• Title/Summary/Keyword: Sulforhodamin B(SRB) assay

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Effect of the various aqua-acupunctures of Hwanggum(黃芩) on proliferation of preadipocyte 3T3-L1 cells (다양(多樣)한 황금약침제제(黃芩藥鍼製劑)가 전지방세포(前脂肪細胞) 3T3-L1의 증식(增殖)에 미치는 영향(影響))

  • Kim, Ho-Kyoung;Kang, Eun-Jung;Ko, Byoung-Seob
    • The Journal of Korean Medicine
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    • v.19 no.1
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    • pp.358-367
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    • 1998
  • These studies were conducted to investigate the effects of the various aqua-aqupunctures of Hwanggum(Scutellariae Radix) on the proliferation of 3T3-Ll cells. They were tested by means of Sulforhodamin B(SRB) assay. The results were summerized as follows: All tested aqua-aqupunctures inhibited the proliferation of preadipose 3T3-Ll cells. In case of the dilution of Hwanggum aqua-aqupuncture(HG), the results were quite opposite. In 1000 times dilution of HG(${\times}1000$), a low concentration increased the proliferation of preadipose 3T3-Ll cells, but the high($100{\mu}l\;and\;200{\mu}l$) concentration inhibited it. These results suggest that Hwanggum aqua-aqupunctures may be used on the obesity induced by the overgrowth of preadipose 3T3-Ll cells.

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Risk assessment on cytotoxicity for benzimidazole fungicides (Benzimidazole계 살균제의 세포독성 평가)

  • Lee, Je-Bong;Sung, Pil-Nam;Jeong, Mi-Hye;Shin, Jin-Sup;Kang, Kyu-Young
    • The Korean Journal of Pesticide Science
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    • v.7 no.3
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    • pp.198-206
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    • 2003
  • To assess potential risk of the benzimidazole fungicides, their cytotoxicities were evaluated. Activities of LDH(Lactic dehydrogenase) in the culture fluid of CHL(chinese hamster lung) fiberoblast cell treated with 4.0, 16.0 or $32.0{\mu}g/mL$ of carbendazim for 24 hours were elevated 2.16, 2.94 and 2.64 folds compared to the control, respectively. DNA synthesis was inhibited by 45% at $2.0{\mu}g/mL$ of carbendazim. Benzimidazole fungicides showed high toxicity to cell and mitochondria of CHL cell by Giemsa and MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide) assay. $IC_{50}$ by the Giemsa assay of thiophanate-methyl, benomyl, carbendazim and captafol were over 125, 1.2, 30.0 and $0.3{\mu}g/mL$, respectively. $IC_{50}$ by the MTT assay of thiophanate-methyl, benomyl, carbendazim and captafol were over 125, 18.7, 20.4 and $2.6{\mu}g/mL$, respectively. Inhibitory concentration of cell median proliferation by SRB (sulforhodamin B) assay for thiophanate-methyl, carbendazim, benomyl, and captafol were 17.4, 5.3, 1.5 and $0.5{\mu}g/mL$, respectively. Accordingly, benzimidazole fungicides inhibited DNA synthesis, mitochondrial function, cell proliferation and induced cell necrosis.

Cytotoxicity of Some Natural Products in Human Cancer Cells (수종의 한국산 야생식물에서의 항암효과 검색)

  • Park, Sung-Hee;Cho, Sung-Ji;Rhee, In-Sun;Lee, Chong-Ock;Kim, Chong-Won;Rhee, In-Ja
    • Korean Journal of Pharmacognosy
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    • v.27 no.4
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    • pp.383-388
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    • 1996
  • The objective of this reseach is to find new antitumoral substances from natural products. Several of natural products have been used as food that were isolated into hexane(Hex.) and/or ethylacetate(EtOAc) extracts. we have tested cytotoxicities of these plants against human solid tumor cells. The cytotoxic activity of these plants were tested using Sulforhodamin B(SRB) assay. Hexane extracts of Chrysanthemum sinense, Allium tubersum. Beta vulgaris, Ixeris dentata have revealed cytotoxicities against five human solid tumor cells, and its cytotoxicities of each cell line were $10-100\;{\mu}l/ml$ ED50 (Effective dose that cause 50% inhibition of cell growth in vitro)

