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Archives of Toxicology

, Volume 90, Issue 2, pp 385–402 | Cite as

Common and distinct mechanisms of induced pulmonary fibrosis by particulate and soluble chemical fibrogenic agents

  • Jie Dong
  • Xiaoqing Yu
  • Dale W. Porter
  • Lori A. Battelli
  • Michael L. Kashon
  • Qiang MaEmail author
Organ Toxicity and Mechanisms

Abstract

Pulmonary fibrosis results from the excessive deposition of collagen fibers and scarring in the lungs with or without an identifiable cause. The mechanism(s) underlying lung fibrosis development is poorly understood, and effective treatment is lacking. Here we compared mouse lung fibrosis induced by pulmonary exposure to prototypical particulate (crystalline silica) or soluble chemical (bleomycin or paraquat) fibrogenic agents to identify the underlying mechanisms. Young male C57BL/6J mice were given silica (2 mg), bleomycin (0.07 mg), or paraquat (0.02 mg) by pharyngeal aspiration. All treatments induced significant inflammatory infiltration and collagen deposition, manifesting fibrotic foci in silica-exposed lungs or diffuse fibrosis in bleomycin or paraquat-exposed lungs on day 7 post-exposure, at which time the lesions reached their peaks and represented a junction of transition from an acute response to chronic fibrosis. Lung genome-wide gene expression was analyzed, and differential gene expression was confirmed by quantitative RT-PCR, immunohistochemistry, and immunoblotting for representative genes to demonstrate their induced expression and localization in fibrotic lungs. Canonical signaling pathways, gene ontology, and upstream transcription networks modified by each agent were identified. In particular, these inducers elicited marked proliferative responses; at the same time, silica preferentially activated innate immune functions and the defense against foreign bodies, whereas bleomycin and paraquat boosted responses related to cell adhesion, platelet activation, extracellular matrix remodeling, and wound healing. This study identified, for the first time, the shared and unique genes, signaling pathways, and biological functions regulated by particulate and soluble chemical fibrogenic agents during lung fibrosis, providing insights into the mechanisms underlying human lung fibrotic diseases.

Keywords

Pulmonary fibrosis Genome-wide gene expression Inflammation Silica Bleomycin Paraquat 

Abbreviations

APC

Anaphase-promoting complex

BAL

Bronchoalveolar lavage

CNT

Carbon nanotubes

Col

Collagen

ECM

Extracellular matrix

FDR

False discovery rate

FN1

Fibronectin

GO

Gene ontology

IL

Interleukin

IPF

Idiopathic pulmonary fibrosis

Lcn2

Lipocalin-2

Mmp

Matrix metalloproteinase

MWCNT

Multi-walled carbon nanotubes

NEK

NIMA (never in mitosis gene a)-related kinase

NF-κB

Nuclear factor-κB

NLRP3

Nucleotide-binding oligomerization domain-like receptor, pyrin domain-containing 3

OPN

Osteopontin

PBS

Phosphate-buffered saline

PGE2

Prostaglandin E2

ROS

Reactive oxygen species

Slpi

Secretory leukocyte peptidase inhibitor

Spp1

Secreted phosphoprotein 1

Timp1

Tissue inhibitor of metalloproteinases 1

TLR

Toll-like receptor

Tnc

Tenascin-C

Notes

Acknowledgments

This work was funded to Q.M. by National Institute for Occupational Safety and Health, Health Effects Laboratory Division.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

Supplementary material

204_2015_1589_MOESM1_ESM.xlsx (89 kb)
Supplementary material 1 (XLSX 89 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg (outside the USA) 2015

Authors and Affiliations

  • Jie Dong
    • 1
  • Xiaoqing Yu
    • 2
  • Dale W. Porter
    • 3
  • Lori A. Battelli
    • 3
  • Michael L. Kashon
    • 4
  • Qiang Ma
    • 1
    Email author
  1. 1.Receptor Biology Laboratory, Toxicology and Molecular Biology Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and HealthCenters for Disease Control and PreventionMorgantownUSA
  2. 2.Department of BiostatisticsYale University School of Public HealthNew HavenUSA
  3. 3.Pathology and Physiology Research Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and HealthCenters for Disease Control and PreventionMorgantownUSA
  4. 4.Biostatistics and Epidemiology Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and HealthCenters for Disease Control and PreventionMorgantownUSA

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