The Protective Effect of Vitamin E and Desferrioxamine on Cultured Cerebral Neurons of Neonatal Mouse Damaged by Ischemic Condition

Lee, So Ra; Oh, Yeon Kyun; Park, Seung Taek

Abstract

PURPOSE: Perinatal asphyxia is an important cause of neurologic morbidity. Experiments in animal models of hypoxic-ischemic brain injury demonstrate that brain damage starts during hypoxia-ischemia. In order to evaluate the ischemic condition-induced neurotoxic effect in view of oxi-dative stress, we examined the cytotoxic effect in cultured cerebral neurons of neonatal mouse.
METHODS
Dissociated cell cultures were prepared from cerebrum of neonatal mouse. Tissues were diced into small pieces and were incubated in phosphate buffered saline at 37degrees C. Isolated cells were resuspended in the medium and plated in poly-L-lysine coated 96 well multichambers at a cell density of 5x104cells/well. Cells were grown in a 5% CO2/95% air atmosphere at 37degrees C. Cytotoxic effects were examined in the cultured cerebral neurons with time interval in the ischemic condition with a 95% nitrogen/5% CO2. And the protective effect of vitamin E and desferrioxamine as an antioxidant was examined by MTT assay and neurofilament enzymeimmunoassay(EIA). Microscopic examinations were also done.
RESULTS
Ischemic condition markedly decreased the cell viability in a time-dependent manner in cultured cerebral neurons. MTT50 value was estimated at 10 minutes, when cerebral neurons were incubated for various time intervals in ischemic condition. Under light microscopy, the number of cells and neurites were decreased when cerebral neurons were cultured for 10 minutes in the ischemic condition. Vitamin E was an effective antioxidant in blocking ischemic condition-induced neurotoxicity, while desferrioxamine was not in these cultures.
CONCLUSION
It is suggested that ischemic conditions are neurotoxic and selective antioxidant such as vitamin E is effective in protecting against the neurotoxicity induced by ischemic condition in cultured cerebral neurons of neonatal mouse.