Skip to main content
SpringerLink
Log in

Effect of anoxia and reoxygenation on antioxidant enzyme activities in immortalized brain endothelial cells

  • Cellular Models
  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

Summary

The effects of anoxia and reoxygenation on major antioxidant enzyme activities were investigatedin vitro in immortalized rat brain endothelial cells (RBE4 cells). A sublethal anoxic period of 12 h was assessed for RBE4 cells using the neutral red uptake test. Anoxia markedly influenced the specific activity of catalase and superoxide dismutase, with no major effect on glutathione peroxidase or glutathione reductase. After 24 h postanoxia, the superoxide dismutase activity modulated by the presence or absence of oxygen returned to control value.

Damage and recovery of RBE4 immortalized rat brain endothelial cells in culture after exposure to free radicals and other oxygen-derived species provides a usefulin vitro model to study anoxia-reoxygenation trauma at the cellular level.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Aebi, H. Catalase. In: Bergmeyer, H. U., ed.. Methods of enzymatic analysis. Vol. 2. New York: Academic Press; 1984:673–684.

    Google Scholar 

  2. Block, E. R.; Patel, J. M.; Sheridan, N. P. Effect of oxygen and endotoxin on lactate dehydrogenase release, 5-hydroxytryptamine uptake, and antioxidant enzyme activities in endothelial cells. J. Cell. Physiol. 122:240–248; 1985.

    Article  PubMed  CAS  Google Scholar 

  3. Buard, A.; Clément, M.; Bourre, J. M. Developmental changes in enzymatic systems involved in protection against peroxidation in isolated rat brain microvessels. Neurosci. Lett. 141:72–74; 1992.

    Article  PubMed  CAS  Google Scholar 

  4. Cazin, M.; Paluszezak, D.; Bianchi, A., et al. Effects of anaerobiosis upon morphology and energy metabolism of alveolar macrophages cultured in gas phase. Eur. Respir. J. 3:1015–1022; 1990.

    PubMed  CAS  Google Scholar 

  5. Chaudière, J.; Gérard, D.; Clément, M., et al. Induction of selenium-glutathione peroxidase by stimulation of metabolic hydrogen peroxide production in vivo. Bioelectrochem. Bioenerg. 18:247–256; 1987.

    Article  Google Scholar 

  6. Dorovini-Zis, K.; Prameya, R.; Bowman, P. Culture and characterization of microvessel endothelial cells derived from human brain. Lab. Invest. 64:425–436; 1991.

    PubMed  CAS  Google Scholar 

  7. Durieu-Trautmann, O.; Bourdoulous, S.; Roux, F., et al. Immortalized rat brain microvessel endothelial cells: II. Pharmacological characterization. In: Drewes, L. R.; Betz, A. L., eds. Frontiers in cerebral vascular biology: transport and its regulation. New York: Plenum Press; 1992:205–210.

    Google Scholar 

  8. Durieu-Trautmann, O.; Federici, C.; Creminon, C., et al. Nitric oxide and endothelin secretion by brain microvessel endothelia cells: regulation by cyclic nucleotides. J. Cell. Physiol. 155:104–111; 1993.

    Article  PubMed  CAS  Google Scholar 

  9. Essman, W. B.; Wollman, S. B. Free radicals, central nervous system processes and brain functions. In: Das, D. K.; Essman, W. B., eds. Oxygen radicals: systemic events and disease processes. Basel, Switzerland: Karger; 1989:172–192.

    Google Scholar 

  10. Flohe, L.; Otting, F. Superoxide dismutase assays. In: Packer, L., ed. Methods in enzymology, oxygen radicals in biological systems. Vol. 105. New York: Academic Press; 1984:93–104.

    Chapter  Google Scholar 

  11. Halliwell, B. Reactive oxygen species and the central nervous system. J. Neurochem. 59:1609–1623; 1992.

    Article  PubMed  CAS  Google Scholar 

  12. Housset, B.; Junod, A. F. Effects of culture conditions and hyperoxia on antioxidant enzymes in pig pulmonary artery and aortic endothelium. Biochim. Biophys. Acta 716:283–289; 1982.

    PubMed  CAS  Google Scholar 

  13. Invittox protocol. The frame modified neutral red uptake cytotoxicity test. The ERCATT/FRAME data bank of in vitro techniques in toxicology. Vol. 3a. 1990.

