Early signs of lung fibrosis after in vitro treatment of rat lung slices with CdCl2 and TGF-β1
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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 CdCl2/TGF-β1. Three days of CdCl2 exposure in combination with TGF-β1 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, α-smooth muscle actin), (2) type I cell injury with loss of type I cell antigens (T1α 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 α-smooth muscle actin, the loss of T1α antigen, and the increase in caveolin-1 immunoreactivity. The explant culture using CdCl2/TGF-β1 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.
KeywordsSlice culture Rat lung Cadmium chloride Fibrosis Aquaporin-5
The authors thank Dr. Stefan Uhlig, Head Division Pulmonary Pharmacology, Research Center Borstel, Germany, and Dr. Jean-Paul Morin, Faculte de Médecine Pharmacie, Universite de Rouen for introducing the slice culture techniques and all advice. The authors also thank Mrs. Petra Peche for excellent technical assistance. The financial support by elbionAG Dresden is acknowledged.
- Fehrenbach H, Kasper M, Tschernig T, Shearman MS, Schuh D, Muller M (1998) Receptor for advanced glycation end products (RAGE) exhibits highly differential cellular and subcellular localisation in rat and human lung. Cell Mol Biol (Noisy-le-grand) 44:1147–1157Google Scholar
- Hoyt DG, Lazo JS (1993) NAD depletion after in vitro exposure of murine lung slices to bleomycin. Biochem Pharmacol 46:1819–1824Google Scholar
- Wright AF, Green TP, Robson RT, Niewola Z, Wyatt I, Smith LL (1987) Specific polyclonal and monoclonal antibody prevents paraquat accumulation into rat lung slices. Biochem Pharmacol 36:1325–1331Google Scholar