Effect of omega‑3 polyunsaturated fatty acids on electric and viscoelastic parameters of erythrocytes in the in vitro experiment in patients with crown disease in the against stage

Kruchinina Margarita Vitalievna, Gromov Andrey Alexandrovich, Generalov Vladimir Mikhailovich, Abaltusova Natalia Vladislavovna, Kruchinina Elina Vladimirovna

The aim of the study was to study changes in the electrical and viscoelastic parameters of erythrocytes in patients with Crohn's disease in the acute stage after exposure to solutions of omega-3 PUFAs (eicosapentaenoic, docosahexaenoic) in vitro experiment. Materials and methods. We examined 41 patients with Crohn's disease (mean age 38 ± 13.05 years) in the acute stage and 53 people in the comparison group, comparable to the main group in terms of gender and age. The electrical and viscoelastic parameters of erythrocytes were studied in an inhomogeneous alternating electric field at four frequencies of 5*104 Hz, 105 Hz, 5*105 Hz and 106 Hz by dielectrophoresis method twice: before and after exposure for 2 hours with 10 μl of eicosapentaenoic (EPA, 20: 5 n-3) and 10 μl of docosahexaenoic (DHA, 22: 6 n-3) omega 3 PUFA (Sigma-Aldrich, USA). Results. The erythrocytes of patients with Crohn's disease in the acute stage differed from the erythrocytes of the control group by an increase in the number of spherocytic, deformed forms of cells with a changed surface with a decrease in the proportion of discocytes (p <0.00001). The amplitude of cell deformation at high frequencies in patients with CD was found to be lower than those for controls at the background of increased summarized indicators of viscosity and rigidity (p <0.00001). In patients with CD, erythrocytes tended to form aggregates and destruction at different frequencies of the electric field (p = 0.014-0.05). The polarizability at frequencies of 106, 0.5 × 106, 105 Hz turned out to be lower, and at a frequency of (0.5 × 104) Hz it was higher in the presence of inflammatory bowel disease compared to the control group (p = 0.0001-0.03). The erythrocytes of patients with Crohn's disease had higher levels of electrical conductivity and lower levels of membrane capacity, speed of cell movement to the electrodes, dipole moment than those in the controls (p = 0.0001-0.0008). The crossover frequency in patients with IBD turned out to be shifted to the high-frequency range. Exposure of erythrocytes of patients with Crohn's disease in vitro for 2 hours with 10 μL solutions of eicosapentaenoic and docosahexaenoic acids led to a significant change in the electrical and viscoelastic parameters of erythrocytes: a decrease in the summarized indicators of rigidity, cell viscosity, electrical conductivity, aggregation and destruction indices at all frequencies of the electric field; an increase in the levels of capacity, dipole moment, the speed of cell movement to the electrodes, polarizability (p = 0.001-0.05). There were revealed tendencies to an increase in the diameter of erythrocytes, the amplitude of cell deformation at high frequencies of the electric field, the proportion of discocytic forms with a decrease in spherocytic, deformed ones. There is a trend towards a shift of the crossover frequency to the low-frequency range. The observed changes in the electrical and viscoelastic parameters of erythrocytes are probably due to the modification of the structure of membranes (membrane signaling platforms) under the influence of omega-3 polyunsaturated fatty acids, changes in the activity of membrane proteins - receptors and enzymes that perform transport functions, and an indirect effect on the cell cytosol. Conclusion. The revealed effects of omega-3 PUFAs on the electrical and viscoelastic parameters of erythrocytes in patients with Crohn's disease in the acute stage may be promising as additional influence on the induction and maintenance of remission of this disease.

Keywords:omega-3 polyunsaturated fatty acids, electrical, viscoelastic parameters of erythrocytes, dielectrophoresis, Crohn's disease, in vitro experiment