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

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야채 및 과일추출물의 항산화작용 (Antioxidant Activity of Vegetables or Fruits Extract in Mice)

  • 허찬;김남이;김현표;허문영
    • 약학회지
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    • 제49권3호
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    • pp.249-254
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    • 2005
  • The ethanol extracts of the mixed vegetables (Bioactive Vegetables, BV) and the mixed fruits (Bioactive Fruits, BF) were evaluated for their in vivo antioxidant activities. Four weeks treatment of oral administration was performed to mice. A $KBrO_3$ as a potent oxidant was used to induce the oxidative stress for in vivo experiment. BV and BF were shown to possess the significant inhibitory effect of lipid peroxidation as measured by the level of malondialdehyde (MDA) formation although the potencies were not higher than those of well-known antioxidants such as vitamin C, trolox and quercetin. Furthermore, BV and BF inhibited DNA damage assessed by single cell gel electrophoresis (comet assay) and reduced the micronucleated reticulocyte (MNRET) formation of peripheral blood. Antioxidants tested also revealed potent inhibitory activities higher than BV and BF. These antigenotoxic activity profiles were similar to that of abovementioned inhibition of lipid peroxidation. Therefore, BV and BF having mild antioxidant activity as functional food candidates may be useful natural antioxidants by the inhibiting of lipid peroxidation and the protecting oxidative DNA and chromosomal damage.

담배연기응축물의 DNA 손상작용과 야채 및 과일추출물의 보호효과 (Antigenotoxicity of Vegetable or Fruit Extract against Cigarette Smoke Condensate)

  • 이형주;허찬;김남이;허문영
    • 약학회지
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    • 제55권3호
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    • pp.251-259
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    • 2011
  • Cigarette smoke condensate (CSC) is known to be carcinogenic compound. CSC contains many organic compounds such as polycyclic aromatic hydrocarbons (PAHs), and heterocyclic amine compounds (HCAs). Reactive oxygen species (ROS) are also generated and induce oxidative DNA damage during the metabolism of CSC. The rat microsome mediated and DNA repair enzyme treated comet assays together with conventional comet assay were performed to evaluate the mechanisms of CSC genotoxicity. The organic extract of CSC induced oxidative and microsome mediated DNA damage. Vitamin C as a model antioxidant reduced DNA damage in endonuclease III treated comet assay. One of flavonoid, galangin as a CYP1A1 inhibitor, reduced DNA damage in the presence of S-9 mixture. The ethanol extracts of the mixed vegetables (BV) or the mixed fruits (BF) showed potent inhibitory effects against CSC induced DNA damage with oxidative DNA lesions and in the prescence of S-9 mixture. These results indicate that BV and BF could prevent CSC-induced cellular DNA damage by inhibiting oxidative stress and suppressing cytochrome P450 in mammalian cells.

대기부유분진추출물로 야기된 DNA 손상에 대한 Quercetin, Vitamin C 및 Trolox 의 보호효과 (Chemopreventive Effect of Quercetin, Vitamin C and Trolox Against the Organic Extract of Airborne Particulate Matter Induced Genotoxicity in A549 Human Lung Carcinoma Cells)

  • 김남이;허문영
    • 약학회지
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    • 제51권4호
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    • pp.239-245
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    • 2007
  • In order to evaluate the genotoxicity of airborne particulate matter extracted with dichloromethane (APE), the rat microsome mediated (S-9) or DNA repair enzyme treated Comet assays were performed using the single cell gel electrophoresis in A549 human lung carcinoma cells. It was found that the cells interacting with APE showed more DNA single-strand breaks relative to untreated cells. The genotoxicity of APE was increased with the treatment of S-9 mixture. Microsome mediated DNA damage was inhibited by CYP1Al inhibitor, quercetin. The APE also showed oxidative DNA damage evaluated by endonuclease III treatment. Oxidative DNA damage of APE was inhibited by antioxidants such as vita- min C and Trolox. We also found that the vegetables or fruits extract may reduce APE-induced genotoxicity by their anti- oxidant activity and CYP1A1 inhibition.

