• Korean Journal of Environmental Agriculture
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• v.16 no.2
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• pp.149-155
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• 1997

This study was carried out to investigate the effects of phenobarbital sodium(PB) and 3-methylcholanthrene(3-MC) on carbofuran cytotoxicity and to develop antitoxic agents based on the effectivness. Experimental groups for carbofuran cytotoxicity were divided into five groups ; medium alone and four treatments of carbofuran (1, 25, 50 and $100{\mu}M)$, and those for compensation effects were divided into six groups ; medium alone, $IC_{50}$ carbofuran and four combinations of carbofuran and PB or 3-MC($IC_{50}$ carbofuran plus 1, 25, 50, $100{\mu}M$ of PB and 3-MC, respectively). After incubation for 48 hrs under the same conditions, MTT(Tetrazolium MTT), NR(Neutral red) and SRB(Sulforhodamine B protein) assay were performed. Fifty percentage inhibition of MTT, NR, and SRB against carbofuran in rat fibroblast cell were 60.7, 82.5 and $87.0{\mu}M$, respectively. At the combination treatments of $IC_{50}$ of carbofuran and $100{\mu}M$ of PB, the significant compensation effects were observed from the results of MTT and NR but not from that of SRB absorbance. And at the combination treatments of $IC_{50}$ of carbofuran and 3-MC, the relatively significant compensation effects were found at $50{\mu}M$ 3-MC from the results of MTT and at $100{\mu}M$ 3-MC from that of NR and SRB absorbances, respectively. From the results of light microscopy, combination treatments of $carbofuran(IC_{50})$ and PB or 3-MC showed good regeneration in carbofuran toxicity of rat fibroblast cells. These results suggest that PB or 3-MC can compensate the cytotoxity of carbofuran insecticide in rat NIH3T3 fibroblast cells.

• Preventive Nutrition and Food Science
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• v.3 no.2
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• pp.169-174
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• 1998

Antimutagenic and anticarcinogenic effects of alginic acids extracted from sea mustard(SM) and sporophyII of sea mustard(SSM) were studied by Salmonella typhimurium assay system and cytotoxicity and transformation tests using C3H/10T1/2 cells, respectively. alginic acid-SM andalginic acid-SSM showed antimutagenic effects on aflatoxin B1(AFB1)and N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) in Salmonella typhimurium TA100 strain. The antimutagenic effect showed concentration dependent manner. At the 2.5mg/plate concentration , alginic acid-SSM exhibited 92% antimutagenicity against AFB1 ,while alginic acid-SM revealed 54% antimutagenictity ,s howing effectiveness of the alginic acid-SSM for the antimutagenicity. Alginic acidSSm also significantly decreased the cytotoxicity induced by 3-methylcholanthrene(MCA) and MNNG in C3H/10T1/2 cells (p<0.05). The type II and type IIItransformation foci formation by MCA and MNNG were also decreased when the alginic acid-SSM was treated, indicating that the alginic acid -SSM reduces the carcinogenesis induced by these carcinogens. The MCA-treated culture produced 10.5foci of type II +III in C3H/10T1/2 cells, however, MCA + 0.2mg/ml alginic acid-SSM treated culture formed only 1.8 foci of the types II + III in C3H/10T1/2 cells, however , MCA+0.2mg/ml alginic acid -SSM treated culture formed only 1.8 foci of the types II+ III(p<0.05). While MNNG-treated culture formed 13.0 foci, MNNG + 0.2mg/ml alginic acid -SSM treated one produced 3.0 foci of type II+III(p<0.05). These results suggest that alginic acid-SSM can effectively prevent the mutagenicities and also decrease cytotoxicity and transformation induced by some carcinogens.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1993.04a
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• pp.96-96
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• 1993

