Prevention of arterial thrombotic diseases has high priority in developed countries. As inappropriate diet is known to enhance the risk for acute thrombotic events, a regular diet with experimentally proven antithrombotic effect might be a beneficial way of prevention. The present study is part of a series of investigations testing fruits and vegetables for antithrombotic activity. The Global Thrombosis Test was used to screen sixteen different apple varieties for antiplatelet and thrombolytic activities. The in vitro effective varieties were further investigated using the laser-induced thrombosis model in mice. In order to investigate the mechanism, hemostatometry and flow-mediated vasodilation test (FMV) were performed. Apple varieties were grouped into subclasses according to their antithrombotic activity. AP-2, AP-13, AP-14 and AP-15 showed significant antithrombotic effect both in vitro and in vivo. AP-8 was antithrombotic in vitro but could not determine in vivo because of the shortage of the sample. The antithrombotic effect was mainly due to activation of endogenous thrombolytic mechanism. The mechanism of such enhanced thrombolysis was investigated using a synthetic inhibitor highly specific to plasmin, polyclonal IgG to t-PA and u-PA, and testing the antithrombotic effect in t-PA knockout mice. Antithrombotic activity was prevented by the synthetic plasmin inhibitor and IgG against t-PA but not against u-PA. There was no antithrombotic activity in t-PA knockout mice. Intake of antithrombotic apple filtrate did not affect FMV and platelet reactivity. The antithrombotic effect was heat stable at 100 °C for 10 min. These results suggest that the mechanism of antithrombotic activity involves an increased t-PA release from the vascular endothelium and an inhibition of plasminogen activator inhibitor (PAI-1) release from activated platelets. The present findings justify including antithrombotic apple varieties in an antithrombotic diet.
1. A.H. Lichtenstein L.J. Appel M. Brands M. Carnethon S. Daniels H.A. Franch B. Franklin P. Kris-Etherton W.S. Harris B. Howard N. Karanja M. Lefevre L. Rudel F. Sacks L. Van Horn M. Winston J. Wylie-Rosett 2006 Summary of American heart association diet and lifestyle recommendations revision Arterioscler Thromb Vasc Biol 26 2186 2191.
2. T.L. Ulbricht D.A. Southgate 1991 Coronary heart disease: seven dietary factors Lancet 338 985 992.
3. S. Renaud M. de Lorgeril 1992 Wine, alcohol, platelets, and the French paradox for coronary heart disease Lancet 339 1523 1526.
4. K.J. Joshipura A. Ascherio J.E. Manson M.J. Stampfer E.B. Rimm F.E. Speizer C.H. Hennekens D. Spiegelman W.C. Willett 1999 Fruit and vegetable intake in relation to risk of ischemic stroke J Am Med Assoc 282 1233 1239.
5. S. Liu J.E. Manson I.M. Lee S.R. Cole C.H. Hennekens W.C. Willett J.E. Buring 2000 Fruit and vegetable intake and risk of cardiovascular disease: the women's health study Am J Clin Nutr 72 922 928.
6. K.J. Joshipura F.B. Hu J.E. Manson M.J. Stampfer E.B. Rimm F.E. Speizer G. Colditz A. Ascherio B. Rosner D. Spiegelman W.C. Willett 2001 The effect of fruit and vegetable intake on risk for coronary heart disease Ann Intern Med 134 1106 1114.
7. L.A. Bazzano J. He L.G. Ogden C.M. Loria S. Vupputuri L. Myers P.K. Whelton 2002 Fruit and vegetable intake and risk of cardiovascular disease in US adults: the first national health and nutrition examination survey epidemiologic follow-up study Am J Clin Nutr 76 93 99.
8. C.P. Ratnatunga S.F. Edmondson G.M. Rees I.B. Kovacs 1982 Highdose aspirin inhibits shear-induced platelet reaction involving thrombin generation Circulation 85 1077 1082.
9. D.A. Gorog I.B. Kovacs 1995 Thrombotic status analyser Thromb Haemostasis 73 514 520.
10. J. Yamamoto T. Yamashita H. Ikarugi T. Taka M. Hashimoto H. Ishii S. Watanabe I.B. Kovacs 2003 Gorog Thrombosis Test: a global in-vitro test of platelet function and thrombolysis Blood Coagul Fibrinolysis 14 31 39.
11. H. Ikarugi T. Yamashita R. Aoki H. Ishii K. Kanki J. Yamamoto 2003 Impairs spontaneous thrombolytic activity in elderly and in habitual smokers, as measured by a new global thrombosis test Blood Coagul Fibrinolysis 14 781 784.
12. J. Yamamoto I.B. Kovacs 2003 Shear-induced in-vitro haemostasis/thrombosis tests: the benefit of using native blood Blood Coagul Fibrinolysis 14 697 702.
13. T. Yamashita A. Sato H. Ikarugi H. Ishii K. Kitamori A. Inoue J. Yamamoto 2005 Significantly reduced spontaneous thrombolytic activity in older men: a possible explanation for the gender differences in risk of acute coronary syndromes Thromb Res 116 127 131.
14. I.B. Kovacs D. Gorog J. Yamamoto 2006 Stimulated spontaneous thrombolysis: a new therapeutic challenge J Thromb Thrombolysis 21 221 227.
