Cigarette smoke condensate induces MMP-12 gene expression in airway-like epithelia

doi:10.1016/j.bbrc.2005.02.144

Biochemical and Biophysical Research Communications

Volume 330, Issue 1, 29 April 2005, Pages 194-203

https://doi.org/10.1016/j.bbrc.2005.02.144 Get rights and content

Abstract

Cigarette smoke (CS)-induced emphysema is attributable to matrix metalloproteinase-12 (MMP-12) in mice, however, a relationship between CS and MMP-12 is absent in human emphysema. Here, we show that cigarette smoke condensate (CSC) induces MMP-12 gene expression in airway-like epithelia through a hydrogen peroxide (H₂O₂)-dependent pathway involving NADPH oxidase, AP-1, and TNF-α. Cigarette smoke condensate-induced H₂O₂ production and MMP-12 gene expression were inhibited by apocynin, a specific inhibitor of NADPH oxidases, while 3-aminobenzamide, an inhibitor of AP-1, attenuated CSC-induced MMP-12 gene expression. Messenger RNAs encoding phagocytic NADPH oxidase components and a homologue of p67phox, p51 (NOXA1), were detected, while mRNA of dual oxidase (Duox)1 was unchanged by CSC. Enbrel, an inhibitor of TNF-α function, reduced CSC-induced H₂O₂ production and MMP-12 expression. These findings provide novel evidence of a direct relationship between CS exposure and MMP-12 in human airway epithelia and suggest several targets for modulation of this potentially pathogenic pathway.

Section snippets

Materials and methods

Derivation of airway-like NHBEC cultures. Differentiation of primary NHBECs (passage 1) into cultures bearing airway-like characteristics, including enhanced mucin production and secretion, has been described [22] and previously performed in this laboratory [6]. Briefly, NHBECs cultured on fibrillar collagen/human fibronectin membranes were maintained at an air-liquid interface for fourteen days in serum-free NHBEC culture medium (50% BEBM, Cambrex, Walkersville, MD; 50% DMEM (high glucose),

Cigarette smoke condensate induces H₂O₂ production in NHBECs through activation of NADPH oxidase

To investigate whether CSC induces oxidant production in airway-like NHBEC cultures, we challenged DHR-loaded, airway-like NHBEC cultures with CSC (0.4 μg/ml) for 60 min. Although the vehicle (DMSO) failed to induce oxidant generation (Fig. 1A), CSC-induced oxidant production in airway-like NHBEC cultures, as indicated by the robust fluorescence observed in CSC-treated cultures (Fig. 1B). Co-challenge of cultures with CSC and catalase significantly reduced DHR-derived fluorescence emission (Fig. 1

Discussion

These findings directly show that a concentrated form of the primary cause of emphysema, namely CS, can regulate the expression of MMP-12, which, based on studies in mice [4], is a protein that could play a major role in CS-induced emphysema in humans. Furthermore, we provide evidence that an H₂O₂-dependent pathway mediates the effect of CSC in this in vitro model of airway bronchial epithelium. Cigarette smoke condensate-induced oxidant generation and MMP-12 expression that were inhibitable by

Acknowledgment

The authors thank Liping Zhang for determining nucleotide sequences of RT-PCR fragments.

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Chronic obstructive pulmonary disease (COPD) is currently the fourth leading cause of death worldwide and, in contrast to the trend for cardiovascular diseases, mortality rates still continue to climb. This increase is in part due to an aging population, being expanded by the “Baby boomer” generation who grew up when smoking rates were at their peak and by people in developing countries living longer. Sadly, there has been a disheartening lack of new therapeutic approaches to counteract the progressive decline in lung function associated with the disease that leads to disability and death. COPD is characterized by irreversible chronic airflow limitation that is caused by emphysematous destruction of lung elastic tissue and/or obstruction in the small airways due to occlusion of their lumen by inflammatory mucus exudates, narrowing and obliteration. These lesions are mainly produced by the response of the tissue to the repetitive inhalational injury inflicted by noxious gases, including cigarette smoke, which involves interaction between infiltrating inflammatory immune cells, resident cells (e.g. epithelial cells and fibroblasts) and the extra cellular matrix. This interaction leads to tissue destruction and airway remodeling with changes in elastin and collagen, such that the epithelial–mesenchymal trophic unit is dysregulated in both the disease pathologies. This review focuses on: 1 – novel inflammatory and remodeling factors that are altered in COPD; 2 – in vitro and in vivo models to understand the mechanism whereby the extra cellular matrix environment in altered in COPD; and 3 – COPD in the context of wound-repair tissue responses, with a focus on the regulation of mesenchymal cell fate and phenotype.

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Cigarette smoke condensate induces MMP-12 gene expression in airway-like epithelia

Abstract

Section snippets

Materials and methods

Cigarette smoke condensate induces H2O2 production in NHBECs through activation of NADPH oxidase

Discussion

Acknowledgment

Am. J. Pathol.

J. Biol. Chem.

Biochem. Biophys. Res. Comm.

Respir. Med.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Free Rad. Biol. Med.

J. Biol. Chem.

FEBS Lett.

FEBS Lett.

J. Biol. Chem.

Cigarette smoking and health

Am. J. Respir. Crit. Care Med.

Oxidants/antioxidants and chronic obstructive pulmonary disease: pathogenesis to therapy

Novartis Found. Symp.

Requirement for macrophage elastase for cigarette smoke-induced emphysema in mice

Science

Oxidative stress and regulation of glutathione synthesis in lung inflammation

Eur. Respir. J.

Regulation of matrix metalloproteinases: an overview

Mol. Cell. Biochem.

Oxidants in cigarette smoke: radicals, hydrogen peroxide, peroxynitrate, and peroxynitrite

Ann. N Y Acad. Sci.

Stable compounds of cigarette smoke induce endothelial superoxide anion production via NADPH oxidase activation

Arterioscler. Thromb. Vasc. Biol.

The respiratory burst oxidase

Disorders of phagocytic cells

Cigarette smoke condensate induces H₂O₂ production in NHBECs through activation of NADPH oxidase