PDF herunterladen
CC BY-NC-ND 4.0 · TH Open 2017; 01(02): e92-e100
DOI: 10.1055/s-0037-1605361
Original Article
Georg Thieme Verlag KG Stuttgart · New York

Induction of Diabetes Abolishes the Antithrombotic Effect of Clopidogrel in Apolipoprotein E–Deficient Mice

A. Sugidachi
1   Rare Disease and LCM Laboratories, Daiichi Sankyo Co., Ltd., Tokyo, Japan
,
K. Ohno
1   Rare Disease and LCM Laboratories, Daiichi Sankyo Co., Ltd., Tokyo, Japan
,
J. A. Jakubowski
2   Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana, United States
,
Y. Ito
1   Rare Disease and LCM Laboratories, Daiichi Sankyo Co., Ltd., Tokyo, Japan
,
A. Tomizawa
1   Rare Disease and LCM Laboratories, Daiichi Sankyo Co., Ltd., Tokyo, Japan
,
M. Mizuno
1   Rare Disease and LCM Laboratories, Daiichi Sankyo Co., Ltd., Tokyo, Japan
› Institutsangaben
Weitere Informationen

Publikationsverlauf

Publikationsdatum:
10. August 2017 (online)

   Lizenzen und Reprints

Abstract

Patients with acute coronary syndrome with diabetes mellitus (DM) exhibit an impaired platelet inhibitory response to clopidogrel which is only partially understood. DM was induced by the administration of streptozotocin (STZ) to 9-week-old mice. The antithrombotic effects of clopidogrel (10 mg/kg/d, orally × 5 days) were determined using a FeCl3-induced thrombosis model employing wild-type (WT), apolipoprotein E (apoE)-deficient, and diabetic apoE-deficient mice at 21 weeks. Antiplatelet effects were determined using flow cytometry. The antithrombotic effects of clopidogrel were similar in WT and apoE-deficient mice but were attenuated in diabetic apoE-deficient mice with the percent inhibition of thrombus area (µm2) by clopidogrel being 85.5% (WT mice), 75.0% (apoE-deficient mice), and 1.9% (diabetic apoE-deficient mice). The time to first occlusion and lumen stenosis also reflected a significant loss of the antithrombotic effects of clopidogrel in diabetic apoE-deficient mice. Ex vivo platelet activation, which was assessed using ADP-induced expression of activated glycoprotein IIb/IIIa, was completely inhibited by clopidogrel in these three groups of mice. In contrast, the effect of clopidogrel on the ex vivo expression of platelet P-selectin induced by protease-activated receptor 4–activating peptide was diminished in diabetic apoE-deficient mice compared with that in WT and apoE-deficient mice. These data suggest that diabetic apoE-deficient mice may serve as a useful model to better understand the impaired responses to clopidogrel in patients with DM, which may partially reflect a reduction of the effect of clopidogrel on thrombin-induced platelet activation.

Keywords

P2Y12 - clopidogrel - diabetes mellitus - platelets - thrombosis - apolipoprotein E–deficient mouse - thrombin

Supplementary Material

 

