Neurochemical Research

, Volume 30, Issue 4, pp 583–589 | Cite as

Neuroprotection and Protein Damage Prevention by Estradiol Replacement in Rat Hippocampal Slices Exposed to Oxygen-Glucose Deprivation

  • Helena Cimarosti
  • Ionara R. Siqueira
  • Lauren L. Zamin
  • Melissa Nassif
  • Rodrigo Balk
  • Rudimar Frozza
  • Carla Dalmaz
  • Carlos Alexandre Netto
  • Christianne Salbego


Here we investigated the effects of estradiol replacement in ovariectomized female rats using hippocampal slices exposed to oxygen-glucose deprivation (OGD). OGD induced lactate dehydrogenase (LDH) release to the incubation medium, what was assumed as a parameter of cellular death. In the estradiol-treated group the LDH release was markedly decreased by 23% as compared to the vehicle-treated group. In attempt to study a possible mechanism by which estradiol acts, we investigated some parameters of oxidative stress. In both vehicle-treated and estradiol-treated groups, OGD significantly increased the free radical production by 34% and 16%, respectively, although no significant differences on total antioxidant capacity were observed. Interestingly, estradiol replacement prevented the significant reduction in tryptophan and tyrosine contents caused by OGD observed in vehicle-treated animals. Our results show that estradiol replacement in ovariectomized female rats decreases cellular susceptibility to an ischemic-like injury and suggest a role for the hormone on protein damage prevention.


Cerebral ischemia estradiol replacement hippocampal slices neuroprotection oxidative stress protein damage 


  1. 1.
    Barret-Connor, E., Bush, T. L. 1991Estrogen and coronary heart disease in womenJAMA26518611867PubMedGoogle Scholar
  2. 2.
    Kannel, W. B., Thom, T. J. 1994

