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. 2021 Jan 11;10(1):47.
doi: 10.3390/biology10010047.

Sub-Fractions of Red Blood Cells Respond Differently to Shear Exposure Following Superoxide Treatment

Marijke Grau  1 , Lennart Kuck  2 , Thomas Dietz  1 , Wilhelm Bloch  1 , Michael J Simmonds  2
Affiliations

Sub-Fractions of Red Blood Cells Respond Differently to Shear Exposure Following Superoxide Treatment

Marijke Grau et al. Biology (Basel). .

Abstract

Red blood cell (RBC) deformability is an essential component of microcirculatory function that appears to be enhanced by physiological shear stress, while being negatively affected by supraphysiological shears and/or free radical exposure. Given that blood contains RBCs with non-uniform physical properties, whether all cells equivalently tolerate mechanical and oxidative stresses remains poorly understood. We thus partitioned blood into old and young RBCs which were exposed to phenazine methosulfate (PMS) that generates intracellular superoxide and/or specific mechanical stress. Measured RBC deformability was lower in old compared to young RBCs. PMS increased total free radicals in both sub-populations, and RBC deformability decreased accordingly. Shear exposure did not affect reactive species in the sub-populations but reduced RBC nitric oxide synthase (NOS) activation and intriguingly increased RBC deformability in old RBCs. The co-application of PMS and shear exposure also improved cellular deformability in older cells previously exposed to reactive oxygen species (ROS), but not in younger cells. Outputs of NO generation appeared dependent on cell age; in general, stressors applied to younger RBCs tended to induce S-nitrosylation of RBC cytoskeletal proteins, while older RBCs tended to reflect markers of nitrosative stress. We thus present novel findings pertaining to the interplay of mechanical stress and redox metabolism in circulating RBC sub-populations.

Keywords: Keywords: blood rheology; nitric oxide; oxidative stress; red blood cell deformability; shear stress conditioning.

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Conflict of interest statement

The authors declare not conflict of interest.

Figures

Figure 1
Figure 1
Markers of oxidative stress in old and young red blood cells (RBCs) after phenazine methosulfate (PMS) and/or shear stress application relative to baseline. (A) Free reactive oxidative and nitrosative species (ROS/RNS) levels were significantly higher in PMS-treated samples compared to the control condition (p < 0.001) but comparable between young and old RBCs. Shear stress application additionally increased ROS/RNS levels in the old PMS-treated RBCs (effect size (ES) = 0.4). (B) Nitrotyrosine staining was not affected by PMS nor by shear stress in young RBCs, but the combination of shear stress and PMS application significantly increased nitrotyrosine signal in old RBCs compared to control and compared to non-sheared samples (p < 0.05). (C) Antioxidant capacity, reflected by Trolox equivalent capacity, was lower in PMS-treated young and old RBCs. Shear stress significantly increased antioxidant capacity of old PMS-treated RBCs (p < 0.05).
Figure 2
Figure 2
RBC deformability responses to PMS and prolonged shear stress exposure. RBC deformability of young RBCs was significantly higher than of old RBCs. (A) Deformability of young RBC was significantly reduced after PMS incubation but was not affected by shear stress. (B) Deformability of old RBCs was significantly reduced by PMS, but increased after shear stress application; the combination of both PMS and shear stress also led to an increase in RBC deformability.
Figure 3
Figure 3
RBC nitric oxide synthase (RBC-NOS) activation and S-nitrosylation of the cytoskeletal spectrins in the RBC sub-fractions after exposure to PMS and prolonged shear stress. (A) RBC-NOS activation, reflected by serine 1177 phosphorylation, significantly decreased in young RBCs after PMS incubation (p < 0.05). Shear stress application following PMS incubation increased RBC-NOS activation in young RBCs (ES = 0.62). In old RBCs, RBC-NOS activation decreased after PMS application but also after shear stress exposure. Shear stress following PMS application led to a significant increase in RBC-NOS activation when compared to shear stress condition solely. S-nitrosylation of (B) α-spectrin and (C) β-spectrin increased in young control and PMS-treated RBCs after shear stress exposure while no such an effect was observed in old RBCs.
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References

    1. Mohandas N., Chasis J.A. Red blood cell deformability, membrane material properties and shape: Regulation by transmembrane, skeletal and cytosolic proteins and lipids. Semin. Hematol. 1993;30:171–192. - PubMed
    1. Simmonds M.J., Meiselman H.J., Marshall-Gradisnik S.M., Pyne M., Kakanis M., Keane J., Brenu E., Christy R., Baskurt O.K. Assessment of oxidant susceptibility of red blood cells in various species based on cell deformability. Biorheology. 2011;48:293–304. doi: 10.3233/BIR-2012-0599. - DOI - PubMed
    1. Mohanty J.G., Nagababu E., Rifkind J.M. Red blood cell oxidative stress impairs oxygen delivery and induces red blood cell aging. Front. Physiol. 2014;5:84. doi: 10.3389/fphys.2014.00084. - DOI - PMC - PubMed
    1. Xiong Y.-L., Xiong Y.-L., Li Y.-J., Tang F.-Z., Wang R.-F., Zhao Y.-J., Wang X. Effects of exhaustive exercise-induced oxidative stress on red blood cell deformability. Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2014;30:289–293. - PubMed
    1. McNamee A.P., Horobin J.T., Tansley G.D., Simmonds M.J. Oxidative Stress Increases Erythrocyte Sensitivity to Shear-Mediated Damage. Artif. Organs. 2018;42:184–192. doi: 10.1111/aor.12997. - DOI - PubMed

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