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Proteolytic Cleavage of the Red Blood Cell Glycocalyx in a Genetic Form of Hypertension

Cellular and Molecular Bioengineering volume 4, pages 678–692 (2011)Cite this article

Abstract

Recent evidence suggests that the spontaneously hypertensive rat (SHR) has an elevated level of proteases, including matrix metalloproteinases (MMPs), involved in cell membrane receptor cleavage. We hypothesize that SHR red blood cells (RBCs) may be subject to an enhanced glycocalyx cleavage compared to the RBCs of the normotensive Wistar-Kyoto (WKY) rats. By direct observation of RBC rouleaux, we found no significant difference in RBC aggregation for unseparated SHR and WKY RBCs. However, lighter SHR RBCs have a greater tendency to aggregate than WKY RBCs when separated by centrifugation. When SHR plasma was mixed with WKY RBCs, SHR plasma proteases cleaved the glycocalyx of WKY RBCs, a process that can be blocked by MMP inhibition. When treated with MMPs, WKY RBCs showed strong aggregation in dextran but not in fibrinogen, indicating that RBC membrane glycoproteins from the inner core of the glycocalyx were cleaved and that dextran was able to bind to the lipid portion of the RBC membrane. In contrast, treatment with amylases produced fibrinogen-induced aggregation with fibrinogen binding to the protein core. As a mechanism to remove old RBCs from the circulation, RBC adhesion to macrophages is reduced by MMP cleavage of the RBC glycocalyx.

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References

  1. Chen, A. Y., F. A. Delano, S. R. Valdez, J. N. Ha, H. Y. Shin, and G. W. Schmid-Schönbein. Receptor cleavage reduces the fluid shear response in neutrophils of the spontaneously hypertensive rat. Am. J. Physiol. Cell Physiol. 299:C1441–C1449, 2010.

    Article  Google Scholar 

  2. Chien, S., and K. Jan. Ultrastructural basis of the mechanism of rouleaux formation. Microvasc. Res. 5:155–166, 1973.

    Article  Google Scholar 

  3. Chien, S., and K. M. Jan. Red cell aggregation by macromolecules: roles of surface adsorption and electrostatic repulsion. J. Supramol. Struct. 1:385–409, 1973.

    Article  Google Scholar 

  4. DeLano, F. A., R. Balete, and G. W. Schmid-Schönbein. Control of oxidative stress in microcirculation of spontaneously hypertensive rats. Am. J. Physiol. Heart Circ. Physiol. 288:H805–H812, 2005.

    Article  Google Scholar 

  5. DeLano, F. A., and G. W. Schmid-Schönbein. Proteinase activity and receptor cleavage: mechanism for insulin resistance in the spontaneously hypertensive rat. Hypertension 52:415–423, 2008.

    Article  Google Scholar 

  6. DeLano, F. A., H. Zhang, E. E. Tran, C. Zhang, and G. W. Schmid-Schönbein. A new hypothesis for insulin resistance in hypertension due to receptor cleavage. Expert Rev. Endocrinol. Metab. 5:149–158, 2010.

    Google Scholar 

  7. Evans, E., and D. Needham. Intrinsic colloidal attraction/repulsion between lipid bilayers and strong attraction induced by non-adsorbing polymers. In: Molecular Mechanisms of Membrane Fusion, edited by S. Ohki, D. Doyle, T.D. Flanagen, S.W. Hui, and E. Mayhew. New York: Plenum Press, pp. 83–100, 1988.

  8. Jan, K. M., and S. Chien. Role of surface electric charge in red blood cell interactions. J. Gen. Physiol. 61:638–654, 1973.

    Article  Google Scholar 

  9. Kleinbongard, P., R. Schulz, T. Rassaf, T. Lauer, A. Dejam, T. Jax, I. Kumara, P. Gharini, S. Kabanova, B. Ozuyaman, H. G. Schnurch, A. Godecke, A. A. Weber, M. Robenek, H. Robenek, W. Bloch, P. Rosen, and M. Kelm. Red blood cells express a functional endothelial nitric oxide synthase. Blood 107:2943–2951, 2006.

