• 약학회지
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• 제46권3호
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• pp.208-212
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• 2002

Dehydroevodiamine HCl (DHED), which is a component separated from Evodia rutaecarpa Bentham, has novel anticholinesterase and antiamnesic activities in the scopolamine-induced amnesia model. Several studies suggest that DHED might be an effective drug for the Alzheimer's disease and the vascular type of dementia. In order to evaluate the mutagenic potential of DHED, Salmonella typhimurium reversion assay, chromosomal aberration test on Chinese hamster lung cells, in vivo micronucleus assay using mouse bone marrow cells, and comet assay were performed. DHED did not increase the number of revertant in the reverse mutation test using Salmonella typhimurium TA1535, TA1537, TA98, TA100. DHED HCl, at concentration of 5 and 10 $\mu\textrm{g}$/mι, increased the number of chromosome aberrated Chinese hamster lung cells with 5 and 10％, respectively. In mouse micronucleus test, no significant increase in the occurrence of micronucleated polychromatic erythrocyte was observed in ICR mice orally administered with DHED. DHED was tested for ability to induce genotoxic effect in L5178Y cells (mouse lymphoma cells) using the single cell gel electrophoresis assay (comet assay). In comet assay, tail moment did not increase in L5178Y cells treated with 10, 100, 300 $\mu$M DHED.

• 한국발생생물학회지:발생과생식
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• 제11권2호
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• pp.127-135
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• 2007

이 연구는 2004년 6월에 전북 김제시 만경면 심포 앞바다에서 채집된 전어를 전남대학교 어류학실험실로 운반하여 습식법으로 인공 수정한 난을 대상으로 난 발생 및 자치어 발육 과정을 관찰하였다. 전어의 산란기는 $3{\sim}6$월이었고, 수정란은 구형의 분리 부성란으로 난경은 $1,14{\sim}1.34\;mm$(평균, 1.21 mm)였다. 수온이 $19.0{\sim}23.0^{\circ}C$(평균 $21.2^{\circ}C$)에서 수정 후 35시간 53분에는 눈에 렌즈가 착색되었고, 심장이 분화되어 꼬리가 난황으로부터 완전히 분리되었으며, 배체 전반부에 흑색 소포가 나타났다. 수정 후 37시간 10분 만에 첫 부화가 시작되었다. 부화 직후 자어의 전장은 $4.26{\sim}5.30\;mm$(평균 4.96 mm)로 난황을 달고 있었고, 입과 항문은 아직 열려 있지 않았으며, 항문은 전장의 80%로 꼬리지느러미의 기저에 약간 앞쪽에 위치하였으며, 근절은 $22{\sim}27$개였고, 눈에는 흑색 소포가 착색되어 있었다. 부화 후 2일째의 전기 자어는 $4.96{\sim}5.74\;mm$(평균 5.24 mm)로 난황이 완전히 흡수되었고, 부화후 16일째 후기 자어는 전장이 $9.66{\sim}10.81\;mm$(평균 9.66 mm)로 꼬리말단이 굽어지기 시작하면서 가슴지느러미, 등지느러미, 꼬리지느러미의 줄기가 형성되었다. 부화 후 53일째의 치어기는 전장이 $27.11{\sim}34.09\;mm$(평균 30.11 mm)로 머리 부분이 현저하게 발달하였고, 등지느러미와 배지느러미는 거의 일직선상에 위치하였다. 이 시기에 모든 지느러미는 정수에 달하였으며, 체형이나 반문이 성어와 닮은 치어기로 이행하였다. 결론적으로 어류는 같은 과 내에서도 흑색 소포의 모양과 위치가 차이가 있다. 그러므로 이 연구는 흑색 소포의 특징과 함께 전어의 초기 발달의 특징을 명확하게 밝히기 위한 연구이다.

