane A3
Biochem Biophys Res Commun
(1988)
M Urakaze et al.
Infusion of fish oil emulsion: effects on platelet aggregation and fatty acid composition in phospholipids of plasma, platelets, and red blood cell membranes in rabbits
Am J Clin Nutr
(1987)

M Urakaze et al.

Infusion of emulsified trieicosapentaenoylglycerol into rabbits—the effects on platelet aggregation, polymorphonuclear leukocyte adhesion, and fatty acid composition in plasma and platelet phospholipids

Thromb Res

(1986)

HJ Krämer et al.

Analysis of 2- and 3-series prostanoids by post-HPLC-ELISA

Anal Biochem

(1993)

A Leaf et al.

Medical progress. Cardiovascular effects of n-3 fatty acids

N Engl J Med

(1988)

C von Schacky et al.

Long-term effects of dietary marine n-3 fatty acids upon plasma and cellular lipids, platelet function and eicosanoid formation in humans

J Clin Invest

(1985)

C Gronn et al.

Dietary n-6 fatty acids inhibit the incorporation of dietary n-3 fatty acids in thrombocyte and serum phospholipids in humans: a controlled dietetic study

Scand. J Clin Lab Invest

(1991)

P Needleman et al.

Triene prostaglandins: prostacyclin and thromboxane biosynthesis and unique biological properties

F Grimminger et al.

Differential vasoconstrictor potencies of free fatty acids in the lung vasculature: 2- versus 3-series prostanoid generation

J Pharmacol Exp Ther

(1993)

N Chetty et al.

Eicosapentaenoic acid interferes with U46619-stimulated formation of inositol phosphates in washed rabbit platelets

Thromb Hemostas

(1989)

Cited by (74)