  • References

  • 1 Levine GN, Bates ER, Blankenship JC. , et al; American College of Cardiology Foundation; American Heart Association Task Force on Practice Guidelines; Society for Cardiovascular Angiography and Interventions. 2011 ACCF/AHA/SCAI Guideline for Percutaneous Coronary Intervention. A report of the American College of Cardiology Foundation/American Heart Association Task Force on Practice Guidelines and the Society for Cardiovascular Angiography and Interventions. J Am Coll Cardiol 2011; 58 (24) e44-e122
    CrossrefPubMedGoogle Scholar
  • 2 Anderson JL, Adams CD, Antman EM. , et al. 2012 ACCF/AHA focused update incorporated into the ACCF/AHA 2007 guidelines for the management of patients with unstable angina/non-ST-elevation myocardial infarction: a report of the American College of Cardiology Foundation/American Heart Association Task Force on Practice Guidelines. J Am Coll Cardiol 2013; 61 (23) e179-e347
    PubMedGoogle Scholar
  • 3 Sugidachi A, Ohno K, Ogawa T, Jakubowski J, Hashimoto M, Tomizawa A. A comparison of the pharmacological profiles of prasugrel and ticagrelor assessed by platelet aggregation, thrombus formation and haemostasis in rats. Br J Pharmacol 2013; 169 (01) 82-89
    CrossrefPubMedGoogle Scholar
  • 4 Wiviott SD, Braunwald E, McCabe CH. , et al; TRITON-TIMI 38 Investigators. Prasugrel versus clopidogrel in patients with acute coronary syndromes. N Engl J Med 2007; 357 (20) 2001-2015
    CrossrefPubMedGoogle Scholar
  • 5 Li YG, Ni L, Brandt JT. , et al. Inhibition of platelet aggregation with prasugrel and clopidogrel: an integrated analysis in 846 subjects. Platelets 2009; 20 (05) 316-327
    CrossrefPubMedGoogle Scholar
  • 6 VAN Giezen JJ, Nilsson L, Berntsson P. , et al. Ticagrelor binds to human P2Y12 independently from ADP but antagonizes ADP-induced receptor signaling and platelet aggregation. J Thromb Haemost 2009; 7 (09) 1556-1565
    CrossrefPubMedGoogle Scholar
  • 7 Wijeyeratne YD, Joshi R, Heptinstall S. Ticagrelor: a P2Y12 antagonist for use in acute coronary syndromes. Expert Rev Clin Pharmacol 2012; 5 (03) 257-269
    CrossrefPubMedGoogle Scholar
  • 8 Ferreiro JL, Angiolillo DJ. New directions in antiplatelet therapy. Circ Cardiovasc Interv 2012; 5 (03) 433-445
    CrossrefPubMedGoogle Scholar
  • 9 Aradi D, Komócsi A, Price MJ. , et al; Tailored Antiplatelet Treatment Study Collaboration. Efficacy and safety of intensified antiplatelet therapy on the basis of platelet reactivity testing in patients after percutaneous coronary intervention: systematic review and meta-analysis. Int J Cardiol 2013; 167 (05) 2140-2148
    CrossrefPubMedGoogle Scholar
  • 10 Angiolillo DJ, Fernandez-Ortiz A, Bernardo E. , et al. Variability in individual responsiveness to clopidogrel: clinical implications, management, and future perspectives. J Am Coll Cardiol 2007; 49 (14) 1505-1516
    CrossrefPubMedGoogle Scholar
  • 11 Bonello L, Tantry US, Marcucci R. , et al; Working Group on High On-Treatment Platelet Reactivity. Consensus and future directions on the definition of high on-treatment platelet reactivity to adenosine diphosphate. J Am Coll Cardiol 2010; 56 (12) 919-933
    CrossrefPubMedGoogle Scholar
  • 12 Price MJ, Coleman JL, Steinhubl SR, Wong GB, Cannon CP, Teirstein PS. Onset and offset of platelet inhibition after high-dose clopidogrel loading and standard daily therapy measured by a point-of-care assay in healthy volunteers. Am J Cardiol 2006; 98 (05) 681-684