    Incidence, prevalence and␣mortality of cardiovascular disease

    Schlant, R.␣C.Alexander, R. W. eds. The heartMcGraw-HillNew York185197
    Google Scholar
  3. 3.
    Wenger, N., Speroff, L., Packard, B. 1993Cardiovascular health and disease in womenN. Engl. J. Med329247256PubMedGoogle Scholar
  4. 4.
    Garcia-Segura, L. M., Azcoitia, I., Don Carlos, L. L. 2001Neuroprotection by estradiolProg. Neurobiol632960PubMedGoogle Scholar
  5. 5.
    Fernandez-Tome, P., Lizasoain, I., Leza, C. L., Lorenzo, P., and Moro, M. A. 1999. Neuroprotective effects of DETA-NONOate, a nitric oxide donor, on hydrogen peroxide-induced neurotoxicity in cortical neurons. Neuropharmacology 38:1307–1315Google Scholar
  6. 6.
    Liao, S. L., Chen, W. Y., Kuo, J. S., Chen, C. J. 2002Estrogen attenuates tumor necrosis factor-α expression to provide ischemic neuroprotection in female ratsNeurosci. Lett330159162PubMedGoogle Scholar
  7. 7.
    Belcredito, S., Vegeto, E., Brusadelli, A., Ghisletti, S., Mussi, P., Ciana, P., Maggi, A. 2001Estrogen neuroprotection: the involvement of Bcl-2 binding protein BNIP2Brain Res. Rev37335342PubMedGoogle Scholar
  8. 8.
    Cimarosti, H., Zamin, L. L., Frozza, R., Nassif, M., Horn, A. P., Tavares, A., Netto, C. A., Salbego, C. 2005Estradiol protects against oxygen and glucose deprivation in rat hippocampal organotypic cultures and activates Akt and inactivates Gsk-3βNeurochem. Res30191199PubMedGoogle Scholar
  9. 9.
    Mendelowitsch, A., Ritz, M. F., Ros, J., Langemann, H., Gratzl, O. 200117β-estradiol reduces cortical lesion size in the glutamate excitotoxicity model enhancing extracellular lactate: a new neuroprotective pathwayBrain Res901230236PubMedGoogle Scholar
  10. 10.
    White, B. C., Sullivan, J. M., DeGracia, D. J., O’Neil, B. J., Neumar, R. W., Grossman, L. I., Rafols, J. A., Krause, G. S. 2000Brain ischemia and reperfusion: molecular mechanisms of neuronal injuryJ. Neurol. Sci179133PubMedGoogle Scholar
  11. 11.
    Cárdenas, A., Moro, M. A., Hurtado, O., Leza, J. C., Lorenzo, P., Castrillo, A., Bodelón, O. G., Boscá, O. G., Lizasoain, I. 2000Implication of glutamate in the expression of nitric oxide synthase after oxygen and glucose deprivation in rat forebrain slicesJ. Neurochem7420412048PubMedGoogle Scholar
  12. 12.
    Moro, M. A., Alba, J., Leza, J. C., Lorenzo, P., Fernandez, A. P., Bentura, M. L., Bosca, L., Rodrigo, J., Lizasoain, I. 1998Neuronal expression of inducible nitric oxide synthase after oxygen and glucose deprivation in rat forebrain slicesEur. J. Neurosci10445446PubMedGoogle Scholar
  13. 13.
    Siqueira, I., Cimarosti, H., Fochesatto, C., Nunes, D. S., Salbego, C., Elisabetsky, E., Netto, C. A. 2004Neuroprotective effects of Ptychopetalum olacoides Bentham (Olacaceae) on oxygen and glucose deprivation induced damage in rat hippocampal slicesLife Sci7518971906PubMedGoogle Scholar
  14. 14.
    Siqueira, I., Cimarosti, H., Fochesatto, C., Salbego, C., Netto, C. A. 2004Age-related susceptibility to oxygen and glucose deprivation damage in rat hippocampal slicesBrain Res1025226230PubMedGoogle Scholar
  15. 15.
    Cimarosti, H., Rodnight, R., Tavares, A., Paiva, R., Valentim, L., Rocha, E., Salbego, C. 2001An investigation of the neuroprotective effect of lithium in organotypic slice cultures of rat hippocampus exposed to oxygen and glucose deprivationNeurosci. Lett3153336PubMedGoogle Scholar
  16. 16.
    Koh, J. Y., Choi, D. W. 1987Quantitative determination of glutamate mediated cortical neuronal injury in cell culture by lactate dehydrogenase efflux assayJ. Neurosci. Methods208390PubMedGoogle Scholar
  17. 17.
    Driver, A. S., Kodavanti, P. R. S., Mundy, W. R. 2000Age-related in reactive oxygen species production in rat brain homogenatesNeurotoxicol. Teratol22175181PubMedGoogle Scholar
  18. 18.
    Lissi, E., Pascual, C., Castillo, M. D. 1992Luminol luminescence induced by 2,2′-azobis-(2-amidinopropane) thermolysisFree Radic. Res. Commun17299311PubMedGoogle Scholar
  19. 19.
