[논문]Study on Genotoxicity of Crocin, a Component of Gardenia Fruit, in Bacterial and Mammalian Cell Systems

Choi, Hae-Yeon; Kim, Youn-Jung; Jeon, Hee-Kyung; Ryu, Jae-Chun

Study on Genotoxicity of Crocin, a Component of Gardenia Fruit, in Bacterial and Mammalian Cell Systems 원문보기

Molecular & cellular toxicology, v.4 no.4, 2008년, pp.285 - 292

Choi, Hae-Yeon (Department of Food and Nutrition, Sookmyung women's University) , Kim, Youn-Jung (Cellular and Molecular Toxicology Laboratory, Korea Institute of Science and Technology) , Jeon, Hee-Kyung (Cellular and Molecular Toxicology Laboratory, Korea Institute of Science and Technology) , Ryu, Jae-Chun (Cellular and Molecular Toxicology Laboratory, Korea Institute of Science and Technology)

Abstract ▼ AI-Helper

Crocin is one of the major components of gardenia fruit and saffron which are widely used as natural food colorants and as traditional Chinese medicines. However, the genotoxicity data on crocin are not sufficient for safety evaluation. The purpose of this study was the examination of the genotoxicity on crocin from gardenia yellow in bacterial and mammalian cells, using various genotoxic battery testing assays and the influence of crocin on methyl methanesulfonate (MMS) and ${H_2}{O_2}$-induced DNA damage in vitro, using single cell gel electrophoresis (comet) assay. From results, no considerable mutagenicity and clastogenicity were seen in bacteria and mammalian cells treated with crocin, by Ames test, chromosomal aberration assay, ${tk}^{+/-}$ gene forward mutation assay and comet assay. And, post-treatment with crocin significantly suppressed ${H_2}{O_2}$-induced DNA damage in a dose-dependent manner. In conclusion, the findings of the present study and other previous observations indicate that crocin has no genotoxic potential. And it showed that crocin clearly repressed the genotoxic potency of ${H_2}{O_2}$. These results suggest that anti-oxidative effects of crocin may be involved in the protective effects of DNA damage.

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제안 방법

Image of 100 randomly selected cells (50 cells from each of two replicate slides) was analysed each sample. All experiments were repeated in an independent test. Measurement was made by image analysis with Komet 3.
In this assay, L5178Y cells were treated for 2 hr with 1,680, 840 and 420 μg/mL or 3,200, 1,600 and 800 μg/mL of crocin in the absence or presence of S9, respectively and subjected to the comet assay in optimal conditions as determined by IWGTP.
In this study, we examined the genotoxicity of crocin from gardenia yellow in bacterial and mammalian cells, using various genotoxic battery testing assays and the influence of crocin on methyl methanesulfonate (MMS) and H₂O₂-induced DNA damage in vitro, using single cell gel electrophoresis (comet) assay.
All experiments were repeated in an independent test. Measurement was made by image analysis with Komet 3.1 (Kinetic Imaging Limited, Liverpool, UK) system, determining the mean tail moment (percentage of DNA in the tail times tail length) of the 50 cells. Differences between the control and the other values were tested for significance using one way of analysis of variance (ANOVA).

대상 데이터

Low melting agarose was a product of Amresco (Solon, OH, USA). Methylmethanesulfonate (MMS, CAS no. 66-27-3) was purchased from Aldrich Chemical Co. (Milwaukee, WI). Trifluorothymidine (TFT), cyclophosphamide and sodium bicarbonate were obtained from Sigma Chemical Co.

데이터처리

1 (Kinetic Imaging Limited, Liverpool, UK) system, determining the mean tail moment (percentage of DNA in the tail times tail length) of the 50 cells. Differences between the control and the other values were tested for significance using one way of analysis of variance (ANOVA).

이론/모형

The classification of aberration types referred to JEMS-MMS³⁰. Aberration frequencies, defined as aberrations observed divided by number of cells counted, were analyzed using Fishers exact test³¹ with Dunnetts adjustment and compared with results from the solvent controls.
Cytotoxicity of crocin was checked by the trypan blue exclusion assay. For the determination of cytotoxicity, 1×10⁵ CHL cells or 1×10⁶ L5178Y cells were treated to various concentrations of crocin in 12 well plate in the absence and presence S9 metabolic activation system for 6 hr or 2 hr, respectively.
After 11-13 days incubation, clones were counted and the colony size distribution was determined. Mutant frequencies were calculated using a statistical package (Mutant^TM; UKEMS, York, UK) in accordance with the UKEMS guidelines.

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