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Molecular Pharmacology

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Research ArticleArticle

Overexpression of Cytochrome P450 CYP2J2 Protects against Hypoxia-Reoxygenation Injury in Cultured Bovine Aortic Endothelial Cells

Baichun Yang, LeRae Graham, Serguei Dikalov, Ronald P. Mason, John R. Falck, James K. Liao and Darryl C. Zeldin
Molecular Pharmacology August 2001, 60 (2) 310-320; DOI: https://doi.org/10.1124/mol.60.2.310
Baichun Yang
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LeRae Graham
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Serguei Dikalov
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Ronald P. Mason
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John R. Falck
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James K. Liao
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Darryl C. Zeldin
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Abstract

CYP2J2 is abundant in human heart and its arachidonic acid metabolites, the epoxyeicosatrienoic acids (EETs), have potent vasodilatory, antiinflammatory and cardioprotective properties. This study was designed to examine the role of CYP2J2 in hypoxia-reoxygenation-induced injury in cultured bovine aortic endothelial cells (BAECs). Early passage BAECs were exposed to 24-h hypoxia followed by 4-h reoxygenation (HR). HR resulted in cell injury, as indicated by significant increases in lactate dehydrogenase (LDH) release and trypan blue stained cells (p < 0.01) and was associated with a decrease in CYP2J2 protein expression. Transfection of BAECs with the CYP2J2 cDNA resulted in increased CYP2J2 expression and arachidonic acid epoxygenase activity, compared with cells transfected with an irrelevant green fluorescent protein (GFP) cDNA. HR induced significant injury in GFP-transfected BAECs, as indicated by increases in LDH release and trypan blue-stained cells (p < 0.01); however, the HR-induced injury was markedly attenuated in CYP2J2-transfected cells (p < 0.01). HR increased cellular 8-iso-prostaglandin F2α (p< 0.05), and decreased eNOS expression, l-arginine uptake and conversion, and nitrite production (p < 0.01) in GFP-transfected BAECs. CYP2J2 transfection attenuated the HR-induced increase in 8-iso-prostaglandin F2α(p < 0.05) and decreased the amount of extracellular superoxide detected by cytochrome creduction under normoxic conditions (p < 0.05) but did not significantly affect HR-induced decreases in eNOS expression,l-arginine uptake and conversion, and nitrite production. Treatment of BAECs with synthetic EETs and/or epoxide hydrolase inhibitors also showed protective effects against HR injury (p < 0.05). These observations suggest: (1) HR results in endothelial injury and decreased CYP2J2 expression; (2) transfection with the CYP2J2 cDNA protects against HR injury; and (3) the cytoprotective effects of CYP2J2 may be mediated, at least in part, by antioxidant effects.

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Molecular Pharmacology: 60 (2)
Molecular Pharmacology
Vol. 60, Issue 2
1 Aug 2001
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Research ArticleArticle

Overexpression of Cytochrome P450 CYP2J2 Protects against Hypoxia-Reoxygenation Injury in Cultured Bovine Aortic Endothelial Cells

Baichun Yang, LeRae Graham, Serguei Dikalov, Ronald P. Mason, John R. Falck, James K. Liao and Darryl C. Zeldin
Molecular Pharmacology August 1, 2001, 60 (2) 310-320; DOI: https://doi.org/10.1124/mol.60.2.310

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Research ArticleArticle

Overexpression of Cytochrome P450 CYP2J2 Protects against Hypoxia-Reoxygenation Injury in Cultured Bovine Aortic Endothelial Cells

Baichun Yang, LeRae Graham, Serguei Dikalov, Ronald P. Mason, John R. Falck, James K. Liao and Darryl C. Zeldin
Molecular Pharmacology August 1, 2001, 60 (2) 310-320; DOI: https://doi.org/10.1124/mol.60.2.310
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