Advertisement

Platelet aggregation is not altered among men with diabetes mellitus

  • Christian KringEmail author
  • Lars M. Rasmussen
  • Jes S. Lindholt
  • Axel C. P. Diederichsen
  • Pernille J. Vinholt
Original Article
  • 17 Downloads

Abstract

Aims

Platelets are pivotal in arterial thrombosis, and platelet hyperresponsiveness may contribute to the increased incidence of cardiovascular events in diabetes mellitus. Consequently, we hypothesized that increased in vitro platelet aggregation responses exist in men with diabetes mellitus.

Methods

The Danish Cardiovascular Screening Trial (DANCAVAS) is a community-based cardiovascular screening trial including men aged 65–74 years. Platelet aggregation was tested using 96-well light transmission aggregometry with thrombin receptor-activating peptide (TRAP), adenosine diphosphate, collagen type 1, arachidonic acid and protease-activated receptor-4 in three concentrations. Further, cardiovascular risk factors and coronary artery calcification (CAC), estimated by CT scans and ankle–brachial index, were obtained.

Results

Included were 720 men aged 65–74 years, 110 with diabetes mellitus. Overall, there was no difference in platelet aggregation among men with versus without diabetes mellitus when adjusting for or excluding platelet inhibitor treatment and men with established cardiovascular disease (CVD). This was true for all agonists, e.g., 10 µM TRAP-induced platelet aggregation of median 69% (IQR 53–75) versus 70% (IQR 60–76) in men with versus without diabetes mellitus. Platelet aggregation did not correlate with HbA1c or CAC. Men with diabetes mellitus displayed higher CAC, median 257 Agatston units (IQR 74–1141) versus median 111 Agatston units (IQR 6–420) in the remaining individuals, p < 0.0001.

Conclusions

Among outpatients with diabetes mellitus, but no CVD and no platelet inhibitor treatment, neither are platelets hyperresponsive in diabetes mellitus, nor is platelet aggregation associated with glycemic status or with the degree of coronary atherosclerosis.

Trial Registration

ISRCTN12157806.

Keywords

Atherosclerosis Platelet aggregation Diabetes mellitus Light transmission aggregometry Coronary artery calcification 

Abbreviations

ADP

Adenosine diphosphate

AU

Agatston units

CAC

Coronary artery calcification

CVD

Cardiovascular disease

HbA1c

Hemoglobin A1c

HDL

High-density lipoprotein

IQR

Interquartile range

LDL

Low-density lipoprotein

LTA

Light transmission aggregometry

PAR-4

Protease-activated receptor-4

PPP

Platelet-poor plasma

PRP

Platelet-rich plasma

TRAP

Thrombin receptor-activating peptide

Notes

Acknowledgements

The authors thank the entire staff of DANCAVAS.

Author contributions

CK contributed to study design, research, analysis and interpretation of the data, and writing of the manuscript. LMR contributed to study design, interpretation of the data and editing of the manuscript. JSL contributed to study design, interpretation of the data and editing of the manuscript. ACPD contributed to study design, interpretation of the data and editing of the manuscript. PJV contributed to study design, data analysis and interpretation of the data, and editing of the manuscript. CK is the guarantor of this work and, as such, had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.

Funding

The study received research support from Odense University Hospital and from the Hansen-Bille Braheske Family foundation.

Compliance with ethical standards

Conflict of interest

The author declares that they have no conflict of interest.

Ethical standards disclosure

The study was approved by the Danish Data Protection Agency (16/18852) and the Regional Scientific Ethical Committees of Southern Denmark (S-20140028). The study was conducted in accordance with the guidelines of the Helsinki Declaration.

Informed consent disclosure

All persons gave written informed consent.

Availability of data and material

The datasets generated and analyzed during the current study are available from the corresponding author on reasonable request.

