• 제목/요약/키워드: An alkaline single cell gel electrophoresis (SCGE) Comet assay

검색결과 6건 처리시간 0.024초

Evaluation of Protective Effects of Houttuynia cordata on H2O2-Induced Oxidative DNA Damage Using an Alkaline Comet Assay in Human HepG2 Cells

  • Hah, Dae-Sik;Kim, Chung-Hui;Ryu, Jae-Doo;Kim, Eui-Kyung;Kim, Jong-Shu
    • Toxicological Research
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    • 제23권1호
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    • pp.25-31
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    • 2007
  • To evaluate the protective effect of Houttuynia cordata on hydrogen peroxide-induced oxidative DNA damage in HepG2 cell line, we used an alkaline single-cell gel electrophoresis (SCGE; comet assay). The DNA damage was analyzed by tail moment (TM) and tail length (TL), which used markers of DNA strand breaks in SCGE. The $100{\mu}g/ml$ of methanolic extract of Houttuynia cordata root showed significant protective effects (p < 0.01) against hydrogen peroxide-induced DNA damage in HepG2 cells and increased cell viability against hydrogen peroxide. The results of this study indicate that Houttuynia cordata root methanol extract acts as a potential antioxidant, and exhibits potential anticancer properties, which may provide a clue to find applications in new pharmaceuticals for oxidative stability.

단세포 겔 전기영동법을 이용한 사람 림프구 DNA 손상에 대한 복숭아씨 추출물의 방사선 방어효과 평가 (Evaluation of protective effect of peach kernel extracts on radiation-induced DNA damage in human blood lymphocytes in the single cell gel electrophoresis assay)

  • 김진규;박태원;이장주;채영규
    • Journal of Radiation Protection and Research
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    • 제24권2호
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    • pp.93-99
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    • 1999
  • Alkaline single cell gel electrophoresis (SCGE) assay는 일명 혜성분석이라고 부르며 in vivo 와 in vitro 에서 많은 화학적, 생물학적인 인자에 의한 DNA 손상을 감지하는데 유용한 기법으로 각각의 세포에서 DNA 단일 가닥 절단과 알칼리에 약한 장소를 평가하는 새로운 방법으로 인정되고 있다. 단세포 겔 전기영동법 (SCGE)을 사용하여 복숭아씨 추출물이 방사선에 의하여 사람 림프구 DNA에 나타나는 손상을 보호하는 지 여부를 평가하였다. 복숭아씨 추출물로 10 분간 전처리한 림프구를 0, 0.1, 0.3, 0.5, 1.0, 2.0 Gy 의 방사선으로 조사하였고 방사선만을 조사한 림프구 실험군과 비교평가하였다. 혜성분석에서 DNA 가닥 절단에 대한 표식인 tail moment의 증가는 감마선에 대해서 뚜렷한 선량-반응 관계를 나타내었으며 각각의 농도별로 복숭아씨 추출물이 처리된 림프구의 DNA 손상은 현저히 감소하였다. 단세포 겔 전기영동법을 통한 평가결과 복숭아씨 추출물은 방사선에 의한 림프구 DNA 손상에 대한 탁월한 방어효과를 나타내었다.

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T-2 toxin을 투여한 닭에서 Comet assay 방법을 이용한 DNA 손상 평가와 독성 (Assessment of DNA Damage using an Alkaline Single Cell Gel Electrophoresis (SCGE) Comet Assay and Toxic Effects in Chickens by T-2 Toxin Treatment)

  • 하대식;허정호;이국천;조명희;김국헌;김충희;류재두;이승환;김곤섭;김의경;김종수
    • Toxicological Research
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    • 제22권2호
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    • pp.75-85
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    • 2006
  • This study was designed to evaluate the possible DNA damaging effects of T-2 toxin using an alkaline single cell gel electrophoresis (SCGE) comet assay and also to investigate toxic effects in chickens. A total of 20 chickens were used in these experiments. Graded concentrations of dietary T-2 toxin (0, 4, 8, and $16{\mu}g/g$ of diet) were given to groups of 5 broiler chickens. In comet assay, The DNA damage was analysed by the tail extent moment (TEM) and tail length (TL), which were used as markers of DNA strand breaks in SCGE. A significant dose-dependent increase in the extent of DNA migration as well as in the percentage of cells with tails was observed after treatment with T-2 toxin (P<0.05). Treatment with the low T-2 toxin ($4{\mu}/g$ of diet) induced a relatively low level of DNA damage in comparison with the high T-2 toxin ($16{\mu}/g$ of diet) group. The growth rate was significantly reduced by concentrations of 8, and $16{\mu}/g$ of diet (P < 0.05). The feed conversion ratio were significantly affected by any concentrations (P < 0.05). The relative weight of the spleen, and lung was decreased by the growth inhibitory concentrations. The bursa of Fabricius, thymus, and kid- ney were decreased in relative weight by concentrations of $16{\mu}/g$ of diet. The relative weight of the liver and heart were unaffected. The hemoglobin (Hb), hematocrit (HCT), and mean corpuscular hemoglobin (MCH) were decreased at concentration of $16{\mu}/g$ of diet. As compared with control chickens, there was no marked change in serum components except uric acid in T-2 treated chickens. All lymphoid tissues retained atrophic and lymphoid cell depletion throughout the three weeks trial.

