Chest
Volume 143, Issue 5, May 2013, Pages 1252-1260
Journal home page for Chest

Original Research
Critical Care
Lumican Regulates Ventilation-Induced Epithelial-Mesenchymal Transition Through Extracelluar Signal-Regulated Kinase Pathway

https://doi.org/10.1378/chest.12-2058Get rights and content

Background

Mechanical ventilation used in patients with acute lung injury can damage pulmonary epithelial cells through production of inflammatory cytokines and excess deposition of the extracellular matrix protein lumican. Lumican participates in macrophage inflammatory protein (MIP)-2 and transforming growth factor-β1 (TGF-β1) signaling during the fibroproliferative phase of acute lung injury, which involves a process of epithelial-mesenchymal transition (EMT). The mechanisms regulating interactions between mechanical ventilation and lung injury are unclear. We hypothesized that lung damage and EMT by high tidal volume (Vt) mechanical stretch causes upregulation of lumican that modulates MIP-2 and TGF-β1 through the extracellular signal-regulated kinase (ERK) 1/2 pathway.

Methods

Male C57BL/6 mice (either wild type or lumican null) aged 3 months and weighing between 25 and 30 g were exposed to low Vt (6 mL/kg) or high Vt (30 mL/kg) mechanical ventilation with room air for 2 to 8 h. Nonventilated mice were used as control subjects.

Results

We found that high Vt mechanical ventilation increased microvascular permeability, neutrophil influx, production of free radicals, MIP-2 and TGF-β1 proteins, positive staining of α-smooth muscle actin and S100A4/fibroblast-specific protein-1, Masson trichrome staining and extracellular collagen, and activation of lumican and ERK1/2 in wild-type mice. Decreased staining of the epithelial marker E-cadherin was also observed. Mechanical stretch-augmented EMT was attenuated with lumican-deficient mice and pharmacologic inhibition of ERK1/2 activity by PD98059.

Conclusions

The data suggest that lumican promotes high Vt mechanical ventilation-induced lung injury and EMT through the activation of the ERK1/2 pathway.

Section snippets

Generation and Maintenance of Lumican-Deficient Mice

Male C57BL/6 (either wild-type or lumican deficient) mice aged 3 months and weighing between 25 and 30 g, were obtained from Chang Gung University Laboratory Animal Center (Taoyuan, Taiwan). Lumican-null mice were generated by targeted gene disruption as previously described.13, 21 All procedures for handling mice conformed to Association of Research for Vision and Ophthalmology guidelines. Statements for the use of animals in research were approved by the Institutional Animal Care and Use

Reduction of VILI in Lumican-Deficient Mice

We measured lung EBD and wet weight-to-dry weight ratio to determine the effects of mechanical ventilation on changes of microvascular permeability and lung water content in VILI (Figs 1A, 1B). MPO assay was used to quantitate total lung neutrophils (Fig 1C). We measured MDA level, which is an aldehydic secondary product of lipid peroxidation used as a marker of oxidative stress9 and TGF-β1 and MIP-2 production, to determine the level of oxidant stress and chemoattractants associated with VILI (

Discussion

High Vts in normal animals have been used to mimic the overdistension of the less-injured and, thus, more-compliant areas of lung found in ARDS. Previous studies demonstrated that hyperexpansion of the lung was the cause of noncardiogenic pulmonary edema in VILI.1, 5, 22, 23 A previous study of an acid-induced lung injury model in mice showed that 2 h of mechanical stretch induced lung fibrogenesis.2 Our previous study of hyaluronan synthase knockout mice showed that 5 h of high Vt mechanical

Conclusions

By using an in vivo mouse model of ARDS, this study demonstrated that high Vt mechanical VILI and EMT are associated with activation of ERK1/2 and the production of MIP-2, TGF-β1, and MDA. This process partly depends on the activation of lumican. Knowledge of the effect of mechanical forces on lumican allows clarification of the pathophysiologic mechanisms regulating the fibroproliferative phase of ARDS that may progress to irreversible pulmonary fibrosis and the need for long-term ventilator

Acknowledgments

Author contributions: Drs Li and Yang had full access to all of the data in the study and take responsibility for the integrity of the data and the accuracy of the data analysis.

Dr Li: contributed to the coordination of the study and review of the manuscript.

Dr Chu: contributed to the coordination of the study and review of the manuscript.