Supplementary material

592_2019_1438_MOESM1_ESM.tif (125 kb)
Supplemental Fig. 1 Coronary artery calcification in patients with and without diabetes mellitus. Coronary artery calcification was determined from low-dose CT scans without contrast of the coronary arteries. Box plots show coronary artery calcification in men with (n = 66) and without (n = 476) diabetes mellitus after exclusion of men treated with platelet inhibitors. Horizontal line indicates median, boxes reflect interquartile range and whiskers are 10th and 90th percentiles. Dots are individual values above the 90th percentile. Men with cardiovascular disease were excluded. p value were generated by Mann–Whitney U test. AU: Agatston units; CAC: coronary artery calcification (TIFF 124 kb)

References

  1. 1.
    Emerging Risk Factors C, Sarwar N, Gao P, Seshasai SR, Gobin R, Kaptoge S, Di Angelantonio E, Ingelsson E, Lawlor DA, Selvin E, Stampfer M, Stehouwer CD, Lewington S, Pennells L, Thompson A, Sattar N, White IR, Ray KK, Danesh J (2010) Diabetes mellitus, fasting blood glucose concentration, and risk of vascular disease: a collaborative meta-analysis of 102 prospective studies. Lancet 375(9733):2215–2222.  https://doi.org/10.1016/S0140-6736(10)60484-9 CrossRefGoogle Scholar
  2. 2.
    Badimon L, Padro T, Vilahur G (2012) Atherosclerosis, platelets and thrombosis in acute ischaemic heart disease. Eur Heart J Acute Cardiovasc Care 1(1):60–74.  https://doi.org/10.1177/2048872612441582 CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Lievens D, von Hundelshausen P (2011) Platelets in atherosclerosis. Thromb Haemost 106(5):827–838.  https://doi.org/10.1160/TH11-08-0592 CrossRefPubMedGoogle Scholar
  4. 4.
    Gum PA, Kottke-Marchant K, Welsh PA, White J, Topol EJ (2003) A prospective, blinded determination of the natural history of aspirin resistance among stable patients with cardiovascular disease. J Am Coll Cardiol 41(6):961–965CrossRefGoogle Scholar
  5. 5.
    Breet NJ, van Werkum JW, Bouman HJ, Kelder JC, Ruven HJ, Bal ET, Deneer VH, Harmsze AM, van der Heyden JA, Rensing BJ, Suttorp MJ, Hackeng CM, ten Berg JM (2010) Comparison of platelet function tests in predicting clinical outcome in patients undergoing coronary stent implantation. JAMA 303(8):754–762.  https://doi.org/10.1001/jama.2010.181 CrossRefPubMedGoogle Scholar
  6. 6.
    Puurunen MK, Hwang SJ, Larson MG, Vasan RS, O’Donnell CJ, Tofler G, Johnson AD (2018) ADP platelet hyperreactivity predicts cardiovascular disease in the FHS (Framingham Heart Study). J Am Heart Assoc.  https://doi.org/10.1161/jaha.118.008522 CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Halushka PV, Lurie D, Colwell JA (1977) Increased synthesis of prostaglandin-E-like material by platelets from patients with diabetes mellitus. N Engl J Med 297(24):1306–1310.  https://doi.org/10.1056/NEJM197712152972402 CrossRefPubMedGoogle Scholar
  8. 8.
    Silberbauer K, Schernthaner G, Sinzinger H, Freyler H (1981) Platelet aggregation and reversible platelet aggregates in type I-diabetes staged by retinal fluorescein angiography. Atherosclerosis 40(1):81–90CrossRefGoogle Scholar
  9. 9.