  14. Joó, F. The blood-brain barrier in vitro: the second decade. Neurochem. Int. 23:499–521; 1993.

    Article  PubMed  Google Scholar 

  15. Jornot, L.; Junod, A. F. Response of human endothelial cell antioxidant enzymes to hyperoxia. Am. J. Respir. Cell Mol. Biol. 6:107–115; 1992.

    PubMed  CAS  Google Scholar 

  16. Kong, X. J.; Fanburg, B. L. Regulation of Cu,Zn-superoxide dismutase in bovine pulmonary artery endothelial cells. J. Cell. Physiol. 153:491–497; 1992.

    Article  PubMed  CAS  Google Scholar 

  17. Niki, E. Antioxidant defenses in eukariotic cells: an overview. In: Poli, G.; Albano, E.; Dianzani, M. U., ed. Free radicals: from basic science to medicine. Basel, Switzerland: Birkhaeuser Verlag; 1993:365–373.

    Google Scholar 

  18. Paglia, D. E.; Valentine, W. N. Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J. Lab. Clin. Med. 70:158–169; 1967.

    PubMed  CAS  Google Scholar 

  19. Preston, E.; Sutherland, G.; Finsten, A. Three openings of the blood-brain barrier produced by forebrain ischemia in the rat. Neurosci. Lett. 149:75–78; 1993.

    Article  PubMed  CAS  Google Scholar 

  20. Rapoport, S. I. Blood-brain barrier in physiology and medicine. New York: Raven Press; 1976:129–152.

    Google Scholar 

  21. Roux, F.; Durieu-Trautmann, O.; Bourre, J. M., et al. Immortalized rat brain microvesel endothelial cells: I. Expression of blood-brain barrier markers during angiogenesis. In: Drewes, L. R.; Betz, A. L., ed. Frontiers in cerebral vascular biology: transport and its regulation. New York: Plenum Press; 1992:201–204.

    Google Scholar 

  22. Roux, F.; Durieu-Trautmann, O.; Chaverot, N., et al. Regulation of gamma-glutamyl transpeptidase and alkaline phosphatase activities in immortalized rat brain microvessel endothelial cells. J. Cell. Physiol. 159:101–113; 1994.

    Article  PubMed  CAS  Google Scholar 

  23. Shaffer, J. B.; Treanor, C. P.; Del Vecchio, P. J. Expression of bovine and mouse endothelial cell antioxidant enzymes following TNF-α exposure. Free Radical Biol. & Med. 8:497–502; 1990.

    Article  CAS  Google Scholar 

  24. Shatos, M. A.; Doherty, J. M.; Stump, D. C., et al. Oxygen radical generated during anoxia followed by reoxygenation reduce the synthesis of tissue-type plasminogen activator and plasminogen activator-inhibitor-1 in human endothelial cell culture. J. Biol. Chem. 33:20443–20448; 1990.

    Google Scholar 

  25. Smith, L. E.; Sweet, E.; Freedman, S., et al. Alterations in endothelial superoxide dismutase levels as a function of growth state in vitro. Invest. Ophthalmol. & Visual Sci. 33:36–41; 1992.

    CAS  Google Scholar 

  26. Starke, P. E.; Farber, J. L. Endogenous defenses against cytotoxicity of hydrogen peroxide in cultured rat hepatocytes. J. Biol. Chem. 260:86–92; 1985.

    PubMed  CAS  Google Scholar 

  27. Tayarani, I.; Cloez, I.; Clément, M., et al. Antioxidant enzymes and related trace elements in aging capillaries and choroid plexus. J. Neurochem. 53:817–824; 1989.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratory of Neurosciences, Building 10-Room 6C103, National Institute of Aging, National Institute of Health, 20892, Bethesda, Maryland

    Olivier Rabin

  2. INSERM U 26, Hôpital Fernand WIDAL, 75010, Paris, France

    Michèle Piciotti, Jean-Marie Bourre & Françoise Roux

  3. Institut Henri Beaufour, 75016, Paris, France

    Katy Drieu

Authors
  1. Olivier Rabin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Michèle Piciotti
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Katy Drieu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jean-Marie Bourre
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Françoise Roux
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rabin, O., Piciotti, M., Drieu, K. et al. Effect of anoxia and reoxygenation on antioxidant enzyme activities in immortalized brain endothelial cells. In Vitro Cell.Dev.Biol.-Animal 32, 221–224 (1996). https://doi.org/10.1007/BF02722949

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02722949

Key words

Search

Navigation