Skip to main content
Log in

Peroxidative damage of the erythrocyte membrane in children with nephrotic syndrome

  • Original Article
  • Published:
Pediatric Nephrology Aims and scope Submit manuscript

Abstract

The structural composition of erythrocyte ghosts was analysed in children affected by steroid-responsive (SRNS) and unresponsive nephrotic syndrome (SUNS). No variation of either intrinsic or extrinsic ghost proteins was found by discontinuous SDS-electrophoresis associated with a very sensitive double staining technique. By contrast, the composition of inner-layer phospholipids — phosphatidyl ethanolamine (PE) and phosphatidyl serine (PS) — was altered in SRNS with minor changes also involving phosphatidic acid, phosphatidyl inositol and lysophosphatidyl choline. Signs of peroxidative damage were present in both SRNS and SUNS ghosts and inside the cells; these included high levels of fluorescent amino-iminopropene derivates of PE and PS, increased intraerythrocytic amounts of malonyldialdehyde and decreased levels of reduced glutathione. Taken together these results support the concept that in SRNS and SUNS erythrocytes are target cells for peroxidative damage. In SRNS peroxidation of membrane lipids results in a marked alteration of the phospholipid composition of erythrocyte ghosts.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Brenner BM, Hostetter TH, Humes HD (1978) Molecular basis of proteinuria of glomerular origin. N Engl J Med 298: 826–833

    Google Scholar 

  2. Rennke HG, Patel Y, Ventakatachalam MA (1978) Effect of molecular charge on glomerular permeability to proteins in the rat: clearance studies using neutral, anionic and cationic horseradish peroxidase. Kidney Int 13: 278–288

    Google Scholar 

  3. Carrie BJ, Salyer WR, Myers BD (1981) Minimal change nephropathy: an electrochemical disorder of the glomerular membrane. Am J Med 70: 262–268

    Google Scholar 

  4. Bridges CR, Myers BD, Brenner BM, Deen WM (1982) Glomerular charge alterations in human minimal change nephropathy. Kidney Int 22: 677–684

    Google Scholar 

  5. Ghiggeri GM, Candiano G, Ginevri F, Gusmano R, Ciardi MR, Perfumo F, Delfino G, Cuniberti C, Queirolo C (1987) Renal selectivity properties towards endogenous albumin in minimal change nephropathy. Kidney Int 32: 69–77

    Google Scholar 

  6. Ghiggeri GM, Candiano G, Queirolo C, Gusmano R, Ginevri F, Bertani T, Remuzzi G (1987) Studies by conventional and “low denaturing” isoelectric focusing on albumin microheterogeneity under normal conditions and in experimental nephrosis. Electrophoresis 8: 215–220

    Google Scholar 

  7. Levin M, Smith C, Walters MDS, Gascoine P, Barrat TM (1985) Steroid-responsive nephrotic syndrome: a generalized disorder of membrane negative charge. Lancet II: 239–242

    Google Scholar 

  8. Boulton-Jones JM, McWilliams C, Chandrachud L (1986) Variation in charge on red cells of patients with different glomerulopathies. Lancet II: 186–188

    Google Scholar 

  9. Feehally J, Samanta A, Kinghom H, Burden AC, Walls J (1986) Red-cell surface charge in glomerular disease. Lancet II: 635

    Google Scholar 

  10. Sewell RF, Brenchley PE (1986) Red-cell surface charge in glomerular disease. Lancet II: 635–636

    Google Scholar 

  11. International Study of Kidney Disease in Children (1978) The nephrotic syndrome in children. Prediction of histopathology from clinical and laboratory characteristics at the time of diagnosis. Kidney Int 13: 160–166

    Google Scholar 

  12. Hanaham DS, Ekholm JE (1974) The preparation of red cell ghosts (membranes). Methods Enzymol 15: 168–172

    Google Scholar 

  13. Laemmli UK (1982) Cleavage of structural proteins during the assembly of the head of bacteriophage T. Nature 277: 680–685

    Google Scholar 

  14. Merril CR, Goldman D, Van Keuren ML (1982) Simplified silver protein detection and image enhancement methods in polyacrylamide gels. Electrophoresis 3: 17–23

    Google Scholar 

  15. Dzandu JK, Deh ME, Barrat DL, Wise GE (1984) Detection of erythrocyte membrane proteins, sialoglycoproteins and lipids in the same polyacrylamide gel using a doublestaining technique. Proc Natl Acad Sci USA 81: 1733–1737

    Google Scholar 

  16. Vaskovsky VE, Kostetsky EY, Vasendin IM (1975) A universal reagent for phospholipid analysis. J Chromatogr 14: 129–141