    Jones RJ, Delamothe AP, Curtis LD, Machin SJ, Betteridge DJ (1985) Measurement of platelet aggregation in diabetics using the new electronic platelet aggregometer. Diabet Med 2(2):105–109CrossRefGoogle Scholar
  10. 10.
    Dinda AK, Kumar R, Saraya AK (1992) Platelet hyperreactivity and vasculopathy in insulin dependent diabetes mellitus. Platelets 3(2):83–86.  https://doi.org/10.3109/09537109209003392 CrossRefPubMedGoogle Scholar
  11. 11.
    Toth L, Szenasi P, Jambor G, Kammerer L, Romics L (1992) Platelet function in male diabetics with and without macrovascular complications. Diabetes Res Clin Pract 15(2):143–148CrossRefGoogle Scholar
  12. 12.
    Raffaelli F, Borroni F, Alidori A, Tirabassi G, Faloia E, Rabini RA, Giulietti A, Mazzanti L, Nanetti L, Vignini A (2015) Effects of in vitro supplementation with Syzygium cumini (L.) on platelets from subjects affected by diabetes mellitus. Platelets 26(8):720–725.  https://doi.org/10.3109/09537104.2014.980797 CrossRefPubMedGoogle Scholar
  13. 13.
    Tan KT, Tayebjee MH, Lim HS, Lip GY (2005) Clinically apparent atherosclerotic disease in diabetes is associated with an increase in platelet microparticle levels. Diabet Med 22(12):1657–1662.  https://doi.org/10.1111/j.1464-5491.2005.01707.x CrossRefPubMedGoogle Scholar
  14. 14.
    Razmara M, Hjemdahl P, Ostenson CG, Li N (2008) Platelet hyperprocoagulant activity in Type 2 diabetes mellitus: attenuation by glycoprotein IIb/IIIa inhibition. J Thromb Haemost 6(12):2186–2192.  https://doi.org/10.1111/j.1538-7836.2008.03185.x CrossRefPubMedGoogle Scholar
  15. 15.
    Hu L, Chang L, Zhang Y, Zhai L, Zhang S, Qi Z, Yan H, Yan Y, Luo X, Zhang S, Wang Y, Kunapuli SP, Ye H, Ding Z (2017) Platelets express activated P2Y12 receptor in patients with diabetes mellitus. Circulation 136(9):817–833.  https://doi.org/10.1161/CIRCULATIONAHA.116.026995 CrossRefPubMedGoogle Scholar
  16. 16.
    Petersen HD, Gormsen J (1978) Platelet aggregation in diabetes mellitus. Acta Med Scand 203(1–2):125–130PubMedGoogle Scholar
  17. 17.
    Porta M, McCraw A, Kohner EM (1982) Inverse relationship between ristocetin co-factor levels and platelet aggregation in insulin dependent diabetes. Thromb Res 25(6):507–512CrossRefGoogle Scholar
  18. 18.
    Jones DB, Davis TM, Bown E, Carter RD, Mann JI, Prescott RJ (1986) Determinants of ADP-induced platelet aggregation in diabetes mellitus. Diabetologia 29(5):291–294CrossRefGoogle Scholar
  19. 19.
    Kutti J, Wadenvik H, Henestam B, Stenstrom G (1986) Evaluation of platelet reactivity in diabetes mellitus. Acta Med Scand 219(2):195–199CrossRefGoogle Scholar
  20. 20.
    Alessandrini P, McRae J, Feman S, FitzGerald GA (1988) Thromboxane biosynthesis and platelet function in type I diabetes mellitus. N Engl J Med 319(4):208–212.  https://doi.org/10.1056/NEJM198807283190404 CrossRefPubMedGoogle Scholar
  21. 21.
    Samos M, Fedor M, Kovar F, Mokan M, Bolek T, Galajda P, Kubisz P, Mokan M (2016) Type 2 diabetes and ADP receptor blocker therapy. J Diabetes Res 2016:6760710.  https://doi.org/10.1155/2016/6760710 CrossRefPubMedGoogle Scholar
  22. 22.