    CrossrefPubMedGoogle Scholar
  • 13 Price MJ. Diabetes mellitus and clopidogrel response variability. J Am Coll Cardiol 2014; 64 (10) 1015-1018
    CrossrefPubMedGoogle Scholar
  • 14 Price MJ, Angiolillo DJ, Teirstein PS. , et al. Platelet reactivity and cardiovascular outcomes after percutaneous coronary intervention: a time-dependent analysis of the Gauging Responsiveness with a VerifyNow P2Y12 assay: Impact on Thrombosis and Safety (GRAVITAS) trial. Circulation 2011; 124 (10) 1132-1137
    CrossrefPubMedGoogle Scholar
  • 15 Stone GW, Witzenbichler B, Weisz G. , et al; ADAPT-DES Investigators. Platelet reactivity and clinical outcomes after coronary artery implantation of drug-eluting stents (ADAPT-DES): a prospective multicentre registry study. Lancet 2013; 382 (9892): 614-623
    CrossrefPubMedGoogle Scholar
  • 16 Brar SS, ten Berg J, Marcucci R. , et al. Impact of platelet reactivity on clinical outcomes after percutaneous coronary intervention. A collaborative meta-analysis of individual participant data. J Am Coll Cardiol 2011; 58 (19) 1945-1954
    CrossrefPubMedGoogle Scholar
  • 17 Price MJ, Baker BA, Jakubowski JA, Li W, Heiselman DE, Angiolillo DJ. Detecting a thienopyridine effect by platelet reactivity assessment and its implications for risk stratification. J Thromb Haemost 2014; 12 (04) 560-563
    CrossrefPubMedGoogle Scholar
  • 18 Gurbel PA, Bliden KP, Hiatt BL, O'Connor CM. Clopidogrel for coronary stenting: response variability, drug resistance, and the effect of pretreatment platelet reactivity. Circulation 2003; 107 (23) 2908-2913
    CrossrefPubMedGoogle Scholar
  • 19 Campo G, Parrinello G, Ferraresi P. , et al. Prospective evaluation of on-clopidogrel platelet reactivity over time in patients treated with percutaneous coronary intervention relationship with gene polymorphisms and clinical outcome. J Am Coll Cardiol 2011; 57 (25) 2474-2483
    CrossrefPubMedGoogle Scholar
  • 20 Price MJ, Murray SS, Angiolillo DJ. , et al; GIFT Investigators. Influence of genetic polymorphisms on the effect of high- and standard-dose clopidogrel after percutaneous coronary intervention: the GIFT (Genotype Information and Functional Testing) study. J Am Coll Cardiol 2012; 59 (22) 1928-1937
    CrossrefPubMedGoogle Scholar
  • 21 Hochholzer W, Trenk D, Fromm MF. , et al. Impact of cytochrome P450 2C19 loss-of-function polymorphism and of major demographic characteristics on residual platelet function after loading and maintenance treatment with clopidogrel in patients undergoing elective coronary stent placement. J Am Coll Cardiol 2010; 55 (22) 2427-2434
    CrossrefPubMedGoogle Scholar
  • 22 Shuldiner AR, O'Connell JR, Bliden KP. , et al. Association of cytochrome P450 2C19 genotype with the antiplatelet effect and clinical efficacy of clopidogrel therapy. JAMA 2009; 302 (08) 849-857
    CrossrefPubMedGoogle Scholar
  • 23 Ferreira IA, Mocking AI, Feijge MA. , et al. Platelet inhibition by insulin is absent in type 2 diabetes mellitus. Arterioscler Thromb Vasc Biol 2006; 26 (02) 417-422
    PubMedGoogle Scholar
  • 24 Kannel WB, McGee DL. Diabetes and glucose tolerance as risk factors for cardiovascular disease: the Framingham study. Diabetes Care 1979; 2 (02) 120-126
    CrossrefPubMedGoogle Scholar
  • 25 Stamler J, Vaccaro O, Neaton JD, Wentworth D. Diabetes, other risk factors, and 12-yr cardiovascular mortality for men screened in the Multiple Risk Factor Intervention Trial. Diabetes Care 1993; 16 (02) 434-444
