ABSTRACT
Reactive oxygen species are known to participate in the regulation of intracellular signaling pathways, including activation of NF-kappaB. Recent studies have indicated that increases in intracellular concentrations of hydrogen peroxide (H(2)O(2)) have anti-inflammatory effects in neutrophils, including inhibition of the degradation of I kappaB alpha after TLR4 engagement. In the present experiments, we found that culture of lipopolysaccharide-stimulated neutrophils and HEK 293 cells with H(2)O(2) resulted in diminished ubiquitination of I kappaB alpha and decreased SCF(beta-TrCP) ubiquitin ligase activity. Exposure of neutrophils or HEK 293 cells to H(2)O(2) was associated with reduced binding between phosphorylated I kappaB alpha and SCF(beta-TrCP) but no change in the composition of the SCF(beta-TrCP) complex. Lipopolysaccharide-induced SCF(beta-TrCP) ubiquitin ligase activity as well as binding of beta-TrCP to phosphorylated I kappaB alpha was decreased in the lungs of acatalasemic mice and mice treated with the catalase inhibitor aminotriazole, situations in which intracellular concentrations of H(2)O(2) are increased. Exposure to H(2)O(2) resulted in oxidative modification of cysteine residues in beta-TrCP. Cysteine 308 in Blade 1 of the beta-TrCP beta-propeller region was found to be required for maximal binding between beta-TrCP and phosphorylated I kappaB alpha. These findings suggest that the anti-inflammatory effects of H(2)O(2) may result from its ability to decrease ubiquitination as well as subsequent degradation of I kappaB alpha through inhibiting the association between I kappaB alpha and SCF(beta-TrCP).
Subject(s)
Acatalasia/metabolism , Acute Lung Injury/metabolism , Hydrogen Peroxide/pharmacology , I-kappa B Proteins/metabolism , Oxidants/pharmacology , SKP Cullin F-Box Protein Ligases/metabolism , Acatalasia/chemically induced , Acatalasia/genetics , Acute Lung Injury/chemically induced , Amitrole/pharmacology , Animals , Catalase/antagonists & inhibitors , Catalase/metabolism , Cells, Cultured , Enzyme Inhibitors/pharmacology , Humans , Kidney/cytology , Lipopolysaccharides/pharmacology , Mice , Mice, Inbred C3H , Mice, Inbred C57BL , Mice, Mutant Strains , NF-KappaB Inhibitor alpha , Neutrophils/cytology , Neutrophils/drug effects , Phosphorylation/drug effects , SKP Cullin F-Box Protein Ligases/genetics , UbiquitinationABSTRACT
The catalase activities in the blood and organs of the acatalasemic (C3H/AnLCsb-Csb) mouse of the C3H strain are lower than those of the normal (C3H/AnLCSa-Csa) mouse. We examined the effects of post low-dose (0.5 Gy) X-ray irradiation which reduced the oxidative damage under carbon tetrachloride-induced hepatopathy in acatalasemic or normal mice. As a result, the 0.5 Gy irradiation after carbon tetrachloride administration decreased the glutamic oxaloacetic and glutamic pyruvic transaminase activity in the acatalasemic mouse blood to a level similar to that of the acatalasemic mouse blood not treated with carbon tetrachloride; this is in contrast to a high-dose (15 Gy) irradiation. In the same manner, pathological disorder was improved by 0.5 Gy irradiation. The fat degeneration in normal mice was quickly reduced, in contrast to acatalasemic mice. These findings suggest that low-dose irradiation after carbon tetrachloride administration accelerates the rate of recovery and that catalase plays an important role in the recovery from hepatopathy induced by carbon tetrachloride, in contrast to high-dose irradiation.
