Early signs of lung fibrosis after in vitro treatment of rat lung slices with CdCl2 and TGF-beta1.
Precision-cut rat lung slices have been employed in combination with an extensive immunohistochemistry of paraffin-embedded slices for monitoring of early pathohistological changes after exposure to CdCl(2)/TGF-beta(1). Three days of CdCl(2) exposure in combination with TGF-beta(1) seem to be sufficient to induce lung injury with alterations similar to changes observed in early lung fibrogenesis: (1) extracellular matrix accumulation and myofibroblast transdifferentiation (Sirius red staining, collagen type IV, alpha-smooth muscle actin), (2) type I cell injury with loss of type I cell antigens (T1alpha antigen, aquaporin-5, RAGE), (3) increased apoptosis of pulmonary cells (active caspase-3, vimentin cleavage product V1 of caspase-9), and (4) activation of microvascular endothelial cells (podocalyxin, caveolin-1). Western blot analysis confirmed the increasing amount of alpha-smooth muscle actin, the loss of T1alpha antigen, and the increase in caveolin-1 immunoreactivity. The explant culture using CdCl(2)/TGF-beta(1) provides a suitable tool for the study of other factors involved in pulmonary pathology including transcription factors, cytokines, and other metabolites involved in early stages of fibrogenesis.
Kasper
M
M
Institute of Anatomy, Medical Faculty, Technical University of Dresden, Fetscherstrasse 74, 01307 Dresden, Germany. michael.kasper@mailbox.tu-dresden.de
Seidel
D
D
Knels
L
L
Morishima
N
N
Neisser
A
A
Bramke
S
S
Koslowski
R
R
D000199:Actins / Q000032:analysis; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D015415:Biomarkers / Q000032:analysis; D019256:Cadmium Chloride / Q000633:toxicity*; D051242:Caveolin 1; D022461:Caveolins / Q000032:analysis; D004195:Disease Models, Animal; D004338:Drug Combinations; D007150:Immunohistochemistry; D008168:Lung / Q000737:chemistry / Q000187:drug effects* / Q000473:pathology; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000032:analysis; D009924:Organ Culture Techniques; D011658:Pulmonary Fibrosis / Q000139:chemically induced* / Q000473:pathology; D051381:Rats; D017208:Rats, Wistar; D016212:Transforming Growth Factor beta / Q000633:toxicity*; D053773:Transforming Growth Factor beta1; D000098853:Podoplanin
D000199:Actins; D015415:Biomarkers; D019256:Cadmium Chloride; D051242:Caveolin 1; D022461:Caveolins; D004338:Drug Combinations; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; C495537:Cav1 protein, rat; C092025:Pdpn protein, rat; D000098853:Podoplanin; C508962:Tgfb1 protein, rat