Nitroprusside inhibits calcium-induced impairment of red blood cell deformability

Authors

  • Viachaslau Barodka,

    Corresponding author
    1. Department of Anesthesiology/Critical Care Medicine, The Johns Hopkins Hospital
    2. Molecular Dynamics Section, National Institute on Aging, National Institutes of Health
    3. The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland
    • Address reprint requests to: Viachaslau Barodka, MD, Department of Anesthesiology & Critical Care Medicine, The Johns Hopkins Hospital, 600 North Wolfe Street, Tower 711, Baltimore, MD 21287; e-mail: vbarodk1@jhmi.edu or Joy G. Mohanty, National Inst. on Aging, Baltimore, MD 21224; email: mohantyj@mail.nih.gov.

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  • Joy G. Mohanty,

    1. Department of Anesthesiology/Critical Care Medicine, The Johns Hopkins Hospital
    2. Molecular Dynamics Section, National Institute on Aging, National Institutes of Health
    3. The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland
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  • Asif K. Mustafa,

    1. Department of Anesthesiology/Critical Care Medicine, The Johns Hopkins Hospital
    2. Molecular Dynamics Section, National Institute on Aging, National Institutes of Health
    3. The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland
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  • Lakshmi Santhanam,

    1. Department of Anesthesiology/Critical Care Medicine, The Johns Hopkins Hospital
    2. Molecular Dynamics Section, National Institute on Aging, National Institutes of Health
    3. The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland
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  • Aoibhinn Nyhan,

    1. Department of Anesthesiology/Critical Care Medicine, The Johns Hopkins Hospital
    2. Molecular Dynamics Section, National Institute on Aging, National Institutes of Health
    3. The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland
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  • Anil K. Bhunia,

    1. Department of Anesthesiology/Critical Care Medicine, The Johns Hopkins Hospital
    2. Molecular Dynamics Section, National Institute on Aging, National Institutes of Health
    3. The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland
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  • Gautam Sikka,

    1. Department of Anesthesiology/Critical Care Medicine, The Johns Hopkins Hospital
    2. Molecular Dynamics Section, National Institute on Aging, National Institutes of Health
    3. The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland
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  • Daniel Nyhan,

    1. Department of Anesthesiology/Critical Care Medicine, The Johns Hopkins Hospital
    2. Molecular Dynamics Section, National Institute on Aging, National Institutes of Health
    3. The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland
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  • Dan E. Berkowitz,

    1. Department of Anesthesiology/Critical Care Medicine, The Johns Hopkins Hospital
    2. Molecular Dynamics Section, National Institute on Aging, National Institutes of Health
    3. The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland
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  • Joseph M. Rifkind

    1. Department of Anesthesiology/Critical Care Medicine, The Johns Hopkins Hospital
    2. Molecular Dynamics Section, National Institute on Aging, National Institutes of Health
    3. The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland
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  • Funded by NIH T32 to VB. This research was also supported in part by the Intramural Research Program of the NIH, National Institute on Aging.
  • [Correction added after online publication 19-Jun-2013: The spelling of author Mustafa has been updated.]

Abstract

Background

Red blood cell (RBC) deformation is critical for microvascular perfusion and oxygen delivery to tissues. Abnormalities in RBC deformability have been observed in aging, sickle cell disease, diabetes, and preeclampsia. Although nitric oxide (NO) prevents decreases in RBC deformability, the underlying mechanism is unknown.

Study Design and Methods

As an experimental model, we used ionophore A23187–mediated calcium influx in RBCs to reduce their deformability and investigated the role of NO donor sodium nitroprusside (SNP) and KCa3.1 (Gardos) channel blockers on RBC deformability (measured as elongation index [EI] by microfluidic ektacytometry). RBC intracellular Ca2+ and extracellular K+ were measured by inductively coupled plasma mass spectrometry and potassium ion selective electrode, respectively.

Results

SNP treatment of RBCs blocked the Ca2+ (approx. 10 μmol/L)-induced decrease in RBC deformability (EI 0.34 ± 0.02 vs. 0.09 ± 0.01, control vs. Ca2+ loaded, p < 0.001; and EI 0.37 ± 0.02 vs. 0.30 ± 0.01, SNP vs. SNP plus Ca2+ loaded) as well as Ca2+ influx and K+ efflux. The SNP effect was similar to that observed after pharmacologic blockade of the KCa3.1 channel (with charybdotoxin or extracellular medium containing isotonic K+ concentration). In RBCs from KCa3.1–/– mice, 10 μmol/L Ca2+ loading did not decrease cellular deformability. A preliminary attempt to address the molecular mechanism of SNP protection suggests the involvement of cell surface thiols.

Conclusion

Our results suggest that nitroprusside treatment of RBCs may protect them from intracellular calcium increase–mediated stiffness, which may occur during microvascular perfusion in diseased states, as well as during RBC storage.

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