Administration of Bleomycin via the Oropharyngeal Aspiration Route Leads to Sustained Lung Fibrosis in Mice and Rats as Quantified by UTE-MRI and Histology
Pulmonary fibrosis can be experimentally induced in small rodents by bleomycin. The antibiotic is usually administered via the intratracheal or intranasal routes. In the present study, we investigated the oropharyngeal aspiration of bleomycin as an alternative route for the induction of lung fibrosis in rats and mice. The development of lung injury was followed in vivo by ultrashort echo time magnetic resonance imaging (UTE-MRI) and by post-mortem analyses (histology of collagen, hydroxyproline determination, and qRT-PCR). In C57BL/6 mice, oropharyngeal aspiration of bleomycin led to more prominent lung fibrosis as compared to intranasal administration. Consequently, the oropharyngeal aspiration route allowed a dose reduction of bleomycin and, therewith, a model refinement. Moreover, the distribution of collagen after oropharyngeal aspiration of bleomycin was more homogenous than after intranasal administration: for the oropharyngeal aspiration route, fibrotic areas appeared all over the lung lobes, while for the intranasal route fibrotic lesions appeared mainly around the largest superior airways. Thus, oropharyngeal aspiration of bleomycin induced morphological changes that were more comparable to the human disease than the intranasal administration route did. Oropharyngeal aspiration of bleomycin led to a homogeneous fibrotic injury also in rat lungs. The present data suggest oropharyngeal aspiration of bleomycin as a less invasive means to induce homogeneous and sustained fibrosis in the lungs of mice and rats.
References
[1]
Gross TJ, Hunninghake GW (2001) Idiopathic pulmonary fibrosis. N Engl J Med 345: 517–525.
[2]
Katzenstein AL, Myers JL (1998) Idiopathic pulmonary fibrosis: clinical relevance of pathologic classification. Am J Respir Crit Care Med 157: 1301–1315.
[3]
Lazenby AJ, Crouch EC, McDonald JA, Kuhn C III (1990) Remodeling of the lung in bleomycin-induced pulmonary fibrosis in the rat. An immunohistochemical study of laminin, type IV collagen, and fibronectin. Am Rev Respir Dis 142: 206–214.
[4]
Brewster CE, Howarth PH, Djukanovic R, Wilson J, Holgate ST, et al. (1990) Myofibroblasts and subepithelial fibrosis in bronchial asthma. Am J Respir Cell Mol Biol 3: 507–511.
[5]
Roche WR, Beasley R, Williams JH, Holgate ST (1989) Subepithelial fibrosis in the bronchi of asthmatics. Lancet 1: 520–524.
[6]
Holgate ST, Davies DE, Lackie PM, Wilson SJ, Puddicombe SM, et al. (2000) Epithelial-mesenchymal interactions in the pathogenesis of asthma. J Allergy Clin Immunol 105: 193–204.
[7]
Jeffery PK (2001) Remodeling in asthma and chronic obstructive lung disease. Am J Respir Crit Care Med 164: S28–38.
[8]
Gong LK, Li XH, Wang H, Zhang L, Chen FP, et al. (2005) Effect of Feitai on bleomycin-induced pulmonary fibrosis in rats. J Ethnopharmacol 96: 537–544.
[9]
Ueda Y, Saito A, Fukuoka Y, Yamashiro Y, Ikeda Y, et al. (1983) Interactions of beta-lactam antibiotics and antineoplastic agents. Antimicrob Agents Chemother 23: 374–378.
[10]
Hay J, Shahzeidi S, Laurent G (1991) Mechanisms of bleomycin-induced lung damage. Arch Toxicol 65: 81–94.
[11]
Snider GL, Hayes JA, Korthy AL (1978) Chronic interstitial pulmonary fibrosis produced in hamsters by endotracheal bleomycin: pathology and stereology. Am Rev Respir Dis 117: 1099–1108.
[12]
Thrall RS, McCormick JR, Jack RM, McReynolds RA, Ward PA (1979) Bleomycin-induced pulmonary fibrosis in the rat: inhibition by indomethacin. Am J Pathol 95: 117–130.
[13]
Degryse AL, Tanjore H, Xu XC, Polosukhin VV, Jones BR, et al. (2010) Repetitive intratracheal bleomycin models several features of idiopathic pulmonary fibrosis. Am J Physiol Lung Cell Mol Physiol 299: L442–452.
[14]
Izbicki G, Segel MJ, Christensen TG, Conner MW, Breuer R (2002) Time course of bleomycin-induced lung fibrosis. Int J Exp Pathol 83: 111–119.
[15]
Moore BB, Hogaboam CM (2008) Murine models of pulmonary fibrosis. Am J Physiol Lung Cell Mol Physiol 294: L152–160.
