• Toxicological Research
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• 제30권3호
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• pp.211-220
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• 2014

Resveratrol has received considerable attention as a polyphenol with various biological effects such as anti-inflammatory, anti-oxidant, anti-mutagenic, anti-carcinogenic, and cardioprotective properties. As part of the overall safety assessment of HS-1793, a novel resveratrol analogue free from the restriction of metabolic instability and the high dose requirement of resveratrol, we assessed genotoxicity in three in vitro assays: a bacterial mutation assay, a comet assay, and a chromosomal aberration assay. In the bacterial reverse mutation assay, HS-1793 did not increase revertant colony numbers in S. typhimurium strains (TA98, TA100, TA1535 and TA1537) or an E. coli strain (WP2 uvrA) regardless of metabolic activation. HS-1793 showed no evidence of genotoxic activity such as DNA damage on L5178Y $Tk^{+/-}$ mouse lymphoma cells with or without the S9 mix in the in vitro comet assay. No statistically significant differences in the incidence of chromosomal aberrations following HS-1793 treatment was observed on Chinese hamster lung cells exposed with or without the S9 mix. These results provide additional evidence that HS-1793 is non-genotoxic at the dose tested in three standard tests and further supports the generally recognized as safe determination of HS-1793 during early drug development.

• Journal of Ginseng Research
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• 제43권4호
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• pp.562-571
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• 2019

Background: Ginsenosides have been widely used clinically for many years and were regarded as very safe. However, a few researches on the toxicities of these kinds of agents showed that some ginsenosides may have side-effect on the rats or dogs. So it is extremely necessary to further clarify the potential toxicity of ginsenosides. This study was carried out to investigate long-term toxicity and genotoxicity of 25-methoxydammarane-3, 12, 20-triol ($25-OCH_3-PPD$), a new derivative of ginsenoside, in beagle dogs. Methods: Twenty-four beagle dogs were divided randomly into four treatment groups and repeatedly orally administered with $25-OCH_3-PPD$ capsule at 60, 120, and 240 mg/kg/day for 91 consecutive days. Ames, micronucleus, and chromosomal aberration tests were established to analyze the possible genotoxicity of $25-OCH_3-PPD$. Results: There was no $25-OCH_3-PPD$einduced systemic toxicity in beagle dogs at any doses. The level of $25-OCH_3-PPD$ at which no adverse effects were observed was found to be 240 mg/kg/day. The result of Ames test showed that there was no significant increase in the number of revertant colonies of $25-OCH_3-PPD$ administrated groups compared to the vehicle control group. There were also no significant differences between $25-OCH_3-PPD$ administrated groups at all dose levels and negative group in the micronucleus test and chromosomal aberration assay. Conclusion: The highest dose level of $25-OCH_3-PPD$ at which no adverse effects were observed was found to be 240 mg/kg per day, and it is not a genotoxic agent either in somatic cells or germs cells. $25-OCH_3-PPD$ is an extremely safe candidate compound for antitumor treatment.

• 방사선산업학회지
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• 제9권4호
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• pp.181-185
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• 2015

Today the biosafety evaluation, a common problem of vital importance, is based on internationally proved test-systems, standards and techniques. The paradigm of biosafety includes multidisciplinary approach, a combination of physical, chemical and biological tests to monitor the environmental level of pollutants and needs to be improved by modern approaches. The genetic risk of environmental pollutions has long been studied by many researchers. In this study, used was the known sensitive plant test-system, clones of plant Tradescantia (spiderwort) able to detect gene mutations (frequency of mutational events and formation of micronuclei) in combination with chemical and, in some instances, with radiological measurements. In addition, male gametophyte generation of fruit trees was applied as bioindicators of genotoxicity. The obtained results did not show any significant increase along with wind direction. As for the male gametophyte assay, the fertility of the investigated fruit-trees near to NPP did not significantly differ from that of the control point. The influence of the NPP on the male generative system of the investigated taxa of fruit trees for the investigated year was not revealed. The system described needs to be expanded by species of interest (human) as there is a difficulty to transfer the revealed dose correlations to humans. The development of this idea includes various levels: population (epidemiological studies), individual, cellular, molecular (DNA), etc.