• Journal of Nutrition and Health
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• 제41권5호
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• pp.391-401
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• 2008

캐피어 원말은 유산균, 효모, 다당 및 여러 영양성분을 다량 함유하고 있으며 장기능 개선 효능을 살펴보기 위하여 3주령의 수컷 흰쥐 (Sprague-Dawley)를 4군으로 나누어 정상대조군 (N군), DSS투여 대조군 (DC군)과 두 군의 캐피어 투여군으로 하여 대조군 사료와 캐피어 원말을 각각 1.5%와 3.0% 혼합한 사료로 3주간 사육하였다. 이 후 DSS투여 대조군과 캐피어 투여군들에 5일간 2% DSS 음용수 동일한 양을 투여하여 경미한 대장염을 유도하였다. 대장염 유도 후 희생하여 소장 단백질 및 DNA 함량, 혈장 및 결장의 조직검사와 결장조직에서 TBARS와 MPO 활성, 혈장 백혈구에서의 DNA 손상 정도를 측정하였다. 실험 자료는 Windows용 SPSS package program version12.0을 이용하여 통계 처리하였고, 네 군간의 평균값의 차이를 검증하기 위하여 일원배치 분산분석 (one-way ANOVA)을 한 후, Duncan's multiple range test로 변인간의 차이를 검증하였다. DSS 투여군 들에서 변 수분함량이 증가하고 음용수 섭취량이 증가하는 경향과 함께 결장의 조직검사 결과 DSS 투여군에서는 염증과 부종 증상을 관찰할 수 있었으며 식이무게의 3% 캐피어 원말 투여군에서는 재생성 변화를 볼 수 있었다. DSS를 투여받은 군들의 소장 점막 단백질 함량은 감소하는 경향을 보였으며 캐피어 3.0%식이섭취한 군들에서는 증가하는 경향을 보였으나 DNA함량에서는 차이를 볼 수 없었다. DSS 투여군에서는 결장조직의 TBARS 값이 유의적으로 증가하였으며 캐피어 투여군에서는 감소하였으나 캐피어 투여 용량에 따른 차이는 보이지 않았다. 혈장 TBARS와 결장조직의 MPO 활성은군 간에 유의한 차이가 없었다. DSS 투여군에서는 혈액 백혈구 DNA의 tail length가 유의하게 증가하였으며 캐피어 투여군에서는 감소하였다. 따라서 약 4주간 캐피어 원말의 투여는 2%의 DSS로 경미한 대장염을 유도한 흰쥐에서 결장 조직의 산화적 스트레스에 대한 저항력을 증가시켜 대장점막을 보호할 수 있는 기능이 있음을 확인할 수 있었다.

• Journal of Nutrition and Health
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• 제49권6호
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• pp.401-410
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• 2016

자화수는 가격이 비싸지 않고 쉽게 만들 수 있으며 환경 친화적인 음용수임에도 불구하고 자화수 섭취의 건강상의 유익에 관해서는 과학적인 문헌이 매우 제한적이다. 본 연구는 유전적 소인에 의해 제2형 당뇨병이 발현되는 모델인 db/db 마우스 6주령 14마리를 당뇨군과 자화수군으로 나누어 각 7마리씩 배치하고, 이형접합체 마우스 10마리를 대조군으로 하여 총 3군으로 나누어 16주 동안 자화수를 투여하였다. 혈당 강하 효과를 보기 위해 공복혈당, 인슐린 농도, 내당능 검사, 당화 헤모글로빈을 측정 하였고, DNA 손상 감소 효과, 항산화 상태를 알아보았다. 당뇨군과 자화수군의 혈당치는 대조군에 비해 유의적으로 높았으며, 당뇨군의 혈당은 자화수 섭취 16주까지 높게 지속되었으나 자화수군의 혈당은 섭취 10주 후부터 유의적으로 감소하였다. 혈중 당화헤모글로빈 함량도 마찬가지로 대조군보다 당뇨군에서 증가하였고 자화수군에서는 유의적으로 감소하였으나 경구 당부하 검사 결과, 혈당 곡선하면적 (area under the curve, AUC)은 당뇨군과 자화수 투여군 사이에 유의적인 차이를 보이지 않았다. 당뇨쥐의 DNA 손상 정도는 세 가지 지표 (DNA in tail, tail length, tail moment) 모두 자화수군이 당뇨군에 비해 유의적으로 감소하였으며 이러한 효과는 혈액과 간에서 모두 나타났다. 적혈구 항산화 효소인 SOD, GSH-Px glutathione peroxidase 활성도는 대조군, 당뇨군, 자화수군 간에 차이를 보이지 않았고, 혈장 총항산화능인 TRAP 수준도 자화수 섭취로 인한 차이를 볼 수 없었다. 결론적으로 자화수 투여가 db/db mouse를 사용한 제2형 당뇨의 혈당 강하를 포함한 glycemic control에 유리한 개선효과를 보일 뿐 아니라 혈액과 간의 DNA 손상 감소효과까지 보임을 확인하였다. 그러나 그 기전에 대해서는 확실하게 규명하지 못하였으므로 앞으로 자화 수음용 효과를 뒷받침할 더욱 깊고 다양한 기전 연구가 수행되어야 할 것이다.

