N Hagimoto, K Kuwano, Y Nomoto, R Kunitake and N Hara
Research Institute for Diseases of the Chest, Faculty of Medicine, Kyushu University, Fukuoka, Japan.

The incidence of apoptosis and the expression of Fas antigen (Fas)/Fas ligand (FasL) mRNA in bleomycin-induced pulmonary fibrosis in mice were examined. Male ICR mice were intratracheally instilled with bleomycin (5 U/kg of body weight). The controls were injected with sterile saline. The animals were anesthetized and killed at 1, 6, and 12 h, and 1, 3, 5, 7, 9, and 14 days after bleomycin instillation. We assessed the incidence of apoptosis in lung tissues by DNA fragmentation on agarose gel electrophoresis, terminal deoxynucleotidyl transferase- mediated dUTP biotin nick end-labeling, and electron microscopy. The expression of Fas and FasL mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). The localization of Fas mRNA was analyzed by in situ hybridization and that of FasL mRNA was analyzed by RT in situ PCR. The results showed that (1) a single instillation of bleomycin leads to the rapid appearance of apoptosis in bronchial and alveolar epithelial cells, which resolves within 1 day, and (2) apoptosis reappears on day 7 and continues for over 14 days after bleomycin instillation. This was accompanied with a progression of fibrosis. Corticosteroid administration completely blocked both apoptosis and fibrosis. The expression of Fas mRNA was upregulated in the alveolar epithelial cells by the bleomycin instillation. FasL mRNA was also upregulated in infiltrating lymphocytes after bleomycin treatment, but not in the control mice. The administration of corticosteroids suppressed the expression of Fas and FasL mRNA as well as apoptosis and fibrosis. Although these results do not show that apoptosis mediated by the Fas/FasL system is directly linked to bleomycin-induced fibrosis, we speculate that excessive apoptosis and the Fas/FasL system play a role in the pathogenesis of bleomycin- induced lung injury.

This article has been cited by other articles:

				M. C. Gongora, H. E. Lob, U. Landmesser, T. J. Guzik, W. D. Martin, K. Ozumi, S. M. Wall, D. S. Wilson, N. Murthy, M. Gravanis, et al. Loss of Extracellular Superoxide Dismutase Leads to Acute Lung Damage in the Presence of Ambient Air: A Potential Mechanism Underlying Adult Respiratory Distress Syndrome Am. J. Pathol., October 1, 2008; 173(4): 915 - 926. [Abstract] [Full Text] [PDF]

				K. Kurosu, Y. Takiguchi, O. Okada, N. Yumoto, S. Sakao, Y. Tada, Y. Kasahara, N. Tanabe, K. Tatsumi, M. Weiden, et al. Identification of Annexin 1 as a Novel Autoantigen in Acute Exacerbation of Idiopathic Pulmonary Fibrosis J. Immunol., July 1, 2008; 181(1): 756 - 767. [Abstract] [Full Text] [PDF]

				M. E. De Paepe, S. Gundavarapu, U. Tantravahi, J. R. Pepperell, S. A. Haley, F. I. Luks, and Q. Mao Fas-Ligand-Induced Apoptosis of Respiratory Epithelial Cells Causes Disruption of Postcanalicular Alveolar Development Am. J. Pathol., July 1, 2008; 173(1): 42 - 56. [Abstract] [Full Text] [PDF]

				C. A Beamer and A. Holian Antigen-Presenting Cell Population Dynamics during Murine Silicosis Am. J. Respir. Cell Mol. Biol., December 1, 2007; 37(6): 729 - 738. [Abstract] [Full Text] [PDF]

				D. Pilling, D. Roife, M. Wang, S. D. Ronkainen, J. R. Crawford, E. L. Travis, and R. H. Gomer Reduction of Bleomycin-Induced Pulmonary Fibrosis by Serum Amyloid P J. Immunol., September 15, 2007; 179(6): 4035 - 4044. [Abstract] [Full Text] [PDF]

				R. Golan-Gerstl, S. B. Wallach-Dayan, G. Amir, and R. Breuer Epithelial Cell Apoptosis by Fas Ligand-Positive Myofibroblasts in Lung Fibrosis Am. J. Respir. Cell Mol. Biol., March 1, 2007; 36(3): 270 - 275. [Abstract] [Full Text] [PDF]

