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J Clin Invest. 1995 Sep; 96(3): 1621–1630.
doi: 10.1172/JCI118201
PMCID: PMC185788
PMID: 7544811

Changes in procoagulant and fibrinolytic gene expression during bleomycin-induced lung injury in the mouse.

M A Olman, N Mackman, C L Gladson, K M Moser, and D J Loskutoff
Author information Copyright and License information PMC Disclaimer

Abstract

Bleomycin-induced lung injury is an established murine model of human pulmonary fibrosis. Although procoagulant molecules (e.g., tissue factor [TF]) and fibrinolytic components (e.g., urokinase [u-PA] and type 1 plasminogen activator inhibitor [PAI-1]) have been detected in alveolar fluid from injured lungs, the origin of these molecules remains unknown. We therefore examined the expression of procoagulant and fibrinolytic components in relation to the distribution of parenchymal fibrin in bleomycin-injured lungs. Extravascular fibrin localized to the alveolar and extracellular matrix in injured lung tissue. Injured lung tissue extracts contained elevated levels of PAI-1 activity and decreased levels of u-PA activity. Whole lung PAI-1 and TF mRNAs were dramatically induced by lung injury. In situ hybridization of injured lungs revealed that PAI-1, u-PA, and TF mRNAs were induced within the fibrin-rich fibroproliferative lesions, primarily in fibroblast-like and macrophagelike cells, respectively, while TF mRNA was also induced in perilesional alveolar cells. Taken together, these observations suggest that the induction of PAI-1 and TF gene expression plays and important role in the formation and persistence of extracellular fibrin in bleomycin injured murine lungs.

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Selected References

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  • Burkhardt A. Alveolitis and collapse in the pathogenesis of pulmonary fibrosis. Am Rev Respir Dis. 1989 Aug;140(2):513–524. [PubMed] [Google Scholar]
  • Crouch E. Pathobiology of pulmonary fibrosis. Am J Physiol. 1990 Oct;259(4 Pt 1):L159–L184. [PubMed] [Google Scholar]
  • Bowden DH. Unraveling pulmonary fibrosis: the bleomycin model. Lab Invest. 1984 May;50(5):487–488. [PubMed] [Google Scholar]
  • Janick-Buckner D, Ranges GE, Hacker MP. Alteration of bronchoalveolar lavage cell populations following bleomycin treatment in mice. Toxicol Appl Pharmacol. 1989 Sep 15;100(3):465–473. [PubMed] [Google Scholar]
  • Lindenschmidt RC, Tryka AF, Godfrey GA, Frome EL, Witschi H. Intratracheal versus intravenous administration of bleomycin in mice: acute effects. Toxicol Appl Pharmacol. 1986 Aug;85(1):69–77. [PubMed] [Google Scholar]
  • Schrier DJ, Kunkel RG, Phan SH. The role of strain variation in murine bleomycin-induced pulmonary fibrosis. Am Rev Respir Dis. 1983 Jan;127(1):63–66. [PubMed] [Google Scholar]
  • Dvorak HF, Senger DR, Dvorak AM, Harvey VS, McDonagh J. Regulation of extravascular coagulation by microvascular permeability. Science. 1985 Mar 1;227(4690):1059–1061. [PubMed] [Google Scholar]
  • Belew M, Gerdin B, Porath J, Saldeen T. Isolation of vasoactive peptides from human fibrin and fibrinogen degraded by plasmin. Thromb Res. 1978 Dec;13(6):983–994. [PubMed] [Google Scholar]
  • Senior RM, Skogen WF, Griffin GL, Wilner GD. Effects of fibrinogen derivatives upon the inflammatory response. Studies with human fibrinopeptide B. J Clin Invest. 1986 Mar;77(3):1014–1019. [PMC free article] [PubMed] [Google Scholar]
  • Bunce LA, Sporn LA, Francis CW. Endothelial cell spreading on fibrin requires fibrinopeptide B cleavage and amino acid residues 15-42 of the beta chain. J Clin Invest. 1992 Mar;89(3):842–850. [PMC free article] [PubMed] [Google Scholar]
  • Limper AH, Roman J. Fibronectin. A versatile matrix protein with roles in thoracic development, repair and infection. Chest. 1992 Jun;101(6):1663–1673. [PubMed] [Google Scholar]
