Abstract
A common factor in equine laminitis, metabolic syndrome and obesity is oxidative stress due to the accumulation of reactive oxygen species (ROS). Several enzymes contribute to the production of ROS including NADPH oxidase (NOX). In the present study, we examined superoxide generation by NOX in equine palmar digital arteries and veins. Palmar digital arteries and veins were isolated from clinically healthy horses euthanized for other reasons. Gene expression of NOX 1, 2 and 4 was detected in both arteries and veins using PCR primers designed from predicted sequences in the Equus caballus (domestic horse) genome. Western blot analysis confirmed protein expression of these isoforms. NOX activity was measured in freshly isolated rings using the superoxide detector lucigenin. The NOX inhibitor diphenylene iodonium (1 μM), as well as the superoxide dismutase mimetic, EUK‐134 (1 μM) significantly inhibited superoxide generation in both arteries and veins suggesting equine digital vessels generate superoxide through NOX. The nitric oxide synthase inhibitor, L‐NAME (10 μM), the mitochondrial inhibitor rotenone (1 μM) and the cyclooxygenase inhibitor indomethacin (1 μM) had no effect on NOX activity. These data suggest that equine palmar digital arteries and veins can generate superoxide through activation of NADPH oxidase. Funded by Auburn University Animal Health & Disease Research Funds.
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