Estimation of hydroxyl radical generation by salicylate hydroxylation method in kidney of mice exposed to ferric nitrilotriacetate and potassium bromate.
Hydroxyl radical (*OH) generation in the kidney of mice treated with ferric nitrilotriacetate (Fe-NTA) or potassium bromate (KBrO3) in vivo was estimated by the salicylate hydroxylation method, using the optimal experimental conditions we recently reported. Induction of DNA lesions and lipid peroxidation in the kidney by these nephrotoxic compounds was also examined. The salicylate hydroxylation method revealed significant increases in the *OH generation after injection of Fe-NTA or KBrO3 in the kidneys. A significant increase in 8-hydroxy-2'-deoxyguanosine in nuclei of the kidney was detected only in the KBrO3 treated mice, while the comet assay showed that the Fe-NTA and KBrO3 treatments both resulted in significant increases in DNA breakage in the kidney. With respect to lipid peroxidation, the Fe-NTA treatment enhanced lipid peroxidation and ESR signals of the alkylperoxy radical adduct. These DNA breaks and lipid peroxidation mediated by *OH were diminished by pre-treatment with salicylate in vivo. These results clearly demonstrated the usefulness of the salicylate hydroxylation method as well as the comet assay in estimating the involvement of *OH generation in cellular injury induced by chemicals in vivo.
Ueno
Shunji
S
Laboratory of Veterinary Public Health, School of Veterinary Medicine, Kitasato University, Higashi 23-35-1, Towada, Japan. ueno@vmas.kitasato-u.ac.jp
Kashimoto
Takashige
T
Susa
Nobuyuki
N
Shiho
Kensuke
K
Seki
Takashi
T
Ito
Nobuhiko
N
Takeda-Homma
Shino
S
Nishimura
Yoshikazu
Y
Sugiyama
Masayasu
M
D000818:Animals; D001241:Aspirin / Q000494:pharmacology; D001959:Bromates / Q000494:pharmacology*; D020552:Comet Assay; D018736:DNA Adducts / Q000032:analysis; D004249:DNA Damage / Q000187:drug effects; D005290:Ferric Compounds / Q000494:pharmacology*; D017665:Hydroxyl Radical / Q000378:metabolism*; D006900:Hydroxylation / Q000187:drug effects; D007668:Kidney / Q000187:drug effects* / Q000378:metabolism; D007677:Kidney Function Tests / Q000379:methods; D015227:Lipid Peroxidation / Q000187:drug effects; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D009571:Nitrilotriacetic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D012459:Salicylates / Q000378:metabolism*
D001959:Bromates; D018736:DNA Adducts; D005290:Ferric Compounds; D012459:Salicylates; C019536:potassium bromate; D017665:Hydroxyl Radical; D006899:Mixed Function Oxygenases; C022619:salicylate 1-monooxygenase; D009571:Nitrilotriacetic Acid; D001241:Aspirin; C020326:ferric nitrilotriacetate