Mechanism of collagen-induced platelet aggregation in patients with impaired carbohydrate tolerance or type 2 diabetes mellitus in combination with arterial hypertension

Introduction. Cardiovascular diseases are increasingly associated with pathologies, such as impaired glucose tolerance and type 2 diabetes mellitus (T2DM). This combination causes several negative phenomena in the body, increasing the likelihood of thrombotic complications and negatively affecting the overall prognosis of the diseases. Platelets are closely related to the onset and spread of thrombosis.

Objective. To study collagen-induced platelet aggregation and to elucidate the mechanism of action through the cyclic adenosine monophosphate (cAMP)-dependent signaling system in patients with impaired carbohydrate tolerance or T2DM in combination with hypertension.

Material and Methods. A cross-sectional study was conducted. The study included 37 patients with hypertension and metabolic disorders aged 40-65 years and 20 healthy volunteers. Serum parameters of carbohydrate metabolism (glucose and glycosylated hemoglobin) were quantitatively determined by the immunoturbidimetric method. Platelet aggregation activity was investigated by turbidimetry with a two-channel laser analyzer. The degree and the rate of platelet aggregation in platelet-rich plasma were determined based on the curves of light transmission and platelet aggregate mean size in the presence of collagen. Intracellular concentrations of cAMP in platelets were increased by preincubation with adenylate cyclase activator, Forskolin (Sigma), and phosphodiesterase type 3 and 5 inhibitor, 3-isobutyl-1-methylxanthine (Sigma), at final concentrations of 100 ^m for 30 min.

Results. An increase in collagen-induced platelet aggregation and size increase of platelet aggregates were observed in patients with impaired carbohydrate tolerance in combination with hypertension. The correlation relationships were detected between the level of glycosylated hemoglobin, degree of platelet aggregation, and size of platelet aggregates in patients of two groups. The correlation relationships were also detected between the size of platelet aggregates and the duration of the disease in patients with type 2 diabetes in combination with hypertension.

Conclusions. The obtained results suggest a dysregulation in the cAMP-mediated signaling system associated with chronic hyperglycemia in study patients.

1. Бондаренко И.З., Ширшина И.А. Механизмы тромбообразования, ассоциированного с сахарным диабетом: что определяет прогноз интервенционного вмешательства? Сахарный диабет. 2013;3:58-63.

2. Kahn N.N., Bauman W.A., Hatcher V.B., Sinha A.K. Inhibition of platelet aggregation and the stimulation of prostacyclin synthesis by insulin in humans. Am. J. Physiol. 1993;265(6):2160-2167.

3. Marcucci R., Gori A.M., Paniccia R., Giusti B., Valente S., Giglioli C. et al. Cardiovascular death and nonfatal myocardial infarction in acute coronary syndrome patients receiving coronary stenting are predicted by residual platelet reactivity to ADP detected by a point-of-care assay. Circulation. 2009;119(2):237-242. DOI: 10.1161/CIRCULATION-AHA.108.812636.

4. Vinik A.I., Erbas T., Park T.S., Nolan R., Pittenger G.L. Platelet dysfunction in type 2 diabetes. Diabetes Care. 2001;24(8):1476-1485. DOI: 10.2337/diacare.24.8.1476.

5. Cambien B., Bergmeier W., Saffaripour S. Antitrombotic activity of TNF-a. J. Clin. Invest. 2003;112(10):1589-1596. DOI: 10.1172/JCI19284.

6. Collet J.-P., Cuisset T., Rang e G., Cayla G., Elhadad S., Pouillot C. et al. Bedside monitoring to adjust antiplatelet therapy for coronary stenting. N. Engl. J. Med. 2012;367(22):P2100-2109. DOI: 10.1056/NEJMoa1209979.

7. Sudic D., Razmara M., Forslund M., Ji Q., Hjemdahl P., Li N. High glucose levels enhance platelet activation: involvement of multiple mechanisms. Br. J. Haematol. 2006;133(3):315-322. DOI: 10.1111/j.1365-2141.2006.06012.x.

8. Kakouros N., Rade J.J., Kourliouros A., Resar J.R. Platelet function in patients with diabetes mellitus from a theoretical to a practical perspective. International Journal of Endocrinology. 2011. DOI: 10.1155/2011/742719.

9. Hunter R.W., Hers I. Insulin/IGF-1 hybrid receptor expression on human platelets: consequences for the effect of insulin on platelet function. J. Thromb. Haemost. 2009;7(12):2123-2130. DOI: 10.1111/j.1538-7836.2009.03637.x.

10. Keating F.K., Sobel B.E., Schneider D.J. Effects of increased concentrations of glucose on platelet reactivity in healthy subjects and in patients with and without diabetes mellitus. Am. J. Cardiol. 2003;92(11):1362-1365. DOI: 10.1016/j.amjcard.2003.08.033.

11. Randriamboavonjy V., Fleming I. Insulin, insulin resistance, and platelet signaling in diabetes. Diabetes Care. 2009;32(4):528-530. DOI: 10.2337/dc08-1942.

12. Tang W.H., Stitham J., Gleim S., Di Febbo C., Porreca E., Fava C. et al. Glucose and collagen regulate human platelet activity through aldose reductase induction of thromboxane. J. Clin. Invest. 2011;121(11):4462-4476. DOI: 10.1172/JCI59291.

13. Livingstone C., McLellan A.R., McGregor M., Wilson A., Connell J.M., Small M. et al. Altered G-protein expression and adenylate cyclase activity in platelets of non-insulin-dependent diabetic (NIDDM) male subjects. Biochim. Biophys. Acta. 1991;1096(2):127-133. DOI: 10.1016/0925-4439(91)90050-j.

14. Neergaard-Petersen S., Hvas A., Grove L.E., Larsen B.S., Gregersen S., Kristensen S.D. Influence of haemoglobin A1c levels on platelet aggregation and platelet turnover in patients with coronary artery disease treated with aspirin. PtciS One. 2015;10(7):e0132629. DOI: 10.1371/journal.pone.0132629.