• 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.

• 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.

• 한국환경성돌연변이발암원학회지
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• 제21권2호
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• pp.89-94
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• 2001

Recently, single cell gel electrophoresis, also known as comet assay, is widely used for the detection and measurement of DNA strand breaks in vitro and in vivo in many toxicological fields such as radiation exposure, human monitoring and toxicity evaluation. As well defined, comet assay is a sensitive, rapid and visual method for the detection of DNA strand breaks in individual cells. Briefly, a small number of damaged cells suspended in a thin agarose gel on a microscope slide were lysed, unwinded, electrophoresed, and stained with a fluorescent DNA binding dye. The electric current pulled the charged DNA from the nucleus such that relaxed and broken DNA fragments migrated further. The resulting images which were subsequently named for their appearance as comets, were measured to determine the extent of DNA damages. However, some variations could be occurred in procedures, laboratories's conditions and kind of cells used. Hence, to overcome and to harmonize these matters in comet assay, International Workshop on Genotoxicity Test Procedure (IWGTP) was held with several topics including comet assay at Washington D.C. on March, 1999. In spite of some consensus in procedures and conditions in IWGTP, there are some problems still remained to be solved. In this respect, we attempted to set the practical optimal conditions in the experimental procedures such as lysis, unwinding, electrophoresis and neutralization conditions and so on. First of all, we determined optimal lysis and unwinding time by using 150 $\mu$M methyl methanesulfonate (MMS) which is usually used concentration. And then, we determined optimal positive control concentrations of benzo(a)pyrene (BaP) and MMS in the presence and absence of S9 metabolic activation system, respectively.

• 대한의생명과학회지
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• 제14권1호
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• pp.13-18
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• 2008

Excessive alcohol consumption is associated with increased risks of many diseases including cancer. Individuals who regularly consume excessive quantities of alcohol have a greater risk of developing head and neck cancers such as esophageal, pharyngeal and oral cavity cancers if they are deficient in ALDH2 expression compared to normal populations. We evaluated lipid peroxidation in Aldh2 +/+ and Aldh2 -/- mice after they had been subjected to acute ethanol exposure. Malondialdehyde(MDA) level in liver tissue was evaluated as a biomarker of oxidative lipid peroxidation. Although the ethanol treatment did not increase the hepatic MDA level both in Aldh2 +/+ mice and in Aldh2 -/- mice, the MDA level was significant higher in the Aldh2 -/- mice than in the Aldh2 +/+ group. The MDA level was also significantly correlated with olive tail moment in blood and the level of 8-OHdG in liver tissue. This is a strong evidence to support our hypothesis that oxidative stress is more intense in Aldh2 -/- mice than in Aldh2 +/+ mice. Our results suggest that ALDH2-deficient individuals may be more susceptible than wild-type ALDH2 individuals to ethanol-mediated liver disease, including cancer.