[16]
Adachi K, Mizoguchi K, Kawarada S, Miyoshi A, Suzuki M, et al. (2010) Effects of erlotinib on lung injury induced by intratracheal administration of bleomycin (BLM) in rats. J Toxicol Sci 35: 503–514.
[17]
Babin AL, Cannet C, Gerard C, Wyss D, Page CP, et al. (2011) Noninvasive assessment of bleomycin-induced lung injury and the effects of short-term glucocorticosteroid treatment in rats using MRI. J Magn Reson Imaging 33: 603–614.
[18]
Gong LK, Li XH, Wang H, Zhang L, Cai Y, et al. (2004) Feitai attenuates bleomycin-induced pulmonary fibrosis in rats. Biol Pharm Bull 27: 634–640.
[19]
Southam DS, Dolovich M, O’Byrne PM, Inman MD (2002) Distribution of intranasal instillations in mice: effects of volume, time, body position, and anesthesia. Am J Physiol Lung Cell Mol Physiol 282: L833–839.
[20]
Foster WM, Walters DM, Longphre M, Macri K, Miller LM (2001) Methodology for the measurement of mucociliary function in the mouse by scintigraphy. J Appl Physiol 90: 1111–1117.
[21]
Rao GV, Tinkle S, Weissman DN, Antonini JM, Kashon ML, et al. (2003) Efficacy of a technique for exposing the mouse lung to particles aspirated from the pharynx. J Toxicol Environ Health A 66: 1441–1452.
[22]
Lakatos HF, Burgess HA, Thatcher TH, Redonnet MR, Hernady E, et al. (2006) Oropharyngeal aspiration of a silica suspension produces a superior model of silicosis in the mouse when compared to intratracheal instillation. Exp Lung Res 32: 181–199.
[23]
De Vooght V, Vanoirbeek JA, Haenen S, Verbeken E, Nemery B, et al. (2009) Oropharyngeal aspiration: an alternative route for challenging in a mouse model of chemical-induced asthma. Toxicology 259: 84–89.
[24]
Morgan DL, Flake GP, Kirby PJ, Palmer SM (2008) Respiratory toxicity of diacetyl in C57BL/6 mice. Toxicol Sci 103: 169–180.
[25]
Babin AL, Cannet C, Gerard C, Saint-Mezard P, Page CP, et al. (2012) Bleomycin-induced lung injury in mice investigated by MRI: model assessment for target analysis. Magn Reson Med 67: 499–509.
[26]
Bosanquet AG (1986) Stability of solutions of antineoplastic agents during preparation and storage for in vitro assays. II. Assay methods, adriamycin and the other antitumour antibiotics. Cancer Chemother Pharmacol 17: 1–10.
[27]
Ble FX, Cannet C, Zurbruegg S, Karmouty-Quintana H, Bergmann R, et al. (2008) Allergen-induced lung inflammation in actively sensitized mice assessed with MR imaging. Radiology 248: 834–843.
[28]
Takahashi M, Togao O, Obara M, van Cauteren M, Ohno Y, et al. (2010) Ultra-short echo time (UTE) MR imaging of the lung: comparison between normal and emphysematous lungs in mutant mice. J Magn Reson Imaging 32: 326–333.
[29]
van Echteld CJ, Beckmann N (2011) A view on imaging in drug research and development for respiratory diseases. J Pharmacol Exp Ther 337: 335–349.
[30]
Zurek M, Boyer L, Caramelle P, Boczkowski J, Cremillieux Y (2012) Longitudinal and noninvasive assessment of emphysema evolution in a murine model using proton MRI. Magn Reson Med 68: 898–904.
[31]
Andrade GB, Riet-Correa F, Montes GS, Battlehner CN, Saldiva PH (1997) Dating of fibrotic lesions by the Picrosirius-polarization method. An application using the lesions of Lechiguana (bovine focal proliferative fibrogranulomatous panniculitis). Eur J Histochem 41: 203–209.
[32]
Bradshaw AD, Baicu CF, Rentz TJ, Van Laer AO, Boggs J, et al. (2009) Pressure overload-induced alterations in fibrillar collagen content and myocardial diastolic function: role of secreted protein acidic and rich in cysteine (SPARC) in post-synthetic procollagen processing. Circulation 119: 269–280.
[33]
Bradshaw AD, Baicu CF, Rentz TJ, Van Laer AO, Bonnema DD, et al. (2010) Age-dependent alterations in fibrillar collagen content and myocardial diastolic function: role of SPARC in post-synthetic procollagen processing. Am J Physiol Heart Circ Physiol 298: H614–622.
[34]
Montes GS, Junqueira LC (1991) The use of the Picrosirius-polarization method for the study of the biopathology of collagen. Mem Inst Oswaldo Cruz 86 Suppl 31–11.
