Physical and Chemical Processes and the Morphofunctional Characteristics of Human Erythrocytes in Hyperglycaemia

Victor V. Revin (Lead / Corresponding author), Natalia A. Klenova, Natalia V. Gromova, Igor P. Grunyushkin, Ilia N. Solomadin, Alexander Y. Tychkov, Anastasia A. Pestryakova, Anna V. Sadykhova, Elvira S. Revina, Ksenia V. Prosnikova, Jean-Christophe Bourdon, Nikolai Zhelev (Lead / Corresponding author)

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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.

Original languageEnglish
Article number606
JournalFrontiers in Physiology
Volume8
DOIs
Publication statusPublished - 30 Aug 2017

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Chemical Phenomena
Physical Phenomena
Hyperglycemia
Hemoglobins
Erythrocytes
Phospholipids
Glycosylation
Membranes
Spherocytes
Interference Microscopy
Oxygen
Methemoglobin
Calpain
Raman Spectrum Analysis
Diglycerides
Erythrocyte Membrane
Hemolysis
Nonesterified Fatty Acids
Confocal Microscopy
Caspase 3

Keywords

  • human erythrocytes
  • hyperglycaemia
  • hemoglobin system
  • membrane lipids
  • proteolytic activity

Cite this

Revin, Victor V. ; Klenova, Natalia A. ; Gromova, Natalia V. ; Grunyushkin, Igor P. ; Solomadin, Ilia N. ; Tychkov, Alexander Y. ; Pestryakova, Anastasia A. ; Sadykhova, Anna V. ; Revina, Elvira S. ; Prosnikova, Ksenia V. ; Bourdon, Jean-Christophe ; Zhelev, Nikolai. / Physical and Chemical Processes and the Morphofunctional Characteristics of Human Erythrocytes in Hyperglycaemia. In: Frontiers in Physiology. 2017 ; Vol. 8.
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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.",
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author = "Revin, {Victor V.} and Klenova, {Natalia A.} and Gromova, {Natalia V.} and Grunyushkin, {Igor P.} and Solomadin, {Ilia N.} and Tychkov, {Alexander Y.} and Pestryakova, {Anastasia A.} and Sadykhova, {Anna V.} and Revina, {Elvira S.} and Prosnikova, {Ksenia V.} and Jean-Christophe Bourdon and Nikolai Zhelev",
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Revin, VV, Klenova, NA, Gromova, NV, Grunyushkin, IP, Solomadin, IN, Tychkov, AY, Pestryakova, AA, Sadykhova, AV, Revina, ES, Prosnikova, KV, Bourdon, J-C & Zhelev, N 2017, 'Physical and Chemical Processes and the Morphofunctional Characteristics of Human Erythrocytes in Hyperglycaemia', Frontiers in Physiology, vol. 8, 606. https://doi.org/10.3389/fphys.2017.00606

Physical and Chemical Processes and the Morphofunctional Characteristics of Human Erythrocytes in Hyperglycaemia. / Revin, Victor V. (Lead / Corresponding author); Klenova, Natalia A.; Gromova, Natalia V.; Grunyushkin, Igor P.; Solomadin, Ilia N.; Tychkov, Alexander Y.; Pestryakova, Anastasia A.; Sadykhova, Anna V.; Revina, Elvira S.; Prosnikova, Ksenia V.; Bourdon, Jean-Christophe; Zhelev, Nikolai (Lead / Corresponding author).

In: Frontiers in Physiology, Vol. 8, 606, 30.08.2017.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Physical and Chemical Processes and the Morphofunctional Characteristics of Human Erythrocytes in Hyperglycaemia

AU - Revin, Victor V.

AU - Klenova, Natalia A.

AU - Gromova, Natalia V.

AU - Grunyushkin, Igor P.

AU - Solomadin, Ilia N.

AU - Tychkov, Alexander Y.

AU - Pestryakova, Anastasia A.

AU - Sadykhova, Anna V.

AU - Revina, Elvira S.

AU - Prosnikova, Ksenia V.

AU - Bourdon, Jean-Christophe

AU - Zhelev, Nikolai

PY - 2017/8/30

Y1 - 2017/8/30

N2 - 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.

AB - 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.

KW - human erythrocytes

KW - hyperglycaemia

KW - hemoglobin system

KW - membrane lipids

KW - proteolytic activity

U2 - 10.3389/fphys.2017.00606

DO - 10.3389/fphys.2017.00606

M3 - Article

C2 - 28912721

VL - 8

JO - Frontiers in Physiology

JF - Frontiers in Physiology

SN - 1664-042X

M1 - 606

ER -