Changes of erythrocyte microrheological characteristics in pre- and post-conditioning models: an in vitro analysis

Abstract

Summary. Introduction. In various diseases and abnormal states, erythrocyte microrheological properties (MPEs) deteriorate, leading to impaired microvascular perfusion. Consequently, restoration of normal MPE represents an important scientific and clinical objective. Aim: to conduct an in vitro study of erythrocyte preand postconditioning in cellular models aimed at improving erythrocyte microrheological characteristics. Materials and Methods. Two groups of participants were enrolled: patients with stage II arterial hypertension (AH group, n = 26) and healthy individuals (control group, n = 17). In vitro experiments modeled sympathoactivation (erythrocytes incubation with phenylephrine, 10 μM) and oxidative stress (erythrocytes incubation with hydrogen peroxide (H2O2), 500 μM) to assess preand postconditioning MPEs effects. Conditioning efficacy was evaluated by changes in the erythrocyte elongation index (EEI), by measure of deformability, and the aggregation index (AI). Results. Compared with healthy controls, patients with AH exhibited a decreased EEI (p < 0.01) and a significantly increased AI (p < 0.01). In vitro, these abnormalities were effectively restored to levels observed in healthy individuals following treatment with the gasotransmitter (GT) donors, sodium nitroprusside (SNP) and sodium hydrosulfide (NaHS). In the sympathoactivation model, pharmacological postconditioning with GTs achieved complete normalization of both AI and EEI (p < 0.01). In the oxidative stress model, both mechanical and pharmacological preconditioning normalized AI and achieved EEI levels exceeding those of untreated control samples. Potential conditioning triggers, associated signaling pathways, and downstream effectors are discussed. Conclusion. Given that a comparative MPEs analysis revealed significant abnormalities in patients with AH compared with healthy individuals, investigating the potential for restoring and preventing these abnormalities through preand postconditioning represents an important scientific and clinical challenge. These findings open a new phase of research into MPEs conditioning, namely the therapeutic application of this technology.

For citation: Muravyov A.V., Volkova E.L., Mikhailov P.V., Tikhomirova I.A., Lugovtsov A.E., Priezzhev A.V., Zamyshlyaev A.V. Changes of erythrocyte microrheological characteristics in preand post-conditioning models: an in vitro analysis. Tromboz, gemostaz i reologiya. 2026;(2):72–80. (In Russ.).

