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. 1985 Jul;76(1):241–247. doi: 10.1172/JCI111953

Rat lung fibroblast collagen metabolism in bleomycin-induced pulmonary fibrosis.

S H Phan, J Varani, D Smith
PMCID: PMC423755  PMID: 2410457

Abstract

Endotracheal bleomycin administration in rats and other animal species causes rapid development of pulmonary fibrosis, characterized by increased lung collagen synthesis and deposition. To clarify the mechanism, lung fibroblasts from bleomycin-treated rats (BRF) were isolated and maintained in tissue culture. They were then compared with those from normal untreated control animals, with respect to several key parameters of collagen metabolism. BRF synthesized collagen at a rate 35-82% above normal rat lung fibroblasts (NRF). This difference did not appear to be due to the selection of a clone by the subculture process. Furthermore, analysis of newly synthesized collagen type composition, revealed a significantly lower ratio of type III to type I collagen. Noncollagenous protein synthesis, however, was not significantly different from normal. Collagenase production and growth rate were also unaffected. BRF, however, was morphologically indistinguishable from NRF, even at the ultrastructural level. Upon further bleomycin (1 microgram/ml) exposure in vitro, BRF could be further stimulated to synthesize collagen at 82% above the rate for untreated BRF. This is comparable to the 90% increase in NRF treated in vitro (compared with untreated NRF). These results would favor the conclusion that bleomycin induces pulmonary fibrosis, by causing directly and/or indirectly lung fibroblasts (or a certain line of lung fibroblasts) to synthesize collagen at a higher rate without any associated increase in growth rate. The data, however, do not rule out the possibility that the fibroblast isolation procedure has selected for a certain population of fibroblasts that may not be typical of the in vivo situation.

