Formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine in rat lung DNA following subchronic inhalation of carbon black.
Chronic high-dose inhalation of carbon black (CB) can produce carcinomas in rat lungs. The mechanisms underlying this response are uncertain. It has been hypothesized that chronic inflammation and cell proliferation may play a role in the development of tumors after high dose, long-term contact of the particles with lung epithelial cells. In this investigation, we analyzed the formation of a known mutagenic lesion [8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dG)] in the lung DNA of rats following subchronic inhalation of CB (Printex-90 and Sterling V). Briefly, female Fischer 344 rats were exposed for 6 h/day, 5 days/week for 13 weeks to 1, 7, and 50 mg/m(3) of Printex-90 (16 nm; specific surface area 300 m(2)/g) and to 50 mg/m(3) of Sterling V CB (70 nm; surface area of 37 m(2)/g). The exposure concentration of Sterling V was selected to be equivalent in terms of retained mass in the lung to the high dose of Printex-90 at the end of exposure. However, in terms of retained particle surface area, the retained lung dose of Sterling V was equivalent to the mid-dose of Printex 90. This design allows comparison of results on the basis of retained particle mass as well as retained particle surface area between the two CB particles. The formation of 8-oxo-dG in the lung DNA was assessed using a reverse phase HPLC system coupled with UV and electrochemical (EC) detection. After 13 weeks of exposure, measurements were made on lung samples obtained at the end of the exposure and a 44-week recovery period in clean air. Lung burdens of CB were determined at both time points as well as differential cell populations from bronchoalveolar lavage fluid (BAL). The results indicate that lung particle overload was achieved after exposure to 7 and 50 mg/m(3) (Printex-90) and 50 mg/m3 (Sterling V) but not at 1 mg/m(3) (Printex-90). Consistent with these results, a significant increase (P < 0.05) in 8-oxo-dG induction was observed following 13 weeks of exposure to 50 mg/m(3) Printex-90 and at 7 and 50 mg/m(3) after the 44-week recovery period. Interestingly, no increase in 8-oxo-dG was observed for Sterling V CB at either time point despite lung particle overload. Although the retained mass dose of Sterling V at the end of exposure was even higher than for Printex 90 (50 mg/m(3) exposure group) (approximately 7.6 vs 4.8 mg), the surface area of the retained Sterling V was similar to that of the retained Printex 90 of the mid-dose exposure (7 mg/m(3)) (approximately 0.2 m(2) in both groups). Since both Sterling V (50 mg/m(3)) and Printex 90 (7 mg/m(3)) did not induce significant increases in 8-oxo-dG in the lung at the end of the 13-week exposure, this finding indicates that a retained large particle mass is not always correlated with similar adverse effects but that particle surface area is a better dose parameter. The lower effect per unit mass dose seen with Sterling V is consistent with earlier studies showing that particle surface area of low toxicity particles is a more appropriate dosemetric for induction of inflammation in the lungs than particle mass (Oberdörster et al., 1994, 2001; Brown et al. 2001; Donaldson et al., 2002). An increase (p < 0.05) in lung lavage neutrophils was observed at 7 mg/m(3) (Printex-90) and 50 mg/m(3) (Printex-90 and Sterling V) at the 13-week exposure period and again at 50 mg/m(3) (Printex-90 and Sterling V, 44-week recovery period). Our current findings suggest that prolonged, high-dose exposure to CB can promote oxidative DNA damage that is consistent with the hypothesis that inflammatory cell-derived oxidants may play a role in the pathogenesis of rat lung tumors following long-term high-dose exposure to CB in rats.
Gallagher
J
J
Epidemiology Biomarker Branch, Human Studies Division, MD-58C, National Health and Environmental Effects Research Laboratory, U.S. Environmental Protection Agency, Research Triangle Park, NC 27711, USA. gallagher.jane@epa.gov
Sams
R
R
Inmon
J
J
Gelein
R
R
Elder
A
A
Oberdörster
G
G
Prahalad
A K
AK
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000280:Administration, Inhalation; D000818:Animals; D001822:Body Burden; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry / Q000166:cytology; D002244:Carbon / Q000008:administration & dosage / Q000633:toxicity*; D002452:Cell Count; D004247:DNA / Q000187:drug effects* / Q000302:isolation & purification; D018736:DNA Adducts / Q000378:metabolism*; D004249:DNA Damage; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000378:metabolism*; D004334:Drug Administration Schedule; D005260:Female; D008168:Lung / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D011916:Rats, Inbred F344; D020127:Recovery of Function
D018736:DNA Adducts; D002244:Carbon; D000080242:8-Hydroxy-2'-Deoxyguanosine; D004247:DNA; D003849:Deoxyguanosine