• Title/Summary/Keyword: MTT and SRB assay

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Genotoxicity and Cytotoxicity in Human Cancer and Normal Cell Lines of the Extracts of Rhododendron brachycarpum D. Don leaves (만병초 잎 추출물의 유전 독성과 사람의 암세포주 등에 대한 세포독성)

  • Byun, Kyoung-Sup;Lee, Young-Woo;Jin, Hyou-Ju;Lee, Mi-Kyoung;Lee, Hyeon-Yong;Lee, Kun-Jae;Heo, Moon-Young;Yu, Chang-Yeon;Lee, Jin-Ha
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.4
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    • pp.199-205
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    • 2005
  • This study was carried out to investigate the effect of 70% ethanol extract and each fraction from Rhododendron brachycarpum D. Don leaves on cytotoxicity, anticancer, genotoxicity and immunological activity in vitro bioassay. Cytotoxicity for human normal cells (HEL299 and Chang) of the samples was shown below 35% in 0.5 mg/ml concentration of samples except aqueous fraction by SRB assay. DNA damage on the Chang cell of the samples alone in comet assay was observed very weak damage activity even in high concentration (1 mg/ml) of the samples. The anticancer effect of the samples on human cancer cell lines (A549, AGS, Hep3B, MCF7) was indicated that the cancer cells were inhibited gradually in proportion to the increase of the concentration of the samples by MTT assay. The growth of the Raji and Jurkat cells were hastened by adding butanol fraction among the samples. In the genotoxicity on $H_2O_2-induced$ DNA damage in Chang cells using alkaline comet assay, most of samples were shown a strong protective activity from DNA OTM values.

The Inhibitory Effects of Trichosanthes kirilowii Root against Cadmium Induced Cytotoxicity (III) (천화분의 카드뮴에 대한 독성억제효과(III))

  • Lee, Jeong-Ho;You, Il-Soo;Kim, Shin-Kee;Lee, Ki-Nam;Chung, Woo-Young;Han, Du-Seok;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.32 no.1 s.124
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    • pp.15-21
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    • 2001
  • This study was conducted to investigate the antitoxic components in the water extract of the roots of Trichosanthes kirilowii (Cucurbitaceae). The results were as follows: Generally, detoxication effects by the water extract of T. kirilowii increased in proportion to the concentrations. Experimental animals were treated with cadmium and T. kirilowii water extract by oral administration. When 40 mg/kg dosage of T. kirilowii extract was administrated it showed the highest antitoxic effects in metallothionein induction. After the water extract treatment, body weights did not increase in proportion to the extract concentrations. These results suggest that T. kirilowii extract increased metallothionein concentration and decreased the toxicity of cadmium in rats. In vitro the antitoxic activity of water extract of T. kirilowii on NIH 3T3 fibroblasts was evaluated by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) and SRB (sulforhodamine B protein) assays. 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 T. kirilowii extract was shown significant antitoxic activity. The number of NIH 3T3 fibroblasts were increased and tend to regenerate. These results suggest that T. kirilowii extract retains a potential antitoxic activity.

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The Inhibitory Effects of the Methanol Extract of Houttuynia cordata $T_{HUNB}$ against Cadmium Induced Cytotoxicity (V) (어성초 메탄올 추출물의 카드뮴에 대한 독성억제효과(V))

  • Lee, Jeong-Ho;Jeong, Seung-Il;You, Il-Soo;Kim, Shin-Kee;Lee, Ki-Nam;Han, Du-Seok;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.32 no.1 s.124
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    • pp.61-67
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    • 2001
  • This study was conducted to investigate the antitoxic agent in methanol extract of Houttuynia cordata $T_{HUNB}$. Detoxication effects By H. cordata $T_{HUNB}$ extract increased in proportion to the extract concentrations. When 40 mg/kg dosage of H. cordata $T_{HUNB}$ extract was administered, it showed the highest antitoxic effects in metallothionein induction. After the extract treatment, body weights generally increased in proportion to the extract concentrations. From the above results, H. cordata $T_{HUNB}$ extract increased metallothionein concentrations and decreased the toxicity of cadmium in rats. In vitro the antitoxic activity of methanol extract of H. cordata $T_{HUNB}$ on NIH 3T3 fibroblasts was evaluated by the MTT {3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide} and SRB (sulforhodamine B protein) assays. The light microscopic study was carried out to observe morphological changes of the treated cells, $10^{-2}\;mg/ml$ Concentrations of H. cordata $T_{HUNB}$ extract was shown significant antitoxic activity. The number of NIH 3T3 fibroblasts were increased and tend to regenerate. These results suggest that H. cordata $T_{HUNB}$ extract retains a potential antitoxic activity.