Paraquat에 의한 사람 임파구 DNA 손상에 대한 환원전리수의 보호효과 (Protective Effect of Electrolyzed Reduced Water on the Paraquat-induced Oxidative Damage of Human Lymphocyte DNA)

  • 박은주;류근걸;이윤배;이종권;이미영
    • Applied Biological Chemistry
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    • 제48권2호
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    • pp.155-160
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    • 2005
  • 본 연구에서는 매우 낮은 음의 산화환원전위를 나타내는 환원전리수가 paraquat에 의한 사람 임파구 DNA의 손상에 미치는 영향을 Comet assay를 사용하여 조사하였다. 환원전리수 처리에 의해서 paraquat에 의해 산화적으로 손상된 DNA가 회복되는 정도를 손상된 DNA로 인한 꼬리부분의 형광광도의 %비율로 나타내었다. Comet assay는 개별 세포의 DNA의 산화적 손상을 측정하는데 널리 사용되고 있다. 사람 임파구에 다양한 농도의 paraquat을 $37^{\circ}C$에서 30분간 처리한 후, 환원전리수를 첨가하여 30분간 반응시켰다. 그 결과 paraquat에 의한 임파구 DNA의 손상은 paraquart 농도증가에 의존적으로 증가하였다. 그러나 환원전리수를 처리한 결과 DNA의 산화적 손상이 paraquat 미처리 대조군 수준으로 거의 다 복구되었다.

Genotoxicity on $21{\alpha}-and\;{\beta}-methylmelianodiol$, a Component of Poncirus trifoliata, in Bacterial and Mammalian Cells

  • Ryu, Jae-Chun;Kim, Youn-Jung;Kim, Mi-Soon;Kim, Min-Ji;Sarma, Sailendra Nath;Lee, Seung-Ho
    • Molecular & Cellular Toxicology
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    • 제1권3호
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    • pp.172-178
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    • 2005
  • [ $21{\alpha}$ ]- and ${\beta}$-Methylmelianodiol were isolated as the inhibitor of IL-5 bioactivity from Poncirus tripoliata. To develope as an anti-septic drug, the genotoxicity of $21{\alpha}\;-and\;{\beta}-methylmelianodiol$ was subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of quantity. Mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay (MOLY), single cell gel electrophoresis (Comet) assay in mammalian cells and Ames reverse mutation assay in bacterial system were used as simplified, inexpensive, short-term in vitro screening tests in our laboratory. These compounds are not mutagenic in S. typhimurium TA98 and TA100 strains both in the presence and absence of metabolic activation. Before performing the comet assay, $IC_{20}$ of $21{\alpha}-methylmelianodiol$ was determined the concentration of $25.51\;{\mu}g/mL\;and\;21.99\;{\mu}g/mL$ with and without S-9, respectively. Also $21{\beta}-methylmelianodiol$ was determined the concentration of $24.15\;{\mu}g/mL\;and\;\;22.46\;{\mu}g/mL$ with and without S-9, respectively. In the comet assay, DNA damage was not observed both $21{\alpha}-methylmelianodiol\;and\;21{\beta}-methylmelianodiol$ in mouse lymphoma cell line. Also, the mutant frequencies in the treated cultures were similar to the vehicle controls, and none of $21{\alpha}\;-and\;{\beta}-methylmelianodiol$ with and without S-9 doses induced a mutant frequency over. twice the background. It is suggests that $21{\alpha}\;-and\;{\beta}-methylmelianodiol$ are non-mutagenic in MOLY assay. The results of this battery of assays indicate that $21{\alpha}\;-and\;{\beta}-methylmelianodiol$ have no genotoxic potential in bacterial or mammalian cell systems. Therefore, we suggest that $21{\alpha}\;-and\;{\beta}-methylmelianodiol$, as the optimal candidates with both no genotoxic potential and IL-5 inhibitory effects must be chosen.

Genotoxicity Study of sophoricoside derivatives in mammalian cells system

  • Yun, Hye-Jung;Kim, Youn-Jung;Kim, Eun-Young;Kim, Young-Soo;Kim, Mi-Kyung;Lee, Seung-Ho;Jung, Sang-Hun;Ryu, Jae-Chun
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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    • pp.178.2-178.2
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    • 2003
  • To develope the novel anti-allergic drug, many sophoricoside derivatives were synthesized. Among these derivatives, JSH-II-3, JSH-Ⅵ-3, JSH-Ⅶ-3, and JSH-Ⅷ-3 were selected and subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of Quantity. Mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay(MOLY) and single cell gel electrophoresis (Comet) assay in mammalian cells were used as HTTS tool in our laboratory. (omitted)

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석류 껍질 추출물이 고지방 고콜레스레롤 식이 급여 C57BL/6J 마우스의 항산화 지표 및 DNA 보호에 미치는 영향 (Effects of Pomegranate Peel (Granati pericarpium) Extracts on the Antioxidant Biomarkers in C57BL/6J Mice Fed a High Fat and Cholesterol Diet)

