• 한국독성학회:학술대회논문집
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• 한국독성학회 2003년도 추계학술대회
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• pp.95-95
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• 2003

The comet assay has been recently validated as a sensitive and specific test system for the quantification of DNA damage. with 11 substances that demonstrated positive results in at least one test among 4 standard short-term genotoxicity tests, and to evaluate its ability to predict rodent carcinogenicity.(omitted)

• Archives of Pharmacal Research
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• 제28권9호
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• pp.1079-1085
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• 2005

The purpose of this study was to investigate the safety of chitosan oligosaccharide and the effects of chitosan oligosaccharide on mercury induced genotoxicity in mice using the micronuclei and chromosome aberration. The micronuclei test was performed by microscopic examination $(\times1,000,\;stained\;using\;a\;May-Grunwald\;solution)$ after administering 0.01, 0.1, and $1\%(10\;mg/mL)$ chitosan oligosaccharide for 7, 60, and 180 days ad libitum in mice. Total micronuclei of 1,000 polychromatic erythrocytes were recorded for each group. There was no difference between the untreated and experimental groups. The intake periods and concentrations of chitosan oligosaccharide did not affect the occurrence of micronuclei in bone marrow cells (P>0.05). The chromosomal aberration test was performed by microscopic examination $({\times}1,000,\;stained\;using\;a\;4\%\;Giemsa\;solution)$ after administering the same concentration of chitosan oligosaccharide to mice, in $F_1,\;F_2,\;F_3$ generations and parents. The frequency of chromosomal aberrations was defined as [Ydr=(D+R)/total number of counted lymphocytes]. Similar to the micronuclei test, there was no difference between the untreated and treated groups. These results showed that the intake periods and concentrations of chitosan oligosaccharide did not affect chromosomal aberrations in bone marrow cells (P>0.05). To investigate the effect of chitosan oligosaccharide on mercury-induced chromosome aberration, mice in each condition were supplied with $^{203}HgCl_2$ and chitosan oligosaccharide ad libitum. Chitosan oligosaccharide significantly inhibited $^{203}HgCl_2-induced$ chromosome aberration in mice. Based on the results of this study, it may be concluded that the chitosan oligosaccharide is a nontoxic material that could be used as a suppressor of heavy metal-induced genotoxicity.

• 한국식품저장유통학회지
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• 제7권1호
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• pp.68-73
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• 2000

These experiments were performed to investigate the safety of two herbs-Houttuynia cordata Thunberg and Lycium chinese Miller-irradiated with gamma-rays in respect of genotoxicity. Water extracts from the 10 kGy gamma-irradiated herbs were examined in two short -term in vitro tests ; (1) Salmonella typhimurium reversion assay (Ames test) in strain TA 98 and Ta100 and (2) Micronuclues test on clutured Chinese hamster ovary(CHO) cells. No mutagenicity was detected in the two assays with or without metabolic activation . From these results , the safety of the herbs irradiated with gamma-rays at practical doses could be revealed in further tests of genotoxicity in vivo, chronic and reproductive toxicity.

• 한국약용작물학회지
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• 제13권6호
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• pp.217-225
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• 2005