    CrossrefPubMedGoogle Scholar
  • 26 Angiolillo DJ, Fernandez-Ortiz A, Bernardo E. , et al. Platelet function profiles in patients with type 2 diabetes and coronary artery disease on combined aspirin and clopidogrel treatment. Diabetes 2005; 54 (08) 2430-2435
    CrossrefPubMedGoogle Scholar
  • 27 Angiolillo DJ. Antiplatelet therapy in diabetes: efficacy and limitations of current treatment strategies and future directions. Diabetes Care 2009; 32 (04) 531-540
    CrossrefPubMedGoogle Scholar
  • 28 Ang L, Palakodeti V, Khalid A. , et al. Elevated plasma fibrinogen and diabetes mellitus are associated with lower inhibition of platelet reactivity with clopidogrel. J Am Coll Cardiol 2008; 52 (13) 1052-1059
    CrossrefPubMedGoogle Scholar
  • 29 Angiolillo DJ, Jakubowski JA, Ferreiro JL. , et al. Impaired responsiveness to the platelet P2Y12 receptor antagonist clopidogrel in patients with type 2 diabetes and coronary artery disease. J Am Coll Cardiol 2014; 64 (10) 1005-1014
    CrossrefPubMedGoogle Scholar
  • 30 Fledderus JO, van Oostrom O, de Kleijn DP. , et al. Increased amount of bone marrow-derived smooth muscle-like cells and accelerated atherosclerosis in diabetic apoE-deficient mice. Atherosclerosis 2013; 226 (02) 341-347
    CrossrefPubMedGoogle Scholar
  • 31 Zhang Y, Wang X, Wang Y, Liu Y, Xia M. Supplementation of cyanidin-3-O-β-glucoside promotes endothelial repair and prevents enhanced atherogenesis in diabetic apolipoprotein E-deficient mice. J Nutr 2013; 143 (08) 1248-1253
    CrossrefPubMedGoogle Scholar
  • 32 Lu Z, Zhang X, Li Y, Jin J, Huang Y. TLR4 antagonist reduces early-stage atherosclerosis in diabetic apolipoprotein E-deficient mice. J Endocrinol 2013; 216 (01) 61-71
    CrossrefPubMedGoogle Scholar
  • 33 Lazaro I, Oguiza A, Recio C. , et al. Targeting HSP90 ameliorates nephropathy and atherosclerosis through suppression of NF-κB and STAT signaling pathways in diabetic mice. Diabetes 2015; 64 (10) 3600-3613
    CrossrefPubMedGoogle Scholar
  • 34 Ohno K, Tomizawa A, Jakubowski JA, Mizuno M, Sugidachi A. Prevention of occlusive arterial thrombus formation by a single loading dose of prasugrel suppresses neointimal hyperplasia in mice. Thromb Res 2015; 136 (06) 1245-1251
    CrossrefPubMedGoogle Scholar
  • 35 Ohno K, Tomizawa A, Mizuno M, Jakubowski JA, Sugidachi A. Prasugrel, a platelet P2Y12 receptor antagonist, improves abnormal gait in a novel murine model of thrombotic hindlimb ischemia. J Am Heart Assoc 2016; 5 (04) e002889
    CrossrefPubMedGoogle Scholar
  • 36 Alexander SP, Mathie A, Peters JA. Guide to receptors and channels (GRAC), 5th edition. Br J Pharmacol 2011; 164 (Suppl. 01) S1-S324
    CrossrefPubMedGoogle Scholar
  • 37 Kilkenny C, Browne W, Cuthill IC, Emerson M, Altman DG. ; NC3Rs Reporting Guidelines Working Group. Animal research: reporting in vivo experiments: the ARRIVE guidelines. Br J Pharmacol 2010; 160 (07) 1577-1579
    CrossrefPubMedGoogle Scholar
  • 38 McGrath JC, Drummond GB, McLachlan EM, Kilkenny C, Wainwright CL. Guidelines for reporting experiments involving animals: the ARRIVE guidelines. Br J Pharmacol 2010; 160 (07) 1573-1576
    CrossrefPubMedGoogle Scholar
  • 39 Curtis MJ, Bond RA, Spina D. , et al. Experimental design and analysis and their reporting: new guidance for publication in BJP. Br J Pharmacol 2015; 172 (14) 3461-3471
    CrossrefPubMedGoogle Scholar
