Bleomycin-induced lung fibrosis model
Objective: To investigate whether Smad7 gene transfer via adenovirus vector can prevent bleomycin-induced lung fibrosis in mice by suppressing TGF-β signaling
This is a Bleomycin-induced lung fibrosis model protocol using mouse as the model organism. The procedure involves 6 procedural steps, 3 materials. Extracted from a 1999 paper published in Journal of Clinical Investigation.
Model and subjects
mouse • C57BL/6 • unknown • Not specified • Not specified
Study window
Estimated timing pending
Core workflow
Bleomycin-induced lung fibrosis induction • Intratracheal injection of recombinant adenovirus • Assessment of type I precollagen mRNA expression
Primary readouts
- Type I precollagen mRNA expression levels
- Hydroxyproline content in lung tissue
- Morphological fibrotic responses in lungs
- Smad2 phosphorylation status
Key equipment and reagents
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Protocol Steps
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Bleomycin-induced lung fibrosis induction
C57BL/6 mice were treated with bleomycin to induce lung fibrosis
Note: Specific dose and administration route for bleomycin not detailed in provided text
View evidence from paper
“C57BL/6 mice with bleomycin-induced lungs received an intratracheal injection”
Intratracheal injection of recombinant adenovirus
Mice with bleomycin-induced lung fibrosis received intratracheal injection of recombinant adenovirus carrying either Smad7 cDNA (treatment group) or Smad6 cDNA (control group)
Note: Timing of injection relative to bleomycin administration not specified
View evidence from paper
“C57BL/6 mice with bleomycin-induced lungs received an intratracheal injection of a recombinant adenovirus carrying mice Smad7 cDNA”
Assessment of type I precollagen mRNA expression
Lung tissue was analyzed for type I precollagen mRNA levels to assess fibrotic response
Note: Smad7-treated mice showed suppression compared to Smad6 controls
View evidence from paper
“These mice demonstrated suppression of type I precollagen mRNA”
Measurement of hydroxyproline content
Lung tissue hydroxyproline content was measured as a marker of collagen deposition
Note: Reduced hydroxyproline content observed in Smad7-treated mice
View evidence from paper
“reduced hydroxyproline content, and no morphological fibrotic responses in the lungs”
Morphological analysis of lung tissue
Lung tissue was examined for morphological fibrotic responses
Note: No morphological fibrotic responses observed in Smad7-treated mice compared to Smad6 controls
View evidence from paper
“no morphological fibrotic responses in the lungs when compared with mice administered adenovirus carrying Smad6 cDNA”
Assessment of Smad2 phosphorylation
Lung tissue was analyzed for Smad2 phosphorylation status to determine TGF-β signaling inhibition
Note: Smad7 expression blocked Smad2 phosphorylation induced by bleomycin
View evidence from paper
“expression of Smad7 transgene blocked Smad2 phosphorylation induced by bleomycin in mouse lungs”