Dr Hung: contributed to the coordination of the study and review of the manuscript.

Dr Kao: contributed to the coordination of the study and review of the

References (33)

  • JD Ricard et al.

    Ventilator-induced lung injury

    Eur Respir J Suppl

    (2003)
  • MS Ludwig

    Proteoglycans and pathophysiology

    J Appl Physiol

    (2007)
  • LF Li et al.

    Serine/threonine kinase-protein kinase B and extracellular signal-regulated kinase regulate ventilator-induced pulmonary fibrosis after bleomycin-induced acute lung injury: a prospective, controlled animal experiment

    Crit Care

    (2008)
  • RJ Fahy et al.

    The acute respiratory distress syndrome: a role for transforming growth factor-beta 1

    Am J Respir Cell Mol Biol

    (2003)
  • MP Keane et al.

    Neutralization of the CXC chemokine, macrophage inflammatory protein-2, attenuates bleomycin-induced pulmonary fibrosis

    J Immunol

    (1999)
  • LF Li et al.

    Lumican expression in diaphragm induced by mechanical ventilation

    PLoS ONE

    (2011)
  • Cited by (28)

    • Epithelial-to-mesenchymal transition and invadopodia markers in breast cancer: Lumican a key regulator

      2020, Seminars in Cancer Biology
      Citation Excerpt :

      In conclusion, the results taken together indicate that lumican was upregulated before EMT of the LECs and that loss of lumican attenuates injury-induced EMT of LECs [115]. Another case where lumican is reported to regulate EMT is the ventilation-induced EMT through extracellular signal-regulated kinase pathway [116]. In acute lung injury, mechanical ventilation is used in patients, since it damages pulmonary epithelial cells through production of inflammatory cytokines and excess deposition of lumican.

    • Potential role of M2 macrophage polarization in ventilator-induced lung fibrosis

      2019, International Immunopharmacology
      Citation Excerpt :

      After confirming depth of anesthesia by absence of response to paw compression, the neck skin of mice were cut, blunt dissection, expose the trachea; mice were then subjected to endotracheal intubation by using a 22G Teflon catheter and the trachea hold with catheter were ligated to prevent air leaks. Mice in model groups were then mechanically ventilated with a small animal ventilator ((55–7040, VentElite; Harvard Apparatus, Holliston, MA, US) for 4 h using VT of 20 mL/kg plus 0 cm H2O positive end-expiratory pressure (PEEP) and fraction of inspiration O2(FiO2) 0.4, respiratory rate 80 breaths/min, inspiratory-expiratory ratio 1:1 [11,12]. During the ventilation period, mice were given pentobarbitone (50 mg/kg) as needed; the cocuronium besylate (0.6 mg/kg, H20130486; MSD Performance Products, Kenilworth, NJ, US) was intraperitonealy administered once per hour to achieve muscle relaxation.

    • MicroRNA-29b Mediates Lung Mesenchymal-Epithelial Transition and Prevents Lung Fibrosis in the Silicosis Model

      2019, Molecular Therapy Nucleic Acids
      Citation Excerpt :

      The proliferation and accumulation of fibroblasts are considered to be vital in the development of pulmonary fibrosis diseases.3 A large proportion of data implicates that epithelial cells undergoing the process of epithelial-mesenchymal transition (EMT) is one source of fibroblasts.4,5 During EMT, epithelial cells gradually lose their epithelial characteristics and transform into a mesenchymal-like cell phenotype, which begins to synthesize the components of ECM, such as collagen I and fibronectin.

    • Complex roles of TGF-b signaling pathways in lung development and bronchopulmonary dysplasia

      2023, American Journal of Physiology - Lung Cellular and Molecular Physiology
    View all citing articles on Scopus

    Reproduction of this article is prohibited without written permission from the American College of Chest Physicians. See online for more details.

    Funding/Support: The study was supported by the National Science Council [98-2314-B-182A-084-MY3 and 101-2314-B-182A-088-MY3], Chang Gung Research Project [3A0711 (to Dr Li) 97-2314-B-182-028-MY2, and 99-2314-B-182-042-MY3 (to Dr Chu)]; National Institutes of Health, National Eye Institute [Grant EY011845], Research to Prevent Blindness; and Ohio Lions Eye Research Foundation (to Dr Kao).

    View full text