Effect of cigarette smoke and its condensates on alveolar epithelial cell injury in vitro

Lannan, S.; Donaldson, Ken; Brown, David M.; MacNee, William

doi:10.1152/ajplung.1994.266.1.l92

Cited by 83 publications

(81 citation statements)

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“…3 Growth of A549 cells on native lung matrix. Lannan et al 1994;Wollmer and Evander 1994;Hoshino et al 2001;Victoni et al 2014). They include down-regulated proliferation, impaired cell attachment, DNA damage and suppressed levels of surfactant proteins.…”

Section: Discussionmentioning

confidence: 99%

Native matrix-based human lung alveolar tissue model in vitro: studies of the reparatory actions of mesenchymal stem cells

et al. 2016

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Studies of lung diseases in vitro often rely on flat, plastic-based monocultures, due to short lifespan of primary cells, complicated anatomy, lack of explants, etc. We hereby present a native 3D model with cues for repopulating epithelial cells. Abilities of mesenchymal stem cells (MSC) to modulate bacterial lipopolysaccharide (LPS) and cigarette smoke-induced injury to pulmonary epithelium were tested in our model. Post-mortem human lung tissue was sliced, cut and decellularized. Resulting matrix pads were reseeded with pulmonary epithelium (A549 line). Markers of the layer integrity and certain secreted proteins in the presence of cigarette smoke extract (CSE) and LPS were assessed via Western blot, ELISA and RT-PCR assays. In parallel, the effects of MSC paracrine factors on exposed epithelial cells were also investigated at gene and protein levels. When cultured on native 3D matrix, A549 cells obtain dual, type I-and II-like morphology. Exposure to CSE and LPS leads to downregulation of several epithelial proteins and suppressed proliferation rate. MSC medium added to the model restores proliferation rate and some of the epithelial proteins, i.e. e-cadherin and beta-catenin. CSE also increases secretion of proinflammatory cytokines by epithelial cells and upregulates transcription factor NFjB. Some of these effects might be counteracted by MSC in our model. We introduce repopulated decellularized lung matrix that highly resembles in vivo situation and is convenient for studies of disease pathogenesis, cytotoxicology and for exploring therapeutic strategies in the human lung context in vitro. MSC paracrine products have produced protecting effects in our model.

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Section: Discussionmentioning

confidence: 99%

Native matrix-based human lung alveolar tissue model in vitro: studies of the reparatory actions of mesenchymal stem cells

et al. 2016

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“…CSC was produced by blowing smoke, generated by a smoking machine using a vacuum syringe system, over 3 ml of phosphate buffered saline (PBS) in a siliconised glass tonometer (Vitalograph, Buckingham, UK). The smoke-generating machine delivers a 37 ml puff of whole cigarette smoke, including particulates, every minute to the tonometer which was rotated gently [14]. The solution was made up fresh on the morning of each experiment and CSC (100%) was filtered L Rahman et aL/FEBS Letters 396 (1996) [21][22][23][24][25] using 0.22 ~tm filter (Millipore S.A., Molsheim, France) to remove large particles and bacteria.…”

Section: Preparation Of Cscmentioning

confidence: 99%

Induction of γ‐glutamylcysteine synthetase by cigarette smoke is associated with AP‐1 in human alveolar epithelial cells

Rahman¹,

Smith

Lawson³

et al. 1996

FEBS Letters

152

117

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Increased levels of glutathione (GSH) occur in the epithelial lining fluid (ELF) of chronic cigarette smokers. Therefore we investigated the effect of cigarette smoke condensate solution (CSC) on GSH synthesis and the regulation of T-glutamylcysteine synthetase (~CS) in human type II alveolar epithelial cells (A549). CSC exposure increased GSH levels, TGCS activity and ~CS heavy subunit (HS) mRNA, as well as increasing DNA binding of the activator protein-1 (AP-1) and the human antioxidant response element (hARE). Transfection of deletion constructs of the ~GCS-HS promoter in a chloramphenicol acetyl transferase (CAT) reporter system revealed that an hARE, present within promoter, is not required tot the CSC mediated induction. We conclude that CSC induction of ~GCS-HS expression is associated with AP-1/AP-I-like responsive elements.

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“…Several studies (Fukano et al, 2006a,b;Hoshino et al, 2001;Lannan et al, 1994) have investigated the effects of smoking machine-generated CSC or whole smoke on cultured cell lines, particularly A549 cells, a transformed cell line with some characteristics of alveolar type II cells.…”

mentioning

confidence: 99%

Effects Of 10 Cigarette Smoke Condensates On Primary Human Airway Epithelial Cells By Comparative Gene And Cytokine Expression Studies

Pickett¹,

Seagrave²,

Boggs³

et al. 2010

D23. Recent Advances on Smoking Health Effects

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Cigarettes vary in tobacco blend, filter ventilation, additives, and other physical and chemical properties, but little is known regarding potential differences in toxicity to a smoker's airway epithelia. We compared changes in gene expression and cytokine production in primary normal human bronchial epithelial cells following treatment for 18 h with cigarette smoke condensates (CSCs) prepared from five commercial and four research cigarettes, at doses of~4 mg/ml nicotine. Nine of the CSCs were produced under a standard International Organization for Standardization smoking machine regimen and one was produced by a more intense smoking machine regimen. Isolated messenger RNA (mRNA) was analyzed by microarray hybridization, and media was analyzed for secreted cytokines and chemokines. Twenty-one genes were differentially expressed by at least 9 of the 10 CSCs by more than twofold, including genes encoding detoxifying and antioxidant proteins. Cytochrome P450, family 1, subfamily A, polypeptide 1 (CYP1A1) and NAD(P)H dehydrogenase, quinone 1 (NQO-1) were selected for validation with quantitative real-time PCR (qRT-PCR) and Western blot analyses. NQO-1 expression determined with microarrays, qRT-PCR, and Western blotting differed among the CSC types, with good correlation among the different assays. CYP1A1 mRNA levels varied substantially, but there was little correlation with the protein levels. For each CSC, the three most induced and three most repressed genes were identified. These genes may be useful as markers of exposure to that particular cigarette type. Furthermore, differences in interleukin-8 secretion were observed. These studies lay the foundation for future investigations to analyze differences in the responses of in vivo systems to tobacco products marketed with claims of reduced exposure or reduced harm.

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Effect of cigarette smoke and its condensates on alveolar epithelial cell injury in vitro

Cited by 83 publications

References 0 publications

Native matrix-based human lung alveolar tissue model in vitro: studies of the reparatory actions of mesenchymal stem cells

Native matrix-based human lung alveolar tissue model in vitro: studies of the reparatory actions of mesenchymal stem cells

Induction of γ‐glutamylcysteine synthetase by cigarette smoke is associated with AP‐1 in human alveolar epithelial cells

Effects Of 10 Cigarette Smoke Condensates On Primary Human Airway Epithelial Cells By Comparative Gene And Cytokine Expression Studies

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