Cell and Tissue Research

, Volume 359, Issue 2, pp 589–603 | Cite as

Aberrant elastin remodeling in the lungs of O2-exposed newborn mice; primarily results from perturbed interaction between integrins and elastin

  • Wenli Han
  • Chunbao Guo
  • Qiutong Liu
  • Benli Yu
  • Zhaoyun Liu
  • Junqing YangEmail author
  • Chun DengEmail author
Regular Article


Excessive localization of elastin from septal tips to alveolar walls is a key feature of bronchopulmonary dysplasia (BPD). The abnormal accumulation of lung elastin, involving the structural and functional interaction of a series of proteins, remains poorly understood. To further investigate the mechanisms accounting for the abnormal accumulation of elastin in the lungs of newborn mice with BPD, we evaluate elastin distribution and its interaction with proteins involved in its aberrant localization, such as integrin αv, fibulin-5 and transforming growth factor β1 (TGF-β1), in lungs of newborn mice exposed to 60 % O2 for 21 days. Lung histology revealed aberrant elastin production and impaired lung septation in O2-exposed lungs, while tropoelastin, integrin αv, fibulin-1, fibulin-2 and fibulin-4 gene expression were elevated. Dual staining image analysis of lung sections revealed that co-localization of integrin αv and elastin increased following O2 exposure with elastin distributed throughout the walls of air spaces rather than at septal tips. Furthermore, integrin αv appeared to be induced initially. Concurrently, increased fibulin-5 and TGF-β1 (which may regulate elastic fiber assembly) expression was detected, which may explain the altered lung elastin deposition and defective septation that are observed during BPD. These data support the hypothesis that excessive and aberrant αv integrin expression was initially induced by hyperoxia; αv integrin then interacted with and recruited elastin. These alterations were accompanied by fibulin-5 deposition and TGF-β1 activation, which may impede normal matrix remodeling, thereby contributing to the pathological pulmonary features of BPD.


Bronchopulmonary dysplasia Integrin αv Elastin Alveolarization 



We thank Prof. Xianqing Jin for providing technical assistance and insightful discussions during the preparation of the manuscript. We thank Dr. Xiaoyong Zhang of the Wistar Institute (USA), who provided medical writing services. This research was supported by the National Natural Science Foundation of China (No. 81270058, 30770950) and by the Chongqing Natural Science Foundation (CSTC, 2009BB6072). Wenli Han, Qiutong Liu and Zhaoyun Liu designed and performed the experiments, analyzed the data and prepared the manuscript. Zhaoyun Liu bred the mice. Chunbao Guo, Junqing Yang and Chun Deng designed the experiments, analyzed the data, evaluated the manuscript and wrote the paper.

Conflicts of Interest

The authors declare that they have no competing interests.