    Bonello L, Tantry US, Marcucci R, Blindt R, Angiolillo DJ, Becker R, Bhatt DL, Cattaneo M, Collet JP, Cuisset T, Gachet C, Montalescot G, Jennings LK, Kereiakes D, Sibbing D, Trenk D, Van Werkum JW, Paganelli F, Price MJ, Waksman R, Gurbel PA, Working Group on High On-Treatment Platelet R (2010) Consensus and future directions on the definition of high on-treatment platelet reactivity to adenosine diphosphate. J Am Coll Cardiol 56(12):919–933.  https://doi.org/10.1016/j.jacc.2010.04.047 CrossRefPubMedGoogle Scholar
  23. 23.
    Angiolillo DJ, Bernardo E, Sabate M, Jimenez-Quevedo P, Costa MA, Palazuelos J, Hernandez-Antolin R, Moreno R, Escaned J, Alfonso F, Banuelos C, Guzman LA, Bass TA, Macaya C, Fernandez-Ortiz A (2007) Impact of platelet reactivity on cardiovascular outcomes in patients with type 2 diabetes mellitus and coronary artery disease. J Am Coll Cardiol 50(16):1541–1547.  https://doi.org/10.1016/j.jacc.2007.05.049 CrossRefPubMedGoogle Scholar
  24. 24.
    Vinholt PJ, Nybo M, Nielsen CB, Hvas AM (2017) Light transmission aggregometry using pre-coated microtiter plates and a Victor X5 plate reader. PLoS ONE 12(10):e0185675.  https://doi.org/10.1371/journal.pone.0185675 CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Paniccia R, Priora R, Liotta AA, Abbate R (2015) Platelet function tests: a comparative review. Vasc Health Risk Manag 11:133–148.  https://doi.org/10.2147/VHRM.S44469 CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Harrison P, Mackie I, Mumford A, Briggs C, Liesner R, Winter M, Machin S, British Committee for Standards in H (2011) Guidelines for the laboratory investigation of heritable disorders of platelet function. Br J Haematol 155(1):30–44.  https://doi.org/10.1111/j.1365-2141.2011.08793.x CrossRefPubMedGoogle Scholar
  27. 27.
    Lordkipanidze M, Lowe GC, Kirkby NS, Chan MV, Lundberg MH, Morgan NV, Bem D, Nisar SP, Leo VC, Jones ML, Mundell SJ, Daly ME, Mumford AD, Warner TD, Watson SP, Genotyping UK, Phenotyping of Platelets Study G (2014) Characterization of multiple platelet activation pathways in patients with bleeding as a high-throughput screening option: use of 96-well Optimul assay. Blood 123(8):e11–22.  https://doi.org/10.1182/blood-2013-08-520387 CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Vinholt PJ, Hvas AM, Nielsen C, Soderstrom AC, Sprogoe U, Fialla AD, Nybo M (2017) Reduced platelet activation and platelet aggregation in patients with alcoholic liver cirrhosis. Platelets.  https://doi.org/10.1080/09537104.2017.1349308 CrossRefPubMedGoogle Scholar
  29. 29.
    Diederichsen AC, Rasmussen LM, Sogaard R, Lambrechtsen J, Steffensen FH, Frost L, Egstrup K, Urbonaviciene G, Busk M, Olsen MH, Mickley H, Hallas J, Lindholt JS (2015) The Danish Cardiovascular Screening Trial (DANCAVAS): study protocol for a randomized controlled trial. Trials 16:554.  https://doi.org/10.1186/s13063-015-1082-6 CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Kvist TV, Lindholt JS, Rasmussen LM, Sogaard R, Lambrechtsen J, Steffensen FH, Frost L, Olsen MH, Mickley H, Hallas J, Urbonaviciene G, Busk M, Egstrup K, Diederichsen AC (2017) The DanCavas pilot study of multifaceted screening for subclinical cardiovascular disease in men and women aged 65–74 years. Eur J Vasc Endovasc Surg 53(1):123–131.  https://doi.org/10.1016/j.ejvs.2016.10.010 CrossRefPubMedGoogle Scholar
  31. 31.