				A. Gazdhar, P. Fachinger, C. van Leer, J. Pierog, M. Gugger, R. Friis, R. A. Schmid, and T. Geiser Gene transfer of hepatocyte growth factor by electroporation reduces bleomycin-induced lung fibrosis Am J Physiol Lung Cell Mol Physiol, February 1, 2007; 292(2): L529 - L536. [Abstract] [Full Text] [PDF]

				A. Shukla, K. M. Lounsbury, T. F. Barrett, J. Gell, M. Rincon, K. J. Butnor, D. J. Taatjes, G. S. Davis, P. Vacek, K. I. Nakayama, et al. Asbestos-Induced Peribronchiolar Cell Proliferation and Cytokine Production Are Attenuated in Lungs of Protein Kinase C-{delta} Knockout Mice Am. J. Pathol., January 1, 2007; 170(1): 140 - 151. [Abstract] [Full Text] [PDF]

				T. Otsuki, Y. Miura, Y. Nishimura, F. Hyodoh, A. Takata, M. Kusaka, H. Katsuyama, M. Tomita, A. Ueki, and T. Kishimoto Alterations of Fas and Fas-Related Molecules in Patients with Silicosis. Experimental Biology and Medicine, May 1, 2006; 231(5): 522 - 533. [Abstract] [Full Text] [PDF]

				S. B. Wallach-Dayan, G. Izbicki, P. Y. Cohen, R. Gerstl-Golan, A. Fine, and R. Breuer Bleomycin initiates apoptosis of lung epithelial cells by ROS but not by Fas/FasL pathway Am J Physiol Lung Cell Mol Physiol, April 1, 2006; 290(4): L790 - L796. [Abstract] [Full Text] [PDF]

				S. K. Frankel, G. P. Cosgrove, S.-I. Cha, C. D. Cool, M. W. Wynes, B. L. Edelman, K. K. Brown, and D. W. H. Riches TNF-{alpha} Sensitizes Normal and Fibrotic Human Lung Fibroblasts to Fas-Induced Apoptosis Am. J. Respir. Cell Mol. Biol., March 1, 2006; 34(3): 293 - 304. [Abstract] [Full Text] [PDF]

				F. Huaux, M. Gharaee-Kermani, T. Liu, V. Morel, B. McGarry, M. Ullenbruch, S. L. Kunkel, J. Wang, Z. Xing, and S. H. Phan Role of Eotaxin-1 (CCL11) and CC Chemokine Receptor 3 (CCR3) in Bleomycin-Induced Lung Injury and Fibrosis Am. J. Pathol., December 1, 2005; 167(6): 1485 - 1496. [Abstract] [Full Text] [PDF]

				J. H. Kim, H. Y. Kim, S. Kim, J.-H. Chung, W. S. Park, and D. H. Chung Natural Killer T (NKT) Cells Attenuate Bleomycin-Induced Pulmonary Fibrosis by Producing Interferon-{gamma} Am. J. Pathol., November 1, 2005; 167(5): 1231 - 1241. [Abstract] [Full Text] [PDF]

				W. E. Lawson, V. V. Polosukhin, G. T. Stathopoulos, O. Zoia, W. Han, K. B. Lane, B. Li, E. F. Donnelly, G. E. Holburn, K. G. Lewis, et al. Increased and Prolonged Pulmonary Fibrosis in Surfactant Protein C-Deficient Mice Following Intratracheal Bleomycin Am. J. Pathol., November 1, 2005; 167(5): 1267 - 1277. [Abstract] [Full Text] [PDF]

				T. R. Martin, N. Hagimoto, M. Nakamura, and G. Matute-Bello Apoptosis and Epithelial Injury in the Lungs Proceedings of the ATS, October 1, 2005; 2(3): 214 - 220. [Abstract] [Full Text] [PDF]

				M. E. De Paepe, Q. Mao, Y. Chao, J. L. Powell, L. P. Rubin, and S. Sharma Hyperoxia-induced apoptosis and Fas/FasL expression in lung epithelial cells Am J Physiol Lung Cell Mol Physiol, October 1, 2005; 289(4): L647 - L659. [Abstract] [Full Text] [PDF]