References 

1. Baskurt O.K., Meiselman H.J. Blood rheology and hemodynamics. Semin Thromb Hemost. 2003;29(5):435–50. DOI: 10.1055/s-2003-44551.
2. Gallagher M.T., Wain R.A.J., Dari S. et al. Non-identifiability of parameters for a class of shear-thinning rheological models, with implications for haematological fluid dynamics. J Biomech. 2019;85:230–8. DOI: 10.1016/j.jbiomech.2019.01.036.
3. Popel A.S., Johnson P.C. Microcirculation and hemorheology. Annu Rev Fluid Mech. 2005;37:43–69. DOI: 10.1146/annurev. fluid.37.042604.133933.
4. Günay C., Kartal H., Demirbaş E. et al. Evaluation of the effects of rosmarinic acid on red blood cell deformability, morphology, and nitric oxide in rat lower limb skeletal muscle ischemiareperfusion injury. Ulus Travma Acil Cerrahi Derg. 2025;31(4):324– 31. DOI: 10.14744/tjtes.2025.25590.
5. Lipowsky H.H. Microvascular rheology and hemodynamics. Microcirculation. 2005;12(1):5–15. DOI: 10.1080/10739680590894966.
6. Anishchenko A.M., Sidekhmenova A.V., Aliev O.I. et al. Relationship of blood viscosity and blood pressure in normotensive and spontaneously hypertensive rats. Rossijskij fiziologicheskij zhurnal imeni I.M. Sechenova. 2024;110(9):1420–9. (In Russ.). DOI: 10.31857/ S0869813924090103.
7. Grau M., Cremer J.M., Schmeichel S. et al. Comparisons of blood parameters, red blood cell deformability and circulating nitric oxide between males and females considering hormonal contraception: a longitudinal gender study. Front Physiol. 2018;9:1835. DOI: 10.3389/fphys.2018.01835.
8. Szijártó I.A., Markó L., Filipovic M.R. et al. Cystathionine γ-lyaseproduced hydrogen sulfide controls endothelial NO bioavailability and blood pressure. Hypertension. 2018;71(6):1210–7. DOI: 10.1161/ HYPERTENSIONAHA.117.10562.
9. Marini E., Rolando B., Sodano F. et al. Comparative study of different H2S donors as vasodilators and attenuators of superoxideinduced endothelial damage. Antioxidants (Basel). 2023;12(2):344. DOI: 10.3390/antiox12020344.
10. Uyuklu M., Meiselman H.J., Baskurt O.K. Role of hemoglobin oxygenation in the modulation of red blood cell mechanical properties by nitric oxide. Nitric Oxide. 2009;21:20–6. DOI: 10.1016/j. niox.2009.03.004.
11. Муравьев А.В. Роль газовых медиаторов (СО, NO и H2S) в регуляции кровообращения: анализ участия микрореологии клеток крови. Регионарное кровообращение и микроциркуляция. 2021;20(1):91–9. DOI: 10.24884/1682-6655-2021-20-1-91-99. Muravyov A.V. The role of gaseous mediators (CO, NO and H2S) in the regulation of blood circulation: analysis of the participation of blood cell microrheology. Regionarnoe krovoobrashchenie i mikrocirkulyaciya. 2021;20(1):91–9. (In Russ.). DOI: 10.24884/1682-6 655-2021-20-1-91-99.
12. Meram E., Yilmaz B.D., Bas C. et al. Shear stress-induced improvement of red blood cell deformability. Biorheology. 2013;50(3–4):165– 76. DOI: 10.3233/BIR-130637.
13. Mc Namee A.P., Horobin J.T., Tansley G.D., Simmonds M.J. Oxidative stress increases erythrocyte sensitivity to shear-mediated damage. Artif Organs. 2018;42(2):184–92. DOI: 10.1111/aor.12997.
14. Grau M., Kuck L., Dietz T. et al. Sub-fractions of red blood cells respond differently to shear exposure following superoxide treatment. Biology (Basel). 2021;10(1):47. DOI: 10.3390/biology10010047.
15. Simmonds M.J., Suriany S., Ponce D., Detterich J.A. Red blood cell mechanical sensitivity improves in patients with sickle cell disease undergoing chronic transfusion after prolonged, subhemolytic shear exposure. Transfusion. 2018;58(12):2788–96. DOI: 10.1111/ trf.14901.
16. Nagarajan S., Raj R.K., Saravanakumar V. et al. Mechanical perturbations trigger endothelial nitric oxide synthase activity in human red blood cells. Sci Rep. 2016;6:26935. DOI: 10.1038/srep26935.
17. Ugurel E., Piskin S., Aksu A.C. et al. From experiments to simulation: shear-induced responses of red blood cells to different oxygen saturation levels. Front Physiol. 2020;10:1559. DOI: 10.3389/ fphys.2019.01559.
18. Sundquist J., Blas S., Hogan J. et al. The alpha 1-adrenergic receptor in human erythrocyte membranes mediates interaction in vitro of epinephrine and thyroid hormone at the membrane Ca(2 + )-ATPase. Cell Signal. 1992;4(6):795–9. DOI: 10.1016/08986568(92)90060-l.
19. Minetti G., Ciana A., Balduini C. Differential sorting of tyrosine kinases and phosphotyrosine phosphatases acting on band 3 during vesiculation of human erythrocytes. Biochem J. 2004;377:489–97. DOI: 10.1042/BJ20031401.
