Identification of Cross-linked 46 KDa Protein in Experimentally Induced Silicotic Nodule in Rat Lung / 대한산업의학회지
Korean Journal of Occupational and Environmental Medicine
;
: 181-187, 2003.
Artigo
em Coreano
| WPRIM
| ID: wpr-213869
ABSTRACT
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.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Plasma
/
Silicose
/
Reagentes de Sulfidrila
/
Ureia
/
Fibrose
/
Imunoglobulina G
/
Cloreto de Sódio
/
Proteínas da Matriz Extracelular
/
Cromatografia
/
Cromatografia de Afinidade
Tipo de estudo:
Estudo diagnóstico
Limite:
Animais
Idioma:
Coreano
Revista:
Korean Journal of Occupational and Environmental Medicine
Ano de publicação:
2003
Tipo de documento:
Artigo
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