Potential conflict of interest: Nothing to report.
Erythropoietin protects the developing brain from hyperoxia-induced cell death and proteome changes†
Article first published online: 9 DEC 2008
Copyright © 2008 American Neurological Association
Annals of Neurology
Volume 64, Issue 5, pages 523–534, November 2008
How to Cite
Kaindl, A. M., Sifringer, M., Koppelstaetter, A., Genz, K., Loeber, R., Boerner, C., Stuwe, J., Klose, J. and Felderhoff-Mueser, U. (2008), Erythropoietin protects the developing brain from hyperoxia-induced cell death and proteome changes. Ann Neurol., 64: 523–534. doi: 10.1002/ana.21471
- Issue published online: 9 DEC 2008
- Article first published online: 9 DEC 2008
- Manuscript Accepted: 20 JUN 2008
- Manuscript Revised: 11 JUN 2008
- Manuscript Received: 16 APR 2008
- German Ministry for Education and Research (BMBF) within the National Genome Research Network (NGFN) program. Grant Number: project no. FK2 01GR0442
- European Commission (Sixth Framework Program). Grant Number: LSHM-CT-2006-036534
- Sanitätsrat Dr Emil Alexander Huebner und Gemahlin-Stiftung. Grant Number: project no. T114/14880/2005/5m
- Rahel Hirsch scholarship from the Charité–Universitätsmedizin Berlin
- Sonnenfeld-Stiftung Berlin
Oxygen toxicity has been identified as a risk factor for adverse neurological outcome in survivors of preterm birth. In infant rodent brains, hyperoxia induces disseminated apoptotic neurodegeneration. Because a tissue-protective effect has been observed for recombinant erythropoietin (rEpo), widely used in neonatal medicine for its hematopoietic effect, we examined the effect of rEpo on hyperoxia-induced brain damage.
Six-day-old C57Bl/6 mice or Wistar rats were exposed to hyperoxia (80% O2) or normoxia for 24 hours and received rEpo or normal saline injections intraperitoneally. The amount of degenerating cells in their brains was determined by DeOlmos cupric silver staining. Changes of their brain proteome were determined through two-dimensional electrophoresis and mass spectrometry. Western blot, enzyme activity assays and real-time polymerase chain reaction were performed for further analysis of candidate proteins.
Systemic treatment with 20,000 IE/kg rEpo significantly reduced hyperoxia-induced apoptosis and caspase-2, -3, and -8 activity in the brains of infant rodents. In parallel, rEpo inhibited most brain proteome changes observed in infant mice when hyperoxia was applied exclusively. Furthermore, brain proteome changes after a single systemic rEpo treatment point toward a number of mechanisms through which rEpo may generate its protective effect against oxygen toxicity. These include reduction of oxidative stress and restoration of hyperoxia-induced increased levels of proapoptotic factors, as well as decreased levels of neurotrophins.
These findings are highly relevant from a clinical perspective because oxygen administration to neonates is often inevitable, and rEpo may serve as an adjunctive neuroprotective therapy. Ann Neurol 2008;64:523–534