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Increased protein glycation in diabetes mellitus is associated with decreased aspirin-mediated protein acetylation and reduced sensitivity of blood platelets to aspirin

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Abstract

Reduced effectiveness of the most common antiplatelet drug, acetylsalicylic acid (ASA, aspirin), in diabetes mellitus has been associated with a lowered platelet sensitivity to ASA and related to glycemic control in diabetic patients. Our objectives were (a) to monitor the chemical background of how chronic hyperglycemia affects platelet response to ASA in diabetes and (b) to study a chemical competition between the amount of bound acetyl residues and the extent of protein glycation in blood platelets. Using whole-blood impedance aggregometry and platelet function analyzer (PFA-100) we observed a reduced platelet response to ASA in diabetic patients (14% vs. 79% for PFA-100 collagen-epinephrine occlusion time) and an association between the index of glycemic control and platelet refractoriness to ASA (rS=−0.378). Impaired platelet response to ASA was related to enhanced platelet protein glycation (3.6±0.4 in diabetes vs. 2.3±0.4 µmol fructosamine/µg protein in control) and reduced incorporation of acetyl residue into proteins of platelets from diabetic patients (47.4±2.0 in control vs. 33.1±0.7 µmol acetyl/µg protein in diabetic subjects). Incubation of blood platelets with increasing concentrations of glucose and ASA under in vitro conditions led to excessive modification in protein amino groups: glucose and ASA competed with each other in the course of nonenzymatic modifications, glycosylation, or acetylation, and their contributions to the occupancy of protein amino groups (R2=0.22 for glucose, R2=0.43 for ASA) were dependent upon the concentrations of glucose and ASA. Overall the effects of high glucose and high ASA on the overall occupancy of protein free amino groups are not additive. While at higher concentrations ASA overcomes the effects of hyperglycemia and retards glycation, high glucose makes acetylation less efficient, and therefore the resultant chemical modification becomes greatly reduced. In conclusion, diminished susceptibility of various platelet proteins and receptors on blood platelet membranes to acetylation and high ambient glucose might underlie the apparently differentiated sensitivity of blood platelets to ASA and determine platelet “insensitivity to aspirin” in diabetic patients

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Abbreviations

AA :

Arachidonic acid

ASA :

Acetylsalicylic acid

[ 14 C]ASA :

Acetyl-1-[14C]salicylic acid

BSA :

Bovine serum albumin

CADP :

Collagen and ADP

CEPI :

Collagen and epinephrine

CS :

Comprehensive score

CT :

Closure time

WBEA :

Whole-blood impedance (electrical) aggregometry

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Acknowledgements

This study was supported by the Polish State Committee for Scientific Research (3P05B10222 and 3P05A08022), the Medical University of Lodz (502-11-698, 502-16-699 and 503-130-2), and a NATO Cooperative Linkage Grant (lst.clg.980106).

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Authors and Affiliations

  1. Department of Hemostasis and Hemostatic Disorders, Medical University of Lodz, ul. Zeromskiego 113, 90-549, Lodz, Poland

    Cezary Watala, Justyna Pluta, Jacek Golanski & Marcin Rozalski

  2. Department of Medical Chemistry, Medical University of Lodz, ul. Morowiecka 6/8, 92-215, Lodz, Poland

    Malgorzata Czyz

  3. Diabetological Unit, University Hospital no. 2, Medical University of Lodz, ul. Zeromskiego 113, 90-549, Lodz, Poland

    Zygmunt Trojanowski

  4. Department of Clinical Pharmacology, Medical University of Lodz, ul. Rewolucji 1905, 27/39, 90-214, Lodz, Poland

    Józef Drzewoski

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  1. Cezary Watala
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  2. Justyna Pluta
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  3. Jacek Golanski
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  4. Marcin Rozalski
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  7. Józef Drzewoski
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Correspondence to Cezary Watala.

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Watala, C., Pluta, J., Golanski, J. et al. Increased protein glycation in diabetes mellitus is associated with decreased aspirin-mediated protein acetylation and reduced sensitivity of blood platelets to aspirin. J Mol Med 83, 148–158 (2005). https://doi.org/10.1007/s00109-004-0600-x

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  • DOI: https://doi.org/10.1007/s00109-004-0600-x

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