강원도 산지의 뽕나무의 잎, 줄기, 뿌리를 건조하여 100% EtOH와 물로 추출하여 얻은 추출물 및 EtOH추출물로부터 용매별 분획물들을 NIH/3T3에 대한 세포독성, comet assay를 이용한 DNA damage손상으로부터 유전 독성, in vitro micronucleus assay를 이용한 유전독성, Ames test를 이용한 돌연변이 유발을 조사하여 다음과 같은 결과를 얻었다. 1) 각 시료들의 NIH/3T3에 대한 세포독성은 0.5mg/ml 농도에서 약 80%의 생육활성을 나타냈다. 2)뽕나무 잎의 1차 추출물들은 comet assay에서 양성 대조구 대비 약 40% 정도의 유전독성을 보였다. 뽕잎의 EtOH 추출물에서 분획한 분획물 시료중 수용성 분획은 거의 독성이 나타나지 않았으나, hexan의 0.25mg/ml 농도의 분획물이 약 50%정도를 보였으며 다른 시료들은 이 보다 약한 독성을 보였다. 또한 뽕나무 줄기의 분획물 시료들 중에서 0.5mg/ml 농도의 chloroform 분획물이 양성 대조구 대비 가장 강한 50%의 유전독성을 보인것 외에 다른 시료들은 그보다 약한 독성을 보였다. 3) In vitro micronucleus assay를 이용한 유전독성은 재료의 1차 추출물은 거의 유전독성이 나타나지 않았고 EtOH 추출물의 재분획물인 시료중에서 0.5mg/ml의 농도의 hexan 및 chloroform 분획물은 MN의 발생 빈도가 음성대조구에 비하여 약 2배의 증가로 나타났다 또 뽕나무 줄기의 분획물 시료들에서 0.5mg/ml의 hekan, chloroform 및 EtOAc 분획물에서 MN이 관찰 되었으나 그중 chloroform 분획물 시료에서 더 많은 MN 발생을 보였다. 4) Ames test를 이용한 돌연변이 실험결과 뽕나무 잎 1차 추출물들은 0.25mg/ml의 농도에서 TA98에 대하여 거의 돌연변이 현상이 없었고, EtOH 추출물로부터 재 분획한 BuOH 및 chloroform분 획물에서는 강한 돌연변이 현상이, 또 5mg/ml의 hekan 분획물에서는 더욱 강한 돌연변이 현상이 관찰되었다. TA100 strain에 대하여는 각 재료의 1차 추출물이나 분획들의 낮은 농도 (1.25mg/ml)에서는 돌연변이 현상이 매우 약하거나 거의 없었고, 0.25mg/ml의 1차 추출물들에서는 매우 약한 돌연변이가 나타났으나 이들의 EtOAc 분획물은 다소 강한 돌연변이 유발성이 관찰 되었다. 그러나 수용성 분획물은 두 균주에 대하여 전혀 돌연변이 유발성을 보이지 않았다. 5) 뽕나무 줄기로부터 조제한 시료중 수용성 분획물과 EtOAc분획물은 IA98에서 전혀 돌연변이 유발성이 없었으나 0.25mg/ml 농도외 물 추출물, EtOH 춘출물, hexan 및 chloroffrm분획물들이 약 40%증가한 복귀변이 콜로니가 관찰되었고, 이보다 농도를 증가시킨 물 추출물 및 BuOH 분획물에서도 같은 수준의 복귀변이 콜로니가 관찰 된 결과로부터 돌연변이 유발성이 있음이 확인되었다 TA100에 대하여 뽕나무 줄기의 수용성 분획물은 전혀 돌연변이가 나타나지 않았고, 0.25mg/ml의 물 추출물, EtOH추출물, hexan 및 BuOH분획물들이 약 10%증가한 약한 돌연변이가 관찰되었다. 그러나 농도를 5 mg/ml로 한 EtOAc 분획물은 26의 돌연변이 유발성을 보였다. 이상의 결과로 부터 뽕나무를 이용한 식용 제품생산을 고려할 때 추출물들의 제조와 선택을 가름하는 자료로서의 활용이 기대되며 앞으로 in vivo test 등 더욱 연구할 필요가 있을 것으로 사료된다.

