The Influence of cytoskeleton-membrane complex of human erythrocytes by osmotic sensitivity under dehydration-rehydration environment

Aims: The aim of this work to investigate the influence of membrane and cytoskeleton by anion transport inhibitors and medium composition on the sensitivity of erythrocytes to posthypertonic lysis (PHL). Method: This work investigates the response of human erythrocytes to transport isotonic medium from hypertonic media depending on the anion composition of rehydrated medium, affected by anion transport inhibitors [4,4'-Diisothiocyano-2,2'-stilbenedisulfonic acid (DIDS)] and dipyridamole. Results: We found that redistribution of ions in the cell during incubation in hypertonic media is the factor contributing to higher PHL. Block of effluent chloride anions through the anion transport inhibitor DIDS and inclusion of sodium ions in the cell at dehydration phase during nigericin treatment is accompanied by increased PHL level. Ionophore nigericin, enhancing transmembrane transport of sodium and potassium ions, has a multidirectional effect on erythrocyte's sensitivity to hypertonic cryohemolysis and post-hypertonic lysis. In case of hypertonic cryohemolysis, nigericin desensitizes cells, while in case of post-hypertonic lysis it increases cell sensitivity. We found that the level of cell hemolysis decreases with increased time of erythrocyte incubation in a hypertonic solution. Whereas using of ionophore (Nigricin) it will be reducing hemolysis level practically to 0 within 10 minutes of pre-incubation. Conclusion: The effect of pH change on osmotic sensitivity of erythrocytes depends on the temperature and osmotic effect. Under conditions of hypertonic cryohemolysis, decrease of pH to acidic values leads to a reduction in the sensitivity of erythrocytes to cooling. In turn, the sensitivity of erythrocytes to post-hypertonic hemolysis increases with pH shift to acidic values. We revealed that hypotonic hemolysis is characterized by a pronounced pH-dependence with maximum hemolysis at pH 5.8. Increase in the osmolarity of pre-incubation medium results in increased level of hemolysis at all pH values in the experiment, and sensitivity of erythrocytes to hypotension increases mainly at acidic pH, although pH-dependence is retained. Change in pH (5.8, 6.6, 7.4) allows to modulate chloride distribution between the cell and its environment.