• 대한약침학회지
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• 제27권1호
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• pp.38-46
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• 2024

Objectives: Genotoxicity is evaluated through a chromosomal aberration test using cultured mammalian cells to determine the toxicity of no-pain pharmacopuncture (NPP), which has recently been used to treat musculoskeletal pain disorders in Korean medical clinical practice. Methods: An initial test was performed to determine the dosage range of the NPP, followed by the main test. In this study, NPP doses of 10.0, 5.0, and 2.5%, and negative and positive controls were tested. An in vitro chromosome aberration test was performed using Chinese hamster lung cells under short-term treatment with or without metabolic activation and under continuous treatment without metabolic activation. Results: Compared with the saline negative control group, NPP did not significantly increase the frequency of chromosomal abnormalities in Chinese hamster lung cells, regardless of the presence or absence of metabolic activation. Additionally, the number of cells with structural chromosomal abnormalities was significantly higher in the positive control group than that in the negative control group that received saline. Conclusion: Based on the above results, the chromosomal abnormality-producing effect of NPP was determined to be negative under these test conditions.

• Environmental Analysis Health and Toxicology
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• 제30권
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• pp.14.1-14.7
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• 2015

Objectives Cigarette smoking is associated with carcinogenesis owing to the mutagenic and genotoxic effects of cigarette smoke. The aim of this study was to evaluate the mutagenic and genotoxic effects of Korean cigarettes using in vitro assays. Methods We selected 2 types of cigarettes (TL and TW) as benchmark Korean cigarettes for this study, because they represent the greatest level of nicotine and tar contents among Korean cigarettes. Mutagenic potency was expressed as the number of revertants per ${\mu}g$ of cigarette smoke condensate (CSC) total particulate matter whereas genotoxic potency was expressed as a concentration-dependent induction factor. The CSC was prepared by the International Organization for Standardization 3308 smoking method. CHO-K1 cells were used in vitro micronucleus (MNvit) and comet assays. Two strains of Salmonella typhimurium (Salmonella enterica subsp. enterica ; TA98 and TA1537) were employed in Ames tests. Results All CSCs showed mutagenicity in the TA98 and TA1537 strains. In addition, DNA damage and micronuclei formation were observed in the comet and MNvit assays owing to CSC exposure. The CSC from the 3R4F Kentucky reference (3R4F) cigarette produced the most severe mutagenic and genotoxic potencies, followed by the CSC from the TL cigarette, whereas the CSC from the TW cigarette produced the least severe mutagenic and genotoxic potencies. Conclusions The results of this study suggest that the mutagenic and genotoxic potencies of the TL and TW cigarettes were weaker than those of the 3R4F cigarette. Further study on standardized concepts of toxic equivalents for cigarettes needs to be conducted for more extensive use of in vitro tests.

• 대한약침학회지
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• 제19권3호
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• pp.213-224
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• 2016

Objectives: Ginseng Rh2+ is enzyme-treated ginseng extract containing high amounts of converted ginsenosides, such as compound k, Rh2, Rg3, which have potent anticancer activity. We conducted general and genetic toxicity tests to evaluate the safety of ginseng Rh2+. Methods: An acute oral toxicity test was performed at a high-level dose of 4,000 mg/kg/day in Sprague-Dawley (SD) rats. A 14-day range-finding study was also conducted to set dose levels for the 90-day study. A subchronic 90-day toxicity study was performed at dose levels of 1,000 and 2,000 mg/kg/day to investigate the no-observed-adverse-effect level (NOAEL) of ginseng Rh2+ and target organs. To identify the mutagenic potential of ginseng Rh2+, we conducted a bacterial reverse mutation test (Ames test) using amino-acid-requiring strains of Salmonella typhimurium and Escherichia coli (E. coli), a chromosome aberration test with Chinese hamster lung (CHL) cells, and an in vivo micronucleus test using ICR mice bone marrow as recommended by the Korean Ministry of Food and Drug Safety. Results: According to the results of the acute oral toxicity study, the approximate lethal dose (ALD) of ginseng Rh2+ was estimated to be higher than 4,000 mg/kg. For the 90-day study, no toxicological effect of ginseng Rh2+ was observed in body-weight changes, food consumption, clinical signs, organ weights, histopathology, ophthalmology, and clinical pathology. The NOAEL of ginseng Rh2+ was established to be 2,000 mg/kg/day, and no target organ was found in this test. In addition, no evidence of mutagenicity was found either on the in vitro genotoxicity tests, including the Ames test and the chromosome aberration test, or on the in vivo in mice bone marrow micronucleus test. Conclusion: On the basis of our findings, ginseng Rh2+ is a non-toxic material with no genotoxicity. We expect that ginseng Rh2+ may be used as a novel adjuvant anticancer agent that is safe for long-term administration.