    Lissi, E., Salim-Hanna, M., Pascual, C., del Castillo, M. D. 1995Evaluation of total antioxidant potential (TRAP) and total antioxidant reactivity from luminol enhanced chemiluminescence measurementsFree Radic. Biol. Med18153158PubMedGoogle Scholar
  20. 20.
    Gusow, K., Szabelski, M., Rzeska, A., Karolczak, J., Sulowska, H., Wiczk, W. 2002Photophysical properties of tyrosine at low pH rangeChem. Phys. Lett362519526Google Scholar
  21. 21.
    Bondy, S. C. 1996Evaluation of free radical-initiated oxidant events within the nervous systemMethods Neurosci30243259Google Scholar
  22. 22.
    Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J. 1951Protein measurement with the folin phenol reagentsJ.␣Biol. Chem193265275PubMedGoogle Scholar
  23. 23.
    Freeman, M. E. 1994. In: Knobil, E. and Neill, J. D. (eds). The physiology of reproduction. New York: Raven Press, pages 613–647Google Scholar
  24. 24.
    Laev, H., Mahadik, S. P., Bonheur, J. L., Hernandez, N., Karpiak, S. E. 1993GM1 ganglioside reduces glutamate toxicity to cortical cells. Lowered LDH release and preserved membrane integrityChem. Neuropathol20229243Google Scholar
  25. 25.
    Segars, J., Driggers, P 2002Estrogen action and cytoplasmic signaling cascades. Part I: membrane-associated signaling complexesTrends Endocrinol. Metab13349354PubMedGoogle Scholar
  26. 26.
    Behl, C. 2002Oestrogen as a neuroprotective hormoneNat. Rev. Neurosci3433442PubMedGoogle Scholar
  27. 27.
    Culmsee, C., Vedder, H., Ravati, A., Junker, V., Otto, D., Ahlemeyer, B., Krieg, J. C., Krieglstein, J. 1999Neuroprotection by estrogens in a mouse model of focal cerebral ischemia and in cultured neurons: evidence for a receptor-independent antioxidative mechanismJ. Cereb. Blood Flow Metab1912631269PubMedGoogle Scholar
  28. 28.
    Kume-Kick, J., Rice, M. E. 1998Estrogen-dependent modulation of rat brain ascorbate levels and ischemia-induced ascorbate lossBrain Res803105113PubMedGoogle Scholar
  29. 29.
    Wang, H., Joseph, J. A. 1999Quantifying cellular oxidative stress by a dichlorofluorescein assay using microplate readerFree Radic. Biol. Med27612616PubMedGoogle Scholar
  30. 30.
    Dean, R. T., Shanlin, F. U., Stocker, R., Davies, M. J. 1997Biochemistry and Pathology of radicals-mediated protein oxidationBiochem. J324118PubMedGoogle Scholar
  31. 31.
    Jones, D. T., Taylor, W. R., Thornton, J. M. 1994A model recognition approach to the prediction of all-helical membrane protein structure and topologyBiochemistry3330383049PubMedGoogle Scholar
  32. 32.
    Von Heijne, G. 1994Membrane proteins: From sequence to structureAnnu. Rev. Biophys. Biomol. Struct23167192PubMedGoogle Scholar
  33. 33.
    Tieleman, D. P., Marrink, S. J., Berendsen, H. J. C. 1997A computer perspective of membranes: molecular dynamics studies of lipid bilayer systemsBiochem. Biophys. Acta1331235270PubMedGoogle Scholar
  34. 34.
    Moosmann, B., Behl, C. 2000Cytoprotective antioxidant function of tyrosine and tryptophan residues in transmembrane proteinsEur. J. Biochem26756875692PubMedGoogle Scholar
  35. 35.
    McCullough, L. D., Hurn, P. D. 2003Estrogen and ischemic neuroprotection: an integrated viewTrends Endocrinol. Metab14228235PubMedGoogle Scholar
  36. 36.
    Bath, P. M. W. and Gray, L. J. 2005. Association between hormone replacement therapy and subsequent stroke: a meta-analysis BMJ, doi: 10.1136/bmj.38331.655347.8FGoogle Scholar
  37. 37.
    Cos, P., De Bruyne, T., Apers, S., Vanden Berghe, D., Pieters, L., Vlietinck, A. J 2003Phytoestrogens: recent developmentsPlanta Med69589599PubMedGoogle Scholar
  38. 38.
    Park, D., Huang, T., Frishman, W. H. 2005Phytoestrogens as cardioprotective agentsCardiol. Rev131317PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2005

Authors and Affiliations

  • Helena Cimarosti
    • 1
  • Ionara R. Siqueira
    • 2
  • Lauren L. Zamin
    • 1
  • Melissa Nassif
    • 1
  • Rodrigo Balk
    • 1
  • Rudimar Frozza
    • 1
  • Carla Dalmaz
    • 1
  • Carlos Alexandre Netto
    • 1
  • Christianne Salbego
    • 1
  1. 1.Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde UFRGS Porto AlegreBrazil
  2. 2.Centro Universitário UNIVATES LajeadoBrazil

Personalised recommendations