study of antiatherosclerotic and endothelioprotective activity of peptide agonists of EPOR/CD131 heteroreceptor

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Abstract

Introduction. The drugs affecting a mitochondrial dysfunction, oxidative stresses, apoptosis and inflammation of the vascular wall, have a high potential for the prevention and treatment of atherosclerotic lesions. In this regard, the use of EPOR/CD131 heteroreceptor agonists which have a similar spectrum of pharmacological effects, is one of the promising strategies in the treatment of cardiovascular diseases.

Materials and Methods. The study was carried out on 68 C57Bl/6J male mice. Atherosclerosis was simulated in transgenic animals with an endotheliospecific knockdown of the Polg gene by simulating a balloon injury and keeping on a Western diet. Then, the studied drugs were injected once every 3 days at the dose of 20 μg/kg for 27 days. On the 28-th day, the animals were euthanized and the area of atherosclerotic plaques was assessed. The gene expression associated with the processes of inflammation, antioxidant protection, apoptosis, and angiogenesis was also determined in the aortic tissues. In addition, the endothelium protective effect of peptides on primary cultures of endothelial cells of wild and transgenic Polg-D257A mice was studied.

Results. No statistically significant effect of drugs on the area of lipid infiltration have been found. However, the studied peptides have significantly reduced the expression of proinflammatory genes (iNos, Icam1, Vcam1, Sele, Il6, Tnfa), the genes associated with angiogenesis (Vegfa, Kdr, and Hif1a), the expression of proapoptic factors; they decreased the Bax/Bcl-2 ratio by more than 1.5 times. In addition, when supplemented with H2O2 in vitro, peptides dose-dependently increased endothelial cell survival.

Conclusion. The erythropoietin-based peptides can be used to improve the functional state of the vascular wall against the background of atherosclerotic lesions and have a depressing effect on pathobiological processes associated with a mitochondrial dysfunction. In addition, the studied peptides have a significant endothelial protective effect in the induction of oxidative stress in vitro.

About the authors

Olesya A. Puchenkova

Belgorod State National Research University

Author for correspondence.
Email: lesya759@yandex.ru
ORCID iD: 0000-0002-7657-0937

6th year student of the Medical Institute

Russian Federation, 85, Pobeda Str., Belgorod, 308015

Sergey V. Nadezhdin

Belgorod State National Research University

Email: sergey_nadezhdin@yahoo.com
ORCID iD: 0000-0002-6249-2464

Candidate of Sciences (Biology), Researcher, the Research Institute of Pharmacology of Living Systems

Russian Federation, 85, Pobeda Str., Belgorod, 308015

Vladislav O. Soldatov

Institute of Gene Biology of the Russian Academy of Sciences

Email: pharmsoldatov@gmail.com
ORCID iD: 0000-0001-9706-0699

Junior Researcher

Russian Federation, 34/5, Vavilov Str., Moscow, 119334

Maxim A. Zhuchenko

Russian Research Center “Kurchatov Institute” – State Science Research Institute of Genetics

Email: maksim.zhuchenko@pharmapark.ru

Candidate of Sciences (Biology), the Head of the Sector

Russian Federation, 1, Academician Kurchatov Square, Moscow, 123098

Diana S. Korshunova

Institute of Gene Biology of the Russian Academy of Sciences

Email: korshunova@genebiology.ru
ORCID iD: 0000-0002-0259-7045

Junior Researcher

Russian Federation, 34/5, Vavilov Str., Moscow, 119334

Marina V. Kubekina

Institute of Gene Biology of the Russian Academy of Sciences

Email: marykumy@gmail.com
ORCID iD: 0000-0002-8834-1111

postgraduate student, junior researcher at the Center for High-Precision Editing and Genetic Technologies for Biomedicine

Russian Federation, 34/5, Vavilov Str., Moscow, 119334

Evgeny N. Korshunov

Institute of Gene Biology of the Russian Academy of Sciences

Email: korshunov@genebiology.ru
ORCID iD: 0000-0001-8170-4656

The head of the vivarium, junior researcher

Russian Federation, 34/5, Vavilov Str., Moscow, 119334

Liliya V. Korokina

Belgorod State National Research University

Email: korokina@mail.ru
ORCID iD: 0000-0001-5402-0697

Candidate of Sciences (Medicine), Assistant Professor, Researcher, the Research Institute of Pharmacology of Living Systems

Russian Federation, 85, Pobeda Str., Belgorod, 308015

Polina A. Golubinskaya

Clinical diagnostic laboratory Voronezh Regional Clinical Ophthalmological Hospital