				S. Hodge, G. Hodge, M. Holmes, and P. N. Reynolds Increased airway epithelial and T-cell apoptosis in COPD remains despite smoking cessation Eur. Respir. J., March 1, 2005; 25(3): 447 - 454. [Abstract] [Full Text] [PDF]

				M. Plataki, A. V. Koutsopoulos, K. Darivianaki, G. Delides, N. M. Siafakas, and D. Bouros Expression of Apoptotic and Antiapoptotic Markers in Epithelial Cells in Idiopathic Pulmonary Fibrosis Chest, January 1, 2005; 127(1): 266 - 274. [Abstract] [Full Text] [PDF]

				S. Bao, Y. Wang, P. Sweeney, A. Chaudhuri, A. I. Doseff, C. B. Marsh, and D. L. Knoell Keratinocyte growth factor induces Akt kinase activity and inhibits Fas-mediated apoptosis in A549 lung epithelial cells Am J Physiol Lung Cell Mol Physiol, January 1, 2005; 288(1): L36 - L42. [Abstract] [Full Text] [PDF]

				H Miyazaki, K Kuwano, K Yoshida, T Maeyama, M Yoshimi, M Fujita, N Hagimoto, R Yoshida, and Y Nakanishi The perforin mediated apoptotic pathway in lung injury and fibrosis J. Clin. Pathol., December 1, 2004; 57(12): 1292 - 1298. [Abstract] [Full Text] [PDF]

				X. Li, R. Shu, G. Filippatos, and B. D. Uhal Apoptosis in lung injury and remodeling J Appl Physiol, October 1, 2004; 97(4): 1535 - 1542. [Abstract] [Full Text] [PDF]

				M. E. De Paepe, Q. Mao, M. Embree-Ku, L. P. Rubin, and F. I. Luks Fas/FasL-mediated apoptosis in perinatal murine lungs Am J Physiol Lung Cell Mol Physiol, October 1, 2004; 287(4): L730 - L742. [Abstract] [Full Text] [PDF]

				H. F. Nadrous, J. H. Ryu, W. W. Douglas, P. A. Decker, and E. J. Olson Impact of Angiotensin-Converting Enzyme Inhibitors and Statins on Survival in Idiopathic Pulmonary Fibrosis Chest, August 1, 2004; 126(2): 438 - 446. [Abstract] [Full Text] [PDF]

				I. Inoshima, K. Kuwano, N. Hamada, M. Yoshimi, T. Maeyama, N. Hagimoto, Y. Nakanishi, and N. Hara Induction of CDK inhibitor p21 gene as a new therapeutic strategy against pulmonary fibrosis Am J Physiol Lung Cell Mol Physiol, April 1, 2004; 286(4): L727 - L733. [Abstract] [Full Text] [PDF]

				M. Mori, H. Morishita, H. Nakamura, H. Matsuoka, K. Yoshida, Y. Kishima, Z. Zhou, H. Kida, T. Funakoshi, S. Goya, et al. Hepatoma-Derived Growth Factor Is Involved in Lung Remodeling by Stimulating Epithelial Growth Am. J. Respir. Cell Mol. Biol., April 1, 2004; 30(4): 459 - 469. [Abstract] [Full Text] [PDF]

				O. Lesur, M. Brisebois, A. Thibodeau, F. Chagnon, D. Lane, and T. Fullop Role of IFN-{gamma} and IL-2 in rat lung epithelial cell migration and apoptosis after oxidant injury Am J Physiol Lung Cell Mol Physiol, January 1, 2004; 286(1): L4 - L14. [Abstract] [Full Text] [PDF]

				R. H. Hastings, R. A. Quintana, R. Sandoval, D. Duey, Y. Rascon, D. W. Burton, and L. J. Deftos Proapoptotic Effects of Parathyroid Hormone-Related Protein in Type II Pneumocytes Am. J. Respir. Cell Mol. Biol., December 1, 2003; 29(6): 733 - 742. [Abstract] [Full Text] [PDF]

				M. Fujita, J. M. Shannon, O. Morikawa, J. Gauldie, N. Hara, and R. J. Mason Overexpression of Tumor Necrosis Factor-{alpha} Diminishes Pulmonary Fibrosis Induced by Bleomycin or Transforming Growth Factor-{beta} Am. J. Respir. Cell Mol. Biol., December 1, 2003; 29(6): 669 - 676. [Abstract] [Full Text] [PDF]

