Effect of cigarette smoke exposure on the modulation of 8-oxo-2'-deoxyguanosine in rat lungs as analyzed by 32P-postlabeling and HPLC-ECD.
Cigarette smoke contains several oxidants and free radicals. In the present study, we examined the formation of 8-oxo-2'-deoxyguanosine (8-oxodG) in the lungs of female Sprague-Dawley rats exposed to side-stream cigarette smoke for 6 h a day, 7 days a week for 1, 2, 4 and 12 weeks in a whole body-exposure system. The samples were analyzed for 8-oxodG by 32P-postlabeling-TLC enrichment and HPLC-ECD techniques to confirm and compare results. Animals were sacrificed 15 h after the cessation of smoke exposure and lung DNA was isolated by phenol/Sevag extractions in the presence of the free radical traps, 8-hydroxyquinoline (6.8 mM) and N-t-butyl-alpha-phenyl nitrone (500 microM) to minimize artifactual formation of 8-oxodG during sample work up. Analysis of lung DNA by 32P-postlabeling-TLC showed 8-oxodG levels (mean +/- SE) of 1.45+/-0.24, 2.68+/-0.65, 2.23+/-0.28 and 2.93+/-0.54 per 106 nucleotides after 1, 2, 4 and 12 weeks of smoke exposure. The respective values in sham-treated rats were 2.76+/-0.19, 3.69+/-0.20, 1.44+/-0.43 and 2.84+/-0.45 per 106 nucleotides, suggesting no significant effect of smoke exposure on tissue levels of 8-oxodG. HPLC-ECD procedure yielded slightly higher values for 8-oxodG in all groups, however, again significant differences between sham and smoke-exposed groups were not detected. It is concluded that the chronic exposure to side-stream cigarette smoke does not enhance the formation of 8-oxodG in rat lungs.
Arif
J M
JM
Department of Preventive Medicine and Environmental Health, University of Kentucky Medical Center, Lexington, KY 40536, USA.
Vadhanam
M V
MV
De Groot
A J
AJ
Van Zeeland
A A
AA
Gairola
C G
CG
Gupta
R C
RC
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000280:Administration, Inhalation; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002855:Chromatography, Thin Layer / Q000379:methods; D004247:DNA / Q000378:metabolism; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000378:metabolism*; D005260:Female; D008168:Lung / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012907:Smoking / Q000009:adverse effects*
D000080242:8-Hydroxy-2'-Deoxyguanosine; D004247:DNA; D003849:Deoxyguanosine