Glycogen synthase kinase 3β and β-catenin pathway is involved in toll-like receptor 4-mediated NADPH oxidase 1 expression in macrophages

Macrophage activation contributes to the pathogenesis of atherosclerosis. In the vascular system, the major source of reactive oxygen species is the NADPH oxidase (Nox) family. Nox1 is induced by lipopolysaccharide (LPS) in macrophages, but the expression mechanism is not fully understood. We found that LPS causes β-catenin accumulation by glycogen synthase kinase 3β (GSK3β) inactivation, and that β-catenin accumulation increases Nox1 expression. LPS induced Nox1 mRNA expression and reactive oxygen species generation in Raw264.7 cells. Using bone marrow-derived macrophages from toll-like receptor 4 mutant mice, we also tested whether LPS-induced Nox1 expression is toll-like receptor 4 dependent. LPS caused GSK3β phosphorylation, induced β-catenin accumulation and increased nuclear translocation. The GSK3β inhibitor LiCl potentiated LPS-induced Nox1 expression in accordance with β-catenin accumulation and nuclear translocation. Conversely, ectopic expression of a constitutively active GSK3β mutant severely attenuated Nox1 expression. These findings identify a novel regulatory pathway controlling Nox1 expression by LPS-stimulated macrophages.