    Agatston AS, Janowitz WR, Hildner FJ, Zusmer NR, Viamonte M Jr, Detrano R (1990) Quantification of coronary artery calcium using ultrafast computed tomography. J Am Coll Cardiol 15(4):827–832CrossRefGoogle Scholar
  32. 32.
    Benjamin EJ, Blaha MJ, Chiuve SE, Cushman M, Das SR, Deo R, de Ferranti SD, Floyd J, Fornage M, Gillespie C, Isasi CR, Jimenez MC, Jordan LC, Judd SE, Lackland D, Lichtman JH, Lisabeth L, Liu S, Longenecker CT, Mackey RH, Matsushita K, Mozaffarian D, Mussolino ME, Nasir K, Neumar RW, Palaniappan L, Pandey DK, Thiagarajan RR, Reeves MJ, Ritchey M, Rodriguez CJ, Roth GA, Rosamond WD, Sasson C, Towfighi A, Tsao CW, Turner MB, Virani SS, Voeks JH, Willey JZ, Wilkins JT, Wu JH, Alger HM, Wong SS, Muntner P, American Heart Association Statistics C, Stroke Statistics S (2017) Heart disease and stroke statistics-2017 update: a Report From the American Heart Association. Circulation 135(10):e146–e603.  https://doi.org/10.1161/cir.0000000000000485 CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Marcucci R, Gori AM, Paniccia R, Giusti B, Valente S, Giglioli C, Buonamici P, Antoniucci D, Abbate R, Gensini GF (2010) High on-treatment platelet reactivity by more than one agonist predicts 12-month follow-up cardiovascular death and non-fatal myocardial infarction in acute coronary syndrome patients receiving coronary stenting. Thromb Haemost 104(2):279–286.  https://doi.org/10.1160/TH10-01-0007 CrossRefPubMedGoogle Scholar
  34. 34.
    Collaboration NCDRF (2016) Worldwide trends in diabetes since 1980: a pooled analysis of 751 population-based studies with 4.4 million participants. Lancet 387(10027):1513–1530.  https://doi.org/10.1016/s0140-6736(16)00618-8 CrossRefGoogle Scholar
  35. 35.
    Yamada K, Mori K, Goto Y (1981) Platelet aggregation and plasma levels of beta-thromboglobulin in diabetes mellitus. Tohoku J Exp Med 135(4):423–429CrossRefGoogle Scholar
  36. 36.
    Lemkes BA, Bahler L, Kamphuisen PW, Stroobants AK, Van Den Dool EJ, Hoekstra JB, Nieuwland R, Gerdes VE, Holleman F (2012) The influence of aspirin dose and glycemic control on platelet inhibition in patients with type 2 diabetes mellitus. J Thromb Haemost 10(4):639–646.  https://doi.org/10.1111/j.1538-7836.2012.04632.x CrossRefPubMedGoogle Scholar
  37. 37.
    Simpson SH, Abdelmoneim AS, Omran D, Featherstone TR (2014) Prevalence of high on-treatment platelet reactivity in diabetic patients treated with aspirin. Am J Med 127(1):95e91–99.  https://doi.org/10.1016/j.amjmed.2013.09.019 CrossRefGoogle Scholar
  38. 38.
    Krasopoulos G, Brister SJ, Beattie WS, Buchanan MR (2008) Aspirin “resistance” and risk of cardiovascular morbidity: systematic review and meta-analysis. BMJ 336(7637):195–198.  https://doi.org/10.1136/bmj.39430.529549.BE CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Sangiorgi G, Rumberger JA, Severson A, Edwards WD, Gregoire J, Fitzpatrick LA, Schwartz RS (1998) Arterial calcification and not lumen stenosis is highly correlated with atherosclerotic plaque burden in humans: a histologic study of 723 coronary artery segments using nondecalcifying methodology. J Am Coll Cardiol 31(1):126–133CrossRefGoogle Scholar
  40. 40.