[35]
Pickering JG, Boughner DR (1991) Quantitative assessment of the age of fibrotic lesions using polarized light microscopy and digital image analysis. Am J Pathol 138: 1225–1231.
[36]
Whittaker P, Kloner RA, Boughner DR, Pickering JG (1994) Quantitative assessment of myocardial collagen with picrosirius red staining and circularly polarized light. Basic Res Cardiol 89: 397–410.
[37]
Zhang H, Sun L, Wang W, Ma X (2006) Quantitative analysis of fibrosis formation on the microcapsule surface with the use of picro-sirius red staining, polarized light microscopy, and digital image analysis. J Biomed Mater Res A 76: 120–125.
[38]
Diaz Encarnacion MM, Griffin MD, Slezak JM, Bergstralh EJ, Stegall MD, et al. (2004) Correlation of quantitative digital image analysis with the glomerular filtration rate in chronic allograft nephropathy. Am J Transplant 4: 248–256.
[39]
Farris AB, Adams CD, Brousaides N, Della Pelle PA, Collins AB, et al. (2011) Morphometric and visual evaluation of fibrosis in renal biopsies. J Am Soc Nephrol 22: 176–186.
[40]
Galli GG, Honnens de Lichtenberg K, Carrara M, Hans W, Wuelling M, et al. (2012) Prdm5 regulates collagen gene transcription by association with RNA polymerase II in developing bone. PLoS Genet 8: e1002711.
[41]
Gold LI, Rahman M, Blechman KM, Greives MR, Churgin S, et al. (2006) Overview of the role for calreticulin in the enhancement of wound healing through multiple biological effects. J Investig Dermatol Symp Proc 11: 57–65.
[42]
Moon JC, Reed E, Sheppard MN, Elkington AG, Ho SY, et al. (2004) The histologic basis of late gadolinium enhancement cardiovascular magnetic resonance in hypertrophic cardiomyopathy. J Am Coll Cardiol 43: 2260–2264.
[43]
Nart P, Williams A, Thompson H, Innocent GT (2004) Morphometry of bovine dilated cardiomyopathy. J Comp Pathol 130: 235–245.
[44]
Ophof R, Maltha JC, Von den Hoff JW, Kuijpers-Jagtman AM (2004) Histologic evaluation of skin-derived and collagen-based substrates implanted in palatal wounds. Wound Repair Regen 12: 528–538.
[45]
Rossi GA, Szapiel S, Ferrans VJ, Crystal RG (1987) Susceptibility to experimental interstitial lung disease is modified by immune- and non-immune-related genes. Am Rev Respir Dis 135: 448–455.
[46]
Schrier DJ, Kunkel RG, Phan SH (1983) The role of strain variation in murine bleomycin-induced pulmonary fibrosis. Am Rev Respir Dis 127: 63–66.
[47]
Shapiro SD, Griffin GL, Gilbert DJ, Jenkins NA, Copeland NG, et al. (1992) Molecular cloning, chromosomal localization, and bacterial expression of a murine macrophage metalloelastase. J Biol Chem 267: 4664–4671.
[48]
Shapiro SD, Kobayashi DK, Ley TJ (1993) Cloning and characterization of a unique elastolytic metalloproteinase produced by human alveolar macrophages. J Biol Chem 268: 23824–23829.
[49]
Morris L, Graham CF, Gordon S (1991) Macrophages in haemopoietic and other tissues of the developing mouse detected by the monoclonal antibody F4/80. Development 112: 517–526.
[50]
Austyn JM, Gordon S (1981) F4/80, a monoclonal antibody directed specifically against the mouse macrophage. Eur J Immunol 11: 805–815.
[51]
England KA, Price AP, Tram KV, Shapiro SD, Blazar BR, et al. (2011) Evidence for early fibrosis and increased airway resistance in bone marrow transplant recipient mice deficient in MMP12. Am J Physiol Lung Cell Mol Physiol 301: L519–526.
[52]
Chua F, Gauldie J, Laurent GJ (2005) Pulmonary fibrosis: searching for model answers. Am J Respir Cell Mol Biol 33: 9–13.
[53]
Corbel M, Caulet-Maugendre S, Germain N, Molet S, Lagente V, et al. (2001) Inhibition of bleomycin-induced pulmonary fibrosis in mice by the matrix metalloproteinase inhibitor batimastat. J Pathol 193: 538–545.
[54]
Takemasa A, Ishii Y, Fukuda T (2012) A neutrophil elastase inhibitor prevents bleomycin-induced pulmonary fibrosis in mice. Eur Respir J 40: 1475–1482.
[55]
Iwasawa T, Ogura T, Sakai F, Kanauchi T, Komagata T, et al.. (2012) CT analysis of the effect of pirfenidone in patients with idiopathic pulmonary fibrosis. Eur J Radiol, in press (DOI 10.1016/j.ejrad.2012.02.014).
