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Fig. 6. Inhibitory effects of ROS on Ipump.
Ipump was recorded in cortical neurons at 60 mV
before and after an oxidant insult. (A) Hydrogen peroxide (0.25 mM) showed a
marked inhibitory effect on Ipump after a few minutes
application. This suppression of the Na+, K+-pump
activity was markedly prevented in the presence of catalase (250 U/ml) or
pyruvate (5 mM) added 5 minutes before and during the exposure. Succinate, on
the other hand, showed no such effect even at high concentration of 20 mM
(n=5-6 for each test). (B) Acute application (10 minutes) of 20 µM
menadione, a stimulator of endogenous production of O2-,
suppressed Ipump; this inhibition was prevented by 25 U/ml
SOD or 5 mM pyruvate but not by 20 mM succinate (n
6 in each
test). (C) Chronic exposure to 8 µM menadione for 9-10 hours also
diminished Ipump; both pyruvate (5 mM) and succinate (20
mM) protected the Na+, K+-pump activity from damage by
the endogenous ROS (n=5-7). (D) Acute (20 minutes) and chronic (15-20
hours) applications of DMNQ (5-20 µM) inhibited Ipump,
pyruvate (5 mM) or succinate (20 mM) antagonized the inhibition in both
conditions (n=5-8).*P<0.05 compared with controls;
#P<0.05 compared with hydrogen peroxide, menadione, or DMNQ
alone.