Влияние глицерина на постгипертонический шок эритроцитов.

Abstract

В роботі досліджено залежності постгіпертонічного гемолізу еритроцитів, оброблених гліцерином, від тривалості інкубування клітин на етапі дегідратації і регідратації, а також від концентрації гліцерину. Встановлено, що визначальним у розвитку гемолізу еритроцитів є етап дегідратації. Якщо при 37°С постгіпертонічний гемоліз еритроцитів підвищується зі зростанням концентрації гліцерину (5, 10, 15%), то при 0ºС високий рівень пошкодження клітин спостерігається при всіх концентраціях гліцерину.---------------------------------------------------------------------------------------------------------------------------------------------В работе исследованы зависимости постгипертонического гемолиза эритроцитов, обработанных глицерином, от продолжительности инкубирования клеток на этапе дегидратации и регидратации, а также от концентрации глицерина. Установлено, что определяющим в развитии гемолиза эритроцитов является этап дегидратации. Если при 37°С постгипертонический гемолиз эритроцитов повышается с ростом концентрации глицерина (5, 10, 15%), то при 0ºС высокий уровень повреждений клеток наблюдается при всех концентрациях глицерина.-------------------------------------------------------------------------------------------------------------------------------------------------The glycerol effect on the sensitivity of human erythrocytes to posthypertonic shock has been studied in the research. Posthypertonic shock was simulated by transferring the erythrocytes from the dehydration medium (1.5 mol/L NaCl) to the rehydration medium (0.15 mol/L NaCl) at 37 and 0°C. The experiment was carried out in two versions. In the first, the erythrocytes previously pretreated with glycerol (20 min, 37°C) were subjected to posthypertonic shock (control). In the second version, pretreated cells with glycerol were also exposed to posthypertonic shock, but glycerol was present in the dehydration medium (1.5 mol/L NaCl). The study of glycerol effect on posthypertonic lysis of human erythrocytes made it possible to reveal that the level of hemolysis of the cells treated with glycerol depends on the duration of their incubation in a dehydration medium. Thus, as the incubation time in the dehydration medium increased from 5 to 20 min, the posthypertonic hemolysis of erythrocytes grew. With a further rise in incubation time of cells, the hemolytic damage growth is insignificant. In the case when erythrocytes, pretreated with glycerol, which were transferred to a dehydration medium containing glycerol, the level of posthypertonic hemolysis of erythrocytes have been increased approximately in 1.7 times. Proceeding from these results, to perform the following experiments, the exposure time of the cells in the dehydration medium was 20 min. Since the statistically significant differences on posthypertonic lysis of erythrocytes with increasing incubation time in the rehydration medium (0.15 mol/L NaCl) were not detected, so in further studies the cells were incubated in rehydration medium for 5 min. Based on the obtained experimental results, a mechanism for the development of posthypertonic lysis of erythrocytes in the presence of glycerol is proposed. So, in the dehydration medium, the erythrocytes pretreated with glycerol are getting shrunk as a result of the water loss, and in the plasmatic membranes there were formed the defects, via those the water enters cells at the rehydration stage. When the erythrocytes reach the critical hemolytic volume, then lysis occurs. In the case of the glycerol presence in the dehydration medium, the total content of intracellular substances becomes higher due to the entrance of extracellular glycerol. Therefore, at the rehydration stage, more water should enter the cell; that is an evidence of a higher posthypertonic lysis level. The usage of different concentrations of glycerol allowed to reveal that at 37°C the posthypertonic lysis level of erythrocytes has not been changed with the use of 5 and 10% glycerol and increase if glycerol concentration is 15%. The addition of glycerol to the dehydration medium leads to an increase of posthypertonic hemolysis of erythrocytes at glycerol concentrations (10 and 15%) approximately in 1.5 times. Apparently, glycerol does not completely come out from the cells during the dehydration stage. This leads to an increase of erythrocytes hemolytic damage at the rehydration stage. At 0°C the high level of posthypertonic lysis of control cells and erythrocytes transferred to the rehydration medium from the dehydration medium containing glycerol (when used throughout the concentration range) is observed. Further studies suggest a more detailed investigation of the effect of temperature and glycerol on the posthypertonic shock of cells hemolytic damage.