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Analysis of Antioxidant Activity and Cytotoxicity against Human Cancer Cell Lines of Extract from Atractylodes rhizoma fermented with Ganoderma lucidum Mycelium (창출과 영지버섯 균사체 발효 추출물의 항산화 활성 및 인체 암세포주에 대한 세포독성 분석)

  • Park, Mi Hye;Kim, Mee Ra
    • The Korean Journal of Food And Nutrition
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    • v.30 no.3
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    • pp.454-463
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    • 2017
  • Ganoderma lucidum has been traditionally used as a medicine for treatment of bronchitis, arthritis, and high blood pressure, and it has been reported to display many biological activities including anticancer and immune activities. Since mushroom mycelium is known to have excellent biological activities together with mushroom fruiting body, studies on biological activities of mushroom mycelium have been actively conducted. Thus, the present study compared the biological activities before and after the cultivation of Ganoderma lucidum mycelium on Atractylodes rhizoma. When the radical scavenging activity was assessed by the DPPH assay, ARGL (ethanol extract of Atractylodes rhizoma mycelium fermented with Ganoderma lucidum) showed radical scavenging activity of 5.58~82.56% at concentrations of $10{\sim}500{\mu}g/assay$, while AR (ethanol extract of Atractylodes rhizoma) showed radical scavenging activity of 5.27~72.08% at the same concentrations. When measured by using the ABTS assay, ARGL showed higher radical scavenging activity than AR, which was consistent with the result obtained by the DPPH assay. In the MTT assay, the cytotoxicity of ARGL against all cell lines was higher than that of AR. In particular, the cytotoxicities of AR and ARGL against Hep3B at a concentration of $400{\mu}g/assay$ were 71.81% and 86.40%, respectively. In addition, the result obtained by the SRB assay was consistent with the result obtained by the MTT assay. According to the results mentioned above, there is a high probability that medicinal herb cultures using mycelium can be used as sources of functional foods since the cytotoxicities against cancer cells and antioxidant activities increased when the mycelium was fermented with Atractylodes rhizoma.

Regenerative Effects of Houttuynia cordata Extracts against Cadmium-induced Cytotoxicity in NIH 3T3 fibroblasts Cultures (VII) (카드뮴 독성으로 손상된 NIH 3T3 섬유모세포에 대한 어성초 추출물의 재생효과(VII))

  • Lee Jeong-Ho;Oh Hyun-Ju;Jeong Seung-Il;Chun Hyun-Ja;Han Do-Seok;Lee Hyun-Ok;Ahn Jong-Woong;Lee Mee-Hee;Baek Seung-Hwa
    • Journal of Society of Preventive Korean Medicine
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    • v.5 no.2
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    • pp.122-128
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    • 2001
  • This study was carried out to evaluate antitoxic effects of Houttuynia cordata extract on cadmium by colorimetric methods. The antitoxic activity ofc in NIH 3T3 fibroblasts was evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) and SRB (sulforhodamine B protein) assays. The light microscopic study was carried out to observe morphological changes of the treated cells. The concentration of 10-2 mg/ml of Houttuynia cordata extract was shown significant antitoxic activity. The number of NIH 3T3 fibroblasts were antitoxic and tend to regenerate. These results suggest that the ethyl acetate extract of Houttuynia cordata retains a potential antitoxic activity.

