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Cell Biology and Toxicology

, Volume 21, Issue 5–6, pp 191–205 | Cite as

Effects of ambient air particles on the endothelin system in human pulmonary epithelial cells (A549)

  • V. Chauhan
  • D. Breznan
  • E. Thomson
  • S. Karthikeyan
  • R. Vincent
Article

Abstract

Inhalation of urban particles results in higher circulating levels of the vasoconstrictor peptide endothelin-1 (ET-1), which may account for the adverse cardiovascular impacts associated with air pollution. The objective of this study was to examine the direct effects of urban particles on the production of ET-1 by human epithelial cells (A549). A549 cells were exposed to TiO2, SiO2, Ottawa urban particulate matter EHC-93, and fractions of the urban particles. The levels of ET-1, interleukin-8 (IL-8) and vascular endothelial growth factor (VEGF) in the culture medium were detected by ELISA. The mRNA levels of preproET-1, endothelin converting enzyme (ECE-1), ETa receptor and ETb receptor, matrix metalloproteinase (MMP-2), tissue inhibitor of MMP (TIMP-2), and heat shock protein (HSP-70) were determined by quantitative real-time RT-PCR. Cluster analysis of the variables identified similarities in the patterns of effects. Cluster I comprised variables that were primarily inhibited by particles: ET-1 and MMP-2 mRNAs, ET-1 and bigET-1 peptides, and cell viability. Clusters II and III comprised variables that were either inhibited or induced, depending on the test material: HSP-70, ETaR and ECE mRNAs, and IL-8 and VEGF proteins. Cluster IV comprised variables that were mainly induced by particle preparations: ETbR and TIMP-2 mRNAs. The decreased expression of preproET-1 in A549 cells suggests that epithelial cells may not be the source of higher pulmonary ET-1 spillover in the circulation measured in vivo in response to inhaled urban particles. However, higher ECE-1 in A549 cells after exposure to particles suggests an increased ability to process bigET-1 into the mature ET-1 peptide, while increased receptor expression implies higher responsiveness. The increased release of IL-8 and VEGF by epithelial cells in response to particles could possibly upregulate ET-1 production in the adjacent pulmonary capillary endothelial cells, with concomitant increased ET-1 spillover in the systemic circulation.

Key Words

cytokines epithelial cells endothelin particulate matter urban dust 

Abbreviations

BSA

bovine serum albumin

ELISA

enzyme-linked immunosorbent assay

ET

endothelin

ETaR

endothelin type A receptor

ECE-1

endothelin converting enzyme

ETbR

endothelin type B receptor

IL-8

interleukin-8

MMP-2

matrix metalloproteinase-2

TIMP

tissue inhibitor of metalloproteinases

PCR

polymerase chain reaction

VEGF

vascular endothelial growth factor

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Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  • V. Chauhan
    • 1
  • D. Breznan
    • 1
  • E. Thomson
    • 1
  • S. Karthikeyan
    • 1
  • R. Vincent
    • 1
    • 2
  1. 1.Safe Environments ProgrammeHealth CanadaOttawaCanada
  2. 2.Environmental Health CentreOttawaCanada

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