Abstract
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.
Highlights
Various physiological, pathological and nutritional conditions such as physical activity, large amounts of sweet food, emotional stress, metabolic syndrome, and diabetes are accompanied by high level of glucose in blood plasma
The low content of phospholipids with polyunsaturated fatty acids (PC, PI, PEA) triggers defatting of membranes and a high correlation of cholesterol/PL, which is accompanied by changes in physicochemical properties, namely, high microviscosity. These findings indicate that hyperglycaemia is followed by a disruption in membrane permeability
Disorder in the metabolic and functional state of human erythrocyte membranes at a high concentration of glucose in the incubation medium occurs primarily due to the general activation of biochemical and physicochemical processes in cells
Summary
Pathological and nutritional conditions such as physical activity, large amounts of sweet food, emotional stress, metabolic syndrome, and diabetes are accompanied by high level of glucose in blood plasma. The high content of glucose in plasma accelerates the probability of non-enzymatic glycosylation of proteins, which induce damage to the cell membrane due to nonspecific aggregation of protein molecules and changes in protein-protein and protein-lipid interactions (Vasilyeva, 2005) Taken together, these changes initiate the rapid aging of cells and the human organism. Glycosylation of erythrocytic membrane proteins and hemoglobin during hyperglycaemia increases adhesion to endothelial cells, resulting in membrane destabilization (change in the asymmetry of membrane phospholipids), changes in viscoelastic properties of cells and their morphology (Riquelme et al, 2005) Taken together, these changes can impair the oxygentransport function of erythrocytes and reduce erythrocyte lifespan. 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
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