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Effects of Herbar Chelidonii on the Cultured Spinal Sensory Neurons Damaged by XO/HX (백굴채(白屈菜)가 손상된 배양척수감각신경세포에 미치는 영향)

  • Shin, Byung-Cheul;Song, Yung-Sun
    • The Journal of Korea CHUNA Manual Medicine
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    • v.2 no.1
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    • pp.143-157
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    • 2001
  • Objectives and Methods : To evaluate the mechanism of oxidative damage by xanthine oxydase(XO) and hypoxanthine(HX)-induced oxygen radicals, MTT assay and NR assay were carried out after the cultured mouse spinal sensory neurons were preincubated for 4 hours with various concentrations of XO/HX. And the amount of total protein. neurofilament EIA. lipid peroxidation and LDH activity were measured, to evaluate the protective effect of Herbar Chelidonii(HC) water extract on cultured spinal sensory neurons damaged by XO/HX. after the cultured mouse spinal sensory neurons were preincubated with various concentrations of HC water extract for 3 hours prior to exposure of XO/HX. Results : XO/HX decreased significantly the survival rate of the cultured mouse sensory neurons by NR assay and MTT assay In proportion to concentration and exposed time. In proportion to concentration and exposed time on cultured spinal sensory neurons, XO/HX showed the quantitative decrease of neurofilament by EIA. the decrease of total protein amount by SRB assay and the Increase of lipid peroxidation as well as LDH. HC showed the quantitative increase of neurofilament and total protein, but showed the decrease of lipid peroxidation and LDH activity against the neurotoxicity of XO/HX. Conclusions : From the above results, it is concluded that XO/HX have a neurotoxic effect on cultured spinal sensory neurons and that the herbs extract, such as HC, prevent the toxicity of XO/HX effectively in that they decrease lipid peroxidation and LDH activity.

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Study on Antioxidant and Cytotoxic Activities in Ethanol Extract from Prunus mume (오매 에탄올 추출물의 항산화 활성 및 항암 활성 연구)

  • Jeon, Yeon-Hee;Kwon, Ji-Eun;Kim, Mee-Ra
    • Journal of the East Asian Society of Dietary Life
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    • v.20 no.5
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    • pp.751-758
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    • 2010
  • Prunus mume is well known contain many functional materials that play beneficial roles in the human body. Studies have found that many organic acids and polyphenol compounds exist in Prunus mume. In this study, content of total polyphenols and flavonoids, antioxidative activity and cytotoxicity of ethanol extract from Prunus mume were measured Contents of total polyphenolic and total flavonoid compounds in ethanol extract from Prunus mume were 16.92 mg/g and 59.95 mg/g, respectively. The $IC_{50}$ of ethanol extract from Prunus mume were $237.72 {\mu}g$/assay and $239.58 {\mu}g$/assay by DPPH and ABTS radical cation scavenging test, respectively. Additionally, ferric ion reducing antioxidant power (FRAP) value of ethanol extract from Prunus mume was 0.94 mM ($FeSO_4$ eq.) by $800 {\mu}g$/assay. Cytotoxicity of Prunus mume against five kinds of cancer cell lines increased as the extract concentration increased Especially, cytotoxicity of the ethanol extract against A-549 (lung cancer line) was higher than that against any other cancer cell line by both MTT and SRB assay. These results show that ethanol extract of Prunus mume has considerably high antioxidative and cytotoxic activities.

Effect of Rhizoma Gastrodiae on Cultured Spinal Motor Neurons Damaged by Oxygen Radicals (천마가 산소자유기로 손상된 생쥐의 배양 척수 운동신경세포에 미치는 영향)

  • Son IL Hong;Lee Jung Hun;kim Sang Su;Lee Kang Chang;Lee Young Mi;Hong Gi Youn;Moon Hyung Bae;Seo Eun A;Han Du Seok;Shin Min Kyo;Song Ho Joan;Park Seung Taeck
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.2
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    • pp.262-266
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    • 2002
  • In order to elucidate the mechanism of cytotoxic effect of oxygen radicals on cultured mouse spinal motor neurons, the neurotoxicity induced by hydrogen peroxide(H₂O₂) was evaluated by MTT assay. The neuroprotective effect of Rhizoma Gastrodiae(RG) against H₂O₂-mediated neurotoxicity was also examined in these cultures by SRB assay. The results were as follows : The value of lethal concentration 50(LC50) of H₂O₂ was estimated at a concentration of 30 uM in these cultures. Cell viability of cultured mouse spinal motor neurons was remarkably decreased by H₂O₂-induced neurotoxicity in a dose- and time-dependent manner. RG was remarkably effective in blocking the neurotoxicity induced by H₂O₂ at a concentration of 120 μM as determined by SRB assay. From above the results, it is suggested that H₂O₂ induce neurotoxicity, and the selective herbal extracted RG was very effective in blocking H₂O₂-mediated neurotoxicity on cultured mouse spinal motor neurons.

