• Environmental Analysis Health and Toxicology
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• 제19권4호
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• pp.335-344
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• 2004

Diesel exhaust particle (<2.5 ${\mu}{\textrm}{m}$, DEP$_{2.5}$) is known to be probarbly carcinogenic (IARC group 2A). DEP$_{2.5}$ contains organic compounds such as polycyclicaromatic hydrocarbon (PAH), heterocyclic compounds, phenols, and nitroarenes. Reactive oxygen species (ROS) are generated by DEP$_{2.5}$ without any biological activation system. Therefore, an alternative mechanism by which DEP$_{2.5}$ could be carcinogenic is known by the generation of oxidative DNA damage. The aim of this study was to investigate genotoxic effects of DEP$_{2.5}$ using single cell gel electrophoresis. In order to evaluate the mechanisms of DEP$_{2.5}$ genotoxicity, the rat micro-some mediated and DNA repair enzyme treated comet assays together with routine comet assay were performed. DEP$_{2.5}$ was collected from diesel engine bus and dichloromethane extract was obtained. The organic extract of DEP$_{2.5}$ revealed DNA damage itself in NIH/3T3 cells. And it showed both oxidative and microsome mediated DNA damages. Vitamin C as an model antioxidant reduced DNA damage in endonuclase III treated comet assay. One of flavonoid, galangin as a CYP1A1 inhibitor reduced DNA damage in the presence of S-9 mixture. Our results show that DEP$_{2.5}$ are genotoxic and a great source of oxidative stress, but antioxidants can significantly reduce oxidative DNA damages. And DEP$_{2.5}$ may contain indirect mutagens which can be inhibited by CYP inhibitors.d by CYP inhibitors.

• 한국자원식물학회지
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• 제22권5호
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• pp.461-466
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• 2009

식물세포에 마(Dioscorea batatas Dence) 추출액의 전처리가 방사선 스트레스에 노출된 배양세포의 활력, 생장 및 핵 DNA 손상에 미치는 영향을 조사하였다. 마의 분획추출물 중 EtOAc 분획추출물을 식물세포에 전처리하고 20 Gy의 방사선에 노출시키면, 마 추출물을 전처리하지 않고 방사선 20 Gy만 처리한 세포보다 세포의 활력과 생체중이 20%이상 증가하였다. Comet 분석에서 꼬리부분의 길이 (T)와 머리부분의 길이 (H)를 측정하여 T/H 비율을 조사하였다. 무처리 세포와 방사선 20 Gy를 처리한 세포의 T/H 비율은 각각 1.05 및 1.68로 나타났고, head DNA 량은 각각 86.7% 및 71.3%로 무처리 세포와 방사선을 처리한 세포간에는 큰 차이를 보여, 방사선에 의한 심각한 핵 DNA 손상을 관찰할 수 있었다. 그러나 마 추출물 중 MeOH, EtOAc 및 n-BuOH 분획추출물을 식물세포에 전처리하고 20 Gy 방사선을 처리하면, T/H 비율은 각각 1.37, 1.01 및 1.10이었고, head DNA량은 81.5%, 87.6% 및 88.7%로 방사선을 처리 하지 않은 무처리 세포 수준으로 회복되었다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 춘계학술발표회
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• pp.74-74
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• 2018

Abeliophyllum distichum is Korea Endemic Plants and its genetic resources found from Korea only. Bioactivities of A. distichum such as antioxidant, anti-cancer, and anti-inflammatory studies have been proved through many researches. Whereas, there are no studies on the biological activity of its callus extracts. In this study, we investigated the antioxidant activities of callus extracts derived from A. distichum and its inhibitory effect on oxidative DNA damage. The antioxidant activities were assessed using radical scavenging assays with DPPH, ABTS, and reducing power assay and the inhibitory effects on oxidative DNA damage were measured using ${\varphi}-174$ RF I plasmid DNA cleavage assay. In addition, callus extracts derived from A. distichum showed high antioxidant acitivties and no cytotoxicity in NIH/3T3. Also, it has significantly suppressed expression of ${\gamma}$-H2AX and p53 protein and mRNA levels in NIH/3T3 cells exposed to oxidative stress. Therefore, the callus extracts derived from A. distichum has potential antioxidant activity that can provide protective effects against the oxidative DNA damage caused by free radicals. This study suggest that it is valuable as cosmetics and medicine for antioxidant and cancer preventive materials.

• 한방비만학회지
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• 제19권1호
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• pp.1-11
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• 2019

Objectives: The purpose of the present study was to evaluate the preventive effects of ethanol extract of Polygalae radix (EEPR) against oxidative stress (hydrogen peroxide, $H_2O_2$)-induced DNA damage and apoptosis in Chang liver cells. Methods: Chang liver cells were pretreated with various concentrations of EEPR and then challenged with 0.5 mM $H_2O_2$. The cell viability and apoptosis were assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and flow cytometry analysis, respectively. The levels of reactive oxygen species (ROS), mitochondrial membrane potentials (MMPs) and adenosine tri-phosphate (ATP) contents were measured. Expression levels of Bcl-2 and Bax were also determined using Western blot analysis. Results: The results showed that the decreased survival rate induced by $H_2O_2$ could be attributed to the induction of DNA damage and apoptosis accompanied by the increased production of ROS, which was remarkably protected by EEPR. In addition, the loss of $H_2O_2$-induced MMPs and ATP contents was significantly attenuated in the presence of EEPR. The inhibitory effect of EEPR on $H_2O_2$-induced apoptosis was associated with up-regulation of Bcl-2 and down-regulation of Bax, thus reducing the Bax/Bcl-2 ratio. Conclusions: Our data prove that EEPR protects Chang liver cells against $H_2O_2$-induced DNA damage and apoptosis by scavenging ROS and thus suppressing the mitochondrial-dependent apoptosis pathway.

