• Title/Summary/Keyword: Sulforhodamine B (SRB) protein assay

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A Study on the Effects of Ramulus et Uncus Uncariae (REUU) on the Cultured Spinal Dorsal Root Ganglion Neurons Damaged by Oxygen Free Radicals (조구등(釣鉤藤)이 산소자유기(酸素自由基)에 의하여 손상(損傷)된 배영척수감각신경절세포(培養脊髓感覺神經節細胞)에 미치는 영향(影響)에 관(關)한 연구(硏究))

  • Kang, Hyung-Won;Park, Jin-Sung
    • Journal of Oriental Neuropsychiatry
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    • v.11 no.1
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    • pp.1-18
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    • 2000
  • To study the effects of Ramulus et Uncus Uncariae (REUU) on oxygen free radical-mediated damage by hydrogen peroxide $(H_{2}O_{2})$ on cultured spinal sensory neurons, in vitro assays such as MTT assay, NR assay, neurofilament enzymeimmuno assay (EIA), sulforhodamine B (SRB) assay, assay for lactate dehydrogenase (LDH) activity and assay for lipid peroxidation were used in cultured spinal dorsal root ganglion neurons derived from mice, Spinal dorsal root ganglion neurons were cultured in media containing various concentrations of $H_{2}O_{2}$ for 5 hours, after which the neurotoxic effect of $H_{2}O_{2}$ was measured by in vitro assay. The protective effect of the herb extract, Ramulus et Uncus Uncariae (REUU) against H2O2-induced neurotoxicity was also examined. The results are as follows. 1. In NR assay and MTT assay, $H_{2}O_{2}$ significantly decreased the cell viability of cultured mouse spinal dorsal root ganglion neurons according to exposure concentration in these cultures. An additional time course study was done on these cultures. 2. Cultured spinal dorsal root ganglion neurons which were exposed to various concentrations of $H_{2}O_{2}$ showed a quantitative decrease of neuronal cells by EIA and of total protein by sulforhodamine B (SRB) assay, while they showed an increase of both lipid peroxidation and LDH activity. 3. The effect of Ramulus et Uncus Uncariae (REUU) on $H_{2}O_{2}$ induced neurotoxicity showed a quantitative increase in both neurofilament and total protein, but showed a decrease of lipid peroxidation and LDH activity. These results suggest that $H_{2}O_{2}$ has a neurotoxic effect on cultured spinal dorsal root ganglion neurons from mice and that the herb extract, Ramulus et Uncus Uncariae (REUU), was very effective in protecting $H_{2}O_{2}$ induced neurotoxicity by decreasing lipid peroxidation and LDH activity.

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Antitumor Evaluation of Cannabidiol and Its Derivatives by Colorimetric Methods

  • Baek, Seung-Hwa;Shin, Ji-Hee;Chung, Woo-Young;Han, Du-Seok
    • Toxicological Research
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    • v.16 no.2
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    • pp.101-107
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    • 2000
  • Cannabidiol derivatives (1, 2 and 3), 5-fluorouracil (4, 5-FU) and adriamycin (5, AM) were tested for their growth inhibitory effects against human tumor cell lines using two different 3-{4,5-dimeth-ylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and sulforhodamine B protein (SRB) assay. The light microscopic study showed morphological changes of the treated cells. Disruptions in cell organelles were determined by colorimetric methods; MTT assay and STB assay. These results suggest that cannabidiol (1, CBD) retains the most growth-inhibitory activity against human tumor cell lines.

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The Inhibitory Effects of Taraxaci Herba against Cadmium induced Cytotoxicity (포공령의 카드뮴에 대한 세포독성 억제효과)

  • Han, Du-Seok;Lee, Ki-Nam;Lee, Jong-Sub;Baek, Seung-Hwa
    • YAKHAK HOEJI
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    • v.42 no.3
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    • pp.307-311
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    • 1998
  • This study was carried out to evaluate antitoxic effects Taraxaci Herba extract against Cadium by calorimetric methods. The antitoxic activity of Taraxaci Herba ex tract in NIH 3T3 fibroblasts was evaluated by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-phenyl-2H-tetrazoliumbromide), NR (Neutral red) and SRB (Sulforhodamine B protein) assay. The light microscopic study was carried out to observe morphological changes of the treated cells. These results were obtained as follows; The concentration of $10^{-2}mg/ml$ of Taraxaci Herba extract was shown significant antitoxic activity. The number of NIH 3T3 fibroblasts were antitoxic and tend to regenerate. These results suggest that Taraxaci Herba extract retains a potential antitoxic activity.