  • Kuhn C, 3rd, Boldt J, King TE, Jr, Crouch E, Vartio T, McDonald JA. An immunohistochemical study of architectural remodeling and connective tissue synthesis in pulmonary fibrosis. Am Rev Respir Dis. 1989 Dec;140(6):1693–1703. [PubMed] [Google Scholar]
  • Drake TA, Morrissey JH, Edgington TS. Selective cellular expression of tissue factor in human tissues. Implications for disorders of hemostasis and thrombosis. Am J Pathol. 1989 May;134(5):1087–1097. [PMC free article] [PubMed] [Google Scholar]
  • Edgington TS, Mackman N, Brand K, Ruf W. The structural biology of expression and function of tissue factor. Thromb Haemost. 1991 Jul 12;66(1):67–79. [PubMed] [Google Scholar]
  • McGee MP, Rothberger H. Tissue factor in bronchoalveolar lavage fluids. Evidence for an alveolar macrophage source. Am Rev Respir Dis. 1985 Mar;131(3):331–336. [PubMed] [Google Scholar]
  • Chapman HA, Stahl M, Allen CL, Yee R, Fair DS. Regulation of the procoagulant activity within the bronchoalveolar compartment of normal human lung. Am Rev Respir Dis. 1988 Jun;137(6):1417–1425. [PubMed] [Google Scholar]
  • Chapman HA, Allen CL, Stone OL. Abnormalities in pathways of alveolar fibrin turnover among patients with interstitial lung disease. Am Rev Respir Dis. 1986 Mar;133(3):437–443. [PubMed] [Google Scholar]
  • Blasi F, Vassalli JD, Danø K. Urokinase-type plasminogen activator: proenzyme, receptor, and inhibitors. J Cell Biol. 1987 Apr;104(4):801–804. [PMC free article] [PubMed] [Google Scholar]
  • Sappino AP, Huarte J, Vassalli JD, Belin D. Sites of synthesis of urokinase and tissue-type plasminogen activators in the murine kidney. J Clin Invest. 1991 Mar;87(3):962–970. [PMC free article] [PubMed] [Google Scholar]
  • Vassalli JD, Sappino AP, Belin D. The plasminogen activator/plasmin system. J Clin Invest. 1991 Oct;88(4):1067–1072. [PMC free article] [PubMed] [Google Scholar]
  • Sawdey MS, Loskutoff DJ. Regulation of murine type 1 plasminogen activator inhibitor gene expression in vivo. Tissue specificity and induction by lipopolysaccharide, tumor necrosis factor-alpha, and transforming growth factor-beta. J Clin Invest. 1991 Oct;88(4):1346–1353. [PMC free article] [PubMed] [Google Scholar]
  • Mackman N, Sawdey MS, Keeton MR, Loskutoff DJ. Murine tissue factor gene expression in vivo. Tissue and cell specificity and regulation by lipopolysaccharide. Am J Pathol. 1993 Jul;143(1):76–84. [PMC free article] [PubMed] [Google Scholar]
  • Idell S, James KK, Gillies C, Fair DS, Thrall RS. Abnormalities of pathways of fibrin turnover in lung lavage of rats with oleic acid and bleomycin-induced lung injury support alveolar fibrin deposition. Am J Pathol. 1989 Aug;135(2):387–399. [PMC free article] [PubMed] [Google Scholar]
  • Chapman HA, Jr, Bertozzi P, Reilly JJ., Jr Role of enzymes mediating thrombosis and thrombolysis in lung disease. Chest. 1988 Jun;93(6):1256–1263. [PubMed] [Google Scholar]
  • Steinert PM, Roop DR. Molecular and cellular biology of intermediate filaments. Annu Rev Biochem. 1988;57:593–625. [PubMed] [Google Scholar]
  • Prendergast GC, Diamond LE, Dahl D, Cole MD. The c-myc-regulated gene mrl encodes plasminogen activator inhibitor 1. Mol Cell Biol. 1990 Mar;10(3):1265–1269. [PMC free article] [PubMed] [Google Scholar]
  • Tso JY, Sun XH, Kao TH, Reece KS, Wu R. Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: genomic complexity and molecular evolution of the gene. Nucleic Acids Res. 1985 Apr 11;13(7):2485–2502. [PMC free article] [PubMed] [Google Scholar]
  • Rickles RJ, Strickland S. Tissue plasminogen activator mRNA in murine tissues. FEBS Lett. 1988 Feb 29;229(1):100–106. [PubMed] [Google Scholar]
  • Keeton M, Eguchi Y, Sawdey M, Ahn C, Loskutoff DJ. Cellular localization of type 1 plasminogen activator inhibitor messenger RNA and protein in murine renal tissue. Am J Pathol. 1993 Jan;142(1):59–70. [PMC free article] [PubMed] [Google Scholar]