    Rumberger JA, Simons DB, Fitzpatrick LA, Sheedy PF, Schwartz RS (1995) Coronary artery calcium area by electron-beam computed tomography and coronary atherosclerotic plaque area. A histopathologic correlative study. Circulation 92(8):2157–2162CrossRefGoogle Scholar
  41. 41.
    Arad Y, Goodman KJ, Roth M, Newstein D, Guerci AD (2005) Coronary calcification, coronary disease risk factors, C-reactive protein, and atherosclerotic cardiovascular disease events: the St. Francis Heart Study. J Am Coll Cardiol 46(1):158–165.  https://doi.org/10.1016/j.jacc.2005.02.088 CrossRefPubMedGoogle Scholar
  42. 42.
    von Hundelshausen P, Weber C (2007) Platelets as immune cells: bridging inflammation and cardiovascular disease. Circ Res 100(1):27–40.  https://doi.org/10.1161/01.RES.0000252802.25497.b7 CrossRefGoogle Scholar
  43. 43.
    Wu MD, Atkinson TM, Lindner JR (2017) Platelets and von Willebrand factor in atherogenesis. Blood 129(11):1415–1419.  https://doi.org/10.1182/blood-2016-07-692673 CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Halvorsen S, Andreotti F, ten Berg JM, Cattaneo M, Coccheri S, Marchioli R, Morais J, Verheugt FW, De Caterina R (2014) Aspirin therapy in primary cardiovascular disease prevention: a position paper of the European Society of Cardiology working group on thrombosis. J Am Coll Cardiol 64(3):319–327.  https://doi.org/10.1016/j.jacc.2014.03.049 CrossRefPubMedGoogle Scholar
  45. 45.
    Sibbing D, Byrne RA, Bernlochner I, Kastrati A (2011) High platelet reactivity and clinical outcome: fact and fiction. Thromb Haemost 106(2):191–202.  https://doi.org/10.1160/TH11-01-0040 CrossRefPubMedGoogle Scholar
  46. 46.
    Tantry US, Bonello L, Aradi D, Price MJ, Jeong YH, Angiolillo DJ, Stone GW, Curzen N, Geisler T, Ten Berg J, Kirtane A, Siller-Matula J, Mahla E, Becker RC, Bhatt DL, Waksman R, Rao SV, Alexopoulos D, Marcucci R, Reny JL, Trenk D, Sibbing D, Gurbel PA, Working Group on On-Treatment Platelet R (2013) Consensus and update on the definition of on-treatment platelet reactivity to adenosine diphosphate associated with ischemia and bleeding. J Am Coll Cardiol 62(24):2261–2273.  https://doi.org/10.1016/j.jacc.2013.07.101 CrossRefPubMedGoogle Scholar
  47. 47.
    Chan MV, Armstrong PC, Warner TD (2018) 96-well plate-based aggregometry. Platelets 29(7):650–655.  https://doi.org/10.1080/09537104.2018.1445838 CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Mathias RA, Kim Y, Sung H, Yanek LR, Mantese VJ, Hererra-Galeano JE, Ruczinski I, Wilson AF, Faraday N, Becker LC, Becker DM (2010) A combined genome-wide linkage and association approach to find susceptibility loci for platelet function phenotypes in European American and African American families with coronary artery disease. BMC Med Genom 3:22.  https://doi.org/10.1186/1755-8794-3-22 CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Italia S.r.l., part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Clinical Biochemistry and PharmacologyOdense University HospitalOdense CDenmark
  2. 2.Centre of Individualized Medicine in Arterial Disease (CIMA)OdenseDenmark
  3. 3.Department of Cardiothoracic and Vascular SurgeryOdense University HospitalOdenseDenmark
  4. 4.Department of CardiologyOdense University HospitalOdenseDenmark

Personalised recommendations