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Treatment with hydrogen molecules prevents RANKL-induced osteoclast differentiation associated with inhibition of ROS formation and inactivation of MAPK, AKT and NF-kappa B pathways in murine RAW264.7 cells

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Abstract

The bone protective effects of the hydrogen molecule (H2) have been demonstrated in several osteoporosis models while the underlying molecular mechanism has remained unclear. Osteoclast differentiation is an important factor related to the pathogenesis of bone-loss related diseases. In this work, we evaluated the effects of incubation with H2 on receptor activator of NFκB ligand (RANKL)-induced osteoclast differentiation. We found that treatment with H2 prevented RANKL-induced osteoclast differentiation in RAW264.7 cells and BMMs. Treatment with H2 inhibits the ability to form resorption pits of BMMs stimulated by RANKL. Treatment with H2 reduced mRNA levels of osteoclast-specific markers including tartrate resistant acid phosphatase, calcitonin receptor, cathepsin K, metalloproteinase-9, carbonic anhydrase typeII, and vacuolar-type H+-ATPase. Treatment with H2 decreased intracellular reactive oxygen species (ROS) formation, suppressed NADPH oxidase activity, down-regulated Rac1 activity and Nox1 expression, reduced mitochondrial ROS formation, and enhanced nuclear factor E2-related factor 2 nuclear translocation and heme oxygenase-1 activity. In addition, treatment with H2 suppressed RANKL-induced expression of nuclear factor of activated T cells c1 and c-Fos. Furthermore, treatment with H2 suppressed NF-κB activation and reduced phosphorylation of p38, extracellular signal-regulated kinase, c-Jun-N-terminal kinase, and protein kinases B (AKT) stimulated with RANKL. In conclusion, hydrogen molecules prevented RANKL-induced osteoclast differentiation associated with inhibition of reactive oxygen species formation and inactivation of NF-κB, mitogen-activated protein kinase and AKT pathways.

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The authors have stated that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Osteology, The 89th Hospital of The Chinese PLA, 256 Beigong West Street, Weifang, 261021, China

    Dong-Zhu Li

  2. Department of Osteology, The 148th Hospital of The Chinese PLA, Zibo, 255300, China

    Qing-Xiang Zhang

  3. Intensive Care Unit, The 89th Hospital of The Chinese PLA, Weifang, 261021, China

    Xiao-Xian Dong

  4. Department of Respiratory Disease, The 88th Hospital of The Chinese PLA, Tai’an, 271000, China

    Huai-Dong Li

  5. Outpatient Clinics, The Air Force Engineering University, Xi’an, 710051, China

    Xin Ma

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  1. Dong-Zhu Li
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Correspondence to Dong-Zhu Li.

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Li, DZ., Zhang, QX., Dong, XX. et al. Treatment with hydrogen molecules prevents RANKL-induced osteoclast differentiation associated with inhibition of ROS formation and inactivation of MAPK, AKT and NF-kappa B pathways in murine RAW264.7 cells. J Bone Miner Metab 32, 494–504 (2014). https://doi.org/10.1007/s00774-013-0530-1

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