  • Olman MA, Marsh JJ, Lang IM, Moser KM, Binder BR, Schleef RR. Endogenous fibrinolytic system in chronic large-vessel thromboembolic pulmonary hypertension. Circulation. 1992 Oct;86(4):1241–1248. [PubMed] [Google Scholar]
  • Erickson LA, Lawrence DA, Loskutoff DJ. Reverse fibrin autography: a method to detect and partially characterize protease inhibitors after sodium dodecyl sulfate--polyacrylamide gel electrophoresis. Anal Biochem. 1984 Mar;137(2):454–463. [PubMed] [Google Scholar]
  • Camiolo SM, Siuta MR, Madeja JM. Improved medium for extraction of plasminogen activator from tissue. Prep Biochem. 1982;12(4):297–305. [PubMed] [Google Scholar]
  • Bernik MB, Kwaan HC. Plasminogen activator activity in cultures from human tissues. An immunological and histochemical study. J Clin Invest. 1969 Sep;48(9):1740–1753. [PMC free article] [PubMed] [Google Scholar]
  • Vassalli JD, Belin D. Amiloride selectively inhibits the urokinase-type plasminogen activator. FEBS Lett. 1987 Apr 6;214(1):187–191. [PubMed] [Google Scholar]
  • Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. [PubMed] [Google Scholar]
  • Schneiderman J, Sawdey MS, Keeton MR, Bordin GM, Bernstein EF, Dilley RB, Loskutoff DJ. Increased type 1 plasminogen activator inhibitor gene expression in atherosclerotic human arteries. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):6998–7002. [PMC free article] [PubMed] [Google Scholar]
  • Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. [PubMed] [Google Scholar]
  • Hoyt DG, Lazo JS. Alterations in pulmonary mRNA encoding procollagens, fibronectin and transforming growth factor-beta precede bleomycin-induced pulmonary fibrosis in mice. J Pharmacol Exp Ther. 1988 Aug;246(2):765–771. [PubMed] [Google Scholar]
  • Harpold MM, Evans RM, Salditt-Georgieff M, Darnell JE. Production of mRNA in Chinese hamster cells: relationship of the rate of synthesis to the cytoplasmic concentration of nine specific mRNA sequences. Cell. 1979 Aug;17(4):1025–1035. [PubMed] [Google Scholar]
  • Gurewich V, Pannell R. Inactivation of single-chain urokinase (pro-urokinase) by thrombin and thrombin-like enzymes: relevance of the findings to the interpretation of fibrin-binding experiments. Blood. 1987 Mar;69(3):769–772. [PubMed] [Google Scholar]
  • Woodcock-Mitchell J, Adler KB, Low RB. Immunohistochemical identification of cell types in normal and in bleomycin-induced fibrotic rat lung. Cellular origins of interstitial cells. Am Rev Respir Dis. 1984 Nov;130(5):910–916. [PubMed] [Google Scholar]
  • Sitrin RG, Brubaker PG, Shellito JE, Kaltreider HB. The distribution of procoagulant and plasminogen activator activities among density fractions of normal rabbit alveolar macrophages. Am Rev Respir Dis. 1986 Mar;133(3):468–472. [PubMed] [Google Scholar]
  • Chapman HA, Jr, Stone OL, Vavrin Z. Degradation of fibrin and elastin by intact human alveolar macrophages in vitro. Characterization of a plasminogen activator and its role in matrix degradation. J Clin Invest. 1984 Mar;73(3):806–815. [PMC free article] [PubMed] [Google Scholar]
  • Idell S, Peterson BT, Gonzalez KK, Gray LD, Bach R, McLarty J, Fair DS. Local abnormalities of coagulation and fibrinolysis and alveolar fibrin deposition in sheep with oleic acid-induced lung injury. Am Rev Respir Dis. 1988 Nov;138(5):1282–1294. [PubMed] [Google Scholar]
  • Idell S, Peters J, James KK, Fair DS, Coalson JJ. Local abnormalities of coagulation and fibrinolytic pathways that promote alveolar fibrin deposition in the lungs of baboons with diffuse alveolar damage. J Clin Invest. 1989 Jul;84(1):181–193. [PMC free article] [PubMed] [Google Scholar]
  • Sitrin RG, Brubaker PG, Fantone JC. Tissue fibrin deposition during acute lung injury in rabbits and its relationship to local expression of procoagulant and fibrinolytic activities. Am Rev Respir Dis. 1987 Apr;135(4):930–936. [PubMed] [Google Scholar]
