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Dependence of NF-κB activation and free radical generation on silica-induced TNF-α production in macrophages

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Abstract

Tumor necrosis factor α (TNFα) plays an important role in the pathogenesis of silicosis and other chronic inflammatory lung diseases. The present study investigates the role nuclear transcription factor κB (NF-κB) and oxygen free radicals in silica-induced TNFα production in primary alveolar macrophages and RAW 264.7 cells. Using electrophoretic mobility shift assay (EMSA) and enzyme-linked immunoadsorbent assay (ELISA), we have demonstrated that silica can induce NF-κB activation and TNFα expression in a dose-dependent manner. Transient transfection assays with a plasmid construct containing NF-κB binding sites linked to a reporter gene further show that silica is able to induce the transcriptional activation of NF-κB-dependent gene. Inhibition of NF-κB activation by SN50, a specific NF-κB blocker, abolishes silica-induced TNFα production. Pretreatment of the cells with catalase (H2O2 scavenger) or deferoxamine (·OH scavenger) effectively inhibits NF-κB and TNFα activation, whereas superoxide dismutase (O2 - scavenger) has an opposite effect. These results indicate that silica-mediated free radical generation and NF-κB activation play important roles in silica-induced TNFα gene expression.

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Authors and Affiliations

  1. Department of Basic Pharmaceutical Sciences, West Virginia University, Health Sciences Center, Morgantown, WV, USA

    Yon Rojanasakul, Liying Wang & Ningli Cheng

  2. Pathology and Physiology Research Branch, National Institute for Occupational Safety and Health, Morgantown, WV, USA

    Jiangping Ye, Fei Chen, Vincent Castranova, Val Vallyathan & Xiannglin Shi

Authors
  1. Yon Rojanasakul
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  2. Jiangping Ye
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  3. Fei Chen
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  4. Liying Wang
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  5. Ningli Cheng
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  6. Vincent Castranova
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  7. Val Vallyathan
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  8. Xiannglin Shi
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Rojanasakul, Y., Ye, J., Chen, F. et al. Dependence of NF-κB activation and free radical generation on silica-induced TNF-α production in macrophages. Mol Cell Biochem 200, 119–125 (1999). https://doi.org/10.1023/A:1007051402840

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