15. I.B. Kovacs J. Yamamoto 2006 Review: Spontaneous thrombolysis. A forgotten determinant of life or death Clin Appl Thromb Hemost 12 358 363.
16. Yamamoto J : Prediction of thrombotic status by a new test and prevention of thrombotic disorders by foods with antithrombotic activity. The 7th TTM Forum Report, 203–220 (Japanese) (2007).
17. J. Yamamoto T. Taka K. Yamada Y. Ijiri M. Murakami Y. Hirata A. Naemura M. Hashimoto T. Yamashita K. Oiwa J. Seki H. Suganuma T. Inakuma T. Yoshida 2003 Tomatoes have natural antithrombotic effects Br J Nutr 90 1031 1038.
18. K. Yamada A. Naemura N. Sawashita Y. Noguchi J. Yamamoto 2004 An onion variety has natural antithrombotic effect as assessed by thrombosis/thrombolysis Thromb Res 114 213 220.
19. A. Naemura T. Mitani Y. Ijiri Y. Tamura T. Yamashita M. Okimura J. Yamamoto 2005 Anti-thrombotic effect of strawberries Blood Coagul Fibrinolysis 16 501 509.
20. J. Yamamoto A. Naemura M. Ura Y. Ijiri T. Yamashita A. Kurioka A. Koyama 2006 Testing various fruits for anti-thrombotic effect: I. Mulberries Platelets 17 555 564.
21. J. Yamamoto A. Naemura Y. Ijiri K. Ogawa T. Suzuki Y. Shimada J.C. Giddings 2008 The antithrombotic effects of carrot filtrates in rats and mice Blood Coagul Fibrinolysis 19 785 792.
22. D. Ichiura A. Naemura M. Ura M. Mori J. Yamamoto 2009 Antithrombotic effect of potato in animal experiments Food 3 SpecialIssue2 8 12.
23. C. Kinugasa A. Naemura K. Hyodo Y. Nakai M. Katsuta J. Yamamoto 2011 Experimental antithrombotic effects of sesame seed whole grains and extracts Blood Coagul Fibrinolysis 22 526 531.
24. K. Hyodo I. Horii M. Nishino J.C. Giddings J. Yamamoto 2001 The antithrombotic effects of onion filtrates in rats and mice Health 3 319 325.
25. Y. Okada Y. Tsuda K. Wanaka M. Tada U. Okamoto S. Okamoto A. Hijikata-Okunomiya G. Bokonyi B. Szende G. Keri 2000 Development of plasmin and plasma kallikrein selective inhibitors and their effects on M1 (Melanoma) and HT29 cell lines Bioorg Med Chem Lett 10 2217 2221.
26. I.B. Kovacs A. Tigyi-Sebes K. Trombitas P. Gorog 1975 Evans blue: an ideal energy-absorbing material to produce intravascular microinjury by He-Ne gas laser Microvasc Res 10 107 124.
27. Y. Ijiri M. Miura M. Hashimoto C. Fukunaga S. Watanabe A. Kubota K. Oiwa T. Okuda J. Yamamoto 2002 A new model to evaluate the diet-induced prothrombotic status, using He-Ne laserinduced thrombogenesis in the carotid artery of apolipoprotein E-deficient and low-density lipoprotein receptor-deficient mice Blood Coagul Fibrinolysis 13 497 504.
28. T. Taka Y. Ohta J. Seki J.C. Giddings J. Yamamoto 2002 Impaired flow-mediated vasodilation in vivo and reduced shear-induced platelet reactivity in vitro in response to nitric oxide in prothrombotic, stroke-prone spontaneously hypertensive rats Pathophysiol Haemost Thromb 32 184 189.
29. Y. Tamura A. Naemura A. Inoue Y. Ijiri J. Seki T. Yada M. Goto M. Shinohara S. Kawashima J.C. Giddings J. Yamamoto 2009 Impaired endothelial function may be due to decreased aortic tetrahydrobiopterin, assessed by a new flow-mediated vasodilation in vivo in hypercholesterolemic/atherogenic mice Blood Coagul Fibrinolysis 20 699 705.
30. K. Taomoto H. Ohnishi Y. Kuga K. Nakashima T. Ichioka Y. Kodama H. Kubota T. Tominaga T. Hirose M. Hayashi C. Kinugasa T. Yamashita J. Yamamoto 2010 Platelet function and spontaneous thrombolytic activity of patients with cerebral infarction assessed by Global Thrombosis Test Pathophysiol Haemost Thromb 37 43 48.
31. G. Hsiao Y. Wang N.H. Tzu T.H. Fong M.Y. Shen K.H. Lin D.S. Chou J.R. Sheu 2005 Inhibitory effects of lycopene on in vitro platelet activation and in vivo prevention of thrombus formation J Lab Clin Med 146 216 226.
32. G.P. Hubbard S. Wolffram R. de Vos A. Bovy J.M. Gibbins J.A. Lovegrove 2006 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 96 482 488.
|Medicine (miscellaneous) Q3|
Cite Score Rank
|General Medicine 199/529 (Q2)|
|Interventional Medicine and Applied Science|
|H-1117 Budapest, Hungary 1516 Budapest, PO Box 245.|
|H-1117 Budapest, Hungary 1516 Budapest, PO Box 245.|
|Chief Executive Officer, Akadémiai Kiadó|
|Abstract Views||Full Text Views||PDF Downloads|