    Article  Google Scholar 

  10. Korbut, R. A., J. Madej, T. Adamek-Guzik, and R. Korbut. Secretory dysfunction of vascular endothelium limits the effect of angiotensin converting enzyme inhibitor quinapril on aggregation of erythrocytes in experimental hypertension. J. Physiol. Pharmacol. 54:397–408, 2003.

    Google Scholar 

  11. Lominadze, D., I. G. Joshua, and D. A. Schuschke. Increased erythrocyte aggregation in spontaneously hypertensive rats. Am. J. Hypertens. 11:784–789, 1998.

    Article  Google Scholar 

  12. Lominadze, D., D. A. Schuschke, I. G. Joshua, and W. L. Dean. Increased ability of erythrocytes to aggregate in spontaneously hypertensive rats. Clin. Exp. Hypertens. 24:397–406, 2002.

    Article  Google Scholar 

  13. Lowe, G. D. Clinical Blood Rheology. Boca Raton: CRC Press, 1988.

    Google Scholar 

  14. Lundin, S., B. Folkow, and B. Rippe. Central blood volume in spontaneously hypertensive rats and Wistar-Kyoto normotensive rats. Acta Physiol. Scand. 112:257–262, 1981.

    Article  Google Scholar 

  15. Milward, E., K. J. Kim, A. Szklarczyk, T. Nguyen, G. Melli, M. Nayak, D. Deshpande, C. Fitzsimmons, A. Hoke, D. Kerr, J. W. Griffin, P. A. Calabresi, and K. Conant. Cleavage of myelin associated glycoprotein by matrix metalloproteinases. J. Neuroimmunol. 193:140–148, 2008.

    Article  Google Scholar 

  16. Mulivor, A. W., and H. H. Lipowsky. Inhibition of glycan shedding and leukocyte-endothelial adhesion in postcapillary venules by suppression of matrixmetalloprotease activity with doxycycline. Microcirculation 16:657–666, 2009.

    Article  Google Scholar 

  17. Nash, J. B., R. B. Wenby, S. O. Sowemimo-Coker, and H. J. Meiselman. Influence of cellular properties on red cell aggregation. Clin. Hemorheol. 7:93–108, 1987.

    Google Scholar 

  18. Pearson, M. J. An Investigation into the Mechanisms of Rouleaux Formation and the Development of Improved Techniques for its Quantitation. London: University of London, 1996.

    Google Scholar 

  19. Piccardoni, P., R. Sideri, S. Manarini, A. Piccoli, N. Martelli, G. de Gaetano, C. Cerletti, and V. Evangelista. Platelet/polymorphonuclear leukocyte adhesion: a new role for SRC kinases in Mac-1 adhesive function triggered by P-selectin. Blood 98:108–116, 2001.

    Article  Google Scholar 

  20. Piomelli, S., and C. Seaman. Mechanism of red blood cell aging: relationship of cell density and cell age. Am. J. Hematol. 42:46–52, 1993.

    Article  Google Scholar 

  21. Rampling, M. W. Rouleaux formation: its causes and consequences. In: Cardiovascular Flow Modelling and Measurement with Application to Clinical Medicine, edited by S. G. Sajjadi, G. B. Nash, and M. W. Rampling. Oxford: Oxford University Press, 1999.

    Google Scholar 

  22. Rampling, M. W., H. J. Meiselman, B. Neu, and O. K. Baskurt. Influence of cell-specific factors on red blood cell aggregation. Biorheology 41:91–112, 2004.

    Google Scholar 

  23. Reitsma, S., D. W. Slaaf, H. Vink, M. A. van Zandvoort, and M. G. oude Egbrink. The endothelial glycocalyx: composition, functions, and visualization. Pflugers Arch 454:345–359, 2007.

    Article  Google Scholar 

  24. Rodrigues, S. F., E. D. Tran, Z. B. Fortes, and G. W. Schmid-Schönbein. Matrix metalloproteinases cleave the beta2-adrenergic receptor in spontaneously hypertensive rats. Am. J. Physiol. Heart Circ. Physiol. 299:H25–H35, 2010.

