DIFFERENT CONCENTRATIONS OF PEROXYNITRITE CAUSE A UNIQUE CELLULAR RESPONSE OF ENDOTHELIOCYTES IN VITRO

Abstract

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.