Applied Information Aspects of Medicine (Prikladnye informacionnye aspekty mediciny)

2070-9277

Voronezh State Medical University named after N.N. Burdenko - The State Budgetary Institution of Higher Professional Education «Voronezh State Medical University named after N.N. Burdenko» of the Ministry of Public Health of the Russian

11574

10.18499/2070-9277-2026-29-1-114-123

The effect of peroxynitrite on tgf- β1 -induced endothelial-mesenchymal transition in primary endothelial cell cultures in vitro

Mzhavanadze

Nina Jansugovna

MD, Associate Professor, Professor of the Department of Cardiovascular, X-ray Endovascular Surgery and Radiation Diagnostics. Leading Researcher at the Interdisciplinary Research and Educational Laboratory Center (MNOLC)nina_mzhavanade@mail.ruStrelnikova

Ekaterina Andreyevna

graduate student of MNOLCekaterina3333@rambler.ruKalinin

Roman

MD, Professor, Head of the Department of Cardiovascular, X-ray Endovascular Surgery, and Radiation Diagnosticsglms.rokptd@mail.ruSuchkov

Igor Aleksandrovich

MD, Professor of the Department of Cardiovascular, X-ray Endovascular Surgery, and Radiation Diagnosticsi.suchkov@rzgmu.ruAbalenikhina

Julia Vladimirovna

Doctor of Medical Sciences, Professor of the Department of Biological Chemistry, Leading Researcher at the MNOLCAbalenihina88@mail.ruKorotkova

Natalya Vasilevna

PhD, Associate Professor of the Department of Biological Chemistry, Senior Researcher at the MNOLCfnv8@yandex.ru

Ryazan State Medical University named after Academician I.P. Pavlov of the Russian Ministry of Health

07042026

291109118

02042026

2026

The aim of the study was to assess the effect of peroxynitrite (ONOO), as a mediator of nitrosative stress (NS), on key indicators of TGF--induced endothelial-mesenchymal transition (EndMT) in primary cultures of human umbilical vein endothelial cells (HUVEC) in vitro. Material and methods. Primary HUVEC cultures at passage 3 were grown in ECGM culture medium for 3 days until a monolayer was formed. EndMT was then induced using recombinant TGF-1 (10 ng/ml) at 37C for 72 hours, after which ischemia-reperfusion was modeled by adding various concentrations of ONOO. The effectiveness of ischemia-reperfusion was confirmed by determining the level of nitric oxide II (NO) using the fluorescent probe DAF FM. At the end of TGF-1 and ONOO treatment, cell lysates were prepared from the primary HUVEC cultures. In the cell lysates, the expression (relative amount) of proteins characterizing EndMT was determined by Western Blot: endothelial markers - cluster of differentiation 31 (CD31) and von Willebrand factor (vWF), as well as mesenchymal markers - fibronectin (FN) and vimentin (VIM). Results. The effectiveness of modeling NS using various concentrations of ONOO was confirmed: exposure to peroxynitrite led to an increase in the level of NO fluorescence labeled with DAF-FM. The NO level in the experimental groups at 1, 10, 50, 100, 250, and 500 M was higher by 28, 63, 71, 87, 122, and 156%, respectively, compared to the control. The combined effect of TGF-1 and peroxynitrite, compared to the isolated effect of TGF-1, was characterized by a statistically significant increase in CD31 expression at concentrations of 1, 10, 50, 100, 250, 500 M by 168.6%, 169.4%, 154.3%, 142.6%, 65.6%, and 57.9%, respectively, and a statistically significant increase in vWF expression at concentrations of 1, 10, 50, and 100 M by 23.1%, 31.3%, 24.4%, 29.4%, and 31.9%, with no change at 500 M. The effect of TGF-1 and peroxynitrite, compared to the isolated effect of TGF-1, was characterized by a statistically significant decrease in FN expression at concentrations of 1, 10, 50, and 500 M by 36%, 25%, 20%, respectively, and a reduction in VIM expression at concentrations of 1, 10, 50, 250, and 500 M by 94.1%, 89.6%, 85.3%, 66.2%, and 78.7%, respectively. Conclusion. The effect of the nitrosative stress mediator peroxynitrite (ONOO) in a TGF--induced EndMT model is associated with increased expression of endothelial markers CD31 and vWF and decreased expression of mesenchymal markers FN and VIM, indicating an inhibitory effect of peroxynitrite on endothelial-mesenchymal transition in vitro.

endothelial-mesenchymal transition (EndMT)transforming factor beta 1 (TGF-β1)nitrosative stressperoxynitrite (ONOO−)von Willebrand factor (vWF)cluster of differentiation 31 (CD31)fibronectin (FN)vimentin (VIM)HUVEC culture.

эндотелиально-мезенхимальный переход (еndothelial-mesenchymal transition, EndMT)трансформирующий фактора бета 1 (TGF-β1)нитрозативный стресспероксинитрит (ONOO−)фактор фон Виллебранда (von Willebrand factor, vWF)кластер дифференцировки 31 (cluster of differentiation 31, CD31)фибронектин (fibronectin, FN)виментин (vimentin, VIM)культура HUVEC.

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