20. Saldanha C., Silva A., Gonçalves S., Martins-Silva J. Modulation of erythrocyte hemorheological properties by band 3 phosphorylation and dephosphorylation. Clin Hemorheol Microcirc. 2007;36(3):183–94.
21. Artmann G.M. Microscopic photometric quantification of stiffness and relaxation time of red blood cells in a flow chamber. Biorheology. 1995;32(5):553–70. DOI: 10.1016/0006-355X(95)00032-5.
22. Ugurel E., Goksel E., Cilek N. et al. Proteomic analysis of the role of the adenylyl cyclase-cAMP pathway in red blood cell mechanical responses. Cells. 2022;11(7):1250. DOI: 10.3390/cells11071250.
23. Ransy C., Vaz C., Lombès A., Bouillaud F. Use of H2O2 to cause oxidative stress, the catalase issue. Int J Mol Sci. 2020;21(23):9149. DOI: 10.3390/ijms21239149.
24. Goksel E., Ugurel E., Nader E. et al. A preliminary study of phosphodiesterases and adenylyl cyclase signaling pathway on red blood cell deformability of sickle cell patients. Front Physiol. 2023;14:1215835. DOI: 10.3389/fphys.2023.1215835.
25. Choi S., Haam C.E., Oh E.Y. et al. Vanillin induces relaxation in rat mesenteric resistance arteries by inhibiting extracellular Ca2 + influx. Molecules. 2022;28(1):288. DOI: 10.3390/molecules28010288.
26. Kobayashi J., Ohtake K., Murata I., Sonoda K. Nitric oxide bioavailability for red blood cell deformability in the microcirculation: a review of recent progress. Nitric Oxide. 2022;129:25–9. DOI: 10.1016/j. niox.2022.09.004.
27. Muravyov A.V., Tikhomirova I.A., Avdonin P.V. et al. Cellular models of erythrocytes for studying the effect of gasotransmitters on their microrheology. J Cell Biotechnol. 2019;5(1):3–10. DOI: 10.3233/ JCB-189009.
28. Simon G., Illyes G. Structural vascular changes in hypertension: role of angiotensin II, dietary sodium supplementation, and sympathetic stimulation, alone and in combination in rats. Hypertension. 2001;37(2):255–60. DOI: 10.1161/01.hyp.37.2.255.
29. Bor-Kucukatay M., Wenby R., Meiselman H., Baskurt O.K. Effects of nitric oxide on red blood cell deformability. Am J Physiol Heart Circ Physiol. 2003;284(5):H1577–84. DOI: 10.1152/ajpheart.00665.2002.
30. Fu G.X., Ji M., Han L. et al. Erythrocyte rheological properties but not whole blood and plasma viscosity are associated with severity of hypertension in older people. Z Gerontol Geriatr. 2017;50(3):233– 8. DOI: 10.1007/s00391-016-1039-8.
31. Wobst J., Rumpf P., Dang T. et al. Molecular variants of soluble guanylyl cyclase affecting cardiovascular risk. Circ J. 2015;79(3):463– 9. DOI: 10.1253/circj.CJ-15-0025.
32. Miraglia E., De Angelis F., Gazzano E. et al. Nitric oxide stimulates human sperm motility via activation of the cyclic GMP/protein kinase G signaling pathway. Reproduction. 2011;141(1):47–54. DOI: 10.1530/REP-10-0151.
33. Bizjak D., Brinkmann C., Bloch W., Grau M. Increase in red blood cellnitric oxide synthase dependent nitric oxide production during red blood cell aging in health and disease: A study on age dependent changes of rheologic and enzymatic properties in red blood cells. PLoS One. 2015;10(4):e0125206. DOI: 10.1371/journal.pone.0125206
34. Coletta C., Papapetropoulos A., Erdelyi K. et al. Hydrogen sulfide and nitric oxide are mutually dependent in the regulation of angiogenesis and endothelium-dependent vasorelaxation. Proc Natl
35. Acad Sci U S A. 2012;109(23):9161–6. DOI: 10.1073/pnas.1202916109. 35. Truss N., Warner T. Gasotransmitters and platelets. Pharmacol Ther. 2011;132(2):196–203. DOI: 10.1016/j.pharmthera.2011.07.001.
36. Piragine E., Citi V., Lawson K. et al. Potential effects of natural H2Sdonors in hypertension management. Biomolecules. 2022;12(4):58.DOI: 10.3390/biom12040581.
37. Muravyov A., Tikhomirova I. Signaling pathways regulating red blood cell aggregation. Biorheology. 2014;51(2–3):135–45.DOI: 10.3233/BIR-140664.
38. Starzyk D., Yilmaz B., Bas C. et al. Shear stress-induced improvement of red blood cell deformability. Biorheology. 2013;50(3–4):165–76. DOI: 10.3233/BIR-130637.
39. Ugurel E., Kisakurek Z.B., Aksu Y. et al. Calcium/protein kinase C signaling mechanisms in shear-induced mechanical responses of red blood cells. Microvasc Res. 2021;135:104124. DOI: 10.1016/j. mvr.2020.104124.
40. Rogers S., Lew V. Up-down biphasic volume response of human red blood cells to PIEZO1 activation during capillary transits. PLoS Comput Biol. 2021;17(3): e1008706. DOI: 10.1371/journal. pcbi.1008706.
41. Cilek N., Ugurel E., Goksel E., Yalcin O. Signaling mechanisms in red blood cells: a view through the protein phosphorylation and deformability. J Cell Physiol. 2024;239(3): e30958. DOI: 10.1002/ jcp.30958.