Comparative Study on the Content and Cytotoxicity of Pseudolaric Acid B in the Five Plant Parts of Pseudolarix kaempferi

  • Nugroho, Agung;Woo, Nam-Tae;Park, Kyoung-Sik;Kwon, Na-Yun;Jung, Woo-Nyung;Lee, Sang Kook;Kim, Dong-Hwa;Park, Hee-Juhn
    • Natural Product Sciences
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    • v.23 no.4
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    • pp.265-269
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    • 2017
  • Pseudolaric acids of Pseudolarix kaempferi (Pinaceae) have been known as diterpenoids with potent anti-fungal-, anti-microbial, and cytotoxic activities. In the present study, the five MeOH extracts were prepared from the five plant part (root bark, stem bark, leaf, the inner part of root, and cone) to find the relation between the concentration of pseudolaric acids and cytotoxicity. Pseudolaric acids B and C were isolated from the root bark of P. kaempferi to use them as standard compounds. The five extracts were tested on cytotoxicity against six cancer cell lines, A549 (lung), HCT116 (colon), MDA-MB-231 (breast), SNU638 (stomach), and SK-hep-1 (liver) by SRB assay, but against K562 (leukemia) by SRB- or MTT assay. HPLC quantification were performed on a Shisheido Capcell PAK C18 column ($5{\mu}m$, $4.6mm{\times}250mm$) using 254 nm wavelength. The cytotoxicity ($IC_{50}$, $0.36{\mu}g/ml$ on K562 cell lines) of the root bark extract was potent and the content (101.1 mg/g extract) of pseudolaric acid B was very high in the root bark. These results suggest that the MeOH extract obtained from the root bark could be developed as the anti-cancer agent with a high quantity of pseudolaric acid B.

Cytotoxicity and Antigenotoxic Effects of Cordyceps militaris Extracts (번데기동충하조(Cordyceps militaris) 추출물의 세포독성 및 유전독성 억제효과)

  • 김미남;최승필;이득식;함승시
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.5
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    • pp.921-927
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    • 2001
  • This study was performed to investigated the effects on the cytotoxicity and antigenotoxicity of Cordyceps militaris extracts on the human cancer cell lines. The ethanol extract and five fractions which were hexane, chloroform, ethylacetate, butanol and aqueous were screened for crytotoxicity on human lung carcinoma(A549). human breast adenocarcinoma (MCF-7) human epitheloid carcinoma(HeLa), human fibrosarcoma(HT1080) human hepatocellular carcinoma(Hep3B), human gastric carcinoma(KATOIII) and chronic myelogenous leukemia(K562) cell by SRB and MTT assays. The results showed that growth inhibition rates of the human cancer cell in the presence of Cordyceps militaris were inhibited with increasing concentration of the extract. The ethanol extract from Cordyceps militaris had strong inhibitory effects in1 mg/mL treatment by SRB assay , showing 89.4%, 85.7%, 72.9% and 65.5% inhibition in HT1080, HeLa, Hep3B and A549, respectively. The treatment of 1 mg/mL hexane fraction by SRB assay had the strongest cytotoxicity with 97.0% on HT1080 followed by MCF-7(92.9%) and HeLA(90.3%). The inhibition ration on KATOIII by MTT assay was much higher in the butanol (83.7%) and aqueous (80.4%) than in the ethanol extract (61.5%) And also, K562 showed similar tendency with KATOIII. The effects of Cordyceps militaris extracts on the frequencies of micronucleated polychromatic erythrocytes (MNPCEs) induced by N-methyl-N-nitro-N-nitrosoguanidime(MNNG) were investigated in the bone-marrow cells of ICR male mice. The amount of 10, 20, 40 and 80 mg/kg of each extract were administered to animals immediately after injection of MNNG, and the exposure time was 36 hours. Significant reductions(p<0.05) with 39.7%, 52.7%, 71.4% and 83.9% were observed in the frequencies of MNPCE when 10, 20, 40 and 80 mg/kg of the hexane fraction of Coryceps militarus extracts were given to the mice.

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