    Google Scholar 

  17. Read SM, Northcate DM (1981) Minimization of variation in the response of different proteins of the Coomassie-Blue dye-binding assay for protein. Anal Biochem 116: 53–64

    Google Scholar 

  18. Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37: 911–917

    Google Scholar 

  19. Ghiggeri GM, Candiano G, Delfino C, Queirolo C, Ginevri F, Perfumo F, Gusmano R (1986) Separation of 9-anthryldiazomethane derivates of fatty acids by HPLC on Fatty Acid Analysis ColumR. Application to albumin-bound fatty acid analysis. J Chromatogr 381: 411–418

    Google Scholar 

  20. Nakaya T, Tomomoto T, Imoto M (1967) The synthesis and the reactions of 9-antryldiazomethane and α-Naphthyldiazomethane. Bull Chem Soc Japn 40: 691–692

    Google Scholar 

  21. Jain SK (1985) In vivo externalization of phosphatidylserine and phosphatidylethanolamine in the membrane bilayer and hypercoagulability by the lipid peroxidation of erythrocytes in rats. J Clin Invest 76: 281–286

    Google Scholar 

  22. Rose HG, Oklander M (1965) Improved procedure for the extraction of lipids from human erythrocytes. J Lipid Res 6: 528–531

    Google Scholar 

  23. Stocks J, Dormandy TL (1971) The autoxidation of human red cell lipids induced by hydrogen peroxide. Br J Haematol 20: 95–111

    Google Scholar 

  24. Dise CA, Goodman DBP (1986) t-Butyl hydroperoxide alters fatty acid incorporation into erythrocyte membrane phospholipid. Biochim Biophys Acta 859: 69–78

    Google Scholar 

  25. Fairbanks G, Steck TL, Wallach DF (1971) Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry 10: 2606–2617

    Google Scholar 

  26. Haest CWM (1982) Interactions between membrane skeleton proteins and the intrinsic domain of the erythrocyte membrane. Biochim Biophys Acta 694: 331–352

    Google Scholar 

  27. Boher MP, Baylis C, Robertson CR, Brenner BM (1972) Mechanisms of the puromycin-induced defects in the transglomerular passage of water and macromolecules. J Clin Invest 60: 152–160

    Google Scholar 

  28. Bertolatus JA, Hunsicker LG (1985) Glomerular sieving of anionic and neutral bovine albumins in proteinuric rats. Kidney Int 28: 467–476

    Google Scholar 

  29. Weening JJ, Rennke HG (1983) Glomerular permeability and polyanion in Adriamycin nephrosis in the rat. Kidney Int 24: 152–159

    Google Scholar 

  30. Bertolatus JA, Almyousef M, Hunsicker LG (1987) Glomerular sieving of high molecular weight proteins in proteinuric rats. Kidney Int 31: 1257–1266

    Google Scholar 

  31. Caufield JP, Farquhar MG (1978) Loss of anionic sites from the glomerular basement membrane in amononucleoside nephrosis. Lab Invest 39: 505–512

    Google Scholar 

  32. Kanwar YS, Jakubowski ML (1984) Unaltered anionic sites of the glomerular basement membrane in aminonucleoside nephrosis. Kidney Int 25: 613–618

    Google Scholar 

  33. Lelongt B, Makino H, Kanwar YS (1987) Status of glomerular proteoglycans in aminonucleoside nephrosis: Kidney Int 31: 1299–1310

    Google Scholar 

  34. Ghiggeri GM, Candiano G, Ginevri F, Mutti A, Bergamaschi E, Alinovi R, Righetti PG (1988) Hydrophobic interaction of Alcian Blue with soluble and erythrocyte membrane proteins. J Chromatogr 452: 347–357

    Google Scholar 

  35. Van Zu J, Dubbelman TM, Van Steveninck J (1985) Peroxide-induced membrane damage in human erythrocytes. Biochim Biophys Acta 818: 38–44

    Google Scholar 

  36. Ozanne P, Francis RB, Meiselman HJ (1983) Red blood cell aggregation in nephrotic children. Kidney Int 23: 519–525

    Google Scholar 

  37. Ghiggeri GM, Ginevri F, Candiano G, Oleggini R, Perfumo F, Queirolo C, Gusmano R (1987) Characterization of cationic albumin in minimal change nephropathy. Kidney Int 32: 547–553

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ginevri, F., Ghiggeri, G.M., Candiano, G. et al. Peroxidative damage of the erythrocyte membrane in children with nephrotic syndrome. Pediatr Nephrol 3, 25–32 (1989). https://doi.org/10.1007/BF00859620

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00859620

Key words

Navigation