Acute glucose overload abolishes Ca2+ oscillation in cultured endothelial cells from bovine aorta: a possible role of superoxide anion.
Effects of acute glucose overload on [Ca2+]i were investigated in cultured endothelial cells from bovine aorta. Application of 0.1 micromol/L ATP elicited an oscillatory increase in [Ca2+]i (Ca2+ oscillation) in Krebs solution containing 11.5 mmol/L glucose. The frequency of Ca2+ oscillation induced by ATP increased in a concentration-dependent manner, ranging between 0.03 and 1 micromol/L. When cells were preincubated with 23 mmol/L glucose-containing Krebs solution (high glucose solution) for 3 hours, 0.1 micromol/L ATP failed to induce Ca2+ oscillation but evoked only a phasic followed by sustained increase in [Ca2+]i. Application of a higher concentration of ATP (10 micromol/L) evoked a transient increase in [Ca2+]i both in control and high glucose-treated cells. However, the falling phase of [Ca2+]i was prolonged in high glucose-treated cells. Thapsigargin (1 micromol/L), an inhibitor of endoplasmic Ca2+-ATPase, induced a transient followed by a sustained increase in [Ca2+]i in control cells. Preincubation with high glucose solution increased the rate of rise of the thapsigargin-induced increase in [Ca2+]i and abolished the sustained increase, suggesting that glucose overload accelerates Ca2+ leak from intracellular store sites and impairs Ca2+ release-activated Ca2+ entry. We found that all of the glucose overload-induced changes in Ca2+ mobilization could be mimicked by xanthine with xanthine oxidase and abolished by superoxide dismutase. These results indicate that acute glucose overload accumulates superoxide anion in bovine aortic endothelial cells, thereby diminishing ATP-induced Ca2+ oscillation through the impairment of Ca2+ homeostasis.
Kimura
C
C
Department of Pharmacology, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Oike
M
M
Ito
Y
Y
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001013:Aorta, Thoracic; D002118:Calcium / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D016166:Free Radical Scavengers / Q000494:pharmacology; D005947:Glucose / Q000494:pharmacology*; D007700:Kinetics; D007811:Lanthanum / Q000494:pharmacology; D009991:Oscillometry; D012430:Ruthenium Red / Q000494:pharmacology; D013482:Superoxide Dismutase / Q000494:pharmacology; D013481:Superoxides / Q000378:metabolism*; D019284:Thapsigargin / Q000494:pharmacology; D013997:Time Factors
D016166:Free Radical Scavengers; D013481:Superoxides; D012430:Ruthenium Red; D019284:Thapsigargin; D007811:Lanthanum; D000255:Adenosine Triphosphate; D013482:Superoxide Dismutase; D005947:Glucose; D002118:Calcium