The aim of this study was to investigate the relationship between the composition and state of erythrocytic lipids and oxygen-binding properties of erythrocyte haemoglobin-derived haematoporphyrin under hypoxia. The results from this study showed that hypoxia causes a profound change in the composition of the lipids that form the bilayer of erythrocyte membranes, as well as in the functional characteristics of erythrocytes. The detected changes correlate with morphological reconstruction of erythrocytes and, consequently, affect the oxygen-transport properties of erythrocytes. The obtained results showed that the lipid phase plays an important role in the functioning of erythrocytes in health as well as in event of development of ischemic heart disease and other pathological processes that accompany hypoxia.

This study shows that membrane-associated cytoskeletal protein structures and the erythrocyte morphology undergo profound changes during hypoxia. Hypoxia also intensified oxidative processes in the lipid phase of the bilayer of red blood cell membranes. Sodium nitroprusside impaired the morphology of red blood cells and altered quantitative and qualitative composition of membrane-skeletal proteins. The findings suggest that hypoxia causes changes at all levels of red blood cell organization, which can cause the functional disorders of hemoglobin oxygen-transporting properties and, eventually, the complete degradation of red blood cells. The use of flavonoids has a protective effect against hypoxia.

Background: This study examines the effect of graduated hyperglycaemia on the state and oxygen-binding ability of hemoglobin, the correlation of phospholipid fractions and their metabolites in the membrane, the activity of proteolytic enzymes and the morphofunctional state of erythrocytes.Methods: Conformational changes in the molecule of hemoglobin were determined by Raman spectroscopy. The structure of the erythrocytes was analyzed using laser interference microscopy (LIM). To determine the activity of NADN-methemoglobinreductase, we used the P.G. Board method. The degree of glycosylation of the erythrocyte membranes was determined using a method previously described by Felkoren et al. Lipid extraction was performed using the Bligh and Dyer method. Detection of the phospholipids was performed using V. E. Vaskovsky method.Results: Conditions of hyperglycaemia are characterized by a low affinity of hemoglobin to oxygen, which is manifested as a parallel decrease in the content of hemoglobin oxyform and the growth of deoxyform, methemoglobin and membrane-bound hemoglobin. The degree of glycosylation of membrane proteins and hemoglobin is high. For example, in the case of hyperglycaemia, erythrocytic membranes reduce the content of all phospholipid fractions with a simultaneous increase in lysoforms, free fatty acids and the diacylglycerol (DAG). Step wise hyperglycaemia in incubation medium and human erythrocytes results in an increased content of peptide components and general trypsin-like activity in the cytosol, with a simultaneous decreased activity of μ-calpain and caspase 3.Conclusions: Metabolic disorders and damage of cell membranes during hyperglycaemia cause an increase in the population of echinocytes and spherocytes. The resulting disorders are accompanied with a high probability of intravascular haemolysis.

The aim of this study was to investigate the effect of hyperoxia, calcium ions and pH value on the composition of major phospholipids in human erythrocyte membranes and erythrocytes' oxygentransport function. To create a model of hyperoxia, we saturated the incubated mixture with oxygen by constant passing of oxygen-air mixture through the incubation medium. To assess the effect of elevated calcium ion concentrations, CaCl 2 was added to the incubation medium. An incubation medium with different pH was used to study the effect of various pH values. Lipids were extracted from erythrocytes and chromatographic separation was carried out in a thin layer of silica gel deposited on a glass plate. The thiobarbituric acid (TBA)-active products and the content of diene conjugates (DC) in erythrocytes were determined. The oxygen-binding capacity of haemoglobin was evaluated using Raman spectroscopy. The obtained results indicated that hyperoxia causes deep changes both in the composition and character of bilayer lipids of erythrocyte membranes, which affects the functional characteristics of erythrocytes, primarily the oxygen-transport properties of erythrocyte haemoglobin. It should be noted that a combination of Ca 2+ ions and change in the pH value intensify the processes associated with disruption of phospholipids' composition. The findings indicate that the lipid phase is one of the key elements in the functioning of erythrocytes in norm as well as during development of various pathological processes.

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