• 한국식품위생안전성학회지
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• 제33권3호
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• pp.207-213
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• 2018

본 연구는 까마귀쪽나무열매추출물(LJF-HE)의 유전독성을 평가하고자 하였다. 유전독성연구는 OECD와 MFDS(Korea Ministry of Food and Drug Safety) 지침에 따라 복귀돌연변이시험, 염색체이상시험, 마우스 골수세포를 이용한 소핵시험을 실시하였다. 세균을 이용한 복귀돌연변이시험은 까마귀쪽나무열매추출물(LJF-HE) 처리군에서 S9 mix 존재유무에 상관없이 복귀돌연변이 콜로니 수는 음성 대조군과 비교하였을 때 증가 양상을 나타내지 않은 반면에 양성 대조물질에서 유발된 복귀돌연변이 콜로니 수는 대사활성계 미적용(S9-) 및 적용(S9+)의 모든 시험 균주에 대하여 음성(용매)대조 값의 2배를 넘어 증가한 것으로 나타났다. 염색체 이상 시험에서 까마귀쪽나무열매추출물(LJF-HE) 처리군에서 모든 세포주의 처리시간 및 S9 mix 존재유무에 상관없이 5%미만의 비정상적인 염색체이상을 나타내었으나, 음성대조군에 비해 유의적인 변화는 없었다. 소핵시험은 까마귀쪽나무열매추출물(LJF-HE) 처리군에서 음성 대조군과 비교하여 소핵을 가진 다염성 적혈구의 증가는 볼 수 없었으며 통계학적인 유의성도 나타나지 않았다. 상기의 결과를 종합하면 까마귀쪽나무열매추출물(LJF-HE)은 유전독성을 유발하지 않는 것으로 판단되어진다.

• 대한화장품학회지
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• 제42권3호
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• pp.211-216
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• 2016

소핵시험은 세포분열 단계 중 간기 세포의 세포질 내 소핵 유무를 조사함으로써 유전독성을 평가하는 시험법이다. 최근 화장품 안전성 평가에 동물실험을 금지하거나 최소화하려는 노력이 확산되고 있어 유전독성 평가에 있어서도 기존의 동물실험이 아닌 새로운 in vitro 시험법이 요구되고 있다. 본 연구에서는 3차원 배양인공피부모델인 KeraSkin$^{TM}$을 이용하여 도포 처치된 물질의 유전독성을 평가하였다. 2종의 유전독성물질인 mitomycin C (MMC)와 methyl methanesulfonate (MMS)는 농도 의존적으로 세포독성과 소핵 형성이 유도된 반면, 대조물질인 4-nitrophenol (4-NP)와 trichloroethylene (TCE)에서는 농도 의존적으로 세포독성은 관찰되었으나 소핵은 형성되지 않았다. 따라서 인공피부모델을 이용한 소핵시험이 화장품과 같은 피부적용물질의 in vitro 유전독성 평가에 유용할 것으로 사료된다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2021년도 춘계학술대회
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• pp.67-67
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• 2021

The genus Glycyrrhiza (Licorice) has been used as an oriental herbal medicine for a long time in Asian countries. Wongam (WG), which is Glycyrrhiza new variety, have been developed to improve limitation of licorice including low productivity, environmental restriction and insufficient components by Korea Rural Development Administration. To using WG as a herbal medicine, it is important to reveal the adverse effects in health. In this study, we evaluated the genotoxicity test of WG extract through in vitro bacterial reverse mutation (AMES) assay, in vitro chromosomal aberration assay and in vivo mouse bone marrow micronucleus assay. When compared with the control, WG extract with or without the S9 mix showed no genotoxicity in the AMES assay up to 5000 ㎍/plate and in the chromosomal aberration assay up to 1100 ㎍/ml. In micronucleus assay, no significant increase in the number of micronucleated polychromatic erythrocytes or in the mean ratio of polychromatic to total erythrocytes up to 5000 mg/kg/day for 2 days. The present study demonstrated that WG extract is safe and reliable herbal medicine since no detectable genotoxic effects at least under the conditions of this study.

