RSS-Feed abonnieren
![](/products/assets/desktop/img/oa-logo.png)
DOI: 10.1055/s-0039-1694028
The Metabolites of the Dietary Flavonoid Quercetin Possess Potent Antithrombotic Activity, and Interact with Aspirin to Enhance Antiplatelet Effects
Funding This work was supported by the Biotechnology and Biological Sciences Research Council (BBSRC, grant GS13-04), British Heart Foundation (PG/15/21/31355, PG/13/93/30593, RG/09/011/28094, and RG/15/2/31224), and Medical Research Council (MR/J002666/1).Publikationsverlauf
20. Dezember 2018
24. Juni 2019
Publikationsdatum:
30. Juli 2019 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00033990/201903/lookinside/thumbnails/10-1055-s-0039-1694028_190030-1.jpg)
Abstract
Quercetin, a dietary flavonoid, has been reported to possess antiplatelet activity. However, its extensive metabolism following ingestion has resulted in difficulty elucidating precise mechanisms of action. In this study, we aimed to characterize the antiplatelet mechanisms of two methylated metabolites of quercetin—isorhamnetin and tamarixetin—and explore potential interactions with aspirin. Isorhamnetin and tamarixetin inhibited human platelet aggregation, and suppressed activatory processes including granule secretion, integrin αIIbβ3 function, calcium mobilization, and spleen tyrosine kinase (Syk)/linker for activation of T cells (LAT) phosphorylation downstream of glycoprotein VI with similar potency to quercetin. All three flavonoids attenuated thrombus formation in an in vitro microfluidic model, and isoquercetin, a 3-O-glucoside of quercetin, inhibited thrombosis in a murine laser injury model. Isorhamnetin, tamarixetin, and quercetin enhanced the antiplatelet effects of aspirin more-than-additively in a plate-based aggregometry assay, reducing aspirin IC50 values by an order of magnitude, with this synergy maintained in a whole blood test of platelet function. Our data provide mechanistic evidence for the antiplatelet activity of two quercetin metabolites, isorhamnetin and tamarixetin, and suggest a potential antithrombotic role for these flavonoids. In combination with their interactions with aspirin, this may represent a novel avenue of investigation for the development of new antithrombotic strategies and management of current therapies.
-
References
- 1 Gibbins JM. Platelet adhesion signalling and the regulation of thrombus formation. J Cell Sci 2004; 117 (Pt 16): 3415-3425
- 2 Bye AP, Unsworth AJ, Gibbins JM. Platelet signaling: a complex interplay between inhibitory and activatory networks. J Thromb Haemost 2016; 14 (05) 918-930
- 3 Nieswandt B, Pleines I, Bender M. Platelet adhesion and activation mechanisms in arterial thrombosis and ischaemic stroke. J Thromb Haemost 2011; 9 (Suppl. 01) 92-104
- 4 Floyd CN, Ferro A. Mechanisms of aspirin resistance. Pharmacol Ther 2014; 141 (01) 69-78
- 5 Hilkens NA, Algra A, Kappelle LJ. , et al; CAT Collaboration. Early time course of major bleeding on antiplatelet therapy after TIA or ischemic stroke. Neurology 2018; 90 (08) e683-e689
- 6 Wang X, Ouyang YY, Liu J, Zhao G. Flavonoid intake and risk of CVD: a systematic review and meta-analysis of prospective cohort studies. Br J Nutr 2014; 111 (01) 1-11