  1. Albertine KH, Jones GP, Starcher BC, Bohnsack JF, Davis PL, Cho SC, Carlton DP, Bland RD (1999) Chronic lung injury in preterm lambs. Disordered respiratory tract development. Am J Respir Crit Care Med 159:945–958PubMedCrossRefGoogle Scholar
  2. Alejandre-Alcázar MA, Kwapiszewska G, Reiss I, Amarie OV, Marsh LM, Sevilla-Pérez J, Wygrecka M, Eul B, Köbrich S, Hesse M, Schermuly RT, Seeger W, Eickelberg O, Morty RE (2007) Hyperoxia modulates TGF-beta/BMP signaling in a mouse model of bronchopulmonary dysplasia. Am J Physiol Lung Cell Mol Physiol 292:L537–L549PubMedCrossRefGoogle Scholar
  3. Asano Y, Ihn H, Yamane K, Jinnin M, Tamaki K (2006) Increased expression of integrin alphavbeta5 induces the myofibroblastic differentiation of dermal fibroblasts. Am J Pathol 168:499–510PubMedCentralPubMedCrossRefGoogle Scholar
  4. Ashour K, Shan L, Lee JH, Schlicher W, Wada K, Wada E, Sunday ME (2006) Bombesin inhibits alveolarization and promotes pulmonary fibrosis in newborn mice. Am J Respir Crit Care Med 173:1377–1385PubMedCentralPubMedCrossRefGoogle Scholar
  5. Benjamin JT, Gaston DC, Halloran BA, Schnapp LM, Zent R, Prince LS (2009) The role of integrin alpha8beta1 in fetal lung morphogenesis and injury. Dev Biol 335:407–417PubMedCentralPubMedCrossRefGoogle Scholar
  6. Blaauboer ME, Boeijen FR, Emson CL, Turner SM, Zandieh-Doulabi B, Hanemaaijer R, Smit TH, Stoop R, Everts V (2014) Extracellular matrix proteins: a positive feedback loop in lung fibrosis? Matrix Biol 34:170–178PubMedCrossRefGoogle Scholar
  7. Blanchevoye C, Floquet N, Scandolera A, Baud S, Maurice P, Bocquet O, Blaise S, Ghoneim C, Cantarelli B, Delacoux F, Dauchez M, Efremov RG, Martiny L, Duca L, Debelle L (2013) Interaction between the elastin peptide VGVAPG and human elastin binding protein. J Biol Chem 288:1317–1328PubMedCentralPubMedCrossRefGoogle Scholar
  8. Bland RD, Mokres LM, Ertsey R, Jacobson BE, Jiang S, Rabinovitch M, Xu L, Shinwell ES, Zhang F, Beasley MA (2007a) Mechanical ventilation with 40 % oxygen reduces pulmonary expression of genes that regulate lung development and impairs alveolar septation in newborn mice. Am J Physiol Lung Cell Mol Physiol 293:L1099–L1110PubMedCrossRefGoogle Scholar
  9. Bland RD, Xu L, Ertsey R, Rabinovitch M, Albertine KH, Wynn KA, Kumar VH, Ryan RM, Swartz DD, Csiszar K, Fong KS (2007b) Dysregulation of pulmonary elastin synthesis and assembly in preterm lambs with chronic lung disease. Am J Physiol Lung Cell Mol Physiol 292:L1370–L1384PubMedCrossRefGoogle Scholar
  10. Bland RD, Ertsey R, Mokres LM, Xu L, Jacobson BE, Jiang S, Alvira CM, Rabinovitch M, Shinwell ES, Dixit A (2008) Mechanical ventilation uncouples synthesis and assembly of elastin and increases apoptosis in lungs of newborn mice. Prelude to defective alveolar septation during lung development? Am J Physiol Lung Cell Mol Physiol 294:L3–L14PubMedCrossRefGoogle Scholar
  11. Bolender RP, Hyde DM, Dehoff RT (1993) Lung morphometry: a new generation of tools and experiments for organ, tissue, cell, and molecular biology. Am J Physiol 265:L521–L548PubMedGoogle Scholar
  12. Brew N, Hooper SB, Allison BJ, Wallace MJ, Harding R (2011) Injury and repair in the very immature lung following brief mechanical ventilation. Am J Physiol Lung Cell Mol Physiol 301:L917–L926PubMedCrossRefGoogle Scholar
