Stroke, Vol 25, 857-862, Copyright © 1994 by American Heart Association
DM Rosenbaum, J Kalberg and JA Kessler
BACKGROUND AND PURPOSE: Studies showing efficacy with free radical
scavengers have been conflicting, and when protection was demonstrated it
was attributed to action at the level of the vascular endothelium. The
purpose of this study was to test the hypotheses that neuronal free radical
formation plays a role in the ischemic cascade and occurs intracellularly
and that free radical scavengers, if taken up intracellularly, will protect
against hypoxic damage. METHODS: A tissue culture model of hypoxia followed
by restoration of oxygen was employed, using both superior cervical ganglia
and hippocampal neurons. Cells were exposed to an anoxic atmosphere of 95%
N2 and 5% CO2 and examined 2 to 24 hours later after restoration of oxygen.
Determination of survival was measured by trypan blue exclusion. Nitroblue
tetrazolium stain was used to assess free radical formation. RESULTS:
Pretreatment with free superoxide dismutase did not decrease cell death
after hypoxia as measured by trypan blue exclusion. However, when
superoxide dismutase was taken up intracellularly under depolarizing
conditions (55 mmol/L KCl in the medium), cell death was decreased
significantly compared with hypoxic controls (28.7 +/- 4.34 versus 40.3 +/-
4.33; P < .03). During hypoxia neurons reduced nitroblue tetrazolium to
form the blue precipitate formazan, and the color change was blocked in
neurons pretreated with superoxide dismutase in depolarizing medium.
Similar findings occurred in both superior cervical ganglia and hippocampal
neurons. CONCLUSIONS: These findings provide evidence to support the role
of neuronal free radical formation in cell death secondary to hypoxia. In
addition, free radical scavengers, if taken up intracellularly, may
partially ameliorate their deleterious effect.
ARTICLES
Superoxide dismutase ameliorates neuronal death from hypoxia in culture
Department of Neurology, Albert Einstein College of Medicine, Bronx, NY 10461.
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