Effect of cigarette smoke exposure on the modulation of 8-oxo-2'-deoxyguanosine in rat lungs as analyzed by 32P-postlabeling and HPLC-ECD

Int J Oncol. 2001 Oct;19(4):763-6. doi: 10.3892/ijo.19.4.763.

Abstract

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.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 8-Hydroxy-2'-Deoxyguanosine
  • Administration, Inhalation
  • Animals
  • Chromatography, High Pressure Liquid / methods
  • Chromatography, Thin Layer / methods
  • DNA / metabolism
  • Deoxyguanosine / analogs & derivatives*
  • Deoxyguanosine / metabolism*
  • Female
  • Lung / drug effects*
  • Lung / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Smoking / adverse effects*

Substances

  • 8-Hydroxy-2'-Deoxyguanosine
  • DNA
  • Deoxyguanosine