간부분 절제술을 하지 않는 비수술적 방법으로서 D-galactosamine을 이용한 중기발암성 시험의 개발을 목적으로 F344 수괵 랫드를 이용하여 본 실험을 수행하였다. 실험 I 에서는 실험방법에 따라 3가지 모델로 구분하고, 각 모델에 처치군과 대조군을 두었다. 모델 1 에서는 실험개시시에 diethylinitrosamine (DEN)을 200 mg/kg body weight로 복강내로 1회 투여하고, 실험개시후 2및 5주에 D-galactosamine을 300 mg/kg body weight로 복강내로 각각 1회 투여하였다. 처치군에는 실험개시후 2주부터 6주간 2-acetylaminofluorene을 0.01%로 혼합한 사료를 급여하였으며, 대조군에는 기초사료를 계속 급여하였다. 모델 2에서는 모델 1의 4주차까지의 처치를 2회 반복하였다. 모델 3은 간부분 절제술을 하는 DEN-PH (diethylnitrosamine-partial hepatectomy) 모델과 같은 방법으로 처치하였다. 사육기간 중 매주 체중 및 사료소비량을 측정하였고, DEN 투여후 8주에 전동물을 부검하여 적출한 간의 중량을 측정하고, glutathione S-transferase placental form (GST-P) 양성 foci에 대한 면역조직화학적 염색표본을 만들어 GST-P 양성 foci의 수 및 면적을 측정하였다. 실험 II에서는 모델 1의 방법으로 phenobarbital(PB), 3-methylcholanthrene (3-MC), n-ethyl-n'-nitro-n-nitrosoguanidine 및 3,3'-diaminobenzidine외 GST-P 양성 foci의 발현정도를 조사하였다. 실험 I의 결과, 모델 1이 정상적인 체중 증가를 보여주었으며, 간조직의 GST-P 양성 foci 의 발현율이 가장 좋았다. GST-P 양성 foci의 면적은 큰것 부터 미상엽, 내측우엽, 외측우엽의 순으로 나타났으나 foci의 수는 모델별로 다르게 나타났다. 실험 II의 PB 투여군과 3-MC 투여군에서 GST-P 양성 foci의 수 및 면적의 유의성 있는 증가가 관찰되었다. 이와 같은 결과로 볼때, 비수술적 방법인 D-galactosamine 을 이용한 중기 발암성 검색법은 간부분 절제술을 이용한 중기발암성 검색법에 비하여 GST-P 양성foci의 발현능력이 동등하거나 더 우수하였으며, 간 및 간이외 장기의 발암물질에 대한 발암성 검색에 보다 유용할 것으로 생각된다.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.3 no.1
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• pp.67-84
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• 1997

Bujeonghangamtang(扶正抗癌湯) has been used for cure of tumor as a traditional medicine without any experimental evidence to support the rational basis for its clinical use. This study was carried out to evaluate the possible therapeutic or antitumoral effects of Bujeonghangamtang extract against tumor, and to carry out some mechanisms responsible for its effect. Some kinds of tumor were induced by the typical application of 3-methylcholanthrene. (MCA) or by the implantation(s.c) of malignant tumor cells such as leukemia cells(3LL cells) or sarcoma cells(Sl80 cells). Treatment of the Bujeonghangamtang water-extract (dailly 1mg／mouse, i. p.) was continued for 7 days prior to tumor induction and after that the treatment was lasted for 20 days. Against squamous cell carcinoma induced by MCA, Bujeonghangamtang decreased not only the frequency of tumor production but also the number and the weight of tumors per tumor bearing mice (TBM). Bujeonghangamtang also significantly suppressed the development of 3LL cell and S180 cell-implanted tumors in occurrence-frequency and their size, and some developed tumors were regressed by the continuous treatment of Bujeonghangamtang extract into TBM. In vitro, treatment of Bujeonghangamtang extract had no effect on the growth of some kinds of cell line such as FsaII, A431 strain but significantly inhibited the proliferation of 3LL, S180 cells and augmented the DNA synthesis of mitogen-activated lymphocytes. Bujeonghangamtang also stimulated the migrative ability of leukocyte, the MIF and IL-2 production of T lymphocytes, but not IL 6 production of B cells. Bujeonghangamtang-administration to mice enhanced NK cells activities. These results demonstrated that Bujeonghangamtang extract exhibited a significant prophylactic benefits against tumors and its antitumor activity was manifested depending on the type of tumor cells. And these results also suggested that effect of Bujeonghangamtang might be chiefly due to nonspecific enhancement of NK cell activities and cell-mediated immune responses.

• Toxicological Research
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• v.3 no.2
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• pp.97-110
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• 1987

The effects of 5 novel hepatotrophic agents, dithiol malonate derivatives (DMDs; DMD1-DMD5), on the liver microsomal lipid peroxidation induced by carbon tetrachloride $(CCl_4)$ and the correlations with the changes of microsomal electron transport system were investigated. All DMDs were found to inhibit the lipid peroxidation induced by $CCl_4$ in mice and rats as well in vitro liver microsomal system. Therefore, each DMD seemed to have direct mode of action on liver microsomes to inhibit the lipid peroxidation. As an ex vivo study, the induced lipid peroxidation by $CCl_4$ and the changes in electron transport system were determined with liver microsomes obtained from rats chronically treated with DMDs for 7 days. The induced lipid peroxide contents in liver microsomal system were lower in DMD1, DMD2 and DMD3 treated group, but higher in DMD4 and DMD5 group when compared to the control group. Cyt. p.450 contents in the microsomes were decreased by the treatment with DMD1, DMD2 and DMD3, but increased significantly by DMD4 with great extent and by DMD5 with less extent. The cyt. p-450 isozymes induced by treatment of DMD4 and DMD5 were identified as 3-methylcholanthrene (MC) type. The NADPH cyt. -C reductase activities of the microsomes treated with DMD1, DMD2, DMD4 and DMD5 were increased in the range of around 20% to 50%, but decreased with DMD3, All DMDs increased dyt. $-b_5$ content and did not alter NAdH-cyt, $-b_5$ reductase activities in the microsomes. In summary, the 5 novel hepatotrophic agents (DMDs) markedly protected against lipid peroxidation induced by $CCl_4$ in vivo and in vitro possibly through the mechanism of direct action on the liver microsomes. The degree of inhibition produced by DMDs on lipid peroxidation induced by $CCl_4$ seemed to coincide rather with cyt. p-450 contents than with other components of liver microsomal electron transport system including NADPH-cyt, -C reductase.

