The study of platelet microparticles and P-selectin expression in patients with the peripheral arterial diseases

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Abstract

BACKGROUND: High platelet reactivity leads to the progression of atherosclerosis and its complications. Activated platelets adhere to the site of endothelium damage on the vessel wall and initiate the formation of an arterial thrombus, followed by acute ischemia of the organ. Biochemical and cellular marker such as platelet microvesicles and P-selectin can be analyzed using flow cytometry, which is based on a specific antigen-antibody interaction. Patients with peripheral arterial disease are at significantly greater risk of cardiovascular and cerebrovascular complications than individuals of the same age and sex. The use of antiplatelet agents is an important part of pathogenetic therapy and prevention of cardiovascular diseases and their complications.

AIM: This study was to evaluate the level of platelet microparticles and P-selectin expression in patients with peripheral arterial disease receiving antiplatelet therapy.

MATERIALS AND METHODS: The study included 49 people, which included three study groups: patients with obliterating disease of the arteries of the lower extremities (n = 14) on the background of long-term use (more than 14 days) of double (75 mg clopidogrel and 100 mg Acetylsalicylic acid) antiplatelet therapy, patients with COVID-19 (n = 15), who made up the positive control group in the determination of microparticles of platelet origin, and healthy volunteers (n = 20) without signs of acute respiratory disease, without a history of cardiovascular and thromboembolic episodes, not taking antiplatelet drugs. The functional activity of platelets was assessed by two methods: using light aggregometry and analysis of P-selectin expression on platelet surface by flow cytometry. The number of platelet’s microparticles in blood plasma was also determined using flow cytometry.

RESULTS: A significant decrease in platelet aggregation was found in patients with peripheral arterial disease taking antiplatelet agents, compared with controls by used light aggregometry. Similar changes were obtained when analyzing the expression of P-selectin on platelets. A higher percentage of platelet’s microparticles with the CD9+CD41+ phenotype was found in patients with severe inflammation compared with peripheral arterial disease patients treated with antiplatelet agents and compared with the healthy controls.

CONCLUSIONS: Thus, our study reflects the consistency of the results of three different laboratory tests in assessing the platelets reactivity in patients with the peripheral arterial diseases taking antiplatelet drugs.

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About the authors

Aleksei I. Ermakov

St. Petersburg City AIDS Center

Author for correspondence.
Email: Ermakovspb@mail.ru
ORCID iD: 0000-0003-3435-5881
SPIN-code: 8921-7251

Head of Clinical Laboratory Department

Russian Federation, Санкт-Петербург

Larisa B. Gaikovaya

North-Western State Medical University named after I.I. Mechnikov

Email: Larisa.Gaikovaya@szgmu.ru
ORCID iD: 0000-0003-1000-1114
SPIN-code: 9424-1076

MD, Dr. Sсi. (Med.), Professor, Head of Sokolovsky Department of Biological and General Chemistry

Russian Federation, Saint Petersburg

Olga V. Sirotkina

Almazov National Medical Research Centre; Petersburg Nuclear Physics Institute named by B.P. Konstantinov of National Research Centre “Kurchatov Institute”

Email: Olga_sirotkina@mail.ru
ORCID iD: 0000-0003-3594-1647
SPIN-code: 1780-5490

Dr. Sci. (Biol.), Professor of Department of Laboratory Medicine and Genetics; Senior Research Associate of Laboratory of Human Molecular Genetics

Russian Federation, Saint Petersburg; Gatchina

Tatiana V. Vavilova

Almazov National Medical Research Centre

Email: Vtv.lab.spb@gmail.com
ORCID iD: 0000-0001-8537-3639
SPIN-code: 9003-6455
Scopus Author ID: 7004477312

MD, Dr. Sci. (Med.), Professor, Head of Department of Laboratory Medicine and Genetics

Russian Federation, Saint Petersburg

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Supplementary files

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2. Fig. 1. The indicator of the degree of aggregation of platelets in the presence of adenosine diphosphate (20 µmol/L). The level of statistical significance is р < 0.0001. PAD — peripheral arterial disease; DAT — double antiplatelet therapy

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3. Fig. 2. P-selectin expression index (percentage of platelets positive for CD62P-PE) in the presence of adenosine diphosphate (20 mmol/L). The level of statistical significance is p < 0.0001. PAD — peripheral arterial disease; DAT — double antiplatelet therapy

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4. Fig. 3. Comparative analysis of microparticles of platelet origin (CD9+CD41+) in patients and healthy volunteers. The level of statistical significance is р = 0,003 for COVID-19 vs PAD, р = 0,005 for COVID-19 vs Controls. PAD — peripheral arterial disease, DAT — double antiplatelet therapy

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