Different concentrations of peroxynitrite cause a unique cellular response of endotheliocytes in vitro

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Introduction. The study of the effect of peroxynitrite, as a product of nitric oxide (II), on the culture of endothelial cells in vitro can contribute to fundamental ideas about the violation of the adhesive function of the endothelium when changing the synthesis of nitric oxide.

The aim of the study. To study the effect of peroxynitrite in various concentrations on the metabolic and migration activity of endotheliocytes, as well as on the expression of endothelium-specific proteins-selectin Р and Е, as an indicator of the adhesive function of the endothelium.

Methods. The cytotoxicity of peroxynitrite in various concentrations was assessed using the MTT test. The migration and proliferative activity of endotheliocytes was studied by a scratch test. The concentration of sP- and sЕ-selectins was determined using the ELISA sandwich method in the supernatant of endotheliocytes culture.

Results. A statistically significant increase in the metabolic activity of endothelial cells was revealed during a 30-minute incubation with peroxynitrite at concentrations of 0,0025, 0,0075 and 0,01 mM. A statistically significant decrease in the migration activity of endothelial cells was revealed after a 30-minute incubation with peroxynitrite at concentrations of 0,0025 and 0,1 mM, both in the first 12 and within 24 hours. A multidirectional effect of peroxynitrite at various concentrations on the level of sP and sE selectins in the cell culture supernatant was revealed: a statistically significant decrease in sE-selectin after 30-minute incubation with peroxynitrite solutions at concentrations of 0,0025, 0,1 and 1,0 mM; as well as a statistically significant increase in sP-selectin during 30-minute incubation with peroxynitrite solutions of 0,0025 and 0,1 mM.

Conclusion. Peroxynitrite for endothelial cells can act as a signaling molecule, stimulating or suppressing mitochondrial and migratory activity, and also as a cytotoxic agent, causing cell death. In addition, at various concentrations it is capable of influencing either the synthesis of endothelium-specific selectin proteins or the proteolytic shedding of the extracellular domains of these proteins, which requires further study.

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作者简介

Natalya Korotkova

Federal State Budgetary Educational Institution of Higher Education “Ryazan State Medical University” Ministry of Health of the Russian Federation

编辑信件的主要联系方式.
Email: fnv8@yandex.ru
ORCID iD: 0000-0001-7974-2450

Associate Professor of the Department of Biological Chemistry, Senior researcher at the Central Research laboratory. PhD.

俄罗斯联邦, Vysokovoltnaya, 9, Ryazan, 390026

Roman Kalinin

Federal State Budgetary Educational Institution of Higher Education “Ryazan State Medical University” Ministry of Health of the Russian Federation

Email: kalinin-re@yandex.ru
ORCID iD: 0000-0002-0817-9573

Head of the Department of cardiovascular, endovascular surgery, and diagnostic radiology. Doctor of medical sciences, Professor.

俄罗斯联邦, Vysokovoltnaya, 9, Ryazan, 390026

Igor Suchkov

Federal State Budgetary Educational Institution of Higher Education “Ryazan State Medical University” Ministry of Health of the Russian Federation

Email: i.suchkov@rzgmu.ru
ORCID iD: 0000-0002-1292-5452

Professor of the Department of cardiovascular, endovascular surgery, and diagnostic radiology. Doctor of medical sciences, Professor.

俄罗斯联邦, Vysokovoltnaya, 9, Ryazan, 390026

Alexander Zakharov

Federal State Budgetary Educational Institution of Higher Education “Ryazan State Medical University” Ministry of Health of the Russian Federation

Email: alexanderzakharov2019@yandex.ru
ORCID iD: 0000-0002-4004-7474

6th–year student of the Faculty of Medicine

俄罗斯联邦, Vysokovoltnaya, 9, Ryazan, 390026

Nina Mzhavanadze

Federal State Budgetary Educational Institution of Higher Education “Ryazan State Medical University” Ministry of Health of the Russian Federation

Email: nina_mzhavanadze@mail.ru
ORCID iD: 0000-0001-5437-1112

Professor the department of cardiovascular, endovascular surgery and diagnostic radiology, Leading Researcher at the central research laboratory. Doctor of Medical Sciences.

俄罗斯联邦, Vysokovoltnaya, 9, Ryazan, 390026

Alexander Nikiforov

Federal State Budgetary Educational Institution of Higher Education “Ryazan State Medical University” Ministry of Health of the Russian Federation

Email: alnik003@yandex.ru
ORCID iD: 0000-0001-9742-4528

Associate professor of the department of pharmacology, Head of the Central Research laboratory. PhD.

俄罗斯联邦, Vysokovoltnaya, 9, Ryazan, 390026

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2. Fig. 1. Optical density of solutions during the MTT test after incubation of endotheliocytes with peroxynitrite solutions of various concentrations for 30 minutes, units of optical density

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3. Fig. 2. The rate of migration activity of endotheliocytes after incubation with peroxynitrite solutions of various concentrations for 30 minutes, mm/h

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4. Fig. 3. The concentration of sP- and sE-selectins in the supernatant of endotheliocyte cell culture after incubation with peroxynitrite solutions of various concentrations for 30 minutes, ng/ml

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5. Fig. 4. Regulation of cell vital activity by proteins Nrf-1,2, PGC-1α, NF-κB)

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