• Title/Summary/Keyword: MTT Assay

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In vitro Effects of the Complex of Korean Citrus junos and Medicinal Herbs on the Cell Growth in CCD-986sk Human Fibroblast Cell Line

  • Park, Kap-Joo
    • Korean Journal of Environmental Biology
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    • v.22 no.1
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    • pp.83-88
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    • 2004
  • In order to investigate whether or not CCD-980sk cell line can be affected by Korean Citrus junos and medicinal herbs, we examined the MTT assay when we treated Korean Citrus junos and medicinal herbs in CCD-986sk human fibroblast cell line. The samples that added complex of grinded extracts of Korean Citrus junos and boiling-water extracts from Korean medicinal herbs were tested toy cell proliferation activity by means of a modification of the MTT assay. Among mixture of Citron 3 (Citron 3, less mellowed citron which was ripened for three months) and boiling-water extracts, the group Citron 3+Phellinus linteus showed significantly strong cell proliferation activity. And among mixture of Citron 4 (Citron 4, completely mellowed citron which was ripened for four months) and boiling-water extracts, the group Citron 4+Cordyceps militaris and Citron 4+Phellinus linteus showed significantly strong cell proliferation activity, respectively. These results suggest that complex of Korean Citrus junos and medicinal herbs could be an excellent candidate toy protection of human skin aging.

In vitro Screening of UVA Phototoxicity Inhibitors using the Natural Products (In vitro 실험법에 의한 천연물 중의 UVA 광독성 억제제 검색)

  • 김현진;김봉희
    • Environmental Analysis Health and Toxicology
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    • v.17 no.3
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    • pp.253-259
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    • 2002
  • The phototoxicity inhibitory activity of 15 natural products having antiinflammatory effect was screened by three in vitro methods: yeast growth inhibition test with Candida albicans, RBC photohemolysis and MTT assay. We induced phototoxic reaction by irradiating UVA (365 nm) on chlorpromazine (CPZ) that has been widely documented as phototoxic agent in clinical and experimental studies and then observed the effects of the natural products after treating them with CPZ. In yeast growth inhibition test, X. stramonium showed the inhibitory effect on the UVA phototoxicity and E. officinalis, Yeast, P. suffruticosa showed phototoxicity inhibitory effect in that their % hemolysis compared with control were 36.14${\pm}$ 2.69, 42.82${\pm}$1.35, 36.41${\pm}$0.48 on UVA. In MTT assay, all tested natural products increased cell viability compared with the control.

Evaluation of the Viability of Rat Periodontal Ligament Cells after Storing at 0℃/2 MPa Condition up to One Week: In Vivo MTT Method

  • Jang, Sun Mi;Cho, Sin-Yeon;Kim, Eui-Seong;Jung, Il-Young;Lee, Seung Jong
    • Journal of Korean Dental Science
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    • v.9 no.1
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    • pp.1-8
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    • 2016
  • Purpose: The aim of this study was to evaluate the rat periodontal ligament cell viability under $0^{\circ}C/2$ MPa condition up to one week using in vivo 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide (MTT) assay. Materials and Methods: As soon as 110 upper molar teeth of rats were extracted, they were stored in Hartman's solution under $0^{\circ}C/2$ MPa condition for 1, 2, 3, 4, and 7 days each. All specimens were treated with in vivo MTT assay and the value of optical density was measured by ELISA reader. These values were statistically analyzed by one-way ANOVA. Result: There was no statistical difference on MTT value between immediate and 1 day storage group. There were statistically significant differences between 1 day and 2 days tsorage, 2 and 3 days storage groups, respectively. Teeth of 34, and 7 days storage groups showed significantly lower MTT valuesc ompared with shorter period storage groups. Conclusion: When the MTT values were substituted in standard curve, 1 day storage group at $0^{\circ}C/2$ MPa condition showed 68% cell viability when compared with immediate group. It dropped to 13% at 2 days, and to less than 5% at 3 days or more.

Cytotoxicity of the Methanol Extract of Crotalariae sessiliflorae L. (농길리 메탄올 추출물의 세포독성)