    Article  Google Scholar 

  25. Rosario, H. S., S. W. Waldo, S. A. Becker, and G. W. Schmid-Schönbein. Pancreatic trypsin increases matrix metalloproteinase-9 accumulation and activation during acute intestinal ischemia-reperfusion in the rat. Am. J. Pathol. 164:1707–1716, 2004.

    Article  Google Scholar 

  26. Schmid-Schönbein, G. W. Analysis of inflammation. Annu. Rev. Biomed. Eng. 8:93–131, 2006.

    Article  Google Scholar 

  27. Schmid-Schönbein, G. W., B. W. Zweifach, F. A. DeLano, and P. C. Chen. Microvascular tone in a skeletal muscle of spontaneously hypertensive rats. Hypertension 9:164–171, 1987.

    Google Scholar 

  28. Spencer, R. P., and H. A. Pearson. The spleen as a hematological organ. Semin. Nucl. Med. 5:95–102, 1975.

    Article  Google Scholar 

  29. Swei, A., F. Lacy, F. A. DeLano, and G. W. Schmid-Schönbein. Oxidative stress in the Dahl hypertensive rat. Hypertension 30:1628–1633, 1997.

    Google Scholar 

  30. Tran, E. D., F. A. DeLano, and G. W. Schmid-Schönbein. Enhanced matrix metalloproteinase activity in the spontaneously hypertensive rat: VEGFR-2 cleavage, endothelial apoptosis, and capillary rarefaction. J. Vasc. Res. 47:423–431, 2010.

    Article  Google Scholar 

  31. Turk, B. E., L. L. Huang, E. T. Piro, and L. C. Cantley. Determination of protease cleavage site motifs using mixture-based oriented peptide libraries. Nat. Biotechnol. 19:661–667, 2001.

    Article  Google Scholar 

  32. Whittingstall, P., K. Tóth, R. B. Wenb, and H. J. Meiselman. Cellular factors in RBC aggregation: effects of autologous plasma and various polymers. In: Hemorheologie et Aggregation Erythrocytair, edited by J. F. Stoltz. Paris: Editions Medicales Internationales, 1994, pp. 21–30.

    Google Scholar 

  33. Wu, K. I., and G. W. Schmid-Schönbein. Nuclear factor kappa B and matrix metalloproteinase induced receptor cleavage in the spontaneously hypertensive rat. Hypertension 57:261–268, 2011.

    Google Scholar 

  34. Zweifach, B. W., S. Kovalcheck, F. De Lano, and P. Chen. Micropressure-flow relationships in a skeletal muscle of spontaneously hypertensive rats. Hypertension 3:601–614, 1981.

    Google Scholar 

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Acknowledgment

This research was supported by NHLBI grant HL 10881.

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No conflicts of interest are declared by the authors.

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Affiliations

  1. Department of Bioengineering, The Institute of Engineering in Medicine, University of California San Diego, La Jolla, CA, 92093-0412, USA

    Cécile Pot, Angela Y. Chen, Jessica N. Ha & Geert W. Schmid-Schönbein

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  1. Cécile Pot
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  2. Angela Y. Chen
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  3. Jessica N. Ha
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  4. Geert W. Schmid-Schönbein
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Corresponding author

Correspondence to Geert W. Schmid-Schönbein.

Additional information

C. Pot and A. Y. Chen contributed equally to this work.

Associate Editors John Shyy and Yingxiao Wang oversaw the review of this article.

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Pot, C., Chen, A.Y., Ha, J.N. et al. Proteolytic Cleavage of the Red Blood Cell Glycocalyx in a Genetic Form of Hypertension. Cel. Mol. Bioeng. 4, 678–692 (2011). https://doi.org/10.1007/s12195-011-0180-0

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  • DOI: https://doi.org/10.1007/s12195-011-0180-0

Keywords

  • Spontaneously hypertensive rat
  • Matrix metalloproteinases
  • Red blood cell aggregation
  • Glycocalyx cleavage
  • Dextran
  • Fibrinogen