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

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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.

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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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Authors and Affiliations

  1. Safe Environments Programme, Health Canada, Ottawa, Ontario, Canada

    V. Chauhan, D. Breznan, E. Thomson, S. Karthikeyan & R. Vincent (Ph.D.)

  2. Environmental Health Centre, Room 103, 0801 Tunney's Pasture, Ottawa, Ontario, Canada, K1A OL2

    R. Vincent (Ph.D.)

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  1. V. Chauhan
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  2. D. Breznan
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  3. E. Thomson
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  4. S. Karthikeyan
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  5. R. Vincent
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Correspondence to R. Vincent.

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Chauhan, V., Breznan, D., Thomson, E. et al. Effects of ambient air particles on the endothelin system in human pulmonary epithelial cells (A549). Cell Biol Toxicol 21, 191–205 (2005). https://doi.org/10.1007/s10565-005-0162-x

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  • DOI: https://doi.org/10.1007/s10565-005-0162-x

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