An in vivo comet assay of multiple organs (liver, kidney and bone marrow) in mice treated with methyl methanesulfonate and acetaminophen accompanied by hematology and/or blood chemistry.
The single cell gel electrophoresis (comet) assay is a simple and effective method for detecting DNA damage in cells with or without the capability of cell division. Methyl methanesulfonate (MMS), as a genotoxic compound that reacts with DNA directly, was confirmed for its DNA damage potential by in vivo comet assay in multiple organs such as liver, kidneys and bone marrow in mice and acetaminophen (APAP), a widely used analgesic drug, was evaluated for whether it possesses DNA damage potential or not. Furthermore, cytotoxicity was verified by hematology and /or blood chemistry simultaneously. Male Crj:CD1(ICR) mice were intraperitoneally once treated with MMS at 50, 100, and 150 mg/kg, and APAP at 12, 60, and 300 mg/kg. These organs were collected at 4 and 24 hr after treatment, and the comet assay was performed concomitantly with hematology and/or blood chemistry. The results showed that MMS induced a significant concentration-dependent increase in the frequency of tailed nuclei (DNA damage), tail moment, % DNA in the tail, and tail length in the liver, kidneys and bone marrow at both time points. With regard to hematology and blood chemistry results, nephrotoxic markers were not changed, but aspartate aminotransferase (AST) and alanine aminotransferase (ALT) increased in the 150 mg/kg-treated group, and bone marrow counts (BMC) decreased in all of the treatment groups 24 hr after treatment. These results suggested that DNA damage observed in the kidneys was due to genotoxicity, not nephrotoxicity. The DNA damage was more severe at 4 hr than 24 hr after treatment. This might indicate that the decrease in DNA damage was due to detoxification, repair of the lesions induced by the treatment, or cell turnover, all of which would reduce cellular damage. On the other hand, APAP induced increases in plasma AST and ALT levels in the highest dose group only, and the DNA damage in the liver increased at the same dose. These results suggest that the in vivo comet assay might be used to detect the DNA damage induced by MMS and the subsequent DNA repair in mouse liver, kidneys and bone marrow. APAP at the highest dose induces DNA damage in liver. Blood chemical results may indicate that the DNA damage by APAP treatment was attributable to hepato-cytotoxicity, because DNA damage and hepato-cytotoxicity were detected at the same doses.
Oshida
Keiyu
K
Toxicology and Pharmacokinetics Laboratories, Pharmaceutical Research Laboratories, Toray Industries, Inc., Kamakura, Kanagawa, Japan.
Iwanaga
Ema
E
Miyamoto-Kuramitsu
Keiko
K
Miyamoto
Yohei
Y
D000082:Acetaminophen / Q000633:toxicity*; D000818:Animals; D015415:Biomarkers / Q000097:blood*; D001772:Blood Cell Count; D001774:Blood Chemical Analysis; D001853:Bone Marrow / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D020552:Comet Assay; D004249:DNA Damage; D003627:Data Interpretation, Statistical; D004305:Dose-Response Relationship, Drug; D007668:Kidney / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008099:Liver / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008297:Male; D008741:Methyl Methanesulfonate / Q000633:toxicity*; D051379:Mice; D008815:Mice, Inbred Strains; D009153:Mutagens / Q000633:toxicity*
D015415:Biomarkers; D009153:Mutagens; D000082:Acetaminophen; D008741:Methyl Methanesulfonate