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Am J Physiol Lung Cell Mol Physiol 280: L1196-L1202, 2001;
1040-0605/01 $5.00
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Vol. 280, Issue 6, L1196-L1202, June 2001

Interleukin-10-mediated inhibition of free radical generation in macrophages

Sujatha Dokka1, Xianglin Shi2, Stephen Leonard2, Liying Wang2, Vincent Castranova2, and Yon Rojanasakul1

1 Department of Basic Pharmaceutical Sciences, West Virginia University, Morgantown 26506; and 2 Pathology and Physiology Research Branch, National Institute for Occupational Safety and Health, Morgantown, West Virginia 26505

Interleukin-10 (IL-10) is a pleiotropic cytokine that controls inflammatory processes by suppressing the production of proinflammatory cytokines that are known to be transcriptionally regulated by nuclear factor-kappa B (NF-kappa B). Although still controversial, IL-10 has been shown to inhibit NF-kappa B activation through a process that involves proteolytic degradation of inhibitory subunit Ikappa B-alpha . What is not known, however, is the mechanism by which IL-10 exerts its effect on Ikappa B-alpha degradation. The present study investigates the possible role of reactive oxygen species (ROS) and their inhibition by IL-10 in NF-kappa B activation and Ikappa B-alpha degradation in macrophages. Treatment of the cells with lipopolysaccharide (LPS) caused activation of NF-kappa B and rapid proteolysis of Ikappa B-alpha as determined by the electrophoretic mobility shift assay, gene transfection, and Western blot. IL-10 pretreatment inhibited both NF-kappa B activation and Ikappa B-alpha degradation. Both of these processes were also inhibited by ROS scavengers, catalase (H2O2 scavenger), and sodium formate (·OH scavenger) but were minimally affected by superoxide dismutase (O<UP><SUB>2</SUB><SUP>−</SUP></UP> scavenger). These results suggests that ·OH radicals, formed by an H2O2-dependent, metal-catalyzed Fenton reaction, play a major role in this process. Electron spin resonance studies confirmed the formation of ·OH radicals in LPS-treated cells. Addition of IL-10 inhibited both Ikappa B-alpha degradation and generation of ·OH radicals in response to LPS stimulation. These results demonstrate, for the first time, direct evidence for the role of IL-10 in ROS-dependent NF-kappa B activation.

nuclear factor-kappa B; tumor necrosis factor-alpha ; oxygen free radicals


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