Evaluation of Oxidative DNA Damage Using an Alkaline Single Cell Gel Electrophoresis (SCGE) Comet Assay, and the Protective Effects of N-Acetylcysteine Amide on Zearalenone-induced Cytotoxicity in Chang Liver Cells

  • Kang, Changgeun;Lee, Hyungkyoung;Yoo, Yong-San;Hah, Do-Yun;Kim, Chung Hui;Kim, Euikyung;Kim, Jong Shu
    • Toxicological Research
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    • 제29권1호
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    • pp.43-52
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    • 2013
  • Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin produced by several species of Fusarium that are found in cereals and agricultural products. ZEN has been implicated in mycotoxicosis in farm animals and in humans. The toxic effects of ZEN are well known, but the ability of an alkaline Comet assay to assess ZEN-induced oxidative DNA damage in Chang liver cells has not been established. The first aim of this study was to evaluate the Comet assay for the determination of cytotoxicity and extent of DNA damage induced by ZEN toxin, and the second aim was to investigate the ability of N-acetylcysteine amide (NACA) to protect cells from ZEN-induced toxicity. In the Comet assay, DNA damage was assessed by quantifying the tail extent moment (TEM; arbitrary unit) and tail length (TL; arbitrary unit), which are used as indicators of DNA strand breaks in SCGE. The cytotoxic effects of ZEN in Chang liver cells were mediated by inhibition of cell proliferation and induction of oxidative DNA damage. Increasing the concentration of ZEN increased the extent of DNA damage. The extent of DNA migration, and percentage of cells with tails were significantly increased in a concentration-dependent manner following treatment with ZEN toxin (p < 0.05). Treatment with a low concentration of ZEN toxin (25 ${\mu}M$) induced a relatively low level of DNA damage, compared to treatment of cells with a high concentration of ZEN toxin (250 ${\mu}M$). Oxidative DNA damage appeared to be a key determinant of ZEN-induced toxicity in Chang liver cells. Significant reductions in cytolethality and oxidative DNA damage were observed when cells were pretreated with NACA prior to exposure to any concentration of ZEN. Our data suggest that ZEN induces DNA damage in Chang liver cells, and that the antioxidant activity of NACA may contribute to the reduction of ZEN-induced DNA damage and cytotoxicity via elimination of oxidative stress.

방사선 유도 DNA 손상에 대한 인진쑥의 방어효과 (Protective Effects of a Herb, Artemisia capillaris, Against Radiation-induced DNA Damage)

  • 조성기;오헌;천의현;정우희;조남정
    • 한국식품영양과학회지
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    • 제33권1호
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    • pp.22-27
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    • 2004
  • 인진쑥의 방사선에 의한 산화적 손상에 대한 DNA 방어효과를 확인하기 위하여 마우스 림프구에서 단세포전기영동법과 CHO 세포에서 미소핵 형성 시험 그리고 마우스의 간과 흉선 조직의 DNA에서 8-OHdG의 형성정도를 관찰하였다. 그 결과 단세포전기영동법 및 미소핵 형성 시험에서는 50 $\mu\textrm{g}$/mL의 농도에서 가장 높은 DNA 손상 억제 효과를 나타내었으며, 8-OHdG 형성 정도는 간과 흉선 모두 30 mg/kg 농도 투여군에서 높은 방어효과를 나타내었다. 이상의 결과에서 인진쑥 추출물은 DNA 상해를 효과적으로 방어하였고 특히, 기존에 알려진 방사선 방호물질에 비하여 독성이 적은 천연물이라는 관점에서 방사선 방어제로 적용이 가능할 것으로 사료된다.

Protective Effects of a Herb, Menthae Herba, against Radiation-induced Oxidative DNA Damage

  • Jo, Sung-Kee;H, Heon-O;Uhee Jung;Kim, Sung-Ho;Byun, Myung-Woo
    • 한국식품저장유통학회:학술대회논문집
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    • 한국식품저장유통학회 2003년도 제23차 추계총회 및 국제학술심포지움
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    • pp.152-152
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    • 2003
  • As utilization of radiation in medicine, industry and biochemical research increases, the protection against radiation damage has become an important issue. Natural products such as herbal medicines are beginning to receive attention as modifiers on the radiation response. In the present study, the protective effect of a herb, Menthae Herba, against radiation-induced DNA damage was evaluated using alkaline single-cell gel electrophoresis (SCGE; comet assay) in the mouse peripheral blood Iymphocytes and the micronucleus formation test in the Chinese hamster ovary (CHO) cells. The tail moment, which was a marker of DNA damage in the SCGE, and the frequency of micronuclei was decreased in groups treated with Mentae Herba extract before exposure to 200 cGy of gamma-ray. We also confirmed its activities to scavenge DPPH and hydroxyl radicals. These experiments demonstrated that Menthae Herba was effective at reducing the radiation-induced damage of DNA and scavenging free radicals. It is plausible that scavenging of free radicals by Menthae Herba may have played an important role in providing the protection against the radiation-induced damage to the DNA. These results indicated that Menthae Herba might be a useful radioprotector and that radical scavenging appears to be one of the mechanisms of radiation protection.

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