  • Idell S, James KK, Levin EG, Schwartz BS, Manchanda N, Maunder RJ, Martin TR, McLarty J, Fair DS. Local abnormalities in coagulation and fibrinolytic pathways predispose to alveolar fibrin deposition in the adult respiratory distress syndrome. J Clin Invest. 1989 Aug;84(2):695–705. [PMC free article] [PubMed] [Google Scholar]
  • Gross TJ, Simon RH, Sitrin RG. Tissue factor procoagulant expression by rat alveolar epithelial cells. Am J Respir Cell Mol Biol. 1992 Apr;6(4):397–403. [PubMed] [Google Scholar]
  • Drake TA, Cheng J, Chang A, Taylor FB., Jr Expression of tissue factor, thrombomodulin, and E-selectin in baboons with lethal Escherichia coli sepsis. Am J Pathol. 1993 May;142(5):1458–1470. [PMC free article] [PubMed] [Google Scholar]
  • Aso Y, Yoneda K, Kikkawa Y. Morphologic and biochemical study of pulmonary changes induced by bleomycin in mice. Lab Invest. 1976 Dec;35(6):558–568. [PubMed] [Google Scholar]
  • Seiffert D, Loskutoff DJ. Kinetic analysis of the interaction between type 1 plasminogen activator inhibitor and vitronectin and evidence that the bovine inhibitor binds to a thrombin-derived amino-terminal fragment of bovine vitronectin. Biochim Biophys Acta. 1991 May 30;1078(1):23–30. [PubMed] [Google Scholar]
  • Liotta LA, Goldfarb RH, Brundage R, Siegal GP, Terranova V, Garbisa S. Effect of plasminogen activator (urokinase), plasmin, and thrombin on glycoprotein and collagenous components of basement membrane. Cancer Res. 1981 Nov;41(11 Pt 1):4629–4636. [PubMed] [Google Scholar]
  • Sato Y, Rifkin DB. Inhibition of endothelial cell movement by pericytes and smooth muscle cells: activation of a latent transforming growth factor-beta 1-like molecule by plasmin during co-culture. J Cell Biol. 1989 Jul;109(1):309–315. [PMC free article] [PubMed] [Google Scholar]
  • Laiho M, Saksela O, Keski-Oja J. Transforming growth factor-beta induction of type-1 plasminogen activator inhibitor. Pericellular deposition and sensitivity to exogenous urokinase. J Biol Chem. 1987 Dec 25;262(36):17467–17474. [PubMed] [Google Scholar]
  • Lund LR, Riccio A, Andreasen PA, Nielsen LS, Kristensen P, Laiho M, Saksela O, Blasi F, Danø K. Transforming growth factor-beta is a strong and fast acting positive regulator of the level of type-1 plasminogen activator inhibitor mRNA in WI-38 human lung fibroblasts. EMBO J. 1987 May;6(5):1281–1286. [PMC free article] [PubMed] [Google Scholar]
  • Ignotz RA, Massagué J. Transforming growth factor-beta stimulates the expression of fibronectin and collagen and their incorporation into the extracellular matrix. J Biol Chem. 1986 Mar 25;261(9):4337–4345. [PubMed] [Google Scholar]
  • Keski-Oja J, Raghow R, Sawdey M, Loskutoff DJ, Postlethwaite AE, Kang AH, Moses HL. Regulation of mRNAs for type-1 plasminogen activator inhibitor, fibronectin, and type I procollagen by transforming growth factor-beta. Divergent responses in lung fibroblasts and carcinoma cells. J Biol Chem. 1988 Mar 5;263(7):3111–3115. [PubMed] [Google Scholar]
  • Ignotz RA, Endo T, Massagué J. Regulation of fibronectin and type I collagen mRNA levels by transforming growth factor-beta. J Biol Chem. 1987 May 15;262(14):6443–6446. [PubMed] [Google Scholar]
  • Broekelmann TJ, Limper AH, Colby TV, McDonald JA. Transforming growth factor beta 1 is present at sites of extracellular matrix gene expression in human pulmonary fibrosis. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6642–6646. [PMC free article] [PubMed] [Google Scholar]
  • Khalil N, Bereznay O, Sporn M, Greenberg AH. Macrophage production of transforming growth factor beta and fibroblast collagen synthesis in chronic pulmonary inflammation. J Exp Med. 1989 Sep 1;170(3):727–737. [PMC free article] [PubMed] [Google Scholar]
  • Raghow R, Lurie S, Seyer JM, Kang AH. Profiles of steady state levels of messenger RNAs coding for type I procollagen, elastin, and fibronectin in hamster lungs undergoing bleomycin-induced interstitial pulmonary fibrosis. J Clin Invest. 1985 Nov;76(5):1733–1739. [PMC free article] [PubMed] [Google Scholar]
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