title: Nrf2 deficiency induces oxidative stress and promotes RANKL-induced osteoclast differentiation.
abstract: Nuclear factor-erythroid 2-related factor 2 (Nrf2) is a redox-sensitive transcription factor that regulates the expression of a variety of antioxidant and detoxification genes through an antioxidant-response element. Nrf2 has been shown to protect several types of cells against the acute and chronic injury that accompanies oxidative stress, but its role in osteoclasts remains unclear. In this study, we investigated the role of Nrf2 in osteoclast (OC) differentiation, a process in which reactive oxygen species (ROS) are generated and then participate, using Nrf2-knockout mice. Receptor activator of nuclear factor κB ligand (RANKL)-induced OC differentiation, actin ring formation, and osteoclastic bone resorption were substantially promoted in Nrf2-deficient OC precursor cells compared to wild-type cells. Under both unstimulated and RANKL-stimulated conditions, Nrf2 loss led to an increase in the intracellular ROS level and the oxidized-to-reduced glutathione ratio and a defect in the production of numerous antioxidant enzymes and glutathione. Moreover, pretreatment with N-acetylcysteine or diphenyleneiodonium significantly reduced the OC differentiation and decreased the intracellular ROS level in both Nrf2-deficient and wild-type cells. Pretreatment with sulforaphane and curcumin also inhibited the OC differentiation by activating Nrf2 in part. Nrf2 deficiency promoted the RANKL-induced activation of mitogen-activated protein kinases, including c-Jun N-terminal kinase, extracellular signal-regulated kinase, and p38; the induction of c-Fos; and the consequent induction of nuclear factor of activated T cells, cytoplasmic 1, a pivotal determinant of OC differentiation. Our results suggest that Nrf2 probably inhibits RANKL-induced OC differentiation by regulating the cellular redox status by controlling the expression of oxidative response genes, findings that might form the basis of a new strategy for treating inflammatory bone diseases.
authors: Department of Life Science and Research Center for Cellular Homeostasis, Ewha Woman's University, Seoul 120-750, Korea.
authors: Seungha
authors: S
authors: Hyeon
authors: Department of Life Science and Research Center for Cellular Homeostasis, Ewha Woman's University, Seoul 120-750, Korea.
authors: Hyojung
authors: H
authors: Lee
authors: Department of Life Science and Research Center for Cellular Homeostasis, Ewha Woman's University, Seoul 120-750, Korea.
authors: Yoohee
authors: Y
authors: Yang
authors: Department of Life Science and Research Center for Cellular Homeostasis, Ewha Woman's University, Seoul 120-750, Korea. Electronic address: jeongw@ewha.ac.kr.
authors: Woojin
authors: W
authors: Jeong
mesh_terms: D000818:Animals; D001862:Bone Resorption; D002454:Cell Differentiation; D004789:Enzyme Activation; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D020935:MAP Kinase Signaling System; D008297:Male; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D051267:NF-E2-Related Factor 2 / Q000172:deficiency* / Q000235:genetics; D016328:NF-kappa B / Q000378:metabolism; D050778:NFATC Transcription Factors / Q000378:metabolism; D010010:Osteoclasts / Q000502:physiology*; D018384:Oxidative Stress; D010544:Peroxidases / Q000378:metabolism; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism; D053245:RANK Ligand / Q000502:physiology*; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D051267:NF-E2-Related Factor 2; D016328:NF-kappa B; D050778:NFATC Transcription Factors; C494005:Nfatc1 protein, mouse; C495636:Nfe2l2 protein, mouse; D016760:Proto-Oncogene Proteins c-fos; D053245:RANK Ligand; D017382:Reactive Oxygen Species; D010544:Peroxidases; D005978:Glutathione
keywords: AP-1; ARE; BMM; DPI; ERK; FBS; Free radicals; GSH; GSSG; JNK; Keap1; Kelch-like ECH-associated protein 1; M-CSF; MAPK; MAPK phosphatase; MEM; MKP; Mitogen-activated protein kinases; N-acetylcysteine; NAC; NAD(P)H:quinone oxidoreductase 1; NADPH; NADPH oxidase; NF-κB; NFATc1; NQO1; Nox; Nrf2; OC; OC differentiation; PBS; Prx; RANKL; ROS; Reactive oxygen species; Redox regulation; SOD; Srx; TNF; TNF receptor-associated factor; TRAF; TRAP; TRAP(+) MNC; TRAP-positive multinucleated cell; Trx; Trx reductase; TrxR; activator protein-1; antioxidant response element; bone marrow-derived macrophage; c-Jun N-terminal kinase; diphenyleneiodonium; extracellular signal-regulated kinase; fetal bovine serum; glutathione; macrophage colony-stimulating factor; minimal essential medium; mitogen-activated protein kinase; nicotinamide adenine dinucleotide phosphate; nuclear factor of activated T cells, cytoplasmic 1; nuclear factor-erythroid 2-related factor 2; nuclear factor-κB; osteoclast; oxidized GSH; peroxiredoxin; phosphate-buffered saline; reactive oxygen species; receptor activator of NF-κB ligand; sulfiredoxin; superoxide dismutase; tartrate-resistant acid phosphatase; thioredoxin; tumor necrosis factor