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Effects of Kuseonwangdogo on the Proliferation of Preadipocyte 3T3-L1 Cells, the Anti-Complementary and the Cytotoxic Effects (구선왕도고가 전지방세포(前脂肪細胞) 3T3-L1의 증식(增殖), 항보체활성(抗補體活性) 및 세포독성(細胞毒性)에 미치는 영향)

  • Choi, Young-Heu;Kim, Ho-Kyoung;Ko, Byoung-Seob;Ju, Young-Sung
    • The Journal of Korean Medicine
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    • v.20 no.3 s.39
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    • pp.105-114
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    • 1999
  • To investigate the anti-complementary and cytotoxic effects of oriental prescription, Kuseonwangdogo, on the proliferation of preadipocyte 3T3- L1 cells, we examined biological effects of Kuseonwangdogo. The results obtained were as follows. 1. After 14 days, the body weight of rats treated with Kuseonwangdogo decreased more than that in the control group (p<0.05). However, the weights of liver, spleen and kidney were unchanged. In serum biochemical test, we examined the level of glucose (GLU) and glutamic pyruvic transaminase (GPT). The levels of GOT and CHOL in serum were decreased remarkably by the administration of Kuseonwangdogo (p<0.05). The haematological examination of the tested group showed significant increment of white blood cells (WBC), hemoglobin concentration (HGB), mean corpuscular hemoglobin (MCH) and monocyte (MO). 2. The effect of Kuseonwangdogo on the proliferation of 3T3-L1 cells was tested by the sulforhodamin B(SRB) assay. The high concentration ($100{\mu}l\;and\;200{\mu}l$) of extracts inhibited the proliferation of 3T3- L1 cells. The p-value was <0.01, respectively. 3. The extract of Kuseonwangdogo showed a potent anti -complementary activity. It was suggested that the active principle may be a kind of polysaccharide molecule. 4. The cytotoxic effects of Kuseonwang dogo and its composing herbs in human liver cells (WRL68) and monkey kidney cells (Vero) were examined by the SRB and 3- (4,5- Dimethylthiazol-2-yl) -2,5 diphenyl-2H- tetrazolium bromide (MTT) assay. Cytotoxic effects were not observed.

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Brine Shrimp Lethality of the Compounds from Phryma leptostachya L.

  • Lee, Sang-Myung;Min, Byung-Sun;Kho, Yung-Hee
    • Archives of Pharmacal Research
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    • v.25 no.5
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    • pp.652-654
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    • 2002
  • Brine shrimp assay-guided fractionation and isolation of the EtOAc soluble fraction of Phryma leptostachya L. (Phrymacaceae) gave two active compounds, phrymarolin II (1) and ursolic acid (2), which were identified by physicochemical and spectroscopic methods. Compound 1 exhibited potent lethality with $LD_{50}$ value of 0.0013 $\mu\textrm{g}$/ml, whereas 2 showed moderate lethality with $LD_{50}$ value of 27.0 $\mu\textrm{g}$/ml against brine shrimp. The cytotoxic activities of 1 and 2 were also evaluated against one murine and five human cancer cell lines employing the sulforhodamin B (SRB) method. Compound 2 exhibited cytotoxic activity against L1210 and SK-MEL-2 cells with $ED_{50}$ values of 3.70 and 9.27 mg/ml, respectively, whereas 1 was devoid of any cytotoxic activity against all cancer cells tested.

Influence of Yeoldahanso-tang on the Hypoxic Damage of Cultured Cerebral Neurons from mouse and SK-N-MC cells (열다한소탕(熱多寒少湯)이 저산소성(低酸素性) 대뇌신경세포(大腦神經細胞) 손상에 미치는 영향(影響))