  13. Bruce MC, Bruce EN, Janiga K, Chetty A (1993) Hyperoxic exposure of developing rat lung decreases tropoelastin mRNA levels that rebound postexposure. Am J Physiol 265:L293–L300PubMedGoogle Scholar
  14. Chow MJ, Turcotte R, Lin CP, Zhang Y (2014) Arterial extracellular matrix: a mechanobiological study of the contributions and interactions of elastin and collagen. Biophys J 106:2684–2692PubMedCrossRefGoogle Scholar
  15. Craig JM, Scott AL, Mitzner W (2013) Elastase-coupled beads as a tool for characterizing localized alveolar tissue destruction associated with the onset of emphysema. J Appl Physiol 114:1637–1644PubMedCentralPubMedCrossRefGoogle Scholar
  16. Dabovic B, Chen Y, Choi J, Vassallo M, Dietz HC, Ramirez F, von Melchner H, Davis EC, Rifkin DB (2009) Dual functions for LTBP in lung development: LTBP-4 independently modulates elastogenesis and TGF-beta activity. J Cell Physiol 219:14–22PubMedCentralPubMedCrossRefGoogle Scholar
  17. Dabovic B, Chen Y, Choi J, Davis EC, Sakai LY, Todorovic V, Vassallo M, Zilberberg L, Singh A, Rifkin DB (2011) Control of lung development by latent TGF-β binding proteins. J Cell Physiol 226:1499–1509PubMedCentralPubMedCrossRefGoogle Scholar
  18. Dabovic B, Robertson IB, Zilberberg L, Vassallo M, Davis EC, Rifkin DB (2014) Function of latent TGF-β binding protein 4 and fibulin 5 in elastogenesis and lung development. J Cell Physiol 229:226–236Google Scholar
  19. de Lurdes PM, Gonçalves C, Rodrigues P, Bairos VA (2006) Quantification by image analysis of the Gallus gallus lung elastic fibres from embryonic to adult birds. Anat Histol Embryol 35:293–298CrossRefGoogle Scholar
  20. Deng C, Wang J, Zou Y, Zhao Q, Feng J, Fu Z, Guo C (2011) Characterization of fibroblasts recruited from bone marrow-derived precursor in neonatal bronchopulmonary dysplasia mice. J Appl Physiol 111:285–294PubMedCrossRefGoogle Scholar
  21. Deslee G, Woods JC, Moore CM, Liu L, Conradi SH, Milne M, Gierada DS, Pierce J, Patterson A, Lewit RA, Battaile JT, Holtzman MJ, Hogg JC, Pierce RA (2009) Elastin expression in very severe human COPD. Eur Respir J 34:324–331PubMedCentralPubMedCrossRefGoogle Scholar
  22. Emery JL, Mithal A (1960) The number of alveoli in the terminal respiratory unit of man during late intrauterine life and childhood. Arch Dis Child 35:544–547PubMedCentralPubMedCrossRefGoogle Scholar
  23. Henderson NC, Arnold TD, Katamura Y, Giacomini MM, Rodriguez JD, McCarty JH, Pellicoro A, Raschperger E, Betsholtz C, Ruminski PG, Griggs DW, Prinsen MJ, Maher JJ, Iredale JP, Lacy-Hulbert A, Adams RH, Sheppard D (2013) Targeting of αv integrin identifies a core molecular pathway that regulates fibrosis in several organs. Nat Med 19:1617–1624PubMedCrossRefGoogle Scholar
  24. Hilgendorff A, Parai K, Ertsey R, Jain N, Navarro EF, Peterson JL, Tamosiuniene R, Nicolls MR, Starcher BC, Rabinovitch M, Bland RD (2011) Inhibiting lung elastase activity enables lung growth in mechanically ventilated newborn mice. Am J Respir Crit Care Med 184:537–546PubMedCentralPubMedCrossRefGoogle Scholar
  25. Hilgendorff A, Parai K, Ertsey R, Juliana Rey-Parra G, Thébaud B, Tamosiuniene R, Jain N, Navarro EF, Starcher BC, Nicolls MR, Rabinovitch M, Bland RD (2012) Neonatal mice genetically modified to express the elastase inhibitor elafin are protected against the adverse effects of mechanical ventilation on lung growth. Am J Physiol Lung Cell Mol Physiol 303:L215–L227PubMedCentralPubMedCrossRefGoogle Scholar
