• Y J Yang, X L Wang, X H Yu, X Wang, M Xie, and C T Liu.
• Division of Neonatology, Department of Pediatrics, Xiang Ya Hospital, Central South University.
• Undersea Hyperbar M. 2008 Mar 1; 35 (2): 113-29.

Background And PurposeStudies suggest that after brain injury, hyperbaric oxygen (HBO2) is neuroprotective by stimulating cell proliferation. We examine whether HBO2 promotes neural stem cells (NSC) to proliferate and differentiate in neonatal hypoxic-ischemic (HI) rats.MethodsSeven-day-old rat pups were subjected to unilateral carotid artery ligation followed by 2 hours of hypoxia (8% O2). HBO2 was administered (2 ATA (atmospheres absolutes), once daily for 7 days) within 3 hours after HI. The proliferating neural stem cells in the subventricular zone (SVZ) and dentate gyrus (DG) were dynamically examined by 5-bromo-2-deoxyuridine (BrdU)/nestin immunofluorescence. Nestin protein was detected by western blot analysis at various time points (from 6 hours to 14 days) after HI. The migrating NSC were examined by BrdU/doublecortin (DCX) immunofluorescence 7 and 14 days after HI. The phenotype of the newborn cells was identified by BrdU/beta-tubulin, BrdU/ glial fibrillary acidic protein (GFAP) and BrdU/O4 (oligodendrocyte marker) immunofluorescence. Myelin basic protein (MBP) was examined by immunohistochemistry and pathological changes of the brain tissue were detected 28 days after HI.ResultsIn neonatal HI rats treated with HBO2, the proliferation of endogenous NSC was observed in the SVZ and DG. Cell numbers peaked 7 days after HI and proliferating NSC migrated to the cerebral cortex at 14 d after HI. Twenty-eight days after HI, an increase in newly generated neurons, oligodendrocytes and MBP was observed in the HBO2 group compared to the untreated and HI-treated rats.ConclusionsThis study suggests that HBO2 treatment may promote neurogenesis of the endogenous NSC in neonatal HI rats, contributing to repair of the injured brain.

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