Role of cytochrome P450-epoxygenase and soluble epoxide hydrolase in the regulation of vascular response
2023, Advances in Pharmacology
The role of cytochrome P450-epoxygenase has been seen in cardiovascular physiology and pathophysiology. The aberration in CYP450-epoxygenase genes occur due to genetic polymorphisms, aging, or environmental factors, that increase susceptibility to cardiovascular diseases (CVDs). The actual role played by the CYP450-epoxygenases is the metabolism of arachidonic acid (AA) and linoleic acid (LA) into epoxyeicosatrienoic acids (EETs) and epoxyoctadecaenoic acid (EpOMEs) metabolites (oxylipins) and others, which is involved in vasodilation and myocardial-protection. But the genetic polymorphisms in CYP450-epoxygenases lose their beneficial cardiovascular effects of oxylipins, and the soluble epoxide hydrolase (sEH) antagonizes beneficial oxylipins into diols. Like sEH converts EETs into dihydroxyeicosatrienoic acid (DHETs), EpOMEs into dihydroxyoctadecaenoic acid (DiHOMEs), and reverses its beneficial effects, and the sEH gene (Ephx2) polymorphisms cause the enzyme to become overactive and convert epoxy-fatty acids into diols, making them vulnerable to CVDs, including hypertension. Other, enzymes like ω-hydroxylases (CYP450-4A11 & CYP450-4F2)-derived oxylipins from AA, ω-terminal-hydroxyeicosatetraenoic acids (19-, 20-HETE), lipoxygenase-derived oxylipins, mid-chain hydroxyeicosatetraenoic acids (5-, 11-, 12-, 15-HETEs), and the cyclooxygenase-derived prostanoids (prostaglandins: PGD₂, PGF_2α; thromboxane: Txs, oxylipins) are involved in vasoconstriction, hypertension, inflammation, and cardiac toxicity. Also, there are significant interactions were seen between adenosine receptors [adenosine A_2A receptor (A_2AAR) and adenosine A₁ receptor (A₁AR)] with CYP450-epoxygenases, ω-hydroxylases, sEH, and their derived oxylipins in the regulation of the cardiovascular response. Moreover, polymorphisms exist in CYP450-epoxygenases, ω-hydroxylases, sEH, and the adenosine receptor genes in populations associated with CVDs. This chapter will discuss the role of oxylipins' interactions with adenosine receptors in cardiovascular function/dysfunction in mice and humans.
Impact of the eicosapentaenoic acid to arachidonic acid ratio on plaque characteristics in statin-treated patients with coronary artery disease
2023, Journal of Clinical Lipidology
Citation Excerpt :
A low eicosapentaenoic acid (EPA)/arachidonic acid (AA) ratio is an indicator of increased risk for cardiovascular events.1,2 Preclinical and clinical studies have demonstrated that EPA stabilizes plaques by inducing an anti-inflammatory response and reducing platelet aggregation.3,4,5 In contrast, AA facilitates plaque instability through the activation of inflammatory processes and platelet activation.6,7
A low eicosapentaenoic acid (EPA)/arachidonic acid (AA) ratio is associated with an increased risk of cardiovascular events in patients with coronary artery disease (CAD).
To clarify the impact of the EPA/AA ratio on the characteristics of non-culprit coronary plaques in statin-treated patients with CAD.
A total of 370 consecutive stable coronary disease patients treated with statins, who underwent percutaneous coronary intervention for the culprit lesion and optical coherence tomography (OCT) imaging of the non-culprit plaque in a culprit vessel were included. The characteristics of non-culprit plaques assessed using OCT were compared between the lower EPA/AA group (EPA/AA <0.4, n = 255) and the higher EPA/AA group (EPA/AA ≥0.4, n = 115).
The prevalence of lipid-rich plaque (58.8 vs. 41.7%, p = 0.003) and plaque with macrophages (56.5 vs. 31.3%, p <0.001) was significantly higher in the lower EPA/AA group than in the higher EPA/AA group. This association was observed even if the LDL-C level was <100 mg/dL. The prevalence of thin-cap fibroatheroma was significantly higher in patients with lower EPA/AA and higher LDL-C (≥100 mg/dL) than in those with higher EPA/AA and lower LDL-C (<100 mg/dL) (odds ratio: 2.750, 95% confidence interval: 1.182-6.988, p = 0.024). An EPA/AA <0.4 was independently associated with a higher prevalence of lipid-rich plaque, plaque with macrophages, and cholesterol crystals.
Lower EPA/AA ratio was associated with higher prevalence of vulnerable characteristics in non-culprit plaques. The present results suggest the importance of EPA/AA ratio on the secondary prevention of CAD.
Crosstalk between adenosine receptors and CYP450-derived oxylipins in the modulation of cardiovascular, including coronary reactive hyperemic response
2022, Pharmacology and Therapeutics
Citation Excerpt :
But, sEH inhibition by using AUDA effects of EpOME/DiHOME ratio improved renal recovery in response to ischemia/reperfusion injury in C57BL/6 mice (Lee et al., 2012) (Table S6). LA gets metabolized through CYP450-epoxygenases to form hydroxyl-LA metabolites (9-, 13-hydroxyoctadecadienoic acids [9-HODE and 13-HODEs]) (Hanif et al., 2016a; Hanif et al., 2017; Konkel & Schunck, 2011; Tam, 2013; Thuresson et al., 2000), and the HODEs were associated with oxidative stress and the 9-HODE was involved in inducing macrophage IL-1β (Folcik & Cathcart, 1994; Honn et al., 1992; Kramer et al., 1996). Interestingly, 9- and 13-HODEs stimulate plasminogen activator inhibitor type-1 involving PPARγ in endothelial cells, and 13-HODE has anti-inflammatory activity with PPARγ-agonist in inflammatory diseases (Altmann et al., 2007; Belvisi & Mitchell, 2009; Emerson & LeVine, 2004; Fritsche, 2008; Marmol et al., 1999; Stoll et al., 1994).