- 7 Borgi L, Muraki I, Satija A, Willett WC, Rimm EB, Forman JP. Fruit and vegetable consumption and the incidence of hypertension in three prospective cohort studies. Hypertension 2016; 67 (02) 288-293
- 8 Wu W, Li R, Li X. , et al. Quercetin as an antiviral agent inhibits influenza A virus (IAV) entry. Viruses 2015; 8 (01) E6
- 9 Srivastava S, Somasagara RR, Hegde M. , et al. Quercetin, a natural flavonoid interacts with DNA, arrests cell cycle and causes tumor regression by activating mitochondrial pathway of apoptosis. Sci Rep 2016; 6: 24049
- 10 Knekt P, Kumpulainen J, Järvinen R. , et al. Flavonoid intake and risk of chronic diseases. Am J Clin Nutr 2002; 76 (03) 560-568
- 11 Lee K-H, Park E, Lee H-J. , et al. Effects of daily quercetin-rich supplementation on cardiometabolic risks in male smokers. Nutr Res Pract 2011; 5 (01) 28-33
- 12 Hubbard GP, Wolffram S, Lovegrove JA, Gibbins JM. Ingestion of quercetin inhibits platelet aggregation and essential components of the collagen-stimulated platelet activation pathway in humans. J Thromb Haemost 2004; 2 (12) 2138-2145
- 13 Oh WJ, Endale M, Park S-C, Cho JY, Rhee MH. Dual roles of quercetin in platelets: phosphoinositide-3-kinase and MAP kinases inhibition, and cAMP-dependent vasodilator-stimulated phosphoprotein stimulation. Evid Based Complement Alternat Med 2012; 2012: 485262
- 14 Nemeth K, Piskula MK. Food content, processing, absorption and metabolism of onion flavonoids. Crit Rev Food Sci Nutr 2007; 47 (04) 397-409
- 15 Asselin J, Gibbins JM, Achison M. , et al. A collagen-like peptide stimulates tyrosine phosphorylation of syk and phospholipase C gamma2 in platelets independent of the integrin alpha2beta1. Blood 1997; 89 (04) 1235-1242
- 16 Kaiser WJ, Holbrook LM, Tucker KL, Stanley RG, Gibbins JM. A functional proteomic method for the enrichment of peripheral membrane proteins reveals the collagen binding protein Hsp47 is exposed on the surface of activated human platelets. J Proteome Res 2009; 8 (06) 2903-2914
- 17 Spyridon M, Moraes LA, Jones CI. , et al. LXR as a novel antithrombotic target. Blood 2011; 117 (21) 5751-5761
- 18 Vaiyapuri S, Roweth H, Ali MS. , et al. Pharmacological actions of nobiletin in the modulation of platelet function. Br J Pharmacol 2015; 172 (16) 4133-4145
- 19 Bye AP, Unsworth AJ, Vaiyapuri S, Stainer AR, Fry MJ, Gibbins JM. Ibrutinib inhibits platelet integrin αIIbβ3 outside-in signaling and thrombus stability but not adhesion to collagen. Arterioscler Thromb Vasc Biol 2015; 35 (11) 2326-2335
- 20 Chan MV, Armstrong PCJ, Papalia F, Kirkby NS, Warner TD. Optical multichannel (optimul) platelet aggregometry in 96-well plates as an additional method of platelet reactivity testing. Platelets 2011; 22 (07) 485-494
- 21 Farndale RW, Sixma JJ, Barnes MJ, de Groot PG. The role of collagen in thrombosis and hemostasis. J Thromb Haemost 2004; 2 (04) 561-573
- 22 Kashino Y, Murota K, Matsuda N. , et al. Effect of processed onions on the plasma concentration of quercetin in rats and humans. J Food Sci 2015; 80 (11) H2597-H2602
- 23 Stopa JD, Neuberg D, Puligandla M, Furie B, Flaumenhaft R, Zwicker JI. Protein disulfide isomerase inhibition blocks thrombin generation in humans by interfering with platelet factor V activation. JCI Insight 2017; 2 (01) e89373
- 24 Maynard DM, Heijnen HF, Horne MK, White JG, Gahl WA. Proteomic analysis of platelet alpha-granules using mass spectrometry. J Thromb Haemost 2007; 5 (09) 1945-1955
