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NADPH oxidase activity is higher in cerebral versus systemic arteries of four animal species: role of Nox2

¹Department of Pharmacology, Monash University, Clayton, Melbourne, Victoria, Australia; and ²Department of Pharmacology and ³Howard Florey Institute, University of Melbourne, Parkville, Melbourne, Victoria, Australia

Submitted 11 September 2008 ; accepted in final form 15 November 2008

We previously reported that NADPH oxidase activity is greater in intracranial cerebral versus systemic arteries of the rat. Here, we first tested whether NADPH oxidase activity is also greater in intracranial cerebral than systemic arteries of three other animal species, i.e., mouse, rabbit, and pig. Second, using Nox2-deficient mice, we evaluated the involvement of Nox2-containing NADPH oxidases in any such regional differences. NADPH-stimulated superoxide (O₂^–) production by basilar, middle cerebral arteries (MCA), and common carotid arteries (CA) and thoracic aorta (AO) from rat, mouse, rabbit, and pig was measured using lucigenin-enhanced chemiluminescence. Basal production of O₂^– and hydrogen peroxide (H₂O₂) by cerebral arteries, AO, and CA from wild-type (WT) and Nox2^–/– mice was measured using L-012-enhanced chemiluminescence and Amplex Red fluorescence, respectively. Western blotting was used to measure Nox2 and SOD1–3 protein expression, and immunofluorescence was used to localize Nox2, in mouse arteries. In rats, WT mice, rabbits, and pigs, NADPH-stimulated O₂^– production by cerebral arteries was up to 40-fold greater than that in AO and CA. In WT mice, basal O₂^– and H₂O₂ production by cerebral arteries was ninefold and 2.5-fold higher, respectively, than that in AO and CA and was associated with 40% greater expression of Nox2 protein. Nox2 immunofluorescence was localized to the endothelium, and to a lesser extent the adventitia, in all mouse arteries and appeared to be more intense in endothelium of MCA than AO or CA. In Nox2^–/– mice, NADPH-stimulated O₂^– production by cerebral arteries was 35% lower than that in WT mice, whereas Nox2 deletion had no significant effect on O₂^– production by AO or CA. Thus NADPH oxidase activity is greater in intracranial cerebral versus systemic arteries of several animal species and is associated with higher cerebrovascular expression and activity of Nox2.

cerebral arteries; reactive oxygen species

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