• Journal of Applied Biological Chemistry
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• 제61권2호
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• pp.109-117
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• 2018

최근까지도, 현대사회의 암 발생률은 급격하게 증가하고 있다. 인체 내부에서 내재적 또는 외재적인 요인에 의해 DNA 손상이 발생되고, 세포는 DNA 손상에 대한 방어기작을 통해 스스로를 방어한다. 또한, 비정상적인 DNA 생성 및 결손된 DNA 가닥의 복원은 노화, 암, 염증 등 다양한 질병으로부터 기인한다. 많은 연구자는 이러한 DNA 손상을 억제하기 위하여 적절한 소재 탐색에 많은 관심을 두고 있으며, 특히 합성화합물의 부작용이 알려지면서, 천연물을 기반으로 한 암 예방적 소재에 대한 연구가 많이 이루어지고 있다. 토복령은 백합과(Liliacese)에 속하는 청미래덩굴(Smilax china L.)의 근경이며, 전통적으로 해독과 종기 등의 치료제로 사용되어왔다. 하지만 토복령의 DNA 손상에 대한 억제 효과에 대한 연구는 미흡하다. 본 논문에서는 토복령의 항산화 효과 및 DNA 손상에 대한 억제 효과를 확인하고, 식물이 포함하는 phenolic 화합물의 활성과 연관 관계를 확인하고자 하였다. 항산화 효과를 확인하기 위해, DPPH 라디칼 및 ABTS 라디칼에 대한 소거 활성을 확인하였다. 토복령 추출물은 DPPH 및 ABTS 라디칼을 효과적으로 제거하였으며, 높은 환원력을 나타냈다. HPLC 분석을 통해 phenolic 화합물을 정량 및 동정하였으며, 항산화 효과와 phenolic 화합물의 연관 관계를 확인하였다. 또한, $OH^-$ 라디칼 및 $Fe^{2+}$으로 유발된 plasmid DNA 손상에 대한 방어 효과를 확인하였다. 세포 수준에서, DNA 손상에 대한 저해 효과는 산화적 스트레스로 유발된 NIH 3T3 세포의 ${\gamma]$-H2AX 및 p53 단백질 발현 저해 활성을 확인하였다. 또한, H2AX 및 p53 mRNA 수준의 저해 활성을 확인하였다. 결론적으로, 토복령 추출물의 phenolic 화합물의 항산화 효과 및 DNA 손상에 대한 억제 효과를 확인하였다.

• Journal of Nutrition and Health
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• 제44권5호
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• pp.366-377
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• 2011

Glutathione S-transferase (GST) is a multigene family of phase II detoxifying enzymes that metabolize a wide range of exogenous and endogenous electrophilic compounds. GSTM1 and GSTT1 gene polymorphisms may account for inter-individual variability in coping with oxidative stress. We investigated the relationships between the level of lymphocyte DNA and antioxidative parameters and the effect on GST genotypes. GSTM1 and GSTT1 were characterized in 301 young healthy Korean adults and compared with oxidative stress parameters such as the level of lymphocyte DNA, plasma antioxidant vitamins, and erythrocyte antioxidant enzymes in smokers and non smokers. GST genotype, degree of DNA damage in lymphocytes, erythrocyte activities of superoxide dismutase, catalase, and glutathione peroxidase (GSH-Px), and plasma concentrations of total radical-trapping antioxidant potential (TRAP), vitamin C, ${\alpha}$- and ${\gamma}$-tocopherol, ${\alpha}$- and ${\beta}$-carotene, and cryptoxanthin were analyzed. Lymphocyte DNA damage assessed by the comet assay was higher in smokers than that in non-smokers, but the levels of plasma vitamin C, ${\beta}$-carotene, TRAP, erythrocyte catalase, and GSH-Px were lower than those of non-smokers (p < 0.05). Lymphocyte DNA damage was higher in subjects with the GSTM1- or GSTT1-present genotype than those with the GSTM1-present or GSTT1- genotype. No difference in erythrocyte antioxidant enzyme activities, plasma TRAP, or vitamin levels was observed in subjects with the GSTM1 or GSTT1 genotypes, except ${\beta}$-carotene. Significant negative correlations were observed between lymphocyte DNA damage and plasma levels of TRAP and erythrocyte activities of catalase and GSH-Px after adjusting for smoking pack-years. Negative correlations were observed between plasma vitamin C and lymphocyte DNA damage only in individuals with the GSTM1-present or GSTT1- genotype. The interesting finding was the significant positive correlations between lymphocyte DNA damage and plasma levels of ${\alpha}$-carotene, ${\beta}$-carotene, and cryptoxanthin. In conclusion, the GSTM1- and GSTT1-present genotypes as well as smoking aggravated antioxidant status through lymphocyte DNA damage. This finding confirms that GST polymorphisms could be important determinants of antioxidant status in young smoking and non-smoking adults. Consequently, the protective effect of supplemental antioxidants on DNA damage in individuals carrying the GSTM1- or GSTT1-present genotypes might show significantly higher values than expected.