  • Kim, Hyoung-Soon;Bae, Young-Chun;Lee, Sang-Min;Kim, Kyung-Yo;Won, Kyoung-Sook;Sihm, Gyue-Hearn;Park, Su-Jeong
    • Journal of Sasang Constitutional Medicine
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    • v.15 no.1
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    • pp.72-89
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    • 2003
  • To elucidate the neuroprotective effect of Yeoldahanso-tang(YHT) on nerve cells damaged by hypoxia, the cytotoxic effects of exposure to hypoxia were determined by XTT(SODIUM3,3'-{I-[(PHENYLAMINO) CARBONYL]-3,4-TETRAZOLIUM}- BIS (4-METHOXY-6-NITRO) BENZENE SULFONIC ACID HYDRATE), NR(Neutral red), MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and SRB(Sulforhodamin B) asssay. The activity of catalase and SOD(Superoxide dismutase) was measured by spectrophometry, and $TNF-{\alpha}$(Tumor cell necrosis $fector-{\alpha}$) and PKC(Protein kinase C) activity was measured after exposure to hypoxia and treatment of YHTWE. Also the neuroprotective effect of YHTWE was researched for the elucidatioion of neuroprotective mechanism. The results were as follows; 1. Hypoxia decreased cell viability measured by XTT, NR assay when cultured cerebral neurons were exposed to 95% N2/5% CO2 for $2{\sim}26$ minutes in these cultures and YHTWE inhibited the decrease of cell viability. 2. H2O2 treatment decreased cell viability measured by MTT, and SRB assay when cultured cerebral neurons were exposed to 1-80 ${\mu}M$ for 6 hours, but YHTWE inhibited the decrease of cell viability. 3. Hypoxia decreased catalase and SOD activity, and also $TNF-{\alpha}$ and PKC activity in these cultured cerebral neurons, but YHTWE inhibited the decrease of the catalase and SOD activity in these cultures. 4. Hypoxia triggered the apoptosis via caspase activation and internucleosomal DNA fragmentation. Also hypoxia stimulate the release of cytochrome c forom mitochondria. YHTWE inhibited the apoptosis via caspase activation induced by hypoxia. From these results, it can be suggested that brain ischemia model induced hypoxia showed neurotoxicity on cultured mouse cerebral neurons, and the YHTWE has the neuroprotective effect in blocking the neurotoxicity induced by hypoxia in cultured mouse cerebral neurons.

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The Effects of Plum Extracts on the Proliferation of Human Epithelial Cell and Human Cervical Cancer Cells (자두 추출물이 인체 상피세포와 자궁경부암세포의 증식에 미치는 효과)

  • Han, Man-Deuk;Kweon, Dur-Han;Kang, Byung-Tae;Lee, Jae-Woo;Yoon, Ok-Hyun
    • Journal of the East Asian Society of Dietary Life
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    • v.17 no.5
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    • pp.710-718
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    • 2007
  • This study was designed to investigate the effects of plum(Prunus salicina Lindl. cultivars 'Oishiwase', 'Formosa', and 'Soldam') extracts on the proliferation as well as inhibition of human epithelial cells(HaCaT), human cervical carcinoma (HeLa, SiHa, and C33A) cells, and human stomach adenocarcinoma(SNU 638) cells. Dried plum was sequentially extracted and fractionated by hexane(KC-01), chloroform(KC-02), ethyl acetate(KC-03), n-butanol(KC-04), water(KC-05), methanol(KC-6), and hot water extract(KC-07). The epithelial and cancer cells were exposed for 48 h to $50{\mu}g/mL$ of plum extract in vitro, and were then analysed by a sulforhodamin B(SRB) staining assay. The methanol extract(KCP-6) of 'Formosa' proliferated not only the HaCaT cells(147.3%), but also the cervical carcinoma C33A cells(167.8%). The ethyl acetate extract of 'Soldam'(KCJ-3) significantly reduced the proliferation rate of the HPV positive conical carcinoma cells, at 61.5% for the SiHa cells and 70.5% for the HeLa cells. In the C33A cells, which are HPV negative cervical carcinoma cells, the hexane fractions of 'Formosa'(KCP-1) and 'Oishiwase'(KCD-1) markedly suppressed proliferation activity at 20.4% and 61.7%, respectively. However, the proliferation rate of the normal epithelial cells(HaCaT cell) was not reduced the proliferation rate by KCJ-3, KCP-1, or KCD-1, There were no significant effects on proliferation of the stomach cancer cells(SNU 638) by any of the extracts or fractions of the plum cultivars. These results suggest that the anti-proliferative effects of the plum cultivars were selective to the cancer cell origin. In conclusion, we found that several plum cultivar extracts, especially, the ethyl acetate fraction of 'Soldam" and the hexane fraction of "Formosa', have anti-proliferative activity toward human cervical carcinoma cells. However, further investigation is needed to assess the molecular mechanisms that mediate the antiproliferation activities of the plum cultivars.

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