Key Laboratory of Functional Small Organic Molecule, Ministry of Education, Key Laboratory of Green Chemistry, Jiangxi Province and College of Chemistry and Chemical Engineering, Jiangxi Normal University, Nanchang, China. Electronic address: [Email]
Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and oxidative stress is usually considered as an important factor to the pathogenesis of various diseases. Inorganic nitrite, previously viewed as a harmful substance in our diet or inert metabolites of endogenous NO, is recently identified as an important biological NO reservoir in vasculature and tissues. Stimulation of a nitrite-NO pathway shows organ-protective effects on oxidative stress and inflammation, but the mechanisms or target are not clear. In this study, the hypothesis that inorganic nitrite attenuated lipopolysaccharide (LPS)-induced oxidative stress in mice and in macrophage cells by modulating NADPH oxidase activity and NO bioavailability were investigated. We showed that nitrite treatment, in sharp contrast with the worsening effect of NO synthases inhibition, significantly attenuated aortic oxidative stress, endothelial dysfunction and mortality in LPS-induced shock in mice. Mechanistically, protective effects of nitrite were abolished by NO scavenger and xanthine oxidase inhibitor, and inhibition of NADPH oxidase with apocynin attenuated LPS-induced oxidative stress similar to that of nitrite. In the presence of nitrite, no further effect of apocynin was observed, suggesting NADPH oxidase as a possible target. In LPS-activated macrophage cells, nitrite reduced NADPH oxidase activity and oxidative stress and these effects of nitrite were also abolished by NO scavenger and xanthine oxidase inhibitor, where xanthine oxidase-mediated reduction of nitrite attenuated NADPH oxidase activity in activated macrophages via a NO-dependent mechanism. In conclusion, these novel findings position NADPH oxidase in the inflammatory vasculature as a prime target for the antioxidant effects of inorganic nitrite, and open a new direction to modulate the inflammatory response.