• 한국산학기술학회논문지
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• 제11권12호
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• pp.4922-4927
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• 2010

오가피 에탄올 추출물이 제초제로 인한 DNA와 적혈구 손상을 억제할 수 있는지 알아보기 위해 코멧 어세이와 용혈반응을 사용하여 오가피 추출물 존재하에서 DNA 산화손상 억제와 적혈구 손상 억제 정도를 측정하였다. 페녹시계 제초제인 2,4-D (2,4-dichlorophenoxyacetic acid), 2,4,5-T (2,4,5-trichlorophenoxyacetic acid) 그리고 바이피리딜계 제초제인 paraquat은 임파구 DNA에 산화적 손상을 유발하였다. 그러나 2,4-D, 2,4,5-T, 혹은 paraquat으로 인한 DNA 산화손상은 오가피 추출물 처리에 의해 시험관에서 억제되었다. 또한 적혈구 손상도 오가피 추출물 처리에 의해 시험관에서 억제되었다.

• Journal of Preventive Medicine and Public Health
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• 제37권4호
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• pp.373-380
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• 2004

Objectives : There has been gradually increasing concern about the adverse health effects of electromagnetic radiation originating from cell phones which are widely used in modern life. Cell phone radiation may affect human health by increasing free radicals of human blood cells. This study has been designed to identify DNA damage of blood cells by electromagnetic radiation caused by cell phone use. Methods : This study investigated the health effect of acute exposure to commercially available cell phones on certain parameters such as an indicator of DNA damage for 14 healthy adult volunteers. Each volunteer during the experiment talked over the cell phone with the keypad facing the right side of the face for 4 hours. The single cell gel electrophoresis assay (Comet assay), which is very sensitive in detecting the presence of DNA strand-breaks and alkali-labile damage in individual cells, was used to assess peripheral blood cells (T-cells, B-cells, granulocytes) from volunteers before and after exposure to cell phone radiation. The parameters of Comet assay measured were Olive Tail Moment and Tail DNA %. Results : The Olive Tail Moment of B-cells and granulocytes and Tail DNA % of B-cells and granulocytes were increased by a statistically significant extent after 4-hour use of a cell phone compared with controls. Conclusion : It is concluded that cell phone radiation caused the DNA damage during the 4 hours of experimental condition. Nonetheless, this study suggested that cell phone use may increase DNA damage by electromagnetic radiation and other contributing factors.

• 한국자원식물학회지
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• 제31권3호
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• pp.228-236
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• 2018

In this study, we evaluated the antioxidant activity and protective effects against oxidative DNA damage of the ethyl acetate fraction from the callus of Abeliophyllum distichum Nakai (ECA). Callus of A. distichum was induced on MS medium containing NAA (1 mg/L) and 2,4-D (1 mg/L), and a sufficient amount was obtained for the extraction by subculture. Acteoside was analyzed and quantified (0.39 mg/g callus) from ECA using the high-performance liquid chromatography-photodiode array detector method. ECA showed very high antioxidative activity as revealed by DPPH and ABTS scavenging assays. The $IC_{50}$ values were 12.4 and $6.8{\mu}g/ml$, respectively. ECA showed protective effects against oxidative DNA damage evaluated by using ${\Psi}X-174$ RF I plasmid DNA. It also inhibited DNA damage by suppressing the oxidative stress-induced protein and mRNA levels of ${\gamma}$-H2AX and p53 in NIH/3T3 cells. In conclusion, ECA protects against oxidative DNA damage through its powerful antioxidant activity.