  • 오상희;양윤형;석대은;김미리
    • 동아시아식생활학회지
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    • 제16권4호
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    • pp.414-420
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    • 2006
  • The present study evaluated the effects of pomegranate peel (Granati pericarpium) extract on the lipid profiles and antioxidant biomarkers in mice fed a high fat and cholesterol diet: the measured biomarkers included the TBARS value, GPx, GR, SOD and GST activities. Body fat depositions were significantly decreased in the group that received pomegranate peel. In addition, the activities of GPx, GST and SOD were significantly higher in the liver and plasma of the pomegranate peel group than in the control group. Also, the pomegranate peel diet decreased lipid peroxidation of the liver and kidney. Alkaline single-cell gel electrophoresis (comet assay) showed that the DNA damage in the plasma of the pomegranate peel group was decreased compared to that of control. The present results show that a diet with added pomegranate peel exerts protective effects against oxidative DNA damage and lipid peroxidation possibly via effects on the free radical levels in mice fed a high fat and cholesterol diet.

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Genotoxicity Study of sophoricoside derivatives in mammalian cells system

  • Yun, Hye-Jung;Kim, Youn-Jung;Kim, Eun-Young;Jung, Sang-Hun;Kim, Youngsoo;Kim, Mi-Kyung;Lee, Seung-Ho;Ryu, Jae-Chun
    • 한국환경독성학회:학술대회논문집
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    • 한국환경독성학회 2003년도 춘계학술대회
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    • pp.185-185
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    • 2003
  • To develope the novel anti-allergic drug, many sophoricoside derivatives were synthesized. Among these derivatives, JSH-II-3, JSH-Ⅵ-3, JSH-Ⅶ-3, and JSH-Ⅷ-3 were selected and subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of quantity. Mouse lymphoma thymidine kinase (tk$\^$+/-/) gene assay (MOLY) and single cell gel electrophoresis (Comet) assay in mammalian cells were used as HTTS tool in our laboratory. In MOLY assay, JSH-Ⅶ-3 at 50 ∼ 6 $\mu\textrm{g}$/ml concentrations was not shown significant mutagenic effect in the absence and presence of S-9 metabolic activation system. However, the concentration of ISH-II-3, 38 $\mu\textrm{g}$/ml, induced increased mutation frequency (MF) in the presence of S-9 metabolic activation system. Also in comet assay, DNA damage was not observed in JSH-Ⅵ-3 and JSH-Ⅶ-3, wherase concentration of 32.8 $\mu\textrm{g}$/ml in JSH-II-3 and 13.9 $\mu\textrm{g}$/ml in JSH-Ⅶ-3 were induced DNA damage in the absence of S-9 metabolic activation system. Therefore, we suggest that JSH-Ⅵ-3 and JSH-Ⅶ-3 have no genotoxic effects but JSH-II-3 and JSH-Ⅷ-3 induce some mutagenicity and DNA strand breaks in mouse lymphoma cell line used this study.

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조선소 배출수 및 주변 하천수의 생물독성 (Biologic Effect of Effluents from Shipyard and the Adjacent Stream Water on Four Cultured Organisms)

  • 서진영;김기범;안준건
    • 한국해양환경ㆍ에너지학회지
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    • 제9권4호
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    • pp.187-192
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    • 2006
  • 조선소에서 배출되는 처리수, 혼합방류수 및 주변 하천수가 양식 생물에 미치는 생물 독성영향을 알아보기 위해 48시간 급성독성과 DNA 손상을 조사하였다. 조사대상 생물종으로는 조선소 주변에서 양식되고 있는 넙치, 조피볼락, 피조개, 멍게가 사용되었으며, 48 시간 노출 후 치사율을 파악하였고, DNA 손상 정도는 Comet assay을 이용하여 측정되었다. 급성독성 실험 결과, 넙치는 장평천에서 치사가 나타났고(26%), 조피볼락은 혼합방류수 1에서 치사가 나타났다(13%). 멍게는 고현천에서 10%의 치사율을 보였고, 피조개는 어느 시료에서도 치사가 나타나지 않았다. 본 연구에 사용되어진 어떠한 시료에서도 실험생물을 50%까지 치사시키는 독성이 나타나지 않아 $LC_{50}$은 계산될 수 없었다. 넙치는 장평천과 혼합방류수에서 대조구보다 유의하게 높은 DNA 손상을 보여주었고, 조피볼락은 장평천에서 유의하게 높은 DNA손상이 나타났다(p<0.05). 멍게는 세탁폐수에서 유의한 DNA손상이 나타났지만, 피조개에서는 모든 처리구에서 DNA 손상을 보이지 않았다. 치사율과 DNA 손상을 고려하였을 때 조선소의 처리수와 혼합방류수보다는 장평천에서 놀은 생물독성을 보여주는 것으로 나타났다.

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