Email: polinapigeon@gmail.com
ORCID iD: 0000-0002-1765-9042

The Head of the Clinical Diagnostic Laboratory

Russian Federation, 22, Revolution of 1905 Str., Voronezh, 394030

Aleksandr L. Kulikov

Belgorod State National Research University

Email: alex-3031@yandex.ru

Researcher, Research Institute of Pharmacology of Living Systems

Russian Federation, 85, Pobeda Str., Belgorod, 308015

Vladimir V. Gureev

Belgorod State National Research University

Email: produmen@yandex.ru
ORCID iD: 0000-0003-1433-1225

Doctor of Sciences (Medicine), Associate Professor, Professor of the Department of Pharmacology and Clinical Pharmacology

Russian Federation, 85, Pobeda Str., Belgorod, 308015

Vladimir V. Pokrovskiy

Belgorod State National Research University

Email: vmpokrovsky@yandex.ru
ORCID iD: 0000-0003-3138-2075

5th year student of the Medical Institute

Russian Federation, 85, Pobeda Str., Belgorod, 308015

Evgeniy A. Patrakhanov

Belgorod State National Research University

Email: pateval7@gmail.com
ORCID iD: 0000-0002-8415-4562

5th year student of the Medical Institute

Russian Federation, 85, Pobeda Str., Belgorod, 308015

Petr R. Lebedev

Belgorod State National Research University

Email: Artkeit@yandex.ru
ORCID iD: 0000-0001-9102-3360

5th year student of the Medical Institute

Russian Federation, 85, Pobeda Str., Belgorod, 308015

Tatyana A. Denisyuk

Kursk State Medical University

Email: denitatyana@yandex.ru
ORCID iD: 0000-0003-0974-4818

Doctor of Sciences (Medicine), Associate Professor of the Department of Pharmacology

Russian Federation, 3, Karl Marx Str., Kursk, 305041

Veronika S. Belyaeva

Belgorod State National Research University

Email: nika.beliaeva@yandex.ru
ORCID iD: 0000-0003-2941-0241

post-graduate student of the Department of Pharmacology and Clinical Pharmacology

Russian Federation, 85, Pobeda Str., Belgorod, 308015

Evgeniya A. Movchan

Belgorod State National Research University

Email: ms.movchan@mail.ru
ORCID iD: 0000-0002-6244-2563

postgraduate student of the Department of Pharmacology and Clinical Pharmacology

Russian Federation, 85, Pobeda Str., Belgorod, 308015

Elizaveta I. Lepetukha

Belgorod State National Research University

Email: lisitsa007@bk.ru

postgraduate student of the Department of Pharmacology and Clinical Pharmacology

Russian Federation, 85, Pobeda Str., Belgorod, 308015

Mikhail V. Pokrovskiy

Belgorod State National Research University

Email: mpokrovsky@yandex.ru
ORCID iD: 0000-0002-2761-6249

Doctor of Sciences (Medicine), Professor of the Department of Pharmacology and Clinical Pharmacology, the Head of the Research Institute of Pharmacology of Living Systems

Russian Federation, 85, Pobeda Str., Belgorod, 308015

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2. Figure 1 – Schematic image and dimensions of the balloon catheter

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3. Figure 2 – Primary monolayer of mouse endothelial cells (a 40× magnification)

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4. Figure 3 – Area of lipid deposits

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5. Figure 4 – Influence of the studied drugs on the relative expression of apoptosis markers

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6. Figure 5 – The expression level of genes Nos2 (iNOS), Icam1, Vcam1 and Sele (E-selectin)

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7. Figure 6 – Expression level of Vegfa (Vegf-A), Kdr (VEGFR-1), and HIF1alpha (HIF-1a)

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8. Figure 7 – Expression level of genes PON2, SOD2

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9. Figure 8 – Influence of H2O2 and tested peptides on the survival of endothelial cells with the Polg-D257A genotype

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Copyright (c) 2020 Puchenkova O.A., Nadezhdin S.V., Soldatov V.O., Zhuchenko M.A., Korshunova D.S., Kubekina M.V., Korshunov E.N., Korokina L.V., Kulikov A.L., Golubinskaya P.A., Pokrovskiy V.V., Patrakhanov E.A., Lebedev P.R., Gureev V.V., Denisyuk T.A., Belyaeva V.S., Movchan E.A., Lepetukha E.I., Pokrovskiy M.V.

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