Comparison of the Toxic Effects of Hydrogen Peroxide and Ozone on Cultured Human Bronchial Epithelial Cells

Gabrielson, Edward; Yu, Xiaoying; Spannhake, E. W.

doi:10.2307/3431921

Cited by 3 publications

(5 citation statements)

References 8 publications

(10 reference statements)

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“…In the pulmonary system, it has been observed that ozone can exacerbate asthma (Mar and Koenig, 2009), causes inflammation in the airway in healthy humans (Chimenti et al, 2009; Alexis et al, 2010), generate reactive oxygen species and cause DNA breaks in vivo in different animal models (Bermúdez et al, 1999; Haney et al, 1999; Bornholdt et al, 2002; Ferng, 2002) and human alveolar epithelial cells (Kosmider et al, 2010). In the current study, we confirmed that ozone induced DNA breaks in HBECs (Gabrielson et al, 1994; Lee et al, 1996) and that the effects of ozone were limited to about 4 h in the conditions studied (Bornholdt et al, 2002). Further, we demonstrated that wortmannin, a DNA repair inhibitor, did enhance the effects of ozone and prolonged the duration of existence of the fragments.…”

Section: Discussionsupporting

confidence: 84%

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l-Arginine promotes DNA repair in cultured bronchial epithelial cells exposed to ozone: involvement of the ATM pathway

et al. 2011

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Ozone may lead to DNA breaks in airway epithelial cells. p-ATM (phosphorylated ataxia telangiectasia mutated) plays a pivotal role in DNA repair. Derivatives of NO (nitric oxide) are regulators of the phosphorylation, and NO is increased under oxidative stress. The present study was aimed to study the effect of NO donor L-arg (L-arginine) on DNA damage repair in human bronchial epithelial cells exposed to ozone and the potential mechanisms involved. HBECs (human bronchial epithelial cells) were cultured with or without ozone (1.5 ppm, 30 min), DNA breaks were measured with a comet assay and agarose gel electrophoresis, cell cycling was determined by flow cytometry and p-ATM was measured by immunofluorescence and Western blot. Data were analysed by ANOVA (analysis of variance). P<0.05 was considered as significant. Ozone induced marked DNA breaks, G1-phase arrest and increased expression of p-ATM in HBECs, while wortmannin reduced the levels of p-ATM induced by ozone; the NO donor, L-arg, minimized the effects of ozone-induced DNA breaks and increased the level of p-ATM, while the NO synthase inhibitor, L-NMMA [N(G)-minomethyl-L-arginine], restrained those effects of L-arg. The effect of L-arg on DNA repair is NO-mediated, and p-ATM is implicated in the processes of DNA repair.

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Section: Discussionsupporting

confidence: 84%

“…Environmental factors, such as ozone, may have tremendous impacts on the function of airway epithelium. For example, ozone exposure can exacerbate asthma (Mar and Koenig, 2009) and lead to DNA damage of the airway epithelial cells (Gabrielson et al, 1994; Lee et al, 1996; Qu et al, 2009; Kosmider et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

l-Arginine promotes DNA repair in cultured bronchial epithelial cells exposed to ozone: involvement of the ATM pathway

et al. 2011

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“…a number of systems, both in vitro and in vivo (McDonald et al, 1993;Gabrielson et al, 1994;Ferng et al, 1997). H 2 O 2 has been shown in the current study to be a potent inducer of SSBs.…”

Section: Discussion and Conclusion Preliminary Studies With H 2 Osupporting

confidence: 51%

“…Hydrogen peroxide (H 2 O 2 ) is a frequently used positive control for DNA damage in the assay, and is a potent inducer of DNA SSBs (McDonald et al, 1993;Gabrielson et al, 1994;Lee et al, 1996). The SSBs caused by oxidants can be used as a biomarker of exposure, as well as an indicator of carcinogenicity/mutagenicity.…”

mentioning

confidence: 99%

Detection of Ozone-Induced Dna Single Strand Breaks in Murine Bronchoalveolar Lavage Cells Acutely Exposed in Vivo

