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Messenger RNA levels of lung extracellular matrix proteins during ozone exposure

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Abstract

Continuous exposure of rats to ozone has been shown to result in lung epithelial damage, inflammation, and subsequent increases in collagen content. The main goal of this study was to identify the earliest time point of altered extracellular matrix protein gene expression by utilizing Northern blot analyses of rat lungs continuously exposed to 1.0 ppm ozone for 14 days. An early increase of steady-state fibronectin mRNA levels was observed at 2 days of exposure, prior to the time point of increased type I collagen mRNA, which was seen at 4 days. This increased level of type I collagen mRNA preceded measurable changes in total lung collagen content, observed at 7 days. In addition, peak levels of the growth-related proto-oncogene c-myc mRNA could be correlated with maximal increases of lung DNA content, although the initial increase in c-myc mRNA preceded measurable changes of total lung DNA. The use of specific cDNA probes for measuring altered gene expression can be useful for defining the early cellular and molecular events in ozone-induced lung injury.

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References

  1. Adachi K, Yamauchi K, Bernaudin JF, Fouret P, Ferrans VJ, Crystal RG (1988) Evaluation of fibronectin gene expression by in situ hybridization. Differential expression of the fibronectin gene among populations of human alveolar macrophages. Am J Pathol 133:193–203

    Google Scholar 

  2. Autor AP, Schmitt SL (1977) Pulmonary fibrosis and paraquat toxicity. In: Autor AP (ed) Biochemical Mechanisms of Paraquat Toxicity. Academic Press, New York, pp 175–186

    Google Scholar 

  3. Bassett DIP, Bowen-Kelly E, Elbon CL, Reichenbaugh SS (1988) Rat lung recovery from 3 days of continuous exposure to 0.75 ppm ozone. J Toxicol Environ Health 25:329–347

    Google Scholar 

  4. Bernard MP, Kolbe M, Weil D, Chu ML (1985) Human cellular fibronectin: comparison of the carboxyl-terminal portion with rat identifies primary structural domains separated by hypervariable regions. Biochemistry 24:2698–2704

    Google Scholar 

  5. Bitterman PB, Rennard SI, Adelberg S, Crystal RG (1983) Role of fibronectin as a growth factor for fibroblasts. J Cell Biol 97:1925–1932

    Google Scholar 

  6. Boorman GA, Schwartz LW, Growth DL (1980) Pulmonary effects of prolonged ozone insult in rats. Lab Invest 43:108–115

    Google Scholar 

  7. Chang L-Y, Huang Y, Stockstill BL, Graham JA, Grose EC, Menache MG, Miller FJ, Costa DL, Crapo JD (1992) Epithelial injury and interstitial fibrosis in the proximal alveolar regions of rats chronically exposed to a simulated pattern of urban ambient ozone. Toxicol Appl Pharmacol 115:241–252

    Google Scholar 

  8. Chirgwin JM, Przbyka AE, MacDonald RJ, Rutter WJ (1979) Isolation of biologically active ribonucleic acids from sources enriched in ribonuclease. Biochemistry 18:5294–5299

    CAS  PubMed  Google Scholar 

  9. Chu ML, Myers JC, Bernard MP, Ding JF, Ramirez F (1982) Cloning and characterization of five overlapping cDNAs specific for the human pro-α1[I] collagen chain. Nucleic Acids Res 10:5925–5931

    Google Scholar 

  10. Crapo JD, Barry BE, Foscue HA, Shelburne J (1980) Structural and biochemical changes in rat lungs occurring during exposures to lethal and adaptive doses of oxygen. Am Rev Respir Dis 122:123–143

    Google Scholar 

  11. Devlin RB, McDonnell WF, Mann R, Becker S, House DE, Schreinemachers D, Koren HS (1991) Exposure of humans to ambient levels of ozone for 6.6 hours causes cellular and biochemical changes in the lung. Am J Respir Cell Biol Mol Biol 4:72–81

    Google Scholar 

  12. Dowdy S, Wearden S (1983) The analysis of variance model. In: Statistics for Research. Wiley, New York, pp 287–304

    Google Scholar 

  13. Evans MJ, Johnson LV, Stephens RJ, Freeman G (1976) Renewal of the terminal bronchiolar epithelium in the rat following exposure to NO2 or O3. Lab Invest 35:246–257

    Google Scholar 

  14. Evans MJ, Johnson LV, Stephens RJ, Freeman G (1976) Cell renewal in the lungs of rats exposed to low levels of ozone. Exp Mol Pathol 24:70–83

    Google Scholar 

  15. Fabisiak JP, Evans JN, Kelley J (1989) Increased expression of PDGF-β (c-sis) mRNA in rat lung precedes DNA synthesis and tissue repair during chronic hyperoxia. Am J Respir Cell Mol Biol 1:181–189