• Herbal Formula Science
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• v.5 no.1
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• pp.147-168
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• 1997

Tish study was carried out to evaluate the possible therapeutic or antitumoral effects of Takrisodokyeum extract against tumor, and immunomodulatory effect. Some kinds of tumor were induced by the typical application of 3-methylcholanthrene (MCA) or by the implantation(s.c) of malignant tumor cells such as leukemia cells(3LL cells) or sarcoma cells(S-I80 and Fas II cells). Treatment of the Takrisodokyeum water-extract(daily 1mg mouse, i.p.) was continued for 7 days prior to tumor induction and after that the treatment was lasted for 15 days. Against squamous cell carcinoma induced by MCA, Takrisodokyeum decreased not only the frequency of tumor production but also the number and the weight of tumors per tumor bearing mice (TBM). Takrisodokyeum also significantly suppressed the development of 3LLcell and S-180 cell by frequency and their size, and some developed tumors were regressed by the continuous treatment of Takrisodokyeum extract into TBM. However, when tumor was induced by FsaII cell-implantation, the growth of implanted cells in mice was delayed by the water extract of Takrisodokyeum until day 7 and then rapid growth ensued. In vitro, treatment of Takrisodokyeum extract had no effect on the growth of some kind of cell lines such as FsaII, A-131 strain but significantly inhibited the proliferation of 3LL, S-180 cells. Takrisodokyeum also stimulated the migrative ability of leucocyte, the MIF and IL 2-production of T lymphocytes, but not IL 6 production of B cells. Takrisodokyeum enhanced Arthus reaction and DTH to sheep erythrocytes, and NK cells activities. These results demonstrated that Takrisodokyeum extract different results according to the type of tumor cells. And these results also suggested that antitumor effect of Takrisodokyeum might be chiefly due to nonspecific enhancement of NK cell activities and cell-mediated immune responses.

• BMB Reports
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• v.32 no.3
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• pp.226-231
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• 1999

In vitro effects of acetone on cytochrome P450 (P450)-dependent benzo(a)pyrene (B(a)P) hydroxylation supported by cumene hydroperoxide (CuOOH) or NADPH/$O_2 $ systems were studied using 3-methylcholanthrene-pretreated rat liver microsomes. The maximal rate of B(a)P hydroxylation at constant concentration ($80\;{\mu}M)$ of the substrate was observed in the presence of $30\;{\mu}M$ CuOOH. However, at concentrations higher than $30\;{\mu}M$ CuOOH the hydroxylation rates were rapidly decreased. In contrast to CuOOH, at a concentration of $200\;{\mu}M$ NADPH, B(a)P hydroxylation rate reached a plateau. At concentrations higher than $200\;{\mu}M$ NADPH, the rates of substrate hydroxylation were maintained at the maximal rate with no inhibition. Acetone at 1% (v/v) enhanced both CuOOH- and NADPH/$O_2$-supported B(a)P hydroxylation at the optimal concentrations of the cofactors. At concentrations higher than 1% (v/v) acetone, substrate hydroxylation was sterero specific under the support of these two cofactors; it was strongly enhanced with $30\;{\mu}M$ CuOOH, but rather inhibited in the $200\;{\mu}M$> NADPH/$0_2 $ system. The lipid peroxidation rate induced during CuOOH-supported P450-dependent B(a)P hydroxylation was increased as CuOOH concentrations were increased. Acetone in the concentration range of 2.5~7.5%(v/v) inhibited lipid peroxidation during CuOOH supported B(a)P hydroxylation. The finding that CuOOH-supported B(a)P hydroxylation is greatly enhanced by acetone suggests that acetone may contribute more to the activation of oxygen (for the insertion of oxygen into the substrate) in the presence of CuOOH than with NADPH/$O_2$. Acetone may also contribute to the partial inhibition of destruction of microsomal membranes by lipid peroxidation.

• Korean Journal of Medicinal Crop Science
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• v.15 no.6
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• pp.437-443
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• 2007

This study was carried out to investigate the effect of dietary supplementation with red ginseng water-extracts on the induction of microsomal cytochrome P-450 in rats. Phenobarbital (PB) and 3-methylcholanthrene (3-MC), P-450 inducers, were administered to 3- or 12-month old rats received red ginseng extracts (25 mg/kg) from 6 weeks to 12 months for 3 days. PB and 3-MC increased levels of P-450, P-450 reductase, ethoxycoumarin O-deethylase, benzphetamine N-demethylase and glutathione-S-transferase in the liver of rats. However, chronic administration of red ginseng significantly reduced these increase of enzyme levels induced by P-450 inducers. Chronic administration of red ginseng did not affect the induction of cytochrome $b_5$ and NADH cytochrome $b_5$ reductase by P-450 inducers. It is suggested that the induction of cytochrome P-450 system in the liver in relation to xenobiotics toxicity can be modulated by long-term supplementation with Korean red ginseng to rats.

• Korean Journal of Food Science and Technology
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• v.30 no.1
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• pp.206-212
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• 1998

This study was undertaken to explore the applicability of glutathione S-transferase (GST) activity as a predictable indicator to monitor chemical pollution in shells and fishes utilized for food. There were some variations in the basal level of GST activity depending on species tested. Ark shells, Anadara satowi, showed the highest normal enzyme activity, followed by catfish and marine mussels, Mytilus coruscus. White clams, Meretrix lusoria, Israeli carp and catfish had lower activity. When A. satowi was exposed to 3-methyl-cholanthrene (3-MC), a prototypic polycyclic aromatic hydrocarbon for 1 week, GST activity decreased by about 30%. This reduction in GST activity induced by 3-MC did not recover until two weeks after the cessation of exposure. GST activity increased in response to 3-MC in most of the other species studied. The GST elevation in M. coruscus attained its maxinum of about 200% at the termination of 3-MC exposure maintaining this level up to 2 weeks, and declined gradually thereafter. 3-MC also induced GST activity in lsraeli carp in a similar fashion to M. coruscus. Phenobarbital induced GST activity both in M. coruscus and lsraeil carp. Other chemicals. such as clofibrate, butylated hydroxyanisole. hexachlorobenzene, and oxolinic acid did not change the enzyme activity significantly in most speciel. Phenol depressed GST activity only in lsraeli carp. These results suggest that the basal level of GST activity is somewhat variable and that the direction of change in response to chemicals seems to be related to its normal activity. The change in enzyme activity can be a preditable indicator of some environmental chemicals such as PAHs and phenol.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.3 no.1
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• pp.169-191
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• 1997

Hyangpaeyangyeongtang, which was first prescribed in the book of Euijonggumgam (醫宗金鑑), has been widely applied to the treatment of scrofula and carbuncle in the neck and tumor as well in the oriental medicine. The Fritillariae roylei bulbus was also used as an anti-tumor medicine. In order to investigate the effect of Hyangpaeyangyeongtang and Fritillariae roylei bulbus extracts on the immune responses to the epithelioma induced by 3-methylcholanthrene and prostaglandin-$F_2{\alpha}$ in mice, the change of epithelioma size was observed and delayed type hypersensitivity, hemagglutinin titer, hemolysin titer, rosette forming cells, natural killer cell activity, lymphocyte transformation, productivity of interleukin-2 and phagocytic activity were also measured. The results were as follows; 1. The epithelioma size was decreased with statistical significance in the Hyangpaeyangyeongtang group at the 14th day, as compared with the control group. 2. Delayed type hypersensitivity was not shown any decrease in the Hyangpaeyangyeongtang group and the Fritillariae roylei bulbus group, as compared with the control group. 3. Hemagglutinin titer was increased with statistical significance in the Hyangpaeyangyeongtang group, but not in the Fritillariae roylei bulbus group as compared with the control group. 4. Hemolysin titer was increased with statistical significance in the Hyangpaeyangyeongtang group, but not in the Fritillariae roylei bulbus group as compared with the control group. 5. Rosette forming cells were not increased in the Hyangpaeyangyeong tang group, but increased with statistical significance in the Fritillariae roylei bulbus group as compared with the control group. 6. Natural killer cell activity at the ratio 100:1 of effector cells to target cells was increased with statistical significance in the Hyangpaeyayeongngtang group and the Fritillariae roylei bulbus group respectively, and the natural killer cell activity at the ratio of 50:1 and 10:1 of effector cells to target cells Hyangpaeyangyeongtang group was increased with statistical significance as compared with the control group. 7. Lymphocyte transformation was increased in the Hyangpaeyangyeong tang group and the Fritillariae roylei bulbus group respectively, as compared with the control group. 8. Productivity of interleukin-2 was increased in the Hyangpaeyangyecing tang group and the Fritillariae roylei bulbus group respectively, as compared with the control group. 9. Phagocytic activity was not shown any decrease in the Hyangpaeyangyeongtang group and the Fritillariae roylei bulbus group, as compared with the control group.