Haney

Connor

Li³

1999

Inhalation Toxicology

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Single-strand breaks (SSBs) in DNA have been used a biomarker of oxidative damage. The comet assay, also known as single-cell gel electrophoresis, was used to investigate the ability of ozone (O(3)) to induce DNA SSBs in murine bronchoalveolar lavage (BAL) cells. The comet assay is more sensitive than other techniques currently utilized for detecting SSBs and requires fewer cells. In the present study, 3 mice were exposed for 3 h to 0.25 ppm of O(3), and 3 to 0.5 ppm of O(3) for 3 h. Two air-exposed mice served as negative controls. All mice were euthanized 3 h after exposure, at which time BAL cells were recovered from the lungs and stained with ethidium bromide. BAL cells recovered from an air-exposed mouse were exposed to various concentrations of H(2)O(2) in vitro for 1 h at 4 degrees C. Excluding cells from the H(2)O(2) group (n = 25), 50 randomly selected BAL cells were graded by comet tail length into 1 of 4 categories: no damage (0 mm), low damage (1-10 mm), medium damage (11-30 mm), and high damage (31 + mm). The nonparametric Wilcoxon rank-sum test was used for statistical analysis, and p values lower than .05 were considered significant. The H(2)O(2) and the 0.25 and 0.5 ppm O3 groups showed statistically significant increases in DNA SSBs as compared to air-exposed controls. The results of this study indicate that (1) O(3) induces DNA strand breaks in murine BAL cells at 0.25 and 0.5 ppm, as evidenced by statistically significant increases in the length of comet tails for O(3)-exposed groups, and (2) the comet assay can be used to assess O(3)-induced SSBs for in vivo exposures. Therefore, it has the potential as a biomarker for in vivo oxidant exposures.

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“…3 Various ALIC-based evaluation systems for gaseous toxicities have previously been reported by several researchers. [5][6][7][8][9][10][11][12] Some of the devices have been applied to evaluate the toxicities for actual environmental gases, such as cigarette smoke, 8,9 diesel exhaust emission, 10 and combustion gas. 11 However, since most devices reported previously are large scale and continuous flow-based systems, several handled liters of sample gases are necessary.…”

Section: Introductionmentioning

confidence: 99%

Development of an in vitro Batch-type Closed Gas Exposure Device with an Alveolar Epithelial Cell Line, A549, for Toxicity Evaluations of Gaseous Compounds

et al. 2008

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To simply evaluate toxicity for various types of exhaust-gas samples collected in various locations, we developed a smallscale (150 mL) batch-type completely closed gas exposure device incorporated with an air-liquid interface culture of a human alveolar epithelial cell line, A549. On the basis of cell viability tests using an acid phosphatase assay after 48 h of gas exposure, the developed device was able to measure clear dose-response relationships for volatile organic and inorganic compounds, such as benzene, trichloroethylene (TCE), acetone, SO2 and NO2 gases, but not CO gas. Although the 50% effective concentration values in the device were much higher than 50% lethal concentration values reported in animal experiments, the tendency of the toxic intensity observed in the former was roughly consistent with that of the acute toxicity in the latter. We further applied the device to evaluate the toxicity of cigarette smoke as an example of actual environmental gases, and successfully measured acute cell death from the gas after 48 h of exposure. The present small device is expected to be one of good tools not only in simultaneously assessing various gaseous chemicals or samples, but also in studying acute toxicity expression mechanisms in human lung epithelia.

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Comparison of the Toxic Effects of Hydrogen Peroxide and Ozone on Cultured Human Bronchial Epithelial Cells

Cited by 3 publications

References 8 publications

l-Arginine promotes DNA repair in cultured bronchial epithelial cells exposed to ozone: involvement of the ATM pathway

l-Arginine promotes DNA repair in cultured bronchial epithelial cells exposed to ozone: involvement of the ATM pathway

Detection of Ozone-Induced Dna Single Strand Breaks in Murine Bronchoalveolar Lavage Cells Acutely Exposed in Vivo

Development of an in vitro Batch-type Closed Gas Exposure Device with an Alveolar Epithelial Cell Line, A549, for Toxicity Evaluations of Gaseous Compounds

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