Lipid-soluble components in cigarette smoke induce mitochondrial production of reactive oxygen species in lung epithelial cells.
Reactive oxygen species (ROS) present in cigarette smoke (CS) are thought to contribute to the development of COPD. Although CS-ROS can hardly enter airway epithelial cells, and certainly not the circulation, systemic levels of ROS have been found to be elevated in COPD patients. We hypothesize that lipophilic components present in CS can enter airway epithelial cells and increase intracellular ROS production by disturbing mitochondrial function. Different airway epithelial cells were exposed to CS extract (CSE), hexane-treated CSE (CSE without lipophilic components), gaseous-phase CS, and water-filtered CS (gaseous-phase CS without ROS). Mitochondrial membrane potential (Deltapsi(m)) and ATP levels were assessed using the bronchial epithelial cell line Beas-2b. ROS generation measured directly by DCF fluorescence and indirectly by measuring free thiol groups (-SH) upon exposure to CS was assessed using lung alveolar epithelial cells devoid of functional mitochondria (A549-rho0), with normal A549 cells serving as controls. In Beas-2b cells, CSE (4 h) caused a dose-dependent decrease in Deltapsi(m) and ATP levels, whereas hexane-treated CSE did not. DCF fluorescence in A549 cells increased in response to CSE, whereas this was not the case in A549-rho0 cells. Exposure of A549 cells to CS resulted in a rapid decrease in free -SH, whereas exposure to ROS-depleted CS only resulted in a delayed decrease. This delayed decrease was less pronounced in A549-rho0 cells. Lipophilic components in CS disturb mitochondrial function, which contributes to increased intracellular generation of ROS. Our results are of importance in understanding the systemic effects of smoking observed in patients with COPD.
Department of Pathology and Medical Biology, Laboratory of Allergology and Pulmonary Diseases, University Medical Center Groningen, University of Groningen, The Netherlands.m.van.der.toorn@med.umcg.nl
Marco
M
van der Toorn
Delaram
D
Rezayat
Henk F
HF
Kauffman
Stephan J L
SJ
Bakker
Rijk O B
RO
Gans
Gerard H
GH
Koëter
Augustine M K
AM
Choi
Antoon J M
AJ
van Oosterhout
Dirk-Jan
DJ
Slebos
D002460:Cell Line; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D005374:Filtration; D005740:Gases; D006801:Humans; D008055:Lipids / Q000737:chemistry*; D008168:Lung / Q000166:cytology*; D008928:Mitochondria / Q000378:metabolism*; D010084:Oxidation-Reduction; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012906:Smoke; D012995:Solubility; D012996:Solutions; D013438:Sulfhydryl Compounds / Q000378:metabolism; D014026:Nicotiana / Q000737:chemistry*; D014867:Water
D005740:Gases; D008055:Lipids; D017382:Reactive Oxygen Species; D012906:Smoke; D012996:Solutions; D013438:Sulfhydryl Compounds; D014867:Water