• 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.

• 대한본초학회지
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• 제32권4호
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• pp.1-8
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• 2017

Objectives : Silkworm is known as immunomodulatory substances and contain various bioactive compounds such as serine, tyrosine and alanine. The aim of this study was to investigated the immunopotentiating activity of combine extract that silkworm and food materials (Eucommia ulmoides, Angelica gigas, Acanthopanax, Allium hookeri, Cinnamomum cassia, Liriope platyphylla, Curcuma longa, Achyranthes japonica, Alpinia oxyphylla, Adenophora triphylla). Methods : Among 10 kinds of food materials, to select food materials with the effect of enhancing the immune function mouse splenocyte proliferation ability was measured by 3-[4,5-dimethylthiazol-2-y]-2,5-diphenyl terazolium bromide (MTT) assay. Then, combine extract of silkworm and food materials were evaluated that mouse splenocyte proliferation ability by EZ-cytox cell viability assay. Morever, cytokines production such as IL-2, IL-4, IL10, IL12, $IFN-{\gamma}$ on mouse T lymphocyte stimulated with concanavalin A (ConA) was measured. Results : Eucommia ulmoides, Acanthopanax, Allium hookeri, Cinnamomum cassia, Liriope platyphylla has high proliferation ability of mouse splenocyte compared with Curcuma longa, Achyranthes japonica, Alpinia oxyphylla, Adenophora triphylla. The silkworm and food material combined extract has a relatively high proliferation ability of mouse splenocyte proliferation when the silkworm and food materials are used as a single material. In particularly, combined extract of silkworm and Cinnamomum cassia was stimulate cytokine production on T lymphocyte such as IL12, $IFN-{\gamma}$. Combined extract of silkworm and Liriope platyphylla was stimulate cytokine production on T lymphocyte such as IL2, IL4, IL10. Conclusion : In conclusion, the combined extract of the silkworm and Cinnamomum cassia or Liriope platyphylla may enhance immune function by regulating mouse splenocyte proliferation and stimulating cytokine production.

• 한국환경성돌연변이발암원학회:학술대회논문집
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• 한국환경성돌연변이발암원학회 2003년도 추계학술대회
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• pp.109-110
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• 2003

• Toxicological Research
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• 제11권2호
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• pp.187-192
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• 1995

The cytotoxic effects of hydrogen peroxide and neuroprotective effects of a variety of agents were investigated in newborn mouse forebrain tissue culture. In our experiments, oxygen radical was generated enzymatically by glucose oxidase and the values were expressed as a percentage of number of living cells by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Cytotoxicity of oxygen radicals was prevented by catalase and (N, N, N', N', -tetrakis (2-pyridylmethyl) ethylenediamine (TPEN), but N-tetra-ot-butyl-phenylnitrone (PBN), and deferoxamine (DFX), failed to show protective effects against oxygen radicals. Antagonists of the N-methyl-D-aspartate (NMDA) receptor, D-2-amino-5-phosphonovaleric acid (APV), 7-chlorokynurenic acid (CKA), and MK801 (a non-competitive NMDA antagonist) were also not effective in blocking neurotoxicity induced by glucose oxidase generated oxygen radicals.

• 동의생리병리학회지
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• 제17권2호
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• pp.447-450
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• 2003

To clerify the cytotoxicity of reactive oxygen species in cultured hippocampal neurons of neonatal mouse, toxic effect was measured by MTT assay after cultured cells were incubated for 3 hours in the media containing 1～40 μM concentrations of H₂O₂. In addition, the protective effect of vitamin E was determined in these cultrures. Cell viability was significantly decreased in a dose-dependent manner after exposure of 10 μM H₂O₂ to cultured mouse hippocampal neurons for 5 hours. In the protective effect of vitamin E, vitamin E prevented the H₂O₂-induced cytotoxicity in these cultures. From these results, it suggests that H₂O₂ has toxic effect in cultured mouse hippocampal neurons and vitamin E has protective effect on the cytotoxicity induced by H₂O₂.