• Journal of Applied Biological Chemistry
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• 제59권3호
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• pp.255-260
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• 2016

활성산소종은 DNA의 손상에서 중요한 역할을 한다. 최근 활성산소를 제어하고 조절하기 위해 천연항산화제를 개발하기 위해 많은 노력이 이루어지고 있다. 느티나무(Zelkova serrate)는 느릅나무과의 식물로 한국 마을 입구에 흔히 심어져 친숙한 식물이다. 하지만 느티나무의 항산화 활성 및 산화적 DNA 손상에 대한 방어효과에 대한 연구는 미흡한 실정이다. 본 연구에서 느티나무 잎의 에틸아세테이트 분획물 및 열수 추출물의 항산화 활성 및 산화적 DNA 손상에 대한 억제활성을 확인하였다. 에틸아세테이트 분획물은 열수 추출물에 비해 DPPH 라디칼 소거활성, ABTS 라디칼 소거활성, $Fe^{2+}$ 킬레이팅 활성 그리고 reducing power에서 높은 항산화 활성을 보였다. 또한, 페놀류 화합물 함량은 각각 에틸아세테이트 분획물은 56.63 mg/g 그리고 열수 추출물은 51.61 mg/g으로 분석됐다. ${\phi}X$-174 RF I plasmid DNA를 이용한 산화적 DNA 손상억제활성은 에틸아세테이트 분획물과 열수 추출물 모두 상당한 방어효과를 나타냈다. 따라서 느티나무 잎의 에틸아세테이트 분획물 및 열수 추출물은 뛰어난 항산화 활성 및 산화적 DNA 손상 억제 효과를 통한 천연 자원으로서의 잠재성을 보였다.

• Molecular & Cellular Toxicology
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• 제4권1호
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• pp.66-71
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• 2008

Long term exposure of Jurkat cells to 2 ATA pressure resulted in the inhibition of cell growth. Under a 2 ATA pressure, the morphological changes in the cells were visualized by electron microscopy. The cells exhibited significant inhibitory responses after three passages. However, short-term exposure study was carried out, 2 ATA pressure may have beneficial effects. The Jurkat cells were exposed to $H_2O_2$ (25 and $50{\mu}M$) in order to induce DNA damage, and then incubated under at either normal pressure or 2 ATA for 1 or 2 hours in order to recover the DNA damage. The extent of DNA damage was determined via Comet assay. More recovery from DNA damage was observed at 2 ATA than at normal pressure. The activity of the DNA repair enzymes, DNA polymerase-$\beta$, was also evaluated at both normal pressure and 2 ATA. The activity of DNA polymerase-$\beta$ was observed to have increased significantly at the 2 ATA than at normal pressure. In conclusion, the effects of hyperbaric pressure from 1 ATA to 2 ATA on biochemical systems can be either beneficial or harmful. Long term exposure to hyperbaric pressure clearly inhibited cell proliferation and caused genotoxic effects, but short-term exposure to hyperbaric pressure proved to be beneficial in terms of bolstering the DNA repair system. The results of the present study have clinical therapeutic application, and might prove to be an useful tool in the study of genotoxicity in the future.

• 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.

• Journal of Forest and Environmental Science
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• 제24권2호
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• pp.61-68
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• 2008

Acrylamide is present as a contaminant in heated food products, predominantly from the precursor asparagine. Nonanchored inter simple sequence repeats (ISSRs) are arbitrary multiloci markers produced by PCR amplification with a microsatellite primer. In order to assess the feasibility of microsatellite primers as markers for DNA damage, the study was conducted on common bean (Phaseolus vulgaris L.) exposed to different concentrations of acrylamide. Polymorphisms were abundant among plant samples treated with acrylamide in comparison to control (untreated one) tested with 4- tri-nucleotide, 2 tetra-nucleotide, and 3- dinucelotide primers. The primer (CCG)4 was the best tested primer to generate polymorphism between the DNA of plants treated or not by acrylamide. Polymorphisms became evident as the presence and absence of DNA fragments in treated samples compared with the untreated one. The highest number of DNA variation on ISSR patterns was observed at the micromollar concentrations of acrylamide. Acrylamide was able to induce DNA damage in non concentration-dependent manner with effectiveness at micromollar concentrations. This study demonstrated that ISSR markers can be highly reliable for identification of DNA damage induced by acrylamide.

• 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.