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Boron Trifluoride Etherate on Silica-A Modified Lewis Acid Reagent (VII). Antitumor Activity of Cannabigerol Against Human Oral Epitheloid Carcinoma Cells

  • Baek, Seung-Hwa;Kim, Young-Ok;Kwag, Jung-Suk;Choi, Kyw-Eun;Jung, Woo-Young;Han, Du-Seok
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.353-356
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    • 1998
  • Geraniol (1), olivetol (2), cannabinoids (3 and 4) and 5-fluorouracil (5) were tested for their growth inhibitory effects against human oral epitheloid carcinoma cell lines (KB) and NIH 3T3 fibrobalsts using two different 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and sulforhodamine B protein (SRB) assay. Cannabigerol (3) exhibited the highest growth-inhibitory activity against the cancer cell lines.

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Development of Anticancer Agents from Korean Medicinal Plants (Part 8). - Cytotoxic Activity of Taraxaci Herba Extract against Human Skin Melanoma Cells - (한국산 생약으로부터 항암물질의 개발(제 8보) - 포공령 추출물이 인체 피부흑색종세포에 미치는 세포독성작용 -)

  • Oh, In-Kio;Yoo, Eun-Ah;Han, Du-Seok;Kang, Kil-Ung;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.29 no.3
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    • pp.198-203
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    • 1998
  • In the present study, we have evaluated cytotoxic effects of Taraxaci herba extract on human skin melanoma cells. The light microscopic study showed morphological changes of the treated cells. Disruptions in cell organelles were determined by calorimetric methods: MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide), NR (Neutral red) and SRB (Sulforhodamine B protein) assay. These results suggest that Taraxaci herba retains a potential antitumor activity.

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Antitumor Evaluation of Epigallocatechin Gallate by Colorimetric Methods (비색분석법에 의한 Epigallocatechin Gallate의 항암효과평가)

  • Baek, Soon Ok;Kim, Il Kwang;Baek, Seung Hwa;Han, Du Seok
    • Journal of the Korean Chemical Society
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    • v.42 no.4
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    • pp.411-415
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    • 1998
  • In the present study, we were evaluated cytotoxic effects of epigallocatechin gallate in human skin melanoma cells such as HTB-69. The light microscopic study showed morphological changes of the treated cells. Disruptions in cell organelles were determined by colorimetric methods; 3-(4,5-dimethyl thiazol-2-yl)-2,5diphenyl-2H-tetrazolium bromide (MTT) assay, neutral red (NR) assay and sulforhodamine B protein (SRB) assay. These results suggest that epigallocatechin gallate retains a potential antitumor activity.

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Development of Anticancer Agents from Korean Medicinal Plants. Part 6. -Cytotoxic Activity of the Ethyl Acetate Soluble Fraction of Lonicerae flos against Human Oral Epitheloid Carcinoma Cells- (한국산 생약으로부터 항암물질의 개발(제6보). -금은화 Ethyl Acetate 가용성 분획의 인체 구강유상피암종세포에 미치는 세포독성작용-)

  • Han, Du-Seok;Baek, Kyong-Hyun;Kim, Young-Ok;Choi, Kyu-Eun;Kwag, Jung-Suk;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.29 no.1
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    • pp.22-27
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    • 1998
  • This study was carried out to develop antitumor agents based on effects of the ethyl acetate soluble fraction of the methanolic extract of Lonicerae flos on human oral epitheloid carcinoma cells. Human oral epitheloid carcinoma cells were cultured in RPMI-1640 media containing 10% fetal bovine serum, antibiotic, and fungizone. After incubation for 24 hrs, the cells were treated with A, B, C, D, and E fractions for 48hrs under the same condition. MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide), NR (Neutral red) and SRB (Sulforhodamine B protein) assay were performed. The light microscopic findings were observed by inverted microscope. In MTT assay, fraction B was shown significant antitumor activity (P<0.001), fraction E was shown significant antitumor activities (P<0.05), but the other fractions were not shown. In NR assay, fraction B was shown significant antitumor activity (P<0.001). In SRB assay, fractions B was shown significant antitumor activities (P<0.01). fractions A and D were shown significant antitumor activities (P<0.05). but the other fractions were not shown. In light microscopy. the fraction B of the ethyl acetate soluble fraction of the methanolic extract of Lonicerae flos showed the highest antitumor activity. These finding suggested that fraction B possessed the most antitumorous agent.