• Molecular & Cellular Toxicology
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• 제2권3호
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• pp.176-184
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• 2006

The controversy on genotoxicity of molinate, an herbicide, has been reported in bacterial system, and in vitro and in vivo mammalian systems. To clarify the genotoxicity of molinate, we performed bacterial gene mutation test, in vitro chromosome aberration and mouse lymphoma $tk^{+/-}$ gene assay, and in vivo micronucleus assay using bone marrow cells and peripheral reticulocytes of mice. In bacterial gene mutation assay, no mutagenicity of molinate ($12-185{\mu}g/plate$) was observed in Salmonella typhimurium TA 98, 100, 1535 and 1537 both in the absence and in the presence of S-9 metabolic activation system. The clastogenicity of molinate was observed in the presence ($102.1-408.2\;{\mu}g/mL$) of metabolic activation system in mammalian cell system using Chinese hamster lung fibroblast. However, no clastogenicity was observed in the absence ($13.6-54.3\;{\mu}g/mL$) of metabolic activation system. It is suggested that the genotoxicity of molinate was derived some metabolites by metabolic activation. Molinate was also subjected to mouse lymphoma L5178Y $tk^{+/-}$ cells using microtiter cloning technique. In the absence of S-9 mixture, mutation frequencies (MFs) were revealed $1.4-1.9{\times}10^{-4}$ with no statistical significance. However, MFs in the presence of metabolic activation system revealed $3.2-3.4{\times}10^{-4}$ with statistical significance (p<0.05). In vivo micronucleus (MN) assay using mouse bone marrow cells, molinate revealed genotoxic potential in the dose ranges of 100-398 mg/kg of molinate when administered orally. Molinate also subjected to acridine orange MN assay with mouse peripheral reticulocytes. The frequency of micronucleated reticulocytes (MNRETs) induced 48 hr after i.p. injection at a single dose of 91, 182 and 363 mg/kg of molinate was dose-dependently increased as $10.2{\pm}4.7,\;14.6{\pm}3.9\;and\;28.6{\pm}6.3\;(mean{\pm}SD\;of\;MNRETs/2,000\;reticulocytes)$ with statistical significance (p<0.05), respectively. Consequently, genotoxic potential of molinate was observed in in vitro mammalian mutagenicity systems only in the presence of metabolic activation system and in vivo MN assay using both bone marrow cells and peripheral reticulocytes in the dose ranges used in this experiment. These results suggest that metabolic activation plays a critical role to express the genotoxicity of molinate in in vitro and in vivo mammalian system.

• Toxicological Research
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• 제24권1호
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• pp.11-15
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• 2008

In this research the genotoxic effect of $Polycan^{TM}$ ${\beta}$-glucans originated from Aureobasidium pullulans SM-2001, was evaluated using the mouse micronucleus test. $Polycan^{TM}$ was administered once a day for 2 days by oral gavage to male ICR mice at doses of 1000, 500 and 250 mg/kg. Cyclophosphamide was used as a known genotoxic agent in a positive control group. The appearance of a micronucleus is used as an index for genotoxic potential. The results obtained indicated that $Polycan^{TM}$ shows no genotoxicity effect up to 1000 mg/kg dosing levels. In addition, it is also considered that there were no problems from cytotoxicity of $Polycan^{TM}$ tested in this study because the polychromatic erythrocyte ratio was detected as > 0.47 in all tested groups.

• Toxicological Research
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• 제25권4호
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• pp.225-230
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• 2009

The genotoxic effects of DHU001, a polyherbal formula were evaluated using the mouse micronucleus test. DHU001 was administered once a day for 2 continuous days by oral gavage to male ICR mice at doses of 2000, 1000 and 500 mg/kg. Cyclophosphamide was used as a known genotoxic agent in a positive control. The appearance of a micronucleus is used as an index for genotoxic potential. In addition, the changes on the total white blood cells and differential counts on the lymphocytes, neutrophils, eosinophils, basophils and monocytes in the prepared blood smears were also conducted to observe the possible immunosuppression. The results indicats that DHU001 showed no genotoxicity effects up to 2000 mg/kg dosing levels and did not influenced on the total white blood cells and differential counts. In addition, it is also considered that there were no problems from cytotoxicity of DHU001 tested in this study because the polychromatic erythrocyte ratio was detected as > 0.41 in all tested groups.