				X. Li, H. Rayford, and B. D. Uhal Essential Roles for Angiotensin Receptor AT1a in Bleomycin-Induced Apoptosis and Lung Fibrosis in Mice Am. J. Pathol., December 1, 2003; 163(6): 2523 - 2530. [Abstract] [Full Text]

				V. Ruiz, R. Ma. Ordonez, J. Berumen, R. Ramirez, B. Uhal, C. Becerril, A. Pardo, and M. Selman Unbalanced collagenases/TIMP-1 expression and epithelial apoptosis in experimental lung fibrosis Am J Physiol Lung Cell Mol Physiol, November 1, 2003; 285(5): L1026 - L1036. [Abstract] [Full Text] [PDF]

				B.D. Uhal Epithelial apoptosis in the initiation of lung fibrosis Eur. Respir. J., September 20, 2003; 22(44_suppl): 7s - 9s. [Full Text] [PDF]

				N. Kaminski, J. A. Belperio, P. B. Bitterman, L. Chen, S. W. Chensue, A. M.K. Choi, S. Dacic, J. H. Dauber, R. M. du Bois, J. J. Enghild, et al. Idiopathic Pulmonary Fibrosis Am. J. Respir. Cell Mol. Biol., September 1, 2003; 29(3): S1 - 105. [Full Text] [PDF]

				W A Dik, R J McAnulty, M A Versnel, B A E Naber, L J I Zimmermann, G J Laurent, and S E Mutsaers Short course dexamethasone treatment following injury inhibits bleomycin induced fibrosis in rats Thorax, September 1, 2003; 58(9): 765 - 771. [Abstract] [Full Text] [PDF]

				L. A. Ortiz, F. Gambelli, C. McBride, D. Gaupp, M. Baddoo, N. Kaminski, and D. G. Phinney Mesenchymal stem cell engraftment in lung is enhanced in response to bleomycin exposure and ameliorates its fibrotic effects PNAS, July 8, 2003; 100(14): 8407 - 8411. [Abstract] [Full Text] [PDF]

				V. E. Kagan, G. G. Borisenko, B. F. Serinkan, Y. Y. Tyurina, V. A. Tyurin, J. Jiang, S. X. Liu, A. A. Shvedova, J. P. Fabisiak, W. Uthaisang, et al. Appetizing rancidity of apoptotic cells for macrophages: oxidation, externalization, and recognition of phosphatidylserine Am J Physiol Lung Cell Mol Physiol, July 1, 2003; 285(1): L1 - L17. [Abstract] [Full Text] [PDF]

				X. Li, H. Zhang, V. Soledad-Conrad, J. Zhuang, and B. D. Uhal Bleomycin-induced apoptosis of alveolar epithelial cells requires angiotensin synthesis de novo Am J Physiol Lung Cell Mol Physiol, March 1, 2003; 284(3): L501 - L507. [Abstract] [Full Text] [PDF]

				T. Fujita, M. Maruyama, J. Araya, K. Sassa, Y. Kawagishi, R. Hayashi, S. Matsui, T. Kashii, N. Yamashita, E. Sugiyama, et al. Hydrogen Peroxide Induces Upregulation of Fas in Human Airway Epithelial Cells via the Activation of PARP-p53 Pathway Am. J. Respir. Cell Mol. Biol., November 1, 2002; 27(5): 542 - 552. [Abstract] [Full Text] [PDF]

				T. Tanaka, M. Yoshimi, T. Maeyama, N. Hagimoto, K. Kuwano, and N. Hara Resistance to Fas-mediated apoptosis in human lung fibroblast Eur. Respir. J., August 1, 2002; 20(2): 359 - 368. [Abstract] [Full Text] [PDF]

				R. G. Crystal, P. B. Bitterman, B. Mossman, M. I. Schwarz, D. Sheppard, L. Almasy, H. A. Chapman, S. L. Friedman, T. E. King Jr., L. A. Leinwand, et al. Future Research Directions in Idiopathic Pulmonary Fibrosis: Summary of a National Heart, Lung, and Blood Institute Working Group Am. J. Respir. Crit. Care Med., July 15, 2002; 166(2): 236 - 246. [Abstract] [Full Text] [PDF]