  • 40 Kawasaki T, Taniguchi M, Moritani Y. , et al. Pharmacological properties of YM-254890, a specific Gαq/11 inhibitor, on thrombosis and neointima formation in mice. Thromb Haemost 2005; 94 (01) 184-192
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 41 Hu L, Fan Z, Du H. , et al. BF061, a novel antiplatelet and antithrombotic agent targeting P2Y12 receptor and phosphodiesterase. Thromb Haemost 2011; 106 (06) 1203-1214
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 42 Tomizawa A, Ohno K, Jakubowski JA, Mizuno M, Sugidachi A. Prasugrel reduces ischaemic infarct volume and ameliorates neurological deficits in a non-human primate model of middle cerebral artery thrombosis. Thromb Res 2015; 136 (06) 1224-1230
    CrossrefPubMedGoogle Scholar
  • 43 Bhatt DL. What makes platelets angry: diabetes, fibrinogen, obesity, and impaired response to antiplatelet therapy?. J Am Coll Cardiol 2008; 52 (13) 1060-1061
    CrossrefPubMedGoogle Scholar
  • 44 Rozalski M, Kassassir H, Siewiera K, Klepacka A, Sychowski R, Watala C. Platelet activation patterns are different in mouse models of diabetes and chronic inhibition of nitric oxide synthesis. Thromb Res 2014; 133 (06) 1097-1104
    CrossrefPubMedGoogle Scholar
  • 45 Kumbhani DJ, Marso SP, Alvarez CA, McGuire DK. State-of-the-art: hypo-responsiveness to oral antiplatelet therapy in patients with type 2 diabetes mellitus. Curr Cardiovasc Risk Rep 2015; 9: 4
    CrossrefPubMedGoogle Scholar
  • 46 Sagel J, Colwell JA, Crook L, Laimins M. Increased platelet aggregation in early diabetes mellitus. Ann Intern Med 1975; 82 (06) 733-738
    CrossrefPubMedGoogle Scholar
  • 47 Honour AJ, Hockaday TD. Increased sensitivity of in vivo platelet aggregation in rabbits after alloxan or streptozotocin. Br J Exp Pathol 1976; 57 (01) 1-10
    PubMedGoogle Scholar
  • 48 Schuette C, Steffens D, Witkowski M. , et al. The effect of clopidogrel on platelet activity in patients with and without type-2 diabetes mellitus: a comparative study. Cardiovasc Diabetol 2015; 14: 15
    CrossrefPubMedGoogle Scholar
  • 49 Sugidachi A, Asai F, Ogawa T, Inoue T, Koike H. The in vivo pharmacological profile of CS-747, a novel antiplatelet agent with platelet ADP receptor antagonist properties. Br J Pharmacol 2000; 129 (07) 1439-1446
    CrossrefPubMedGoogle Scholar
  • 50 Gachet C, Stierlé A, Cazenave JP. , et al. The thienopyridine PCR 4099 selectively inhibits ADP-induced platelet aggregation and fibrinogen binding without modifying the membrane glycoprotein IIb-IIIa complex in rat and in man. Biochem Pharmacol 1990; 40 (02) 229-238
    CrossrefPubMedGoogle Scholar
  • 51 Jakubowski JA, Zhou C, Egan B. , et al. Modification of the VerifyNow® P2Y12 test BASE channel to accommodate high levels of P2Y12 antagonism. Platelets 2011; 22 (08) 619-625
    CrossrefPubMedGoogle Scholar
  • 52 Wiviott SD, Braunwald E, Angiolillo DJ. , et al; TRITON-TIMI 38 Investigators. Greater clinical benefit of more intensive oral antiplatelet therapy with prasugrel in patients with diabetes mellitus in the trial to assess improvement in therapeutic outcomes by optimizing platelet inhibition with prasugrel-Thrombolysis in Myocardial Infarction 38. Circulation 2008; 118 (16) 1626-1636
    CrossrefPubMedGoogle Scholar
  • 53 James S, Angiolillo DJ, Cornel JH. , et al; PLATO Study Group. Ticagrelor vs. clopidogrel in patients with acute coronary syndromes and diabetes: a substudy from the PLATelet inhibition and patient Outcomes (PLATO) trial. Eur Heart J 2010; 31 (24) 3006-3016
    CrossrefPubMedGoogle Scholar