Acute Glucose Overload Abolishes Ca²⁺ Oscillation in Cultured Endothelial Cells From Bovine Aorta

A Possible Role of Superoxide Anion

From the Department of Pharmacology, Faculty of Medicine, Kyushu University, Fukuoka, Japan.

Correspondence to Masahiro Oike, MD, PhD, Department of Pharmacology, Faculty of Medicine, Kyushu University, Fukuoka 812-8582, Japan. E-mail moike{at}pharmaco.med.kyushu-u.ac.jp

Abstract—Effects of acute glucose overload on [Ca²⁺]_i were investigated in cultured endothelial cells from bovine aorta. Application of 0.1 µmol/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 µmol/L. When cells were preincubated with 23 mmol/L glucose–containing Krebs solution (high glucose solution) for 3 hours, 0.1 µmol/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 µmol/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 µmol/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.

Key Words: endothelium • Ca²⁺ • oscillation • superoxide • glucose overload

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