• Journal of Nutrition and Health
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• 제36권3호
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• pp.255-261
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• 2003

Food irradiation has been steadily increasing in many countries in line with increasing international trade and concerns about naturally occurring harmful contaminants in food. Although irradiation provides an excellent safeguard for the consumer by destroying almost 100% of harmful bacteria, it is necessary to ensure the safety of irradiated foods. This study was performed to investigate the effect of an irradiated diet on lipid peroxidation in the plasma, liver, small intestinal mucosa, and lymphocyte DNA damage in mice. Eight-week old ICR mice were assigned to two groups to receive either non-irradiated or irradiated (10 kGy) diets containing 20.38% fish powder and 6.06% sesame seeds for 4 weeks. The resulting changes in the degrees of lipid peroxidation were evaluated based on the level of plasma and liver thiobarbituric acid reactive substance (TBARS), transmission electron micrograph of jejunal mucosa, and free radical-induced oxidative DNA damage in lymphocytes, as measured by alkaline comet assay (single cell gel electrophoresis). The peroxide values of the gamma irradiated diet were measured every week, and the sample for comet assay was taken at the end of the four week experimental period. There was no significant difference in food efficiency ratio between the two groups. The peroxide values of the diet were immediately increased to 35.5% after gamma irradiation and kept on increasing during storage. After 4 weeks, no differences in tissue or plasma TBARS value were observed between the two groups, but epithelial cells of jejumum showed osmiophillic laminated membranous structures, considered as myelin figures,. The oxidative DNA damage expressed as tail moment (TM) increased 30% in the blood lymphocytes of the mice fed the irradiated diet. In conclusion, the comet assay sensitively detected differences in lymphocyte DNA damage after feeding with the irradiated diet for 4 weeks. However, in order to ensure the safety of irradiated foods, it would be more useful to conduct a long-term feeding regimen using an irradiated diet and examine the level of lipid peroxidation and the state of oxidative stress in a greater range of organs.

• Journal of Nutrition and Health
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• 제40권8호
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• pp.701-707
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• 2007

This study was to investigate lipid peroxidation, antioxidant enzyme activity and DNA damage after exercise, and the protective effect of garlic against exercise-induced oxidative stress. Male Sprague-Dawley rats(4 weeks old) were randomly divided into three groups of 6 rats each; control group(Con) without garlic and exercise, Ex group with exercise alone, and Ex-G group with 2% garlic and exercise. For 4 weeks, rats were given diets containing 15% corn oil and 1% cholesterol with or without garlic. The swimming was selected as a model for exercise performance. Rats swam for 40 min a day, for 5 days a week. Group Ex and Ex-G showed significant lowering in body weight gain and fat accumulation compared to control. No significant changes were observed in levels of plasma cholesterol and triglyceride among three groups, demonstrating that exercise and garlic had no effects on changes of blood lipid. This finding of blood lipid seems to be due to higher plant sterol content in corn oil. The DNA tail moment of lymphocytes showed greater tendency in Ex and Ex-G than in control, but garlic supplements failed to suppress DNA damages. Compared to control, Ex had higher plasma TBARS which was lowered to the control's level in Ex-G with 2% garlic supplementation(p<0.05). Ex-G led to a higher hepatic superoxide dismutase(SOD) activity than control and Ex(p<0.05). Activity of hepatic catalase was also increased in Ex-G, while in Ex it was significantly low(p<0.05). It seemed that TBARS levels were related to the activities of SOD and catalase, and that garlic contributed to increasing the enzyme activities and led to decrease of TBARS. These results demonstrate that lipid peroxidation and DNA damage occur as a consequences of oxidative stress after exercise, and that antioxidant defense against oxidative stress could be enhanced by garlic supplementation through the induction of antioxidant enzymes. However, further investigations should be done on the garlic effect on DNA damage.