  26. Hilgendorff A, Reiss I, Ehrhardt H, Eickelberg O, Alvira CM (2014) Chronic lung disease in the preterm infant. Lessons learned from animal models. Am J Respir Cell Mol Biol 50:233–245PubMedGoogle Scholar
  27. Joss-Moore LA, Wang Y, Yu X, Campbell MS, Callaway CW, McKnight RA, Wint A, Dahl MJ, Dull RO, Albertine KH, Lane RH (2011) IUGR decreases elastin mRNA expression in the developing rat lung and alters elastin content and lung compliance in the mature rat lung. Physiol Genomics 43:499–505PubMedCentralPubMedCrossRefGoogle Scholar
  28. Kuang PP, Goldstein RH, Liu Y, Rishikof DC, Jean JC, Joyce-Brady M (2003) Coordinate expression of fibulin-5/DANCE and elastin during lung injury repair. Am J Physiol Lung Cell Mol Physiol 285:L1147–L1152PubMedGoogle Scholar
  29. Kumarasamy A, Schmitt I, Nave AH, Reiss I, van der Horst I, Dony E, Roberts JD Jr, de Krijger RR, Tibboel D, Seeger W, Schermuly RT, Eickelberg O, Morty RE (2009) Lysyl oxidase activity is dysregulated during impaired alveolarization of mouse and human lungs. Am J Respir Crit Care Med 180:1239–1252PubMedCrossRefGoogle Scholar
  30. Lohmann P, Luna RA, Hollister EB, Devaraj S, Mistretta TA, Welty SE, Versalovic J (2014) The airway microbiome of intubated premature infants: characteristics and changes that predict the development of bronchopulmonary dysplasia. Pediatr Res 76:294–301Google Scholar
  31. Madurga A, Mižíková I, Ruiz-Camp J, Vadász I, Herold S, Mayer K, Fehrenbach H, Seeger W, Morty RE (2014) Systemic hydrogen sulfide administration partially restores normal alveolarization in an experimental animal model of bronchopulmonary dysplasia. Am J Physiol Lung Cell Mol Physiol 306:L684–L697PubMedCrossRefGoogle Scholar
  32. Mariani TJ, Sandefur S, Pierce RA (1997) Elastin in lung development. Exp Lung Res 23:131–145PubMedCrossRefGoogle Scholar
  33. Mascaretti RS, Mataloun MM, Dolhnikoff M, Rebello CM (2009) Lung morphometry, collagen and elastin content: changes after hyperoxic exposure in preterm rabbits. Clinics (Sao Paulo) 64:1099–1104CrossRefGoogle Scholar
  34. McKenna S, Michaelis KA, Agboke F, Liu T, Han K, Yang G, Dennery PA, Wright CJ (2014) Sustained hyperoxia-induced NF-κB activation improves survival and preserves lung development in neonatal mice. Am J Physiol Lung Cell Mol Physiol 306:L1078–L1089PubMedCrossRefGoogle Scholar
  35. Merrilees MJ, Ching PS, Beaumont B, Hinek A, Wight TN, Black PN (2008) Changes in elastin, elastin binding protein and versican in alveoli in chronic obstructive pulmonary disease. Respir Res 9:41PubMedCentralPubMedCrossRefGoogle Scholar
  36. Miao M, Reichheld SE, Muiznieks LD, Huang Y, Keeley FW (2013) Elastin binding protein and FKBP65 modulate in vitro self-assembly of human tropoelastin. Biochemistry 52:7731–7741PubMedCrossRefGoogle Scholar
  37. Mokres LM, Parai K, Hilgendorff A, Ertsey R, Alvira CM, Rabinovitch M, Bland RD (2010) Prolonged mechanical ventilation with air induces apoptosis and causes failure of alveolar septation and angiogenesis in lungs of newborn mice. Am J Physiol Lung Cell Mol Physiol 298:L23–L35PubMedCentralPubMedCrossRefGoogle Scholar
  38. Pierce RA, Albertine KH, Starcher BC, Bohnsack JF, Carlton DP, Bland RD (1997) Chronic lung injury in preterm lambs: disordered pulmonary elastin deposition. Am J Physiol 272:L452–L460PubMedGoogle Scholar