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Selected References

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  1. Absher M., Hildebran J., Trombley L., Woodcock-Mitchell J., Marsh J. Characteristics of cultured lung fibroblasts from bleomycin-treated rats. Comparisons with in vitro exposed normal fibroblasts. Am Rev Respir Dis. 1984 Jan;129(1):125–129. doi: 10.1164/arrd.1984.129.1.125. [DOI] [PubMed] [Google Scholar]
  2. Adamson I. Y., Bowden D. H. Bleomycin-induced injury and metaplasia of alveolar type 2 cells. Relationship of cellular responses to drug presence in the lung. Am J Pathol. 1979 Aug;96(2):531–544. [PMC free article] [PubMed] [Google Scholar]
  3. Bateman E. D., Turner-Warwick M., Adelmann-Grill B. C. Immunohistochemical study of collagen types in human foetal lung and fibrotic lung disease. Thorax. 1981 Sep;36(9):645–653. doi: 10.1136/thx.36.9.645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bitterman P. B., Rennard S. I., Hunninghake G. W., Crystal R. G. Human alveolar macrophage growth factor for fibroblasts. Regulation and partial characterization. J Clin Invest. 1982 Oct;70(4):806–822. doi: 10.1172/JCI110677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brody J. S., Kaplan N. B. Proliferation of alveolar interstitial cells during postnatal lung growth. Evidence for two distinct populations of pulmonary fibroblasts. Am Rev Respir Dis. 1983 Jun;127(6):763–770. doi: 10.1164/arrd.1983.127.6.763. [DOI] [PubMed] [Google Scholar]
  6. Clark J. G., Starcher B. C., Uitto J. Bleomycin-induced synthesis of type I procollagen by human lung and skin fibroblasts in culture. Biochim Biophys Acta. 1980 Aug 13;631(2):359–370. doi: 10.1016/0304-4165(80)90309-8. [DOI] [PubMed] [Google Scholar]
  7. Clore J. N., Cohen I. K., Diegelmann R. F. Quantitation of collagen types I and III during wound healing in rat skin. Proc Soc Exp Biol Med. 1979 Jul;161(3):337–340. doi: 10.3181/00379727-161-40548. [DOI] [PubMed] [Google Scholar]
  8. Gabbiani G., Hirschel B. J., Ryan G. B., Statkov P. R., Majno G. Granulation tissue as a contractile organ. A study of structure and function. J Exp Med. 1972 Apr 1;135(4):719–734. doi: 10.1084/jem.135.4.719. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ichihashi M., Shinkai H., Takei M., Sano S. Analysis of the mechanism of bleomycin-induced cutaneous fibrosis in mice. J Antibiot (Tokyo) 1973 Apr;26(4):238–242. doi: 10.7164/antibiotics.26.238. [DOI] [PubMed] [Google Scholar]
  10. Ishizuka M., Takayama H., Takeuchi T., Umezawa H. Activity and toxicity of bleomycin. J Antibiot (Tokyo) 1967 Jan;20(1):15–24. [PubMed] [Google Scholar]
  11. Juva K., Prockop D. J. Modified procedure for the assay of H-3-or C-14-labeled hydroxyproline. Anal Biochem. 1966 Apr;15(1):77–83. doi: 10.1016/0003-2697(66)90249-1. [DOI] [PubMed] [Google Scholar]
  12. Keogh B. A., Crystal R. G. Alveolitis: the key to the interstitial lung disorders. Thorax. 1982 Jan;37(1):1–10. doi: 10.1136/thx.37.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Korn J. H. Fibroblast prostaglandin E2 synthesis. Persistence of an abnormal phenotype after short-term exposure to mononuclear cell products. J Clin Invest. 1983 May;71(5):1240–1246. doi: 10.1172/JCI110873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Madri J. A., Furthmayr H. Collagen polymorphism in the lung. An immunochemical study of pulmonary fibrosis. Hum Pathol. 1980 Jul;11(4):353–366. doi: 10.1016/s0046-8177(80)80031-1. [DOI] [PubMed] [Google Scholar]
  15. Neilson E. G., Jimenez S. A., Phillips S. M. Cell-mediated immunity in interstitial nephritis. III. T lymphocyte-mediated fibroblast proliferation and collagen synthesis: an immune mechanism for renal fibrogenesis. J Immunol. 1980 Oct;125(4):1708–1714. [PubMed] [Google Scholar]
  16. Neville D. M., Jr Molecular weight determination of protein-dodecyl sulfate complexes by gel electrophoresis in a discontinuous buffer system. J Biol Chem. 1971 Oct 25;246(20):6328–6334. [PubMed] [Google Scholar]
  17. Peterkofsky B., Diegelmann R. Use of a mixture of proteinase-free collagenases for the specific assay of radioactive collagen in the presence of other proteins. Biochemistry. 1971 Mar 16;10(6):988–994. doi: 10.1021/bi00782a009. [DOI] [PubMed] [Google Scholar]
  18. Phan S. H., Thrall R. S. Inhibition of bleomycin-induced pulmonary fibrosis by cobra venom factor. Am J Pathol. 1982 Apr;107(1):25–28. [PMC free article] [PubMed] [Google Scholar]
  19. Phan S. H., Thrall R. S., Ward P. A. Bleomycin-induced pulmonary fibrosis in rats: biochemical demonstration of increased rate of collagen synthesis. Am Rev Respir Dis. 1980 Mar;121(3):501–506. doi: 10.1164/arrd.1980.121.3.501. [DOI] [PubMed] [Google Scholar]
  20. Phan S. H., Thrall R. S., Williams C. Bleomycin-induced pulmonary fibrosis. Effects of steroid on lung collagen metabolism. Am Rev Respir Dis. 1981 Oct;124(4):428–434. doi: 10.1164/arrd.1981.124.4.428. [DOI] [PubMed] [Google Scholar]
  21. Phan S. H., Ward P. A. Generation of biologic activity from the purified alpha-chain of C5. J Immunol. 1979 Dec;123(6):2735–2740. [PubMed] [Google Scholar]
  22. Postlethwaite A. E., Kang A. H. Induction of fibroblast proliferation by human mononuclear leukocyte-derived proteins. Arthritis Rheum. 1983 Jan;26(1):22–27. doi: 10.1002/art.1780260104. [DOI] [PubMed] [Google Scholar]
  23. Reiser K. M., Last J. A. Pulmonary fibrosis in experimental acute respiratory disease. Am Rev Respir Dis. 1981 Jan;123(1):58–63. doi: 10.1164/arrd.1981.123.1.58. [DOI] [PubMed] [Google Scholar]
  24. Schrier D. J., Phan S. H., McGarry B. M. The effects of the nude (nu/nu) mutation on bleomycin-induced pulmonary fibrosis. A biochemical evaluation. Am Rev Respir Dis. 1983 May;127(5):614–617. doi: 10.1164/arrd.1983.127.5.614. [DOI] [PubMed] [Google Scholar]
  25. Seyer J. M., Hutcheson E. T., Kang A. H. Collagen polymorphism in idiopathic chronic pulmonary fibrosis. J Clin Invest. 1976 Jun;57(6):1498–1507. doi: 10.1172/JCI108420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sterling K. M., Jr, Harris M. J., Mitchell J. J., Cutroneo K. R. Bleomycin treatment of chick fibroblasts causes an increase of polysomal type I procollagen mRNAs. Reversal of the bleomycin effect by dexamethasone. J Biol Chem. 1983 Dec 10;258(23):14438–14444. [PubMed] [Google Scholar]
  27. Sykes B. C. The separation of two soft-tissue collagens by covalent chromatography. FEBS Lett. 1976 Jan 15;61(2):180–185. doi: 10.1016/0014-5793(76)81032-0. [DOI] [PubMed] [Google Scholar]
  28. Thrall R. S., Phan S. H., McCormick J. R., Ward P. A. The development of bleomycin-induced pulmonary fibrosis in neutrophil-depleted and complement-depleted rats. Am J Pathol. 1981 Oct;105(1):76–81. [PMC free article] [PubMed] [Google Scholar]
  29. Wahl S. M., Wahl L. M., McCarthy J. B. Lymphocyte-mediated activation of fibroblast proliferation and collagen production. J Immunol. 1978 Sep;121(3):942–946. [PubMed] [Google Scholar]

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