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Papers In Press, published online ahead of print April 7, 2004
J. Biol. Chem, 10.1074/jbc.M402920200
Submitted on March 16, 2004
Revised on March 31, 2004
Accepted on April 6, 2004

Reoxygenation-induced constriction in murine coronary arteries: role of endothelial NADPH oxidase (gp91phox) and intracellular superoxide

John Q. Liu, Igor N. Zelko, and Rodney J. Folz

Medicine, Duke University Medical Center, Durham, NC 27710

Corresponding Author: john.liu{at}duke.edu

Previous work suggests that superoxide mediates hypoxia-reoxygenation (H/R)-induced constriction of isolated mouse coronary arteries (CA). To determine the source of superoxide overproduction during H/R, we studied CA obtained from transgenic (Tg) mice overexpressing human Cu/Zn SOD and mice lacking gp91phox, using an in vitro vascular ring bioassay. We found that under normoxic conditions, CA isolated from wild type (wt) mice, Cu/Zn SOD Tg mice and gp91phox knockout (KO) mice had similar contractile responses to U46619 and hypoxia, and similar dilation responses to acetylcholine (ACh). In wt CA, 30 minutes of hypoxia (1% O2) followed by reoxygenation (16% O2) resulted in further coronary vasoconstriction (internal diameter (ID) from 105 ± 11 to 84.5 ± 17.9 µm), whereas this response was completely blocked in both Cu/Zn SOD Tg and gp91phox KO CA (104.3 ± 10.5 to 120.7 ± 14 µm and 143.3 ± 15.3 to 172.7 ± 12.5 µm, respectively, p<0.01). Furthermore, we show that H/R enhances the generation of superoxide radicals in wt CA (25.8 ± 0.7 RLU /s), whereas Cu/Zn SOD Tg CA (12.2 ± 0.8 RLU/s, p<0.01) and gp91phox CA (12.5 ± 0.9 RLU /s, p<0.01) show reduced levels. These results demonstrate that H/R-induced vasoconstriction is mediated by intracellular superoxide overproduction via endothelial NADPH oxidase gp91phox. Therefore, increasing endogenous levels of Cu/Zn SOD in CA may provide a novel cardio-protective strategy for maintaining coronary perfusion under conditions of H/R.


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