TY - JOUR
T1 - Endothelial cell respiration is affected by the oxygen tension during shear exposure
T2 - Role of mitochondrial peroxynitrite
AU - Jones, Charles I.
AU - Han, Zhaosheng
AU - Presley, Tennille
AU - Varadharaj, Saradhadevi
AU - Zweier, Jay L.
AU - Ilangovan, Govindasamy
AU - Alevriadou, B. Rita
PY - 2008/7
Y1 - 2008/7
N2 - Cultured vascular endothelial cell (EC) exposure to steady laminar shear stress results in peroxynitrite (ONOO-) formation intramitochondrially and inactivation of the electron transport chain. We examined whether the "hyperoxic state" of 21% O2, compared with more physiological O2 tensions (PO2), increases the shear-induced nitric oxide (NO) synthesis and mitochondrial superoxide (O 2.-) generation leading to ONOO- formation and suppression of respiration. Electron paramagnetic resonance oximetry was used to measure O2 consumption rates of bovine aortic ECs sheared (10 dyn/cm2, 30 min) at 5%, 10%, or 21% O2 or left static at 5% or 21% O2. Respiration was inhibited to a greater extent when ECs were sheared at 21% O2 than at lower PO2 or left static at different PO2. Flow in the presence of an endothelial NO synthase (eNOS) inhibitor or a ONOO- scavenger abolished the inhibitory effect. EC transfection with an adenovirus that expresses manganese superoxide dismutase in mitochondria, and not a control virus, blocked the inhibitory effect. Intracellular and mitochondrial O2.- production was higher in ECs sheared at 21% than at 5% O2, as determined by dihydroethidium and MitoSOX red fluorescence, respectively, and the latter was, at least in part, NO-dependent. Accumulation of NO metabolites in media of ECs sheared at 21% O2 was modestly increased compared with ECs sheared at lower PO2, suggesting that eNOS activity may be higher at 21% O 2. Hence, the hyperoxia of in vitro EC flow studies, via increased NO and mitochondrial O2.- production, leads to enhanced ONOO- formation intramitochondrially and suppression of respiration.
AB - Cultured vascular endothelial cell (EC) exposure to steady laminar shear stress results in peroxynitrite (ONOO-) formation intramitochondrially and inactivation of the electron transport chain. We examined whether the "hyperoxic state" of 21% O2, compared with more physiological O2 tensions (PO2), increases the shear-induced nitric oxide (NO) synthesis and mitochondrial superoxide (O 2.-) generation leading to ONOO- formation and suppression of respiration. Electron paramagnetic resonance oximetry was used to measure O2 consumption rates of bovine aortic ECs sheared (10 dyn/cm2, 30 min) at 5%, 10%, or 21% O2 or left static at 5% or 21% O2. Respiration was inhibited to a greater extent when ECs were sheared at 21% O2 than at lower PO2 or left static at different PO2. Flow in the presence of an endothelial NO synthase (eNOS) inhibitor or a ONOO- scavenger abolished the inhibitory effect. EC transfection with an adenovirus that expresses manganese superoxide dismutase in mitochondria, and not a control virus, blocked the inhibitory effect. Intracellular and mitochondrial O2.- production was higher in ECs sheared at 21% than at 5% O2, as determined by dihydroethidium and MitoSOX red fluorescence, respectively, and the latter was, at least in part, NO-dependent. Accumulation of NO metabolites in media of ECs sheared at 21% O2 was modestly increased compared with ECs sheared at lower PO2, suggesting that eNOS activity may be higher at 21% O 2. Hence, the hyperoxia of in vitro EC flow studies, via increased NO and mitochondrial O2.- production, leads to enhanced ONOO- formation intramitochondrially and suppression of respiration.
KW - Endothelium
KW - Mitochondria
KW - Reactive oxygen species
KW - Shear stress
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U2 - 10.1152/ajpcell.00549.2007
DO - 10.1152/ajpcell.00549.2007
M3 - Article
C2 - 18480296
AN - SCOPUS:52749091387
SN - 0363-6143
VL - 295
JO - American Journal of Physiology - Cell Physiology
JF - American Journal of Physiology - Cell Physiology
IS - 1
ER -