Impairment of endothelial nitric oxide production by acute glucose overload

We examined the effects of acute glucose overload (pretreatment for 3 h wit h 23 mM D-glucose) on the cellular productivity of nitric oxide (NO) in bov ine aortic endothelial cells (BAEC). We had previously reported (Kimura C, Oike M, and Ito Y. Circ Res, 82: 677-685, 1998) that glucose overload impai rs Ca2+ mobilization due to an accumulation of superoxide anions (O-2(-)) i n BAEC. In control cells, ATP induced an increase in NO production, assesse d 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 dismutas e, but not by catalase or deferoxamine. Furthermore, pyrogallol, an O-2(-) donor, also attenuated ATP-induced increase in DAF-2 fluorescence. In contr ast, a nonspecific intracellular Ca2+ concentration increase induced by the Ca2+ ionophore A-23187, which depletes the intracellular store sites, elev ated DAF-2 fluorescence in both control and high D-glucose-treated cells in Ca2+-free solution. These results indicate that glucose overload impairs N O production by the O-2(-)-mediated attenuation of Ca2+ entry.