- 25 Takagi J, Petre BM, Walz T, Springer TA. Global conformational rearrangements in integrin extracellular domains in outside-in and inside-out signaling. Cell 2002; 110 (05) 599-11
- 26 Lee D, Fong KP, King MR, Brass LF, Hammer DA. Differential dynamics of platelet contact and spreading. Biophys J 2012; 102 (03) 472-482
- 27 Stefanini L, Roden RC, Bergmeier W. CalDAG-GEFI is at the nexus of calcium-dependent platelet activation. Blood 2009; 114 (12) 2506-2514
- 28 Hollman PC, de Vries JH, van Leeuwen SD, Mengelers MJ, Katan MB. Absorption of dietary quercetin glycosides and quercetin in healthy ileostomy volunteers. Am J Clin Nutr 1995; 62 (06) 1276-1282
- 29 Paulke A, Eckert GP, Schubert-Zsilavecz M, Wurglics M. Isoquercitrin provides better bioavailability than quercetin: comparison of quercetin metabolites in body tissue and brain sections after six days administration of isoquercitrin and quercetin. Pharmazie 2012; 67 (12) 991-996
- 30 Arts IC, Sesink AL, Faassen-Peters M, Hollman PC. The type of sugar moiety is a major determinant of the small intestinal uptake and subsequent biliary excretion of dietary quercetin glycosides. Br J Nutr 2004; 91 (06) 841-847
- 31 García Rodríguez LA, Martín-Pérez M, Hennekens CH, Rothwell PM, Lanas A. Bleeding risk with long-term low-dose aspirin: a systematic review of observational studies. PLoS One 2016; 11 (08) e0160046
- 32 Baigent C, Blackwell L, Collins R. , et al; Antithrombotic Trialists' (ATT) Collaboration. Aspirin in the primary and secondary prevention of vascular disease: collaborative meta-analysis of individual participant data from randomised trials. Lancet 2009; 373 (9678): 1849-1860
- 33 Hubbard GP, Wolffram S, de Vos R, Bovy A, Gibbins JM, Lovegrove JA. Ingestion of onion soup high in quercetin inhibits platelet aggregation and essential components of the collagen-stimulated platelet activation pathway in man: a pilot study. Br J Nutr 2006; 96 (03) 482-488
- 34 Kundu SK, Heilmann EJ, Sio R, Garcia C, Davidson RM, Ostgaard RA. Description of an in vitro platelet function analyzer--PFA-100. Semin Thromb Hemost 1995; 21 (Suppl. 02) 106-112
- 35 Hubbard GP, Stevens JM, Cicmil M. , et al. Quercetin inhibits collagen-stimulated platelet activation through inhibition of multiple components of the glycoprotein VI signaling pathway. J Thromb Haemost 2003; 1 (05) 1079-1088
- 36 Boly R, Gras T, Lamkami T. , et al. Quercetin inhibits a large panel of kinases implicated in cancer cell biology. Int J Oncol 2011; 38 (03) 833-842
- 37 Böhl M, Tietze S, Sokoll A. , et al. Flavonoids affect actin functions in cytoplasm and nucleus. Biophys J 2007; 93 (08) 2767-2780
- 38 Yang Y, Oh J-M, Heo P. , et al. Polyphenols differentially inhibit degranulation of distinct subsets of vesicles in mast cells by specific interaction with granule-type-dependent SNARE complexes. Biochem J 2013; 450 (03) 537-546
- 39 Hong Y-J, Mitchell AE. Metabolic profiling of flavonol metabolites in human urine by liquid chromatography and tandem mass spectrometry. J Agric Food Chem 2004; 52 (22) 6794-6801
- 40 Lee J, Ebeler SE, Zweigenbaum JA, Mitchell AE. UHPLC-(ESI)QTOF MS/MS profiling of quercetin metabolites in human plasma postconsumption of applesauce enriched with apple peel and onion. J Agric Food Chem 2012; 60 (34) 8510-8520