				H. Matsuoka, T. Arai, M. Mori, S. Goya, H. Kida, H. Morishita, H. Fujiwara, I. Tachibana, T. Osaki, and S. Hayashi A p38 MAPK inhibitor, FR-167653, ameliorates murine bleomycin-induced pulmonary fibrosis Am J Physiol Lung Cell Mol Physiol, July 1, 2002; 283(1): L103 - L112. [Abstract] [Full Text] [PDF]

				G. S. Filippatos, N. Gangopadhyay, O. Lalude, N. Parameswaran, S. I. Said, W. Spielman, and B. D. Uhal Regulation of apoptosis by vasoactive peptides Am J Physiol Lung Cell Mol Physiol, October 1, 2001; 281(4): L749 - L761. [Abstract] [Full Text] [PDF]

				T. Betsuyaku, G. L. Griffin, M. A. Watson, and R. M. Senior Laser Capture Microdissection and Real-Time Reverse Transcriptase/ Polymerase Chain Reaction of Bronchiolar Epithelium after Bleomycin Am. J. Respir. Cell Mol. Biol., September 1, 2001; 25(3): 278 - 284. [Abstract] [Full Text] [PDF]

				S. Sleijfer Bleomycin-Induced Pneumonitis Chest, August 1, 2001; 120(2): 617 - 624. [Abstract] [Full Text] [PDF]

				T. Maeyama, K. Kuwano, M. Kawasaki, R. Kunitake, N. Hagimoto, and N. Hara Attenuation of bleomycin-induced pneumopathy in mice by monoclonal antibody to interleukin-12 Am J Physiol Lung Cell Mol Physiol, June 1, 2001; 280(6): L1128 - L1137. [Abstract] [Full Text] [PDF]

				F. Takahashi, K. Takahashi, T. Okazaki, K. Maeda, H. Ienaga, M. Maeda, S. Kon, T. Uede, and Y. Fukuchi Role of Osteopontin in the Pathogenesis of Bleomycin-Induced Pulmonary Fibrosis Am. J. Respir. Cell Mol. Biol., March 1, 2001; 24(3): 264 - 271. [Abstract] [Full Text] [PDF]

				Y. KITAMURA, S. HASHIMOTO, N. MIZUTA, A. KOBAYASHI, K. KOOGUCHI, I. FUJIWARA, and H. NAKAJIMA Fas/FasL-dependent Apoptosis of Alveolar Cells after Lipopolysaccharide-induced Lung Injury in Mice Am. J. Respir. Crit. Care Med., March 1, 2001; 163(3): 762 - 769. [Abstract] [Full Text] [PDF]

				K. Kuwano, R. Kunitake, T. Maeyama, N. Hagimoto, M. Kawasaki, T. Matsuba, M. Yoshimi, I. Inoshima, K. Yoshida, and N. Hara Attenuation of bleomycin-induced pneumopathy in mice by a caspase inhibitor Am J Physiol Lung Cell Mol Physiol, February 1, 2001; 280(2): L316 - L325. [Abstract] [Full Text] [PDF]

				M. E. De Paepe, L. P. Rubin, C. Jude, A. M. Lesieur-Brooks, D. R. Mills, and F. I. Luks Fas ligand expression coincides with alveolar cell apoptosis in late-gestation fetal lung development Am J Physiol Lung Cell Mol Physiol, November 1, 2000; 279(5): L967 - L976. [Abstract] [Full Text] [PDF]

				D. , D. , , and D. Nitric Oxide-dependent Activation of p53 Suppresses Bleomycin-induced Apoptosis in the J. Exp. Med., September 18, 2000; 192(6): 857 - 870. [Abstract] [Full Text] [PDF]

				K. AOSHIBA, S. YASUI, J. TAMAOKI, and A. NAGAI The Fas/Fas-Ligand System Is Not Required for Bleomycin-induced Pulmonary Fibrosis in Mice Am. J. Respir. Crit. Care Med., August 1, 2000; 162(2): 695 - 700. [Abstract] [Full Text] [PDF]