• 한국식품영양학회지
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• 제20권2호
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• pp.240-244
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• 2007

In this study, we found that the methanolic extract of Areca catechu repressed expression of the tyrosinase gene in B16 mouse melanoma cells containing a tyrosinase promoter. Extract concentrations of 100 ${\mu}$g/ml and 500 ${\mu}$g/ml exhibited tyrosinase gene expression rates of aproximately 62% and 48%, respectively, compared to the control. The fraction layers consisting of ethyl acetate, butyl alcohol, and water showed repressive effects on the tyrosinase gene. In particular, the butyl alcohol fraction highly repressed at 100 ${\mu}$g/ml and 500 ${\mu}$g/ml. In the MTT assay, the methanolic extracts exhibited very low cytotoxicities at 1 ${\mu}$g/ml, 10 ${\mu}$g/ml, and 100 ${\mu}$g/ml.

• 대한한방피부미용학회지
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• 제1권1호
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• pp.5-15
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• 2005

The purpose of this study was to investigate the effect of the aqueous extract of the Astragalus membranaceous(AM). AM showed inhibitory effect on the tyrosinase activity using L-tyrosine as a substrate. Tyrosinase plays an important role in the process of melanin polymer biosynthesis. In vitro AM extract(1mg/ml) inhibited melanin biosynthesis and are useful for the material used in cosmetics. B16 mouse melanoma cells were cultured in different concentrations. The non-cytotoxicity of the plant extracts was confirmed by MTT assay.

• 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.

• Journal of Microbiology and Biotechnology
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• 제23권4호
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• pp.527-533
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• 2013

Polyclonal antibodies labeled with a tracer have been commonly used as secondary antibodies in immunochemical assays to quantify the concentration of antibody-antigen complexes. The majority of these antibodies conjugated with a tracer are commercially available, with the exception of few untouched targets. This study focused on the production and application of mouse anti-quail IgY as an intermediate antibody to link between quail egg yolk IgY and goat anti-mouse IgG-HRP as primary and secondary antibodies, respectively. Subsequently, the produced mouse anti-quail IgY was labeled with horseradish peroxidase (HRP) and its efficiency on enzyme linked immunosorbent assay (ELISA) was compared with that of commercial rabbit anti-chicken IgY-HRP. As an intermediate antibody, mouse anti-quail IgY was successfully produced with good affinity and sensitivity (1:10,000) to the primary and secondary antibodies. Subsequently, mouse anti-quail IgY was effectively conjugated with HRP enzyme, resulting in a secondary antibody with good sensitivity (1:10,000) to quail anti-V. parahaemolyticus and V. vulnificus IgY. The detection limit was $10^5$ CFU/ml for both V. parahaemolyticus and V. vulnificus. The efficiency of the produced conjugate to detect quail IgY on ELISA was comparable to that of the commercial rabbit anti-chicken IgY-HRP, and hence the produced and labeled mouse anti-quail IgY-HRP can be used as a secondary antibody to detect any antibody produced in quail.

• 동의생리병리학회지
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• 제20권4호
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• pp.896-901
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• 2006

Zingiberis rhizoma(ZB) has been used to treat a various condition and disease in traditional oriental medicine. The present study was conducted to evaluate the immunomodulatory effect of aqueous-extracted ZB(ZBE) on methotrexate (MTX)-induced immune suppression in mouse spleen cells. In spleen cell proliferation assay, ZBE enhanced mitogenic activity in mouse spleen cells. In RT-PCR, ZBE induced IL-2, IFNr and IL-6 cytokine gene expression in mouse spleen cells. In spite of MTX treatment, IL-2, IFNr and IL-6 gene expressions sustained in MTX treated spleen cells. CD45R/B220, pan B marker was slightly increased in ZBE treated mouse spleen cells. IL-6, B cell tropical cytokine, production was induced by ZBE-treated mouse spleen cells and IL-6 production was sustained on MTX-ZBE co-cultured cells. ZBE administration enhanced suNival of S-180 bearing mouse. These data indicate that ZBE has a protective effect of immune suppression caused by MTX, and ZBE may be enhance cellular and humoral function by regulate cytokine gene expression as well as the mitogenic effect on spleen cells.