- 41 Datta A, Huber F, Boettiger D. Phosphorylation of beta3 integrin controls ligand binding strength. J Biol Chem 2002; 277 (06) 3943-3949
- 42 Wu Y, Span LM, Nygren P. , et al. The tyrosine kinase c-Src specifically binds to the active integrin αIIbβ3 to initiate outside-in signaling in platelets. J Biol Chem 2015; 290 (25) 15825-15834
- 43 Guglielmone HA, Agnese AM, Núñez Montoya SC, Cabrera JL. Anticoagulant effect and action mechanism of sulphated flavonoids from Flaveria bidentis. Thromb Res 2002; 105 (02) 183-188
- 44 Choi JH, Kim KJ, Kim S. Comparative effect of quercetin and quercetin-3-O-β-D-glucoside on fibrin polymers, blood clots, and in rodent models. J Biochem Mol Toxicol 2016; 30 (11) 548-558
- 45 Guerrero JA, Navarro-Nuñez L, Lozano ML. , et al. Flavonoids inhibit the platelet TxA(2) signalling pathway and antagonize TxA(2) receptors (TP) in platelets and smooth muscle cells. Br J Clin Pharmacol 2007; 64 (02) 133-144
- 46 Wright B, Moraes LA, Kemp CF. , et al. A structural basis for the inhibition of collagen-stimulated platelet function by quercetin and structurally related flavonoids. Br J Pharmacol 2010; 159 (06) 1312-1325
- 47 Bergmeier W, Stefanini L. Novel molecules in calcium signaling in platelets. J Thromb Haemost 2009; 7 (Suppl. 01) 187-190
- 48 Doucet MS, Jougleux JL, Poirier SJ. , et al. Identification of peracetylated quercetin as a selective 12-lipoxygenase pathway inhibitor in human platelets. Mol Pharmacol 2019; 95 (01) 139-150
- 49 Yeung J, Tourdot BE, Fernandez-Perez P. , et al. Platelet 12-LOX is essential for FcγRIIa-mediated platelet activation. Blood 2014; 124 (14) 2271-2279
- 50 Dufour C, Dangles O. Flavonoid-serum albumin complexation: determination of binding constants and binding sites by fluorescence spectroscopy. Biochim Biophys Acta 2005; 1721 (1–3): 164-173
- 51 Rivera J, Lozano ML, Navarro-Núñez L, Vicente V. Platelet receptors and signaling in the dynamics of thrombus formation. Haematologica 2009; 94 (05) 700-711
- 52 Feng W, Valiyaveettil M, Dudiki T. , et al. β3 phosphorylation of platelet αIIbβ3 is crucial for stability of arterial thrombus and microparticle formation in vivo. Thromb J 2017; 15 (01) 22
- 53 Day AJ, Gee JM, DuPont MS, Johnson IT, Williamson G. Absorption of quercetin-3-glucoside and quercetin-4′-glucoside in the rat small intestine: the role of lactase phlorizin hydrolase and the sodium-dependent glucose transporter. Biochem Pharmacol 2003; 65 (07) 1199-1206
- 54 Srivastava P. Optimization of antiplatelet/antithrombotic therapy for secondary stroke prevention. Ann Indian Acad Neurol 2010; 13 (01) 6-13
- 55 Al-Fayez M, Cai H, Tunstall R, Steward WP, Gescher AJ. Differential modulation of cyclooxygenase-mediated prostaglandin production by the putative cancer chemopreventive flavonoids tricin, apigenin and quercetin. Cancer Chemother Pharmacol 2006; 58 (06) 816-825
- 56 Wang H, Nair MG, Strasburg GM, Booren AM, Gray I, Dewitt DL. Cyclooxygenase active bioflavonoids from Balaton tart cherry and their structure activity relationships. Phytomedicine 2000; 7 (01) 15-19
- 57 Guerrero JA, Lozano ML, Castillo J, Benavente-García O, Vicente V, Rivera J. Flavonoids inhibit platelet function through binding to the thromboxane A2 receptor. J Thromb Haemost 2005; 3 (02) 369-376