• 한방재활의학과학회지
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• 제31권1호
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• pp.59-79
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• 2021

Objectives This study was performed to observe the genotoxic effect of the ChondroT. Methods To evaluate the genotoxicity of ChondroT, an experiment of bacterial reverse mutation test, in vitro mammalian chromosomal aberration test and mammalian erythrocyte micronucleus test in mouse was conducted. Results TA98, TA100 and TA1537 strains in the absence of metabolic activation system (S9 mix), the number of revertant colonies being greater than 2-fold of the respective negative control value. Both in -S9 mix and +S9 mix, the frequencies of aberration cells with structural aberration and numerical aberrations of chromosome were less than 5%. There was no increase of polychromatic erythrocyte with one or more micronuclei at any dose of test substance compared to the negative control group (p<0.05). Conclusions In TA98, TA100 and TA1537 strains in the absence of metabolic activation system (S9 mix), the number of revertant colonies was greater than 2-fold of the respective negative control value, showing positive results. ChondroT was considered to be non-clastogenic to Chinese hamster lung (CHL/IU) cells under the present experimental condition. and ChondroT was determined not to induce an increased frequency of micronuclei in the bone marrow cells of male ICR mice under the present experimental condition.

• 한국식품영양과학회지
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• 제31권5호
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• pp.910-916
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• 2002

생약재의 식품ㆍ생물 산업적 이용증대에 따라 생약재의 안전한 위생화 기술이 요구되고 있다 본 연구에서는 생약재의 위생화 기술로서 방사선 조사기법의 활용 가능성을 검토하기 위하여, 감마선을 조사한 생약재 3종에 대한 유전독성학적 안전성을 평가하고자 하였다. 공시 재료는 오염유기체 완전 구제선량인 10 kGy의 감마선을 조사시킨 황기, 백출 및 승마로 하였으며, 각각의 열수 추출물의 유전독성을 in vitro 시험으로 평가하였다. 유전독성 평가는 Salmonella typhimurium TA98 및 TA100 균주를 이용한 복귀 돌연변이 시험(Ames test)과 Chiilese hamster ovary(CHO) 세포를 이용한 in vitro 소핵 유발 시험으로 시행하였다. 각각의 시험은 59 nix를 첨가한 대사 활성화 시스템과 첨 가하지 않은 비활성화 시스템으로 구분하여 실시하였으며, 시료의 최고 처리 농도는 복귀돌연변이 시험에서는 5mg/plate로, 소핵유발시험에서는 50%의 세포증식 억제를 나타내는 농도(1 mg/mL)로 하였다. 복귀 돌연변이 시험 결과 대사 활성화 및 비활성화의 경우 모두에서 각 시료에 의한 복귀변이 집락수의 증가를 인정할 수 없었으며, 각 용량단계에서 감마선 조사군과 비조사군 간의 차이도 볼 수 없었으므로 음성으로 판정하였다. 소핵 유발시험에서도 음성 대조군 및 감마선 조사군과 비조사군 모두 각 용량 단계에서 세포 내에 생성된 소핵의 빈도가 3% 이하로 나타남에 따라, 시료에 의한 소핵의 유발을 인정할 수 없었으므로 음성으로 판정하였다 따라서 감마선이 조사된 각각의 시료는 직접 및 간접 돌연변이원으로 작용하지 않으며 세포유전 독성을 나타내지 않음을 확인할 수 있었다. 향후, 생체내 유전독성 시험, 만성독성 시험 및 생식독성 시험 등의 추가적인 in vivo실험이 행하여진다면 감마선 조사 생약재의 안전성을 보다 명확히 밝힐 수 있을 것으로 생각된다.

• Molecular & Cellular Toxicology
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• 제1권3호
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• pp.179-188
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• 2005

To develop the novel anti-allergic drug, many sophoricoside derivatives were synthesized. Among these derivatives, JSH-II-3, VI-3, VII-3, VIII-3, VII-20 and VII-20 (sodium salt) were selected and subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of quantity. Single cell gel electrophoresis (Comet) assay, mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay (MOLY), chromosomal aberration 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. Through the primary screening using the comet assay, we could choose the first candidates of sophoricoside derivatives with no genotoxic potentials as JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt). Also JSH-VII-3, VII-20 and VII-20 (sodium salt) are non-mutagenic in MOLY assay, while JSH-II-3 is mutagenic at high concentration with the presence of metabolic activation system in both comet assay and MOLY assay. The selected derivatives (JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt) are not mutagenic in S. typhimurium TA98 and TA100 strains both in the presence and absence of metabolic activation. From results of chromosomal aberration assay, 6 h treatment of JSH-VI-3, VII-3 and VII-20 (sodium salt) were not revealed clastogenicity both in the presence and absence of S-9 mixture. Therefore, we suggests that JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt), as the optimal candidates with both no genotoxic potential and IL-5 inhibitory effects must be chosen. To process the development into new anti-inflammatory drug of these derivatives, further investigation will need.

• 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.