Formation of intracellular free radicals in guinea pig airway epithelium during in vitro exposure to ozone.
In vivo exposure to ozone (O3) has been shown to cause airway epithelial damage and lipid peroxidation. The oxidation of polyunsaturated fatty acids has been shown to produce hydrogen peroxide and aldehydes with reactive oxygen species (ROS) as intermediates. These products of ozonation may react with other bioorganic molecules and cause cellular damage. To assess the production of ROS, confluent primary cultures of guinea pig airway epithelial cells were grown on Costar membrane with a liquid-air interface and exposed to 0.2, 0.4, and 0.6 ppm O3. The concentrations of intracellular ROS during the exposure were monitored using the fluorescent dye dihydrorhodamine-123. The intracellular concentration of ROS increased immediately upon the commencement of the O3 exposure and persisted until the end of the exposure period (up to 1 hr). The concentration of ROS increased with increasing O3 concentration. To determine the species of ROS produced during O3 exposure, airway epithelial cells were perfused with dimethyl sulfoxide (DMSO), sodium formate (hydroxyl radical scavengers), NaN3 (catalase inhibitor), or diethyl-dithio carbamate (DEDC, superoxide dismutase inhibitor) prior to and during the exposure period and the fluorescent intensity was monitored continuously. While both DMSO and sodium formate decreased the concentration of ROS, DEDC and NaN3 had no effect. We concluded that hydroxyl radicals instead of H2O2 or superoxide anions were produced immediately following the commencement of O3 exposure in guinea pig airway epithelial cells in an exposure concentration-dependent fashion.
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Department of Environmental Medicine, New York University Medical Center, Tuxedo 10987, USA. chenl@charlotte.med.nyu.edu
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D000818:Animals; D001980:Bronchi / Q000187:drug effects* / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D006168:Guinea Pigs; D017665:Hydroxyl Radical / Q000378:metabolism*; D008297:Male; D010083:Oxidants, Photochemical / Q000633:toxicity*; D010126:Ozone / Q000633:toxicity*; D017382:Reactive Oxygen Species / Q000378:metabolism
D010083:Oxidants, Photochemical; D017382:Reactive Oxygen Species; D017665:Hydroxyl Radical; D010126:Ozone