• Title/Summary/Keyword: MTT assay

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Studies on the Anticancer Effect of Broussonetia kazinoki Extracts (닥나무(Broussonetia kazinoki) 추출물의 항암효과에 관한 연구)

  • 민경진;정승희;구성자
    • Korean journal of food and cookery science
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    • v.15 no.3
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    • pp.231-237
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    • 1999
  • The anticancer effect of the bark of Broussonetia kazinoki root extracts (hexane. chloroform, ethylacetate, butanol, aqueous) were studied. The cytotoxicity by MTT assay and inhibitory effect on the growth of sarcoma 180 cells were tested in vitro. The reduction rate of the tumor formation and spleen/body weight rate on BALB/c mouse were tested in vivo. From the tests, each fraction showed the cytotoxic effect against the sarcoma 180 cells. In addition, as the concentration of the fractions increased, cytotoxic effect tendency increased as well. The cytotoxic rate of the hexane, chloroform, ethylacetate, butanol and aqueous fractions showed by 58.7%, 40.1%, 75.7%, 52.6% and 62.7% respectively after testing by MTT assay system. And sarcoma 180 cells were incubated for 6 days at 37$^{\circ}C$ with various concentrations of each fraction. As the incubation days go on, the number of cells increased, while the inhibition rate on the growth of sarcoma 180 cells were decreased. Especially the ethylacetate fraction at the concentration of 1.0 mg/ml strongly inhibited the growth of sarcoma 180 cells by 74% compared with the control for a day 37$^{\circ}C$ The hexane, chloroform, ethylacetate, butanol and aqueous fractions inhibited on the growth of sarcoma 180 cells by 31%, 19%, 60%, 30% and 42% respectively, when sarcoma 180 cells has been incubated for 6 days at 37$^{\circ}C$. The each fraction exhibited the antitumor effect in vivo. The ethylacetate fraction reduced the tumor formation by 41% compared with the control, when sarcoma 180 cells were injected subcutaneously into the left groin of BALB/c mice. Also spleen/body weight rate of ethylacetate fraction was increased by 2.10% compared with the control (1.08%). And it is considered that there would be no toxic effect caused by each fraction of body weight and organ as there was on more changes in mouse' weight compared with the control.

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Effects of DSG on Osteoblastic Cell from Rat Calvariae in the Presence of Dexamethasone (단치소요산가미방이 Dexamethasone 처리한 랫드의 두개골 세포에 미치는 영향)

  • Park, Jong-Hyeong;Hwang, Gwi-Seo
    • Journal of Society of Preventive Korean Medicine
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    • v.10 no.2
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    • pp.19-30
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    • 2006
  • It is well known that glucocorticoid may induce osteoporosis as its side effect in long-term therapy. The inhibition of osteoblast by glucocorticoid is also recognized as its action mechanism of decreased bone formation. In this study, the effect of DSG, Danchisoyosangamibang, on the differentiation and function of osteoblastic cells was investigated. The osteoblastic cells were isolated from rat calvariae using collagenase treatment. The cell counting, enzyme activity assay, MTT assay, collagen content assay were done to determine the cell proliferation, intracellular alkaline phosphatase (ALP) activity, bone martrix production, and cell apoptosis. DSG enhanced the cell proliferation after the culture for 10 days. ALP activity and total protein synthesis, and intracelluar collagen synthesis were increased time dependently when the cells were treated with DSG in the presence of dexamethasone. And, DSG restored calvarial cell function decreased by dexamethasone.

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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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Effects of JY on Osteoblastic Cell from Rat Calvariae in the Presence of Glucocorticoid (자혈양근탕(滋血養筋湯)이 부신피질호르몬에 의해 억제된 조골세포 기능에 미치는 영향)

  • Choi, Jeong-Sin;Hwang, Gwi-Seo
    • Journal of Society of Preventive Korean Medicine
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    • v.12 no.2
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    • pp.197-206
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    • 2008
  • The inhibition of osteoblast by glucocorticoid is recognized as its action mechanism of decreased bone formation. In this study, the effect of JY, Jahyulyangkeuntang, on the differentiation and mineralization of osteoblastic cells was investigated in the presence of dexamethasone. The cell counting, enzyme activity assay, MTT assay, collagen content assay were done to determine the cell proliferation, alkaline phosphatase(ALP) activity, bone martrix production, and cell apoptosis. JY enhanced the cell proliferation after the culture for 10 days. ALP activity and total protein synthesis, and intracellular collagen synthesis were increased when the cells were treated with JY. And JY restored calvarial cell function decreased by dexamethasone.

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Antioxidant Activity and Protective Effects of Extracts from Sambucus williamsii var. coreana on t-BHP Induced Oxidative Stress in Chang cells (접골목 추출물에 의한 항산화 활성이 정상 간세포의 t-BHP 유발 산화스트레스에 미치는 영향)

  • Kim, Kitae
    • The Journal of the Society of Korean Medicine Diagnostics
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    • v.17 no.3
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    • pp.275-286
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    • 2013
  • In the present study, antioxidant activity and protective effect of extracts from Sambucus williamsii var. coreana stems (SWC) were evaluated on tert-butyl hydroperoxide (t-BHP) induced oxidative stress in human liver (Chang) cells. Antioxidant activities of the SWC extracts were determined by various radical scavenging activities, such as DPPH, ferric reducing antioxidant power (FRAP), 2,2'-azinobis-(3-ethybenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity and oxