• KSBB Journal
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• 제15권4호
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• pp.331-336
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• 2000

여름과 가을에 수확한 연진쑥을 열수 빛 에탄올로 추출하 여 B(a)P의 변이원성에 대한 억제효과를 SOS Chromotest로 시험한 결과 여름에 수확한 시료의 물 추출물에서 강한 억제 효과를 나타내었다. 따라서 에탄올 가용성 분획과 불용성 분 획으로 분리하였으며 분획별 돌연변이 억제효과는 불용성 분 획에서 더 높게 나타났다. 불용성 분획은 SOS Chromotest외 A Ames test에서 정확한 용량의존성 억제효과를 나타내었고, 50% 돌연변이 억제농도 $(IC_{50}$는 E. coli PQ37에 대하여는 $200{\mu}g/assay$, S. typhimurium TA98에 대하여는 $800{\mu}b/plate$ TAIOO에서는 $600{\mu}g/plate$이었다. 그러나 세포내.외 항돌연변이 효과를 비교한 결과 세포내 항돌연변이 효과는 나타내지 않았다 따라서 인진쑥 물 추출물의 돌연변이 억제효과는 d desmutagenic effect에 의한 것으로 확인되었다 HPLC를 사용 하여, B(a)P의 변이원성에 주된 효소인 cytochrome P-4S0 1A1에 의해 대사되어지는 aflatoxin Mj 농도를 측정한 결 과 AFM1의 형성이 크게 감소되었다. B(a)P의 변이원성에 대 한 인진쑥 불추출물의 항돌연변이 효과는 아마도 B(a)P를 ultimate mutagen으로 대사시키는 cytochrome P-4S0 1A1 효소계를 저해하여 나타나는 것으로 해석된다.

• 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.

• 한국연초학회지
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• 제13권1호
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• pp.19-26
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• 1991

Ames test using special strains which are histidine requiring mutant of Salmonella typhimufium , is widely utilized as short-term bioassay to evaluate the genotoxicity of chemicals. This method requires the liver microsome(5-9 fraction) to provide mammalian metabolism of the compounds, Therefore, the mutagenic potency of the chemicals is affected by not only the intrinsic properties of them but also the efficiency of the in vitro microsomal activation system. For this reason, the complex mixtures such as environmental pollutants from occupational sources, natural products or cigarette smoke condensates(CSC) , might be often appeared the false results. Induction of sister chromatid exchange(SCE) in cultured cells is known as another sensitive and powerful tool for the measurement of genotoxicity and the method has also an advantage which is able to apply to the in vitro and in vivo systems. In the present study, the inducibilities of revertant colonies in tester strain TA98 and SCE in human Iymphocytes by positive controls and total particulated materials(TPM) obtained from various brand(domastic and imported) cigarettes were compared in order to investigate whether the results in Ames test are in agreement with those in SCE analysis. The mutagenic activities of well known mutagens such as B(a)P showed excellent dose-response in the both methods although the induction mechanism was different each other, but cyclophosphamide resulted such effect only in SCE analysis. Most TPM tested showed a similar pattern in the mutagenic activities in those methods. However, only two(one imported brand and one domestic sample cigarettes) among the TMP obtained from various cigarettes appeared the higher induction in SCE than Ames test.

• 생명과학회지
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• 제15권2호
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• pp.215-219
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• 2005

감궁탕의 돌연변이원성을 유무를 알아보기 위해 Salmonella typhimurium TA 98 및 TA100을 이용한 돌연변이 원성 실험에서도 감궁탕은 어느 균에서도 돌연변이원성을 나타내지 않았으며, 이는 S-9 mixture 의해 감궁탕이 대사가 된 후에도 이와 유사한 경향을 나타내었다. 또한, SOS umu test의 경우에서도 $\beta-galactosidase$활성에는 별다른 영향을 미치지 않는 것으로 보아 감궁탕은 돌연변이원성을 일으키지 않는 것으로 판정되었으며 S-9 mixture처리에 의한 대사 후에도 이와 유사한 실험결과가 나타났다. 따라서 감궁탕은 그 자체 및 대사 후에도 DNA에 별다른 영향을 미치지 못하는 비교적 안전한 생약처방으로 여겨진다.

• 한국환경성돌연변이발암원학회지
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• 제8권1호
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• pp.13-21
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• 1988

Mutagenic actions of aflatoxin B$_1$ (AFB$_1$) in the presence of various concentrations of L-ascorbIc acid (AA) in Salmonella typhimurium strains TA 100 and TA98 were studied. Spontaneous revertants per plate of the tester strains TA100 and TA98 were 121-125 and 25-30 with or without S9 mix, respectively. The negative controls used in the study did not show any mutagenesis in the tester strains. AFB$_1$ revealed strong mutagenicity at the dose levels of 0.05, 0.1 and 0.25 ${\mu}$g/plate with metabolic activation system in both strains. However, it showed a toxic effect when the levels were more than 0.5 ${\mu}$g/plate. When lower concentrations of AA (5-20 ${\mu}$g/plate) were added to AFB$_1$ in the Ames assay system with S9 mix the mutagenic action of AFB$_1$ decreased in both strains. About 70-90% of mutagenicity of AFB$_1$ disappeared in strain TA100 when 20${\mu}$g of AA was added to 0.05 ${\mu}$g of AFB$_1$. The inhibitory effect was greatly increased by the addition of higher concentrations of AA to AFB$_1$ in TA100 strain. The mutagenicity of AFB$_1$ was completely inhibited when 100 ${\mu}$g and 500 ${\mu}$g of AA were added to 0.05 ${\mu}$g and 0.1 ${\mu}$g of AFB$_1$, respectively, However, this protective effect of AA on AFB$_1$ mediated mutagenesis was less effective in TA98 strain than that in TA100.

• Toxicological Research
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• 제17권
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• pp.319-321
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• 2001

Taihu Lake is a major water source for part of Yangtze Delta, which is one of the most urbanized and economically prosperous areas in China. In last couple of decades, some parts of the lake were highly polluted due to eutrophication. This study analyzed dioxin-like potential and mutagenic potential in surface water samples from Taihu Lake. The samples were prepared by XAD-2 resin procedure. A batch of biological assays, including dioxin-like potential microassay with the rat hepatocyte cell line H411E, and Ames test was employed in the research. Results showed that jour water samples have high content of dioxin-like biological potential, the highest activity TEQ to 2,3,7,8-TCDD was 48 pg/ι in sample 1. The mutagenic effect with reading-frame shifting mechanism was confirmed in 3 of 4 samples. The effective sewage treatment facilities and reliable monitoring surveillance system are urgently needed for this area.

• 한국환경성돌연변이발암원학회지
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• 제22권2호
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• pp.106-111
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• 2002

AG6O, the complex of acriflavine and guanosine, has been shown to possess the synergistic antitumorigenic activity in the previous paper (J. Pharm. Pharmacol. 1997, 49:216). In this study, we have investigated the genotoxic properties of AG60 using in vitro and in vivo system such as Ames bacterial reversion test, chromosomal aberration assay and micronucleus assay. In Ames reverse mutation test, AG60 treatment at the dose range up to 250 $\mu\textrm{g}$/plate caused the dose-independent random induction of the mutagenic colony formation in S. typhimurium TA98, TA100, TA1537, and E. coli WP2uvrA, while any mutagenic effect of AG60 wasn't observed in S. typhimurium TA1535. Any significant chromosomal aberration wasn't observed in chinese hamster lung (CHL) fibroblast cells incubated with PBS or AG60 at the concentrations of 2.5, 5, 10 $\mu\textrm{g}$/$m\ell$ for 24 hours without but even with 59 metabolic activation system for 6 hours. In vivo ICR mice, the intramuscular injection of AG60 at the doses of 7.15, 14.3, and 28.6 mg/kg did not induce the frequency of micronucleus formation. However, mitomycin C, as one of the positive controls at the dose of 2 mg/kg caused the 8.4% induction in the frequency of micronucleus and 24% increase in the chromosomal aberration.

• 생약학회지
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• 제36권2호통권141호
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• pp.93-96
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• 2005

Mutagenicity of Gamgung-tang (GGT) was tested using in vitro S-9 mixture in vitro host-mediated assay with Salmonella typhimurium. In the previous reports, GGT was tested for the safety using Ames(-S-9), Bacillus subtilis Rec, and umu gene expression mutagenicity tests. Mutagenic activity in any assays we tested was not found. In this report, we further investigated safety of GGT after metabolic activation in vivo. Ames test with S-9 mixture and host-mediated assay with Salmonella typhimurium TA98 were used to identify metagenic property of GGT. GGT was administered 3 times with i.m. to Balb/c mice did not induced mutagenic effect in Salmonella typhimurium TA98 recovered from the liver after 3.5h with i.p. treatment. Over the entire dose range $(3{\sim}150mg/mouse)$ tested no toxicity was detected to the bacterial cells. These results suggest that there was no DNA damage and mutagenicity by GGT.

• 한국수산과학회지
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• 제19권2호
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• pp.127-135
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• 1986

D-glucose-glycine계로 부터 조제한 Maillard 반응생성물을 한외여과로 각 분자량별로 분획(분자량 1,000이하, $1,000{\sim}5,000$, 5,000이상)하고, 투석에 의하여 비투석성 melanoidin을, 오존처리에 의하여 오존처리 melanoidin을 각각 얻었다. 이들 각 시료를 아미노산 및 단백질의 가열분해 유래의 변이원성 물질인 TrP-P-1, TrP-P-2, Glu-P-1, Glu-P-2 및 IQ에 각각 작용($37^{\circ}C$, 30분) 시켜서, 변이원성억제효과를 검토하였다. 그 결과, Maillard반응생성물의 변이원성억제효과는 반응생성물의 분자량의 크기에 비례하여 높게 나타났다. Maillard 반응생성물의 환원력 및 항산화력 또한 분자량이 큰 획분일수록 크게 나타났다. 그러나, Sodium borohydride로 melanoidin을 환원시켰을 때, melanoidin의 변이원성억제효과 및 환원력이 감소하였다. 또한, Trp-P-1의 일부가 melanoidin 분자중에 흡착되는 것이 밝혀졌고, 카르 보닐화합물(diacetyl 및 glyceraldehyde)로 이들 변이원성물질의 아미노기를 수식함으로써 변이원성물질의 변이원활성이 크게 저하하였다. 따라서, Maillard 반응생성물 즉 melanoidin의 변이원성억제효과는 melanoidin의 환원력 및 항산화능을 비롯하여 정전기적인 흡착 및 melanoidin 분자중의 카르보닐기에 기인한다고 추찰된다.

• 한국환경독성학회:학술대회논문집
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• 한국환경독성학회 2003년도 춘계학술대회
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• pp.183-184
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• 2003

Sophoricoside was isolated as the inhibitor of IL-5 bioactivity from Sophora japonica (Leguminosae). It has been reported to have an anti-inflammatory effect on rat paw edema model. To develop as an anti-allergic drug, genotoxicity of sophoricoside was investigated in bacterial and mammalian cell system such as Ames bacterial test, chromosomal aberration assay, Comet assay and MOLY assay. In Ames test, sophoricoside of 5000 ∼ 313 $\mu\textrm{g}$/plate concentrations was not shown significant mutagenic effect in Salmonella typhimurium TA98, TA100, TA1535 and TA1537 strains. The cytotoxicity (IC$\_$50/ and IC$\_$20/) of sophoricoside was determined above the concentration of 5000 $\mu\textrm{g}$/ml in Chinese hamster lung (CHL) fibroblast cell and L5178Y mouse lymphoma cell line. At concentrations of 5000, 2500 and 1250 $\mu\textrm{g}$/ml, this compound was not induced chromosomal aberration in CHL fibroblast cell in the absence and presence of S-9 metabolic activation system. Also in comet assay, DNA damage was not observed in L5178Y cell line. Also in MOLY assay, sophoricoside of 5000 ∼ 313 $\mu\textrm{g}$/ml concentrations was not shown significant mutagenic effect in absence of S-9 metabolic activation system. However, the higher concentration of 5000 and 2500 $\mu\textrm{g}$/ml of sophoricoside induced the increased mutation frequency (MF) in the presence of S-9 metabolic activation system. From these results, no genotoxic effects of sophoricoside observed in bacterial systems whereas, genotoxic effects observed in mammalian cell systems in the presence of metabolic activation system. These results suggested that the metabolite(s) of sophoricoside can cause some genotoxic effects in mammalian cells.