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The antiproliferative activity of cannabidiol ethyl ethers against human ora epitheloid carcinoma cells

  • Baek, Seung-Hwa;Kang, Kil-Ung;Chung, Soon-Ryang;Kim, Hyung-Min;Chung, Woo-Young;Han, Du-Seok
    • Advances in Traditional Medicine
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    • v.1 no.1
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    • pp.45-54
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    • 2000
  • Cannabidiol derivatives (1, 2 and 3), and 5-fluorouracil (4, 5-FU) were tested for their growth inhibitory effects against human oral epitheloid carcinoma cell lines (KB) using two different 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and sulforhodamine B protein (SRB) assay. These compounds showed a potent inhibitory activity in vitro in the micromolar range against KB cell lines. In general, the antitumor activity of these compounds (1, 2, 3 and 4) was in a dose-dependent over the micromolar concentration ranges from $1\;{\mu}M\;to\;100\;{\mu}M$. The comparison of $IC_{50}$ values of these compounds in tumor cell lines shows that their susceptibility to these compounds decreases in the following order: CBD > 5-FU > CBDME > CBDDE by the MTT assay and SRB assay. Cannabidiol derivatives (1, 2 and 3), and 5-FU were tested for their cytotoxic effects on NIH 3T3 fibroblasts using two different MTT assay and SRB assay. These compounds exhibited potent cytotoxic activities in vitro in the micromolar range against NIH 3T3 fibroblasts. In general, the cytotoxic activities of these compounds (1, 2, 3 and 4) were in a dose-dependent over the micromolar concentration range $1\;{\mu}M\;to\;100\;{\mu}M$. The comparison of $CD_{50}$ values of these compounds on NIH 3T3 fibroblasts shows that their susceptibility to these compounds decreases in the following order; CBD > 5-FU > CBDDE > CBDME by MTT assay, CBD > 5-FU > CBDME > CBDDE by SRB assay. These results suggest that cannabidiol (1, CBD) retains the most growth-inhibitory activity against KB cell lines.

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Development of Antitoxic Agents from Korean Medicinal Plants (Part 3) Repaired Effects of Methanol Fraction of Perilla Frutescens on 3T3 Fibroblast Treated with Cadmium (한국산 생약으로부터 해독물질의 개발(제3보) 카드뮴으로 처리한 3T3섬유모세포에서 소엽 메탄올 분획의 수복효과)

  • 한두석;유현경;곽정숙;백승화
    • Toxicological Research
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    • v.11 no.2
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    • pp.229-234
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    • 1995
  • This study was carried out to evaluate the cytotoxicity of cadmium on 3T3 fibroblast and to develop the antidote on 3T3 fibroblast which was injuried by $IC_{50}$ of cadmium. The groups for repaired effects were divided into 7 groups such as medium alone treated group. Cadmium treated $IC_{50}$ groups and 5 experimental groups $(IC_{50}$ cadmium plus $10^{-4}$ concentration of each methanol fraction). After incubation for 48 hrs in the same conditions, MTT (tetrazolium MTT), NR (neutral red) and SRB (sulforhodamine B protein) assay were measured. Light microscopic observations were also investigated. The ethyl acetate fraction of Perilla frutescens showed significantly repaired effect against cadmium cytotoxiclty and this fraction inhibited critical cell regeneration in light microscopy.

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Development of Anticancer Agents from Korean Medicinal Plants. Part 7. Cytotoxic Activity of the Chloroform soluble Fraction of Perrila frutescens Against Human Oral Epitheloid Carcinoma Cells (한국산 생약으로 부터 항암물질의 개발 (제7보), 소엽의 Chloroform 가용성 분획이 인체 구강유상피암종세포에 미치는 세포독성작용)

  • Han, Du-Seok;Kim, Young-Il;Choi, Kyw-Eun;Kwag, Jung-Suk;Baek, Seung-Hwa
    • Environmental Mutagens and Carcinogens
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    • v.18 no.1
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    • pp.37-42
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    • 1998
  • In the present study, we have evaluated cytotoxic effects of the chloroform soluble fraction of the methanolic extract of Perilla frutescens in human oral epitheloid carcinoma cells. The light microscopic study showed morphological changes of the treated cells. Cell membrane damaging activity was measured by the lactate dehydrogenase (LDH) assay and disruptions in cell organelles were determined by 3-(4,5-dime-thylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), neutral red (NR) and sulforhodamine protein B (SRB) of colorimetric assay. These results suggest that Perilla frutescens retains a potential antitumor activity.

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