    Google Scholar 

  16. Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13

    CAS  PubMed  Google Scholar 

  17. Frankel FR, Steeger JR, Damiano VV, Sohn M, Oppenheim D, Weinbaum G (1991) Induction of unilateral pulmonary fibrosis in the rat by cadmium chloride. Am J Respir Cell Mol Biol 5:385–394

    Google Scholar 

  18. Freeman G, Stephens RJ, Coffin DL, Stara JF (1973) Changes in dogs' lungs after long-term exposure to ozone. Arch Environ Health 26:209–216

    Google Scholar 

  19. Freeman G, Juhos LT, Furiosi NJ, Mussenden R, Stephens RJ, Evans MJ (1974) Pathology of pulmonary disease from exposure to interdependent ambient gases (nitrogen dioxide and ozone). Arch Environ Health 29:203–210

    Google Scholar 

  20. Gunning P, Ponte P, Okayama H, Engel J, Blau H, Kedes L (1983) Isolation and characterization of full-length cDNA clones for human alph-, beta-, and gamma-actin mRNAs: skeletal but not cytoplasmic actins have an amino-terminal cysteine that is subsequently removed. Mol Cell Biol 3:787–795

    Google Scholar 

  21. Hesterberg TW, Last JA (1981) Ozone-induced acute pulmonary fibrosis in rats. Am Rev Respir Dis 123:47–52

    Google Scholar 

  22. Hesterberg TW, Gerriets JE, Reiser KM, Jackson AC, Cross CE, Last JA (1981) Bleomycin induced pulmonary fibrosis: correlation of biochemical, physiological, and histological changes. Toxicol Appl Pharmacol 60:360–367

    Google Scholar 

  23. Hoyt DG, Lazo JS (1988) Alterations in pulmonary mRNA encoding procollagens, fibronectin, and transforming growth factor-B precedes bleomycin-induced pulmonary fibrosis in mice. J Pharmacol Exper Ther 246:765–771

    Google Scholar 

  24. Kukinen M, Vaheri A, Roberts PJ, Stenman S (1980) Sequential appearance of fibronectin and collagen in experimental granulation tissue. Lab Invest 43:47–51

    Google Scholar 

  25. Last JA, Greenberg DB (1980) Ozone-induced alterations in collagen metabolism of rat lungs. II. Long-term exposures. Toxicol Appl Pharmacol 55:108–114

    Google Scholar 

  26. Last JA, Reiser KM, Tyler WS, Rucker RB (1984) Long term consequences of exposure to ozone. I. Lung collagen content. Toxicol Appl Pharmacol 72:111–118

    Google Scholar 

  27. Lazenby AJ, Crouch EC, McDonald C, Kuhn C (1990) Remodeling of the lung in bleomycin-induced pulmonary fibrosis in the rat. Am Rev Respir Dis 142:206–214

    Google Scholar 

  28. McMaster GK, Carmichael GG (1977) Analysis of single and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci 74:4835–4838

    Google Scholar 

  29. Munro HN, Fleck A (1966) The determination of nucleic acids. Methods Biochem Anal 14:114–176

    Google Scholar 

  30. Persson H, Hennighausen L, Taub R, DeGrado W, Leder P (1984) Antibodies to human c-myc oncogene product: evidence of an evolutionary conserved protein induced during cell proliferation. Science 225:687–693

    Google Scholar 

  31. Pickrell JA, Hahn FF, Rebar AH, Horoda RA, Henderson RF (1987) Changes in collagen metabolism and proteinolysis after repeated inhalation exposure to ozone. Exp Mol Path 46:159–167

    Google Scholar 

  32. Raghow R, Lurie S, Seyer JM, Kang AH (1985) 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 76:1733–1739

    Google Scholar 

  33. Raghow R, Irish P, Kang AH (1989) Coordinate regulation of transforming growth factor- B gene expression and cell proliferation in hamster lungs undergoing bleomycin-induced pulmonary fibrosis. J Clin Invest 84:1836–1842

    Google Scholar 

  34. Shoji S, Rickard KA, Ertl RF, Robbins RA, Linder JL, Rennard SI (1989) Bronchial epithelial cells produce lung fibroblast chemotactic factor: fibronectin. Am J Respir Cell Mol Biol 1:13–20

    Google Scholar 

  35. Stegemann H, Stadler K (1967) Determination of hydroxyproline. Clin Chem Acta 18:207–273

    Google Scholar 

  36. Wright ES, Kehrer JP, White DM, Smiler KL (1988) Effects of chronic exposure to ozone on collagen in rat lung. Toxicol Appl Pharmacol 92:445–452

    Google Scholar 

  37. Yamauchi K, Martinet Y, Crystal RG (1987) Modulation of fibronectin gene expression in human mononuclear phagocytes. J Clin Invest 80:1720–1727

    Google Scholar 

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Choi, A.M.K., Elbon, C.L., Bruce, S.A. et al. Messenger RNA levels of lung extracellular matrix proteins during ozone exposure. Lung 172, 15–30 (1994). https://doi.org/10.1007/BF00186166

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