350 rub
Journal №7 for 2013 г.
Article in number:
The ways for realization of human lymphocyte apoptosis, induced by hydrogen peroxide
Keywords: apoptosis p53 protein DNA comet CD95 caspase-3 hydrogen peroxide cytoplasmic calcium cytochrome с DNA fragmentation
Authors:
V.G. Artyukhov, M.S. Trubitsyna, M.A. Nakvasina, O.V. Lidokhova, S.V. Ryazantsev
Abstract:
It is researched the markers of apoptosis indicators: the number of apoptotic cells, fragmentation and the degree of DNA damage, the level of the membrane death receptor (Fas) expression, the functional activity of the effector caspase-3, p53 concentrations, cytochrome c and calcium ions in the cytosol - after 1-24 hours since exposure of hydrogen peroxide at concentration 10-6 mol/l on human peripheral blood lymphocytes. It was found, that after 20 hours exposure of hydrogen peroxide on human lymphocytes DNA molecules are fragmented. Number of apoptotic cells was thus 28.5% (in control - 6.3%). Using the DNA comet assay, it was revealed, that DNA damage (strand breaks) are detected immediately after the H2O2 addition to lymphocytes suspension (comet C1) and reach a maximum at 6 h after exposure with these agents (Comet C2 and C3). The largest volume of DNA damage comes at 6 h after H2O2 treatment of lymphocytes. It was revealed, that the addition of hydrogen peroxide to lymphocytes suspension causes a statistically significant p53 concentration increase of 42% as compared to intact cells. It was shown, that increase of the Fas-receptor expression level in lymphocytes after exposure to hydrogen peroxide is associated not only with the unmasking previously hidden Fas-receptor molecules of lymphocyte membranes, but with the new molecules synthesis/ Statistically significant increase in the level of caspase-3 activity after 2 and 4 hours of incubation for cells, treated with hydrogen peroxide, as compared with the control. It is made conclusion about participation of receptor caspase-dependent and p53-dependent pathways in the lymphocyte apoptosis realization, induced by hydrogen peroxide. It was found, that increasing the level of calcium in the cells cytosol, treated with hydrogen peroxide, caused by the release of Ca2+ from intracellular stores as a result of phosphoinositide cells communication mechanisms component activation. It was put forward an idea about the relationship between the change in the level of calcium and initiation of programmed cell death in human lymphocytes in conditions of hydrogen peroxide influence.
Pages: 7-16
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