  • Han Du-Seok;Chung Woo-Young;Park Myung-Oh;Shin Min-Kyo;Oh Hyun-Ju;Baek Seung-Hwa
    • Journal of Society of Preventive Korean Medicine
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    • v.5 no.1
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    • pp.144-150
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    • 2001
  • The cytotoxic activity of Cratalariae sessiliflorae on cultured NIH 3T3 fibroblasts and human oral epithelioid carcinoma cells (KB) were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide (MTT) colorimetric method These fractions of methanol extract of Cratalariae sessiliflorae showed inhibitory effect in vitro in the milligram range against KB cell lines. In general, the antitumor activities of these fractions were does-dependent over the milligram range. The comparison of IC50 values of these fractions in tumor cell lines showed that their susceptibility to these fractions decrease in the following order: Fr. 4> Fr. 6> Fr. 10> Fr. 2> Fr. 11> Fr. 3> Fr. 8> Fr. 7> Fr. 9> Fr. 1> Fr. 5 by the MTT assay. These fractions were tested for their cytotoxic effects on NIH 3T3 fibroblasts using MTT assay. They exhibited potent cytotoxic activities in vitro in the milligram range against NIH 3T3 fibroblasts. In general, the cytotoxic activities of these fractions were does-dependent over the milligram range. The comparison of CD50 values of these fractions in NIH 313 fibroblasts shows that their susceptibility to these fractions in decrease the following order: Fr. 10> Fr. 9> Fr. 2 = Fr. 4> Fr. 8> Fr. 11> Fr. 1 = Fr. 7> Fr. 3> Fr. 5 = Fr. 6 by the MTT assay. These results suggests that fraction 5 has the most growth - inhibitory activity against KB cell lines.

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Antioxidant Activity and Cytotoxic Effect of an Ethanol Extract from Seoritae (서리태 에탄올 추출물의 항산화 활성 및 암세포 증식 억제 효과)

  • Jeon, Yeon-Hee;Won, Ji-Hye;Kwon, Ji-Eun;Kim, Mee-Ra
    • Korean journal of food and cookery science
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    • v.27 no.3
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    • pp.1-10
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    • 2011
  • The antioxidant activity and cytotoxic effect of an ethanol extract from Seoritae were analyzed to develop new functional food materials. The antioxidant activity of Seoritae was determined by measuring electron donating ability with 1,1-diphenyl-2picrylhydrazyl (DPPH) and 2-2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) assays, as well as the ferric reducing antioxidant power (FRAP) assay. The cytotoxic effect of the Seoritae ethanol extract was measured with the 3-(4,5-dimethylthiazol-2-yl)-2,5-dipheltetrazolium (MTT) and sulforhodamine B (SRB) assays. As a result, the electron donating abilities of Seoritae against the DPPH and ABTS radicals were 63.75% and 87.68% at 500 ${\mu}g$/assay, respectively. The $IC_{50}$ values of Seoritae in the DPPH and ABTS assays were 385.39 ${\mu}g$/assay (128.46 ${\mu}g/mL$) and 209.39 ${\mu}g$/assay (51.83 ${\mu}g/mL$). Additionally, the FRAP value of Seoritae was 0.84 $FeSO_4$ eq. mM at 800 ${\mu}g$/assay. The total amounts of polyphenols and flavonoids, which indicate the antioxidant capability of Seoritae extract were 1.65 mg/g and 0.59 mg/g, respectively. Moreover, Seoritae extract showed a high cytotoxic effect of up to 81% against human cancer cells, particularly A-549 and HeLa cells. The growth inhibition rate of Seoritae extract against A-549 and HeLa cells was up to 76.48% and 75.67% in the MTT assay, and 78.98% and 80.54% in the SRB assay, respectively. The results of this study suggest that an ethanol extract of Seoritae is a potentially good natural antioxidant.

A Continuous Spectrophotometric Assay for NADPH-cytochrome P450 Reductase Activity Using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide

  • Yim, Sung-Kun;Yun, Chul-Ho;Ahn, Tae-Ho;Jung, Heung-Chae;Pan, Jae-Gu
    • BMB Reports
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    • v.38 no.3
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    • pp.366-369
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    • 2005
  • NADPH-cytochrome P450 reductase (CPR) transfers electrons from NADPH to cytochrome P450 and also catalyzes the one-electron reduction of many drugs and foreign compounds. Various spectrophotometric assays have been performed to examine electron-accepting properties of CPR and its ability to reduce cytochrome $b_5$, cytochrome c, and ferricyanide. In this report, reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) by CPR has been assessed as a method for monitoring CPR activity. The principle advantage of this substance is that the reduction of MTT can be assayed directly in the reaction medium by a continuous spectrophotometric method. The electrons released from NADPH by CPR were transferred to MTT. MTT reduction activity was then assessed spectrophotometrically by measuring the increase of $A_{610}$. MTT reduction followed classical Michaelis-Menten kinetics ($K_m\;=\;20\;{\mu}M$, $k_{cat}\;=\;1,910\;min^{-1}$). This method offers the advantages of a commercially available substrate and short analysis time by a simple measurement of enzymatic activity of CPR.

