Comparative investigation of multiple organs of mice and rats in the comet assay

Sekihashi, Kaoru; Yamamoto, Ayumu; Matsumura, Yukie; Ueno, Shunji; Watanabe-Akanuma, Mie; Kassie, Fekadu; Knasmüller, Siegfried; Tsuda, Shuji; Sasaki, Yu

doi:10.1016/s1383-5718(02)00034-7

Cited by 138 publications

(74 citation statements)

References 17 publications

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“…To obtain nuclei, the homogenate was centrifuged at 700 g for 10 min at 08C, and the precipitate was resuspended in chilled homogenizing buffer at 0.5 g=mL and allowed to settle; precipitated clumps were then removed. Doses and times were selected based on the preliminary studies as well as literature-reported values (Sasaki et al, 1997a(Sasaki et al, , 1997bTsuda et al, 2000;Sekihashi et al, 2002).…”

Section: Treatment and Organ Preparationmentioning

confidence: 99%

Protective Effect of Crocus sativus Stigma Extract and Crocin (trans-crocin 4) on Methyl Methanesulfonate–Induced DNA Damage in Mice Organs

Hosseinzadeh

Abootorabi

Sadeghnia

2008

DNA and Cell Biology

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This study was designed to examine the effect of aqueous extract of Crocus sativus stigmas (CSE) and crocin (trans-crocin 4) on methyl methanesulfonate (MMS)-induced DNA damage in multiple mice organs using comet assay. Adult male NMRI mice in different groups were treated with either physiological saline (10 mL=Kg, intraperitoneal [ip]), CSE (80 mg=Kg, ip), crocin (400 mg=Kg, ip), MMS (120 mg=Kg, ip), and CSE (5, 20, and 80 mg=Kg, ip) 45 min prior to MMS administration or crocin (50, 200, and 400 mg=Kg, ip) 45 min prior to MMS administration. Mice were scarified about 3 h after each different treatment, and the alkaline comet assay was used to evaluate the effect of these compounds on DNA damage in different mice organs. The percent of DNA in the comet tail (% tail DNA) was measured. A significant increase in the % tail DNA was seen in nuclei of different organs of MMS-treated mice. In control groups, no significant difference was found in the % tail DNA between CSE-or crocin-pretreated and saline-pretreated mice. The MMS-induced DNA damage in CSEpretreated mice (80 mg=Kg) was decreased between 2.67-fold (kidney) and 4.48-fold (lung) compared to those of MMS-treated animals alone ( p < 0.001). This suppression of DNA damage by CSE was found to be depended on the dose, which pretreatment with CSE (5 mg=Kg) only reduced DNA damage by 6.97%, 6.57%, 7.27%, and 9.90% in liver, lung, kidney, and spleen, respectively ( p > 0.05 as compared with MMS-treated group). In the same way, crocin also significantly decreased DNA damage by MMS (between 4.69-fold for liver and 6.55-fold for spleen, 400 mg=Kg), in a dose-dependent manner. These data indicate that there is a genoprotective property in CSE and crocin, as revealed by the comet assay, in vivo.

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Section: Treatment and Organ Preparationmentioning

confidence: 99%

Protective Effect of Crocus sativus Stigma Extract and Crocin (trans-crocin 4) on Methyl Methanesulfonate–Induced DNA Damage in Mice Organs

Hosseinzadeh

Abootorabi

Sadeghnia

2008

DNA and Cell Biology

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“…The alkaline comet assay is increasingly used in industrial in vitro genotoxicity testing (Rojas et al, 1999;Hartmann et al, 2001) and has also been used an important tool to evaluate the genotoxic potential of compounds in vivo (Rojas et al, 1999;Sekihashi et al, 2002). Our comet assay results for peripheral blood leukocytes from blood taken from 12-day old female and male Swiss mice 4 h and 24 h after treatment are summarized in Table 1.…”

Section: Resultsmentioning

confidence: 99%

Evaluation of the in vivo mutagenic potential of hydroalcoholic extracts of the northern highbush blueberry (Vaccinium corymbosum L. Ericales, Ericaceae) on peripheral blood cells of Swiss mice (Mus musculus Rodentia, Muridae)

2008

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The northern highbush blueberry (Vaccinium corymbosum L. Ericales, Ericaceae) is very rich in anthocyanins, natural pigments which have strong antioxidant properties and potential health benefits, resulting in the worldwide use the blueberry as a medicinal plant. We investigated the mutagenic potential of simple hydroalcoholic extracts of V. corymbosum acutely administrated by gavage to Swiss mice at doses of 1 g kg -1 , 1.5 g kg -1 and 2 g kg -1 . Peripheral blood cells were collected 4 h and 24 h post-gavage and assessed by the alkaline comet assay, with further blood samples being collected at 48 h and 72 h for assessment using the micronucleus (MN) assay. Our results show that the V. corymbosum extracts did not induce any statistically significant increase in the average amount of DNA damage in peripheral blood leukocytes. However, we did record a significant increase in the frequency of micronucleated polychromatic erythrocytes at the three doses tested.

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“…The comet assay is a sensitive, reliable and rapid method for the detection of double-and single-strand DNA breaks, alkali-labile sites and delayed repair sites in individual eukaryotic cells, and is an important tool for evaluating the in vitro and in vivo genotoxic potential of compounds (Rojas et al, 1999;Tice et al, 2000;Sekihashi et al, 2002). Our comet assay results are shown in Tables 1, 2 and 3, where the female and male results for the different concentrations of extract and the N-nitroso-N-ethylurea (ENU) and cyclophosphamide (CP) positive controls are compared with the negative control (water).…”

Section: Resultsmentioning

confidence: 99%

Clastogenicity of Piper cubeba (Piperaceae) seed extract in an in vivo mammalian cell system

et al. 2007

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The plant Piper cubeba is widely distributed in tropical and subtropical regions and is used medically for various purposes but has not yet been evaluated for genotoxicity. We used male and female Swiss mice and Wistar rats and the comet assay and micronucleus test to investigate the mutagenic potential of a crude extract of P. cubeba seeds. The rodents were administered 0.5 g kg -1 , 1.0 g kg -1 and 1.5 g kg -1 of the extract by gavage. For the Swiss mice, peripheral blood was collected 24 h after treatment for the comet assay, and at 48 and 72 h for the micronucleus test. For the Wistar rats, peripheral blood and hepatic cells were collected for the comet assay and bone marrow cells were collected for the micronucleus test 24 h after treatment. At 1.5 g kg -1 , the highest dose tested, the extract induced a statistically significant increase in both the mean number of micronucleated polychromatic erythrocytes and the level of DNA damage in the rodent cell types analyzed. Under our experimental conditions, the P. cubeba seed extract was genotoxic in vivo when administered orally to mice and rats.

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Comparative investigation of multiple organs of mice and rats in the comet assay

Cited by 138 publications

References 17 publications

Protective Effect of Crocus sativus Stigma Extract and Crocin (trans-crocin 4) on Methyl Methanesulfonate–Induced DNA Damage in Mice Organs

Protective Effect of Crocus sativus Stigma Extract and Crocin (trans-crocin 4) on Methyl Methanesulfonate–Induced DNA Damage in Mice Organs

Evaluation of the in vivo mutagenic potential of hydroalcoholic extracts of the northern highbush blueberry (Vaccinium corymbosum L. Ericales, Ericaceae) on peripheral blood cells of Swiss mice (Mus musculus Rodentia, Muridae)

Clastogenicity of Piper cubeba (Piperaceae) seed extract in an in vivo mammalian cell system

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