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Am J Physiol Endocrinol Metab 280: E171-E178, 2001;
0193-1849/01 $5.00
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Vol. 280, Issue 1, E171-E178, January 2001

Impairment of endothelial nitric oxide production by acute glucose overload

Chiwaka Kimura, Masahiro Oike, Tetsuya Koyama, and Yushi Ito

Department of Pharmacology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812 - 8582, Japan

We examined the effects of acute glucose overload (pretreatment for 3 h with 23 mM D-glucose) on the cellular productivity of nitric oxide (NO) in bovine aortic endothelial cells (BAEC). We had previously reported (Kimura C, Oike M, and Ito Y. Circ Res, 82: 677-685, 1998) that glucose overload impairs Ca2+ mobilization due to an accumulation of superoxide anions (O2-) in BAEC. In control cells, ATP induced an increase in NO production, assessed by diaminofluorescein 2 (DAF-2), an NO-sensitive fluorescent dye, mainly due to Ca2+ entry. In contrast, ATP-induced increase in DAF-2 fluorescence was impaired by glucose overload, which was restored by superoxide dismutase, but not by catalase or deferoxamine. Furthermore, pyrogallol, an O2- donor, also attenuated ATP-induced increase in DAF-2 fluorescence. In contrast, a nonspecific intracellular Ca2+ concentration increase induced by the Ca2+ ionophore A-23187, which depletes the intracellular store sites, elevated DAF-2 fluorescence in both control and high D-glucose-treated cells in Ca2+-free solution. These results indicate that glucose overload impairs NO production by the O2--mediated attenuation of Ca2+ entry.

calcium; superoxide; diaminofluorescein 2


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