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Original Article
- Identification of Cross-linked 46 KDa Protein in Experimentally Induced Silicotic Nodule in Rat Lung
- You Mie Kim, Young Jin Kim, Soo Young Lee
- Korean Journal of Occupational and Environmental Medicine 2003;15(2):181-187. Published online June 30, 2003
- DOI: https://doi.org/10.35371/kjoem.2003.15.2.181
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Abstract
PDF - OBJECTIVES
This study was conducted in order to understand the cellular events associated with silica-induced pathogenesis of the rat lung.
METHODS
Silicosis was induced by an intratracheal instillation of 50 mg of silica (SiO2, 0.15 - 10 micrometer) suspended in 500 microliter of a sterile saline solution in Sprague-Dawley rats weighing 200g. Silicotic nodules were excised from the rat lungs 4 weeks after silica instillation, then boiled for 4 days at 110 degrees in solution containing 2% SDS, 10 M urea and 40 mM DTT. The insoluble cellular encapsulates were electrophoresed on 4-12 % gradient SDS-PAGE, and the amino acid composition was analyzed. Affinity chromatographies of the homogenate supernatants of the control lung, silicotic nodule, and normal rat plasma were performed using rabbit IgG, anti-rat, cross-linked protein from the silicotic nodule. The amounts of N epsilon-(gamma-glutamyl) lysine cross-linked in the control lungs and silicotic nodules were determined using HPLC analysis.
RESULTS
The remaining cross-linked protein was insoluble in the 10 M urea and 40 mM sulfhydryl reagents even under prolonged boiling conditions. The encapsulate revealed the retention of silica particles within the protein whose amino acid composition showed a high percentage of alanine, leucine and glycine. A 46 KDa protein was identified as a cross-linked protein in the silicotic nodule by affinity chromatography. The level of N epsilon-(gamma-glutamyl) lysine dipeptide in the nodule digest was prominently increased compared with that in the control lung.
CONCLUSIONS
Transglutaminase (TGase)-catalyzed cross-linking appears to be involved in the silicotic nodule formation, and the 46 KDa protein may be cross-linked to itself and other extracellular matrix proteins during fibrosis and the formation of eventually insoluble nodule. -
Citations
Citations to this article as recorded by
- Inhibition of extracellular tissue transglutaminase activity attenuates development of pulmonary fibrosis in a novel rabbit model
Thomas H. Thatcher, Margaret A. T. Freeberg, Sarah V. Camus, Linghong Huang, John Atkinson, Shannon H. Lacy, Wade Narrow, Robert M. Kottmann, Khoa Huynh, Jinze Liu, Valentina Robila, Timothy S. Johnson, Patricia J. Sime
American Journal of Physiology-Lung Cellular and Molecular Physiology.2026; 331(5): L421. CrossRef
- Inhibition of extracellular tissue transglutaminase activity attenuates development of pulmonary fibrosis in a novel rabbit model
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Original Article
- Identification of Plasma Coagulation Factor XIII, Transglutaminase 3 and N epsilon-(gamma-glutamyl) lysine cross-Link in the Silicotic Nodule by Immunohistochemistry
- You Mie Kim, Young Jin Kim, Soo Young Lee
- Korean Journal of Occupational and Environmental Medicine 2003;15(2):173-180. Published online June 30, 2003
- DOI: https://doi.org/10.35371/kjoem.2003.15.2.173
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Abstract
PDF
- OBJECTIVES
This study was performed to examine the immunohistochemical distribution of TGase 1, 2, 3, coagulation factor XIII and N epsilon-(gamma-glutamyl) lysine cross-link in the silicotic nodules formed after an intratracheal instillation of the silica.
METHODS
The immunohistochemical examinations used antibodies against TGase 1, 2, 3, coagulation factor XIII and N epsilon-(gamma-glutamyl) lysine isopeptide in the silicotic nodules induced after an intratracheal instillation of 50 mg of size fractionated, crystalline silica.
RESULTS
A high level of TGase 3 was related to the severity of fibrosis in silicotic nodules and extracellular coagulation factor XIII was detected around the nodules. Expressions of both membrane-bound TGase 1 and TGase 2 were barely detected in the nodules although high expressions were detected in the intact lung. Formation of N epsilon-(gamma-glutamyl) lysine cross-links was increased in severe fibrotic nodules.
CONCLUSIONS
TGase 3 might contribute to the eventual stone-like fibrosis via formation of N epsilon-(gamma-glutamyl) lysine cross-links. Futhermore, coagulation factor XIII plays a role in the formation of a provisional matrix which results in fibrogenesis during silicotic nodule formation.
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