Cigarette smoke extract induces oxidative stress and apoptosis in human lung fibroblasts.
Cigarette smoke is a mixture of chemicals having direct and/or indirect toxic effects on different lung cells. We investigated the effect of cigarette smoke on human lung fibroblasts (HFL-1) oxidation and apoptosis. Cells were exposed to various concentrations (1, 5, and 10%) of cigarette smoke extract (CSE) for 3 h, and oxidative stress and apoptosis were assessed by fluorescence-activated cell sorting and confocal laser fluorescence microscopy. Both oxidative stress and apoptosis exhibited a dose-response relationship with CSE concentrations. Lung fibroblasts also showed marked DNA fragmentation at the Comet assay after exposure to 10% CSE. Coincubation of HLF-1 cells with N-acetylcysteine (1 mM) during CSE exposure significantly reduced oxidative stress, apoptosis, and DNA fragmentation, whereas preincubation (3 h) with the glutathione-depleting agent buthionine sulfoximine (125 microM) produced a significant increase of oxidative stress. Cigarette smoke is a potent source of oxidative stress, DNA damage, and apoptosis for HFL-1 cells, and we speculate that this could contribute to the development of pulmonary emphysema in the lungs of smokers.
Carnevali
Stefano
S
Pulmonary Unit, Cardiothoracic Department, Pharmacology Unit, University of Pisa, 56124 Pisa, Italy. carnevali.stefano@unimo.it
Petruzzelli
Stefano
S
Longoni
Biancamaria
B
Vanacore
Renato
R
Barale
Roberto
R
Cipollini
Monica
M
Scatena
Fabrizio
F
Paggiaro
Pierluigi
P
Celi
Alessandro
A
Giuntini
Carlo
C
D000111:Acetylcysteine / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D001562:Benzimidazoles; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D020552:Comet Assay; D004249:DNA Damage; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D016166:Free Radical Scavengers / Q000494:pharmacology; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D008168:Lung / Q000166:cytology*; D018613:Microscopy, Confocal; D018384:Oxidative Stress / Q000187:drug effects*; D012907:Smoking / Q000009:adverse effects*
D001562:Benzimidazoles; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D016166:Free Radical Scavengers; D019328:Buthionine Sulfoximine; D005978:Glutathione; C017807:bisbenzimide ethoxide trihydrochloride; D000111:Acetylcysteine