Platelets Express Activated P2Y12 Receptor in Patients With Diabetes Mellitus

Liang Hu; Lin Chang; Yan Zhang; Lili Zhai; Shenghui Zhang; Zhiyong Qi; Hongmei Yan; Yan Yan; Xinping Luo; Si Zhang; Yiping Wang; Satya P Kunapuli; Hongying Ye; Zhongren Ding

doi:10.1161/CIRCULATIONAHA.116.026995

Platelets Express Activated P2Y₁₂ Receptor in Patients With Diabetes Mellitus

Circulation. 2017 Aug 29;136(9):817-833. doi: 10.1161/CIRCULATIONAHA.116.026995. Epub 2017 Jun 21.

Authors

Affiliations

¹ From Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China (L.H., L.C., Y.Z., L.Z., Shenghui Z., Si Z., Z.D.); Division of Cardiovascular Disease (Z.Q.), Division of Endocrinology and Metabolism (H.Y.), Huashan Hospital, Fudan University, Shanghai, China; Department of Endocrinology and Metabolism (H.Y.), Division of Cardiovascular Disease (Y.Y.), Zhongshan Hospital, Fudan University, Shanghai, China; Department of Pharmacology I, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China (L.H., Y.W.); Department of Physiology and Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA (S.P.K.). Dr Shenghui Zhang is presently at Department of Hematology, Wenzhou Key Laboratory of Hematology, First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.
² From Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China (L.H., L.C., Y.Z., L.Z., Shenghui Z., Si Z., Z.D.); Division of Cardiovascular Disease (Z.Q.), Division of Endocrinology and Metabolism (H.Y.), Huashan Hospital, Fudan University, Shanghai, China; Department of Endocrinology and Metabolism (H.Y.), Division of Cardiovascular Disease (Y.Y.), Zhongshan Hospital, Fudan University, Shanghai, China; Department of Pharmacology I, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China (L.H., Y.W.); Department of Physiology and Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA (S.P.K.). Dr Shenghui Zhang is presently at Department of Hematology, Wenzhou Key Laboratory of Hematology, First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China. dingzr@fudan.edu.cn.

PMID: 28637879
DOI: 10.1161/CIRCULATIONAHA.116.026995

Abstract

Background: Platelets from patients with diabetes mellitus are hyperactive. Hyperactivated platelets may contribute to cardiovascular complications and inadequate responses to antiplatelet agents in the setting of diabetes mellitus. However, the underlying mechanism of hyperactivated platelets is not completely understood.

Methods: We measured P2Y₁₂ expression on platelets from patients with type 2 diabetes mellitus and on platelets from rats with diabetes mellitus. We also assayed platelet P2Y₁₂ activation by measuring cAMP and VASP phosphorylation. The antiplatelet and antithrombotic effects of AR-C78511 and cangrelor were compared in rats. Finally, we explored the role of the nuclear factor-κB pathway in regulating P2Y₁₂ receptor expression in megakaryocytes.

Results: Platelet P2Y₁₂ levels are 4-fold higher in patients with type 2 diabetes mellitus compared with healthy subjects. P2Y₁₂ expression correlates with ADP-induced platelet aggregation (r=0.89, P<0.01). P2Y₁₂ in platelets from patients with diabetes mellitus is constitutively activated. Although both AR-C78511, a potent P2Y₁₂ inverse agonist, and cangrelor have similar antiplatelet efficacy on platelets from healthy subjects, AR-C78511 exhibits more powerful antiplatelet effects on diabetic platelets than cangrelor (aggregation ratio 36±3% versus 49±5%, respectively, P<0.05). Using a FeCl₃-injury mesenteric arteriole thrombosis model in rats and an arteriovenous shunt thrombosis model in rats, we found that the inverse agonist AR-C78511 has greater antithrombotic effects on GK rats with diabetes mellitus than cangrelor (thrombus weight 4.9±0.3 mg versus 8.3±0.4 mg, respectively, P<0.01). We also found that a pathway involving high glucose-reactive oxygen species-nuclear factor-κB increases platelet P2Y₁₂ receptor expression in diabetes mellitus.

Conclusions: Platelet P2Y₁₂ receptor expression is significantly increased and the receptor is constitutively activated in patients with type 2 diabetes mellitus, which contributes to platelet hyperactivity and limits antiplatelet drug efficacy in type 2 diabetes mellitus.

Keywords: AR-C78511; P2Y12; P2Y12 receptor antagonist; antiplatelet; antithrombotic; cangrelor; inverse agonist; type 2 diabetes mellitus.

MeSH terms

Adenosine Monophosphate / analogs & derivatives
Adenosine Monophosphate / pharmacology
Animals
Blood Platelets / metabolism*
Cell Adhesion Molecules / genetics
Cell Adhesion Molecules / metabolism
Cell Line
Chlorides / toxicity
Cyclic AMP / analysis
Diabetes Mellitus, Experimental / metabolism
Diabetes Mellitus, Experimental / pathology
Diabetes Mellitus, Type 2 / metabolism
Diabetes Mellitus, Type 2 / pathology*
Disease Models, Animal
Drug Inverse Agonism
Ferric Compounds / toxicity
Fibrinolytic Agents / pharmacology
Fibrinolytic Agents / therapeutic use
Humans
Male
Microfilament Proteins / genetics
Microfilament Proteins / metabolism
NF-kappa B / metabolism
Phosphoproteins / genetics
Phosphoproteins / metabolism
Platelet Aggregation / drug effects
Platelet Aggregation Inhibitors / pharmacology
Platelet Aggregation Inhibitors / therapeutic use
Rats
Rats, Wistar
Reactive Oxygen Species / metabolism
Receptors, Purinergic P2Y12 / metabolism*
Thrombosis / chemically induced
Thrombosis / drug therapy
Thrombosis / pathology

Substances

Cell Adhesion Molecules
Chlorides
Ferric Compounds
Fibrinolytic Agents
Microfilament Proteins
NF-kappa B
Phosphoproteins
Platelet Aggregation Inhibitors
Reactive Oxygen Species
Receptors, Purinergic P2Y12
vasodilator-stimulated phosphoprotein
Adenosine Monophosphate
cangrelor
Cyclic AMP
ferric chloride