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Department of Pharmacology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan
To investigate the possible
cellular mechanisms of the ischemia-induced impairments of cerebral
microcirculation, we investigated the effects of hypoxia/reoxygenation
on the intracellular Ca2+ concentration
([Ca2+]i) in bovine brain microvascular
endothelial cells (BBEC). In the cells kept in normal air, ATP elicited
Ca2+ oscillations in a concentration-dependent manner. When
the cells were exposed to hypoxia for 6 h and subsequent
reoxygenation for 45 min, the basal level of
[Ca2+]i was increased from 32.4 to 63.3 nM,
and ATP did not induce Ca2+ oscillations.
Hypoxia/reoxygenation also inhibited capacitative Ca2+
entry (CCE), which was evoked by thapsigargin
(
[Ca2+]i-CCE: control, 62.3 ± 3.1 nM;
hypoxia/reoxygenation, 17.0 ± 1.8 nM). The impairments of
Ca2+ oscillations and CCE, but not basal
[Ca2+]i, were restored by superoxide
dismutase and the inhibitors of mitochondrial electron transport,
rotenone and thenoyltrifluoroacetone (TTFA). By using a superoxide
anion (O2
)-sensitive luciferin derivative MCLA, we
confirmed that the production of O2 was induced by
hypoxia/reoxygenation and was prevented by rotenone and TTFA. These
results indicate that hypoxia/reoxygenation generates O2 at mitochondria and impairs some Ca2+
mobilizing properties in BBEC.
superoxide anion; adenosine 5'-triphosphate; capacitative calcium entry; mitochondria
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