				R. Wang, O. Ibarra-Sunga, L. Verlinski, R. Pick, and B. D. Uhal Abrogation of bleomycin-induced epithelial apoptosis and lung fibrosis by captopril or by a caspase inhibitor Am J Physiol Lung Cell Mol Physiol, July 1, 2000; 279(1): L143 - L151. [Abstract] [Full Text] [PDF]

				M. Behnia, K. A. Robertson, and W. J. Martin II Lung Infections : Role of Apoptosis in Host Defense and Pathogenesis of Disease Chest, June 1, 2000; 117(6): 1771 - 1777. [Abstract] [Full Text] [PDF]

				A. Mishra, N. A. Doyle, and W. J. Martin II Bleomycin-Mediated Pulmonary Toxicity . Evidence for a p53-Mediated Response Am. J. Respir. Cell Mol. Biol., May 1, 2000; 22(5): 543 - 549. [Abstract] [Full Text]

				E. Marchi, W. Liu, and V. C. Broaddus Mesothelial cell apoptosis is confirmed in vivo by morphological change in cytokeratin distribution Am J Physiol Lung Cell Mol Physiol, March 1, 2000; 278(3): L528 - L535. [Abstract] [Full Text] [PDF]

				K. Kuwano and N. Hara Signal Transduction Pathways of Apoptosis and Inflammation Induced by the Tumor Necrosis Factor Receptor Family Am. J. Respir. Cell Mol. Biol., February 1, 2000; 22(2): 147 - 149. [Full Text]

				S. HASHIMOTO, A. KOBAYASHI, K. KOOGUCHI, Y. KITAMURA, H. ONODERA, and H. NAKAJIMA Upregulation of Two Death Pathways of Perforin/Granzyme and FasL/Fas in Septic Acute Respiratory Distress Syndrome Am. J. Respir. Crit. Care Med., January 1, 2000; 161(1): 237 - 243. [Abstract] [Full Text]

				R. Wang, A. Zagariya, E. Ang, O. Ibarra-Sunga, and B. D. Uhal Fas-induced apoptosis of alveolar epithelial cells requires ANG II generation and receptor interaction Am J Physiol Lung Cell Mol Physiol, December 1, 1999; 277(6): L1245 - L1250. [Abstract] [Full Text] [PDF]

				C. L. Zanella, C. R. Timblin, A. Cummins, M. Jung, J. Goldberg, R. Raabe, T. R. Tritton, and B. T. Mossman Asbestos-induced phosphorylation of epidermal growth factor receptor is linked to c-fos and apoptosis Am J Physiol Lung Cell Mol Physiol, October 1, 1999; 277(4): L684 - L693. [Abstract] [Full Text] [PDF]

				K. Okudela, T. Ito, H. Mitsui, H. Hayashi, N. Udaka, M. Kanisawa, and H. Kitamura The Role of p53 in Bleomycin-Induced DNA Damage in the Lung : A Comparative Study with the Small Intestine Am. J. Pathol., October 1, 1999; 155(4): 1341 - 1351. [Abstract] [Full Text] [PDF]

				N. Hagimoto, K. Kuwano, M. Kawasaki, M. Yoshimi, Y. Kaneko, R. Kunitake, T. Maeyama, T. Tanaka, and N. Hara Induction of Interleukin-8 Secretion and Apoptosis in Bronchiolar Epithelial Cells by Fas Ligation Am. J. Respir. Cell Mol. Biol., September 1, 1999; 21(3): 436 - 445. [Abstract] [Full Text]

				D. W Kamp and S. A Weitzman The molecular basis of asbestos induced lung injury Thorax, July 1, 1999; 54(7): 638 - 652. [Full Text]

				A. N. Liu, A. Z. Mohammed, W. R. Rice, D. T. Fiedeldey, J. S. Liebermann, J. A. Whitsett, T. J. Braciale, and R. I. Enelow Perforin-Independent CD8+ T-Cell-Mediated Cytotoxicity of Alveolar Epithelial Cells Is Preferentially Mediated by Tumor Necrosis Factor-alpha . Relative Insensitivity to Fas Ligand Am. J. Respir. Cell Mol. Biol., May 1, 1999; 20(5): 849 - 858. [Abstract] [Full Text]