Effects of Jangwon-hwan(Zhuangyuan-wan) Water Extract against Xanthine Oxidase / Hypoxanthine-induced Neurotoxicity in the Cultured Mouse Cerebral Cortical Neurons (장원환이 XO/HX에 의해 손상된 대뇌피질 신경세포에 미치는 영향)

  • 김영수;권강범;민영기;조현익;박준배;이호섭;류도곤
    • The Journal of Korean Medicine
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    • v.20 no.4
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    • pp.3-10
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    • 2000
  • In order to elucidate the toxic mechanism of neurotoxical damage and neuroprotective effect of Jangwon-hwan(Zhuangyuan-wan) water extract, this experiment was performed. Neurotoxic effects of xanthine oxidase/hypoxanthine(XO/HX) were examined by MTT and NR assay, neuroprotective effects of Jangwon-hwan(Zhuangyuan-wan) water extract were examined by neurofilament enzymeimmuno assay(EIA). XO/HX induced an increase in cell viability, and a decrease in the amount of neurofilament on cultured mouse cerebral cortical neurons in dose-dependent manner. In neuroprotective effect of herb medicine, Jangwon-hwan(Zhuangyuan-wan) water extract increased the amount of neurofilament on cultured mouse cerebral cortical neurons damaged by XO/HX. From the results, it is suggested that XO/HX showed toxic effect in cultured mouse cerebral cortical Neurons and Jangwon-hwan(Zhuangyuan-wan) water extract is very effective in the prevention of neurotoxicity induced by XO/HX.

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Analysis of Antioxidant effects of Embelia scandens and Cornus hongkongensis

  • Oh, Jee-Min;Lee, Sang-Soo;Kim, June-Hyun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.78-78
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    • 2019
  • The aim of the study was to determine the antioxidant activities of the plants with origin of Vietnam. The Embelia scandens (Lour.) Mez which is a species of plant in the family Myrsinaceae and Cornus hongkongensis Hemsl., which is a species of plant in the family Cornaceae were tested for antioxidant activities. Samples were prepared using 95% ethanol using DPPH assay for assessing the antioxidant activity. Ascorbic acid was used for positive control for DPPH assay. DPPH assay experiment showed that extracts of the Embelia scandens (Lour.) Mez and Cornus hongkongensis Hemsl. might have anti-oxidant activity 4.77 times and 5.65 times higher, respectively, compared to control. To determine the cell toxicity, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used. MTT assay experiment showed that Embelia scandens (Lour.) Mez might have 13.1% more toxicity whereas Cornus hongkongensis Hemsl. might have 47.3% less toxicity compared to control. Taken together, these experiments showed that Cornus hongkongensis Hemsl. extracts might have significantly higher antioxidant activities and relatively lower toxicity, compared to control.

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Screening of Korean Medicinal and Food Plants with Antitumoral Activity (국내 약용 및 식용식물중 항종양활성 식물탐색)

  • Chung, Ill-Min;Kim, Kwang-Ho;Ahn, Joung-Kuk;Ahn, Jong-Seog;Ahn, Soon-Cheol
    • Korean Journal of Medicinal Crop Science
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    • v.7 no.1
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    • pp.37-44
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    • 1999
  • This study was conducted to screen antitumoral activity by in vitro bioassay method using 60 Korean medicinal and food plants extracted by 80% EtOH. Antitumor activity test was applied by the PKC(protein kinase C) and antibleb formation, PLC (Phospholipase C), and colorimetric tetrazolium assay (MTT assay) methods. Chenopodjum album and black Glycine max showed high antitumoral activity by 73.5% and 81.0%, respectively, against PKC by bleb-forming assay and PKC enzyme assay on human chronic leukemia K562 cell. Black Glycine max also showed 91.2% antitumoral activity in the PLC method and the lowest $IC_{50}$ $value(4.7{\mu}g/ml)$ by MTT method against P-338 cell line. In the effect of the concentration treatment on antitumoral test, the more concentration indicated the more activity value.

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Protective Effect of Neuronal Cell on Glutamate-induced Oxidative Stress from Viola mandshurica Extracts (Glutamate에 의한 산화적 스트레스로부터 신경세포를 보호하는 제비꽃 추출물의 영향)

  • Lee, Mi-Ra;Han, Chang-Suk;Han, Dong-Youl;Park, Eun-Ju;Lee, Seung-Cheol;Park, Hae-Ryong
    • Applied Biological Chemistry
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    • v.51 no.1
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    • pp.79-83
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    • 2008
  • The present study describes glutamate which is known as excitatory neurotransmitter is related with oxidative damages and the Viola mandshurica extracts. Showed protective effects against glutamate-induced cytotoxicity. The protective effect of antioxidant on the glutamate treated N18-RE-I05 cells was determined by a MTT reduction assay. The neuroprotective effect of methanol, ethanol, and acetone extracts from V. mandshurica against glutamate-induced cytotoxicity was assessed by the results of an MTT reduction assay. Among the three extracts, the acetone extract showed the highest protective effect by the results of an lactate dehydrogenase release assay. Therefore, these results suggest that V. mandshurica extracts could be a new potential candidate against glutamate-induced oxidative stress.