  39. Puthawala K, Hadjiangelis N, Jacoby SC, Bayongan E, Zhao Z, Yang Z, Devitt ML, Horan GS, Weinreb PH, Lukashev ME, Violette SM, Grant KS, Colarossi C, Formenti SC, Munger JS (2008) Inhibition of integrin alpha(v)beta6, an activator of latent transforming growth factor-beta, prevents radiation-induced lung fibrosis. Am J Respir Crit Care Med 177:82–90PubMedCentralPubMedCrossRefGoogle Scholar
  40. Qiu L, Deng C, Fu Z, Guo C (2011) The role of transforming growth factors beta1 and beta3 in pre- and post-natal pulmonary surfactant development. Cell Biol Int 35:287–292PubMedCrossRefGoogle Scholar
  41. Rich CB, Carreras I, Lucey EC, Jaworski JA, Buczek-Thomas JA, Nugent MA, Stone P, Foster JA (2003) Transcriptional regulation of pulmonary elastin gene expression in elastase-induced injury. Am J Physiol Lung Cell Mol Physiol 285:L354–L362PubMedGoogle Scholar
  42. Roman J, Little CW, McDonald JA (1991) Potential role of RGD-binding integrins in mammalian lung branching morphogenesis. Development 112:551–558PubMedGoogle Scholar
  43. Sakurai R, Villarreal P, Husain S, Liu J, Sakurai T, Tou E, Torday JS, Rehan VK (2013) Curcumin protects the developing lung against long-term hyperoxic injury. Am J Physiol Lung Cell Mol Physiol 305:L301–L311PubMedCentralPubMedCrossRefGoogle Scholar
  44. Sideek MA, Menz C, Parsi MK, Gibson MA (2014) LTBP-2 competes with tropoelastin for binding to fibulin-5 and heparin, and is a negative modulator of elastinogenesis. Matrix Biol 34:114–123PubMedCrossRefGoogle Scholar
  45. Thibeault DW, Mabry SM, Ekekezie II, Truog WE (2000) Lung elastic tissue maturation and perturbations during the evolution of chronic lung disease. Pediatrics 106:1452–1459PubMedCrossRefGoogle Scholar
  46. Turino GM, Ma S, Lin YY, Cantor JO, Luisetti M (2011) Matrix elastin: a promising biomarker for chronic obstructive pulmonary disease. Am J Respir Crit Care Med 184:637–641PubMedCrossRefGoogle Scholar
  47. Wang H, Jafri A, Martin RJ, Nnanabu J, Farver C, Prakash YS, MacFarlane PM (2014) Severity of neonatal hyperoxia determines structural and functional changes in developing mouse airway. Am J Physiol Lung Cell Mol Physiol 307:L295–L301Google Scholar
  48. Wendel DP, Taylor DG, Albertine KH, Keating MT, Li DY (2000) Impaired distal airway development in mice lacking elastin. Am J Respir Cell Mol Biol 23:320–326PubMedCrossRefGoogle Scholar
  49. Yanagisawa H, Schluterman MK, Brekken RA (2009) Fibulin-5, an integrin-binding matricellular protein: its function in development and disease. J Cell Commun Signal 3:337–347PubMedCentralPubMedCrossRefGoogle Scholar
  50. Yi M, Jankov RP, Belcastro R, Humes D, Copland I, Shek S, Sweezey NB, Post M, Albertine KH, Auten RL, Tanswell AK (2004) Opposing effects of 60 % oxygen and neutrophil influx on alveologenesis in the neonatal rat. Am J Respir Crit Care Med 170:1188–1196PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  1. 1.Department of Neonatology, Children’s Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and DisordersChongqing Medical UniversityChongqingChina
  2. 2.Department of PharmacologyChongqing Medical UniversityChongqingChina
  3. 3.Department of Hepatology and Liver transplantation Center, Children’s Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and DisordersChongqing Medical UniversityChongqingChina

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