DUAL PROAPOPTOTIC AND PRONECROTIC EFFECT OF HYDROGEN PEROXIDE ON HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS
A.D. Nadeev,1,* I.V. Kudryavtsev,2,3 M.K. Serebriakova,2 P.V. Avdonin,4 V.P. Zinchenko,1 N.V. Goncharov 5,6
1 Institute of Cell Biophysics RAS, Pushchino, 142290,
2 Institute of Experimental Medicine, St. Petersburg, 197376,
3 Far East Federal University, Vladivostok, 690091,
4 N.K. Koltsov Institute of Developmental Biology RAS, Moscow, 119334,
5 Research Institute of Hygiene, Occupational Pathology and Human Ecology, St. Petersburg, 188663,
and 6 I.M. Sechenov Institute of Evolutionary Physiology and Biochemistry RAS, St. Petersburg, 194223;
* e-mail: madeev1987@gmail.com
The ratio of early apoptosis and late apoptosis (necrosis) in the cultured human umbilical vein endothelial cells was estimated after exposure to hydrogen peroxide
(H2O2) in vitro trying to keep them close to the physiological conditions (high cell density, high serum content, H2O2
concentration not over 500 µM). Cell viability was assessed using flow cytometry and simultaneous staining with fluorescent dyes PO-PRO-1 to detect early apoptotic cells, and
DRAQ7 to detect late apoptotic and necrotic cells. The data obtained suggest that the primary mechanism of cytotoxic response is apoptosis. The critical concentration of
H2O2 causing the death of the cell population in a dense monolayer is 250 µM. Lower concentrations of H2O2 (up to 200 µM)
cause death of individual cells; however, viability of endothelial cell population is retained, and response to calcium activating agonists does not change compared with control
cells.
Key words:
hydrogen peroxide, endothelium, cytotoxicity, apoptosis, necrosis
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