Hypoxic ischaemic hypothermia promotes neuronal differentiation and inhibits glial differentiation from newly generated cells in the SGZ of the neonatal rat brain

Abstract

Hypothermia is a potential therapy for cerebral hypoxic ischaemic injury in adults and neonates. The mechanism of the neuroprotective effects of hypothermia after hypoxia-ischaemia (HI) in the developing rat brain remains unclear. In this research, 7-day-old rats underwent left carotid artery ligation followed by the administration of 8% oxygen for 2 h. These rats were divided into hypothermic (rectal temperature, 32-33 °C for 24 h) and normothermic (36-37 °C for 24 h) groups immediately after HI. All rats were given 50 mg/kg/day 5-bromodeoxyuridine (BrdU) intraperitoneally at 4-6 days and sacrificed at 1 or 2 weeks after HI. We found a significant decrease in infarct volume and the neuron loss were also detected in the subgranular zone (SGZ) in the hypothermic group at 7 and 14 days after HI compared with the normothermic group. BrdU immunopositive cells were reduced greatly in the hypothermic group compared with the normothermic group. Hypothermia did not change the number of nestin-labelled cells in the ipsilateral SGZ at 1 and 2 weeks after HI. The differentiation of newly generated cells was assessed by double immunolabelling of BrdU with glial fibrillary acidic protein (GFAP), O4 or Neuronal Nuclei (NeuN). The ratio of BrdU(+)-GFAP(+) or BrdU(+)-O4(+) to total BrdU(+) staining decreased dramatically, but the ratio of BrdU(+)-NeuN(+) to total BrdU(+) staining increased significantly in the hypothermic group compared to the normothermic group at 2 and 6 weeks after HI. These results suggest that the reduction in neuron loss observed after mild hypothermia may be associated with enhanced neuronal differentiation and decreased glial differentiation in the SGZ after HI. These observations are noteworthy for clinical hypothermia therapy following cerebral HI injury during the perinatal period.