Abstract
Introduction. Studying the effect of the nitric oxide (II) donor, S-nitrosoglutathione, on the relative amount of selectin P in HUVEC cell lysates in vitro may contribute to fundamental understanding of the effect of the nitric oxide system on the adhesive function of the endothelium.
Objective: To evaluate the effect of nitric oxide (II) donor S-nitrosoglutathione (GSNO) on the activity of mitochondrial endotheliocyte oxidoreductases, the relative amount of selectin P and nitric oxide metabolites in HUVEC cell lysates, and the absolute amount of sP-selectin (soluble) in a conditioned medium.
Material and methods. The mitochondrial dehydrogenase activity of HUVEC cells was evaluated using an MTT test. The relative amount of P-selectin in endotheliocyte lysates was determined by the Western blot method, the absolute amount of the soluble selectin fragment (sP) in the cellular supernatant was determined by the sandwich ELISA method, the level of persistent nitric oxide (II) metabolites was determined by the method based on the reduction of nitrates to nitrites in the presence of vanadium chloride and subsequent spectrophotometric detection using Griss reagent.
Results. The study found that upon joint incubation of the primary HUVEC cell line with nitrosoglutathione solutions in various concentrations, the development of nitrosative stress is noted, as evidenced by an increase in its markers – persistent nitric oxide (II) metabolites in cell lysates. Under conditions of nitrosative stress, there is a decrease in the activity of mitochondrial endotheliocyte oxidoreductases, which may be due to the negative effects of excess nitric oxide or its toxic products. With a three-hour exposure of nitrosoglutathione to endotheliocytes at a low concentration of 10 mkM, there is a statistically significant increase in the relative amount of P-selectin; at high concentrations of 100-500 mkM, there is a statistically significant decrease in the relative amount of P-selectin.
Conclusion. The results obtained in this study indicate the effect of nitric oxide (II) on the functional response of the endothelium by quantifying the cell adhesion molecule P-selectin.
Authors and affiliations
N.V. Korotkova, R.E. Kalinin, I.A. Suchkov, Yu.V. Abalenikhina, N.D. Mzhavanadze, A.A. Nikiforov, D.I. Fomina
Ryazan State Medical University, Vysokovoltnaya str., 9, Ryazan, 390026, Russian Federation
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