• 방사선산업학회지
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• 제17권3호
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• pp.257-264
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• 2023

This study was conducted to evaluate the applicability of the Comet assay, which is widely used for the identification of irradiated meats, to detect irradiated beef cut, ground beef, and Tteokgalbi during freezing storage. Gamma-irradiation significantly increased the DNA damage in frozen beef cut and ground beef samples. Among those, DNA nuclei of samples irradiated with absorbed doses of 1kGy or more showed typical comet-shaped damage, convincing that the samples were irradiated. Meanwhile, DNA nuclei in non-irradiated beef cut and ground beef samples were also damaged according to storage time. In particular, since the damage of DNA nuclei in the non-irradiated samples frozen for three months was similar to that of samples irradiated with a dose of 0.5 kGy, it was considered difficult to detect whether these samples were irradiated by Comet assay analysis. Likewise, gamma-irradiation of Tteokgalbi increased DNA damage. However, significant damage to DNA nuclei was observed even in the non-irradiated samples. Therefore, the application of the analysis method to determine whether the Tteokgalbi sample was irradiated was not appropriate. In conclusion, these results suggest that Comet assay could be limitedly applied only to fresh meat with a short storage period and minimal processing.

• Food Science and Biotechnology
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• 제17권2호
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• pp.343-348
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• 2008

In this study, antioxidant activity of methanol extract of Glycyrrhiza uralensis Fisch (GUE) against $H_2O_2$-induced DNA damage in human leucocytcs and cell death in PC12 cells was determined. The effect of GUE on $H_2O_2$-induced DNA damage in human leucocytcs was evaluated by the comet assay, where GUE ($1-50\;{\mu}g/mL$) was a dose dependent inhibitor of DNA damage induced by $H_2O_2$. The protective effect of GUE against $H_2O_2$-induced damage on PC12 cells was investigated by MTT reduction assay and lactate dehydrogenase release assay. A marked reduction in cell survival induced by $H_2O_2$ was significantly prevented by $1-50\;{\mu}g/mL$ of GUE. The enzyme activity of caspase-3 was elevated in $H_2O_2$-treated PC12 cells, while preincubation with GUE for 30 min inhibited $H_2O_2$-induced caspase-3 activation in a dose-dependent manner. In conclusion, GUE ameliorates $H_2O_2$-induced DNA damage in human leucocytes and has neuroprotective effect by preventing cell death in PC12 cell, suggesting that GU may be a potential candidate for novel therapeutic agents for neuronal diseases associated with oxidative stress.

• 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.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2004년도 International Meeting of the Microbiological Society of Korea
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• pp.47-50
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• 2004

The defenses against free radical damage include specialized repair enzymes that correct oxidative damages in DNA, and detoxification systems such as superoxide dismutases. These defenses may be coordinated genetically as global responses. We hypothesized that the expression of the SOD and the DNA repair genes would inhibit DNA damage under oxidative stress. Therefore, the protection of E. coli mutants deficient in SOD and DNA repair genes $(sod^-\;xth^-\;and\;nfo^-)$ was demonstrated by transforming the mutant strain with a plasmid pYK9 which encoded Photobacterium leiognathi CuZnSOD and human AP endonuclease. The results show that survival rates were increased in $sod^+\;xth^-\;nfo^+$ cells compared to $sod^-\;xth^-\;ap^+,\;sod^-\;xth^-\;ap^-,\;and\;sod^+\;xth^-\;ap^-$ cells under oxidative stress generated from 0.1 mM Paraquat or 3 mM $H_2O_2$. The data suggested that, at least, SOD and DNA repair enzymes may have collaborate protection and repair of the damaged DNA. Additionally, both enzymes are required for protection against free radicals.

• 한국환경성돌연변이발암원학회지
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• 제19권1호
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• pp.28-33
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• 1999

The SNF2 family includes proteins from a variety of species with roles I cellular processes such as transcriptional regulation, recombination and various types of DNA repair. Several proteins with unknown function are also included in this family. Here, we report the cloning and characterization of hrp 2+ gene (helicase related gene from S. pombe) which was isolated by PCR amplication using the conserved domain of SNF2 motifs within the ERCC6 gene which encodes a protein involved in DNA excision repair. The hrp2+ gene was isolated by screening with yeast S. pombe genomic library. The isolated cloned contained 6.5 kb insert DNA. Southern blot analysis confirmed that S. pombe chromosome contains the same DNA as hrp2+ gene and this gene exists as a single copy in S. pombe genome. The 4.7 kb transcript of mRNA was identified by Northern blot. To examined the transcriptional regulation of hrp2+ gene, DNA damaging agents were treated. These results indicated that the hrp2+ gene may not be directly involved in DNA replication, but may be involved in damage response pathway.

• 한국환경성돌연변이발암원학회지
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• 제25권2호
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• pp.71-75
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• 2005

Green tea and their major constituents such as catechins are famous materials for their anti-oxidative and anti-carcinogenic activity, but many compounds with reducing power can promote the oxidation in their oxidized form or in the presence of metal ion. We investigated the pro-oxidative effect of the ethanol extract equivalent up to 30mg of dried weight of green tea leaves in four in vitro systems which could be used for detecting DNA damage. Although ethanol extract of green tea did not show significant mutagenicity in Salmonella typhimurium TA102, which is sensitive strain to oxidative stress, it degraded deoxyribose extensively in the presence of $FeCl_3-EDTA$ complex, promoted 8-oxoguanine formation in the live bacteria cell, Salmonella typhimurium TAI04, and cleaved super coiled DNA strand with the help of copper ion. It suggested that green tea, famous anti-oxidative material, can be pro-oxidant according to the condition of extraction or metal existence.