Adenosine is a ubiquitous endogenous nucleoside or autacoid that affects the cardiovascular system through the activation of four G-protein coupled receptors: adenosine A₁ receptor (A₁AR), adenosine A_2A receptor (A_2AAR), adenosine A_2B receptor (A_2BAR), and adenosine A₃ receptor (A₃AR). With the rapid generation of this nucleoside from cellular metabolism and the widespread distribution of its four G-protein coupled receptors in almost all organs and tissues of the body, this autacoid induces multiple physiological as well as pathological effects, not only regulating the cardiovascular system but also the central nervous system, peripheral vascular system, and immune system. Mounting evidence shows the role of CYP450-enzymes in cardiovascular physiology and pathology, and the genetic polymorphisms in CYP450s can increase susceptibility to cardiovascular diseases (CVDs). One of the most important physiological roles of CYP450-epoxygenases (CYP450-2C & CYP2J2) is the metabolism of arachidonic acid (AA) and linoleic acid (LA) into epoxyeicosatrienoic acids (EETs) and epoxyoctadecaenoic acid (EpOMEs) which generally involve in vasodilation. Like an increase in coronary reactive hyperemia (CRH), an increase in anti-inflammation, and cardioprotective effects. Moreover, the genetic polymorphisms in CYP450-epoxygenases will change the beneficial cardiovascular effects of metabolites or oxylipins into detrimental effects. The soluble epoxide hydrolase (sEH) is another crucial enzyme ubiquitously expressed in all living organisms and almost all organs and tissues. However, in contrast to CYP450-epoxygenases, sEH converts EETs into dihydroxyeicosatrienoic acid (DHETs), EpOMEs into dihydroxyoctadecaenoic acid (DiHOMEs), and others and reverses the beneficial effects of epoxy-fatty acids leading to vasoconstriction, reducing CRH, increase in pro-inflammation, increase in pro-thrombotic and become less cardioprotective. Therefore, polymorphisms in the sEH gene (Ephx2) cause the enzyme to become overactive, making it more vulnerable to CVDs, including hypertension. Besides the sEH, ω-hydroxylases (CYP450-4A11 & CYP450-4F2) derived metabolites from AA, ω terminal-hydroxyeicosatetraenoic acids (19-, 20-HETE), lipoxygenase-derived mid-chain hydroxyeicosatetraenoic acids (5-, 11-, 12-, 15-HETEs), and the cyclooxygenase-derived prostanoids (prostaglandins: PGD₂, PGF_2α; thromboxane: Txs, oxylipins) are involved in vasoconstriction, hypertension, reduction in CRH, pro-inflammation and cardiac toxicity. Interestingly, the interactions of adenosine receptors (A_2AAR, A₁AR) with CYP450-epoxygenases, ω-hydroxylases, sEH, and their derived metabolites or oxygenated polyunsaturated fatty acids (PUFAs or oxylipins) is shown in the regulation of the cardiovascular functions. In addition, much evidence demonstrates polymorphisms in CYP450-epoxygenases, ω-hydroxylases, and sEH genes (Ephx2) and adenosine receptor genes (ADORA1 & ADORA2) in the human population with the susceptibility to CVDs, including hypertension. CVDs are the number one cause of death globally, coronary artery disease (CAD) was the leading cause of death in the US in 2019, and hypertension is one of the most potent causes of CVDs. This review summarizes the articles related to the crosstalk between adenosine receptors and CYP450-derived oxylipins in vascular, including the CRH response in regular salt-diet fed and high salt-diet fed mice with the correlation of heart perfusate/plasma oxylipins. By using A_2AAR^−/−, A₁AR^−/−, eNOS^−/−, sEH^−/− or Ephx2^−/−, vascular sEH-overexpressed (Tie2-sEH Tr), vascular CYP2J2-overexpressed (Tie2-CYP2J2 Tr), and wild-type (WT) mice. This review article also summarizes the role of pro-and anti-inflammatory oxylipins in cardiovascular function/dysfunction in mice and humans. Therefore, more studies are needed better to understand the crosstalk between the adenosine receptors and eicosanoids to develop diagnostic and therapeutic tools by using plasma oxylipins profiles in CVDs, including hypertensive cases in the future.
Lipid Mediators in Cardiovascular Physiology and Disease
2022, Cardiovascular Signaling in Health and Disease
Efficacy and Safety of Omega-3 Fatty Acids in the Prevention of Cardiovascular Disease: A Systematic Review and Meta-analysis
2022, Cardiovascular Drugs and Therapy
Icosapent ethyl for reduction of persistent cardiovascular risk: a critical review of major medical society guidelines and statements
2022, Expert Review of Cardiovascular Therapy

View all citing articles on Scopus

^☆: We thank Dr. J. Mollenhauer and Prof. K. Brune, Erlangen (Germany) for supplying the monoclonal anti-Tx antibody, F. Michnacs for excellent technical assistance and Dr. R. Snipes for thorough linguistic editing of the manuscript.

^∗: This manuscript includes portions of a doctoral thesis by J. Stevens.

View full text

Research paperFish oil fatty acids and human platelets: Dose-dependent decrease in dienoic and increase in trienoic thromboxane generation☆

Abstract

Prog Lipid Res

Lancet

Prostaglandins

Biochim Biophys Acta

J Lipid Res

Prostagl Leukotr Essent Fatty Acids

J Biol Chem

Am J Clin Nutr

Biochem Biophys Res Commun