Smad3 mediates cigarette smoke extract (CSE) induction of VEGF release by human fetal lung fibroblasts

Maha Farid, Nobuhiro Kanaji, Masanori Nakanishi, Yoko Gunji, Joel Michalski, Shunichiro Iwasawa, Jun Ikari, Xingqi Wang, Hesham E Basma, Amy J. Nelson, Xiang-de Liu, Stephen I. Rennard

Research output: Contribution to journalArticle

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Abstract

Cigarette smoke is the major cause of chronic obstructive pulmonary disease (COPD), yet pathogenic mechanisms are not fully understood. Vascular endothelial growth factor (VEGF) is one of the major regulators of endothelial cell survival and is believed to play a role in the pathogenesis of COPD. Fibroblasts are a significant source of VEGF in the lungs; however the effect of cigarette smoke exposure on VEGF release by fibroblasts is not fully understood. We hypothesized that cigarette smoke-induced disturbed VEGF release by human lung fibroblasts is a potential pathogenic mechanism that could contribute to COPD. Cigarette smoke extract (CSE) was prepared by modification of the methods of Carp and Janoff (American Review of Respiratory Disease, 1978). Human fetal lung fibroblasts (HFL-1) were exposed to different concentrations of CSE and for different durations. VEGF release into the media was measured using ELISA. TGF-β1 receptor (TβR1)/Smad3 as a potential pathway for CSE modulated VEGF release was also investigated using biochemical analyses and siRNA inhibition of Smad3 and siRNA and pharmacologic inhibition of TβR1. CSE induced VEGF release by HFL-1 in concentration and time dependent manner. This was confirmed in two additional types of primary human fetal lung fibroblasts. CSE induced Smad3 phosphorylation and nuclear translocation in HFL-1 cells. Silencing of Smad3 by siRNA not only eliminated the stimulatory effect of CSE on VEGF release but also inhibited baseline VEGF production. Suppression of TβR1 by the pharmacological inhibitor (SB431542) markedly reduced VEGF release by HFL-1 in response to CSE and this effect was confirmed by TβR1 siRNA. In contrast, nicotine inhibited VEGF release by HFL-1 in a dose and time dependent manner. Our findings indicate that CSE stimulates Smad3-mediated VEGF release by lung fibroblasts. Nicotine does not account for the CSE stimulation of VEGF in HFL-1. The ability of lung fibroblasts to produce VEGF may play a role in pathogenesis of cigarette smoke induced lung disease.

Original languageEnglish (US)
Pages (from-to)126-134
Number of pages9
JournalToxicology Letters
Volume220
Issue number2
DOIs
StatePublished - Jul 4 2013

Fingerprint

Fibroblasts
Smoke
Tobacco Products
Vascular Endothelial Growth Factor A
Lung
Pulmonary diseases
Small Interfering RNA
Chronic Obstructive Pulmonary Disease
Nicotine
human VEGFA protein
Carps
Phosphorylation
Endothelial cells
Lung Diseases
Cell Survival
Endothelial Cells
Enzyme-Linked Immunosorbent Assay
Pharmacology

Keywords

  • Cigarette smoke
  • Lungs
  • Smad3
  • VEGF

ASJC Scopus subject areas

  • Toxicology

Cite this

Farid, M., Kanaji, N., Nakanishi, M., Gunji, Y., Michalski, J., Iwasawa, S., ... Rennard, S. I. (2013). Smad3 mediates cigarette smoke extract (CSE) induction of VEGF release by human fetal lung fibroblasts. Toxicology Letters, 220(2), 126-134. https://doi.org/10.1016/j.toxlet.2013.04.011

Smad3 mediates cigarette smoke extract (CSE) induction of VEGF release by human fetal lung fibroblasts. / Farid, Maha; Kanaji, Nobuhiro; Nakanishi, Masanori; Gunji, Yoko; Michalski, Joel; Iwasawa, Shunichiro; Ikari, Jun; Wang, Xingqi; Basma, Hesham E; Nelson, Amy J.; Liu, Xiang-de; Rennard, Stephen I.

In: Toxicology Letters, Vol. 220, No. 2, 04.07.2013, p. 126-134.

Research output: Contribution to journalArticle

Farid, M, Kanaji, N, Nakanishi, M, Gunji, Y, Michalski, J, Iwasawa, S, Ikari, J, Wang, X, Basma, HE, Nelson, AJ, Liu, X & Rennard, SI 2013, 'Smad3 mediates cigarette smoke extract (CSE) induction of VEGF release by human fetal lung fibroblasts', Toxicology Letters, vol. 220, no. 2, pp. 126-134. https://doi.org/10.1016/j.toxlet.2013.04.011
Farid, Maha ; Kanaji, Nobuhiro ; Nakanishi, Masanori ; Gunji, Yoko ; Michalski, Joel ; Iwasawa, Shunichiro ; Ikari, Jun ; Wang, Xingqi ; Basma, Hesham E ; Nelson, Amy J. ; Liu, Xiang-de ; Rennard, Stephen I. / Smad3 mediates cigarette smoke extract (CSE) induction of VEGF release by human fetal lung fibroblasts. In: Toxicology Letters. 2013 ; Vol. 220, No. 2. pp. 126-134.
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