Activation of NF-kappaB-dependent gene expression by silica in lungs of luciferase reporter mice.
Occupational exposure to crystalline silica is associated with the development of pulmonary inflammation and silicosis, yet how silica initiates pulmonary fibrosis and which cell types are involved are unclear. In studies here, we hypothesized that silica particles interact initially with pulmonary epithelial cells and alveolar macrophages (AMs) to cause transcriptional activation of nuclear factor (NF)-kappaB-regulated genes encoding inflammatory cytokines. Exposure of NF-kappaB luciferase reporter mice intratracheally to silica or lipopolysaccharide (LPS), but not the nonfibrogenic particle titanium dioxide (TiO(2)), increased immunoreactivity of luciferase protein in bronchiolar epithelial cells and AMs. Ribonuclease protection assays revealed significant (P < or = 0.05) increases in mRNA levels of inducible nitric oxide synthase, tumor necrosis factor-alpha, macrophage inflammatory protein-2, macrophage chemotactic protein-1 (MCP-1), interferon-gamma, interleukin (IL)-6, and IL-12 in lung homogenates of reporter mice after exposures to silica or LPS. Immunoreactivity of MCP-1 in these animals was localized to AMs and epithelial cells. These data are the first to show activation of NF-kappaB in situ by fibrogenic particles in pulmonary epithelial cells and AMs. Increased expression of NF-kappaB-related inflammatory cytokines by these cell types, which first encounter silica after inhalation, may be critical to the initiation of silica-associated lung diseases, thus providing a rationale for focusing on NF-kappaB in preventive and therapeutic strategies.
Hubbard
Andrea K
AK
Department of Pharmaceutical Sciences, University of Connecticut, Storrs 06269, USA.
Timblin
Cynthia R
CR
Shukla
Arti
A
Rincón
Mercedes
M
Mossman
Brooke T
BT
D000818:Animals; D001980:Bronchi / Q000166:cytology / Q000276:immunology; D018932:Chemokine CCL2 / Q000378:metabolism; D016207:Cytokines / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects / Q000276:immunology; D017930:Genes, Reporter; D008070:Lipopolysaccharides; D008156:Luciferases / Q000235:genetics; D008168:Lung / Q000502:physiology*; D016676:Macrophages, Alveolar / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D016328:NF-kappa B / Q000378:metabolism*; D011014:Pneumonia / Q000139:chemically induced / Q000276:immunology / Q000503:physiopathology; D012333:RNA, Messenger / Q000032:analysis; D020545:Respiratory Mucosa / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D012822:Silicon Dioxide / Q000494:pharmacology*; D012829:Silicosis / Q000276:immunology / Q000503:physiopathology*; D014025:Titanium / Q000494:pharmacology
D018932:Chemokine CCL2; D016207:Cytokines; D008070:Lipopolysaccharides; D016328:NF-kappa B; D012333:RNA, Messenger; C009495:titanium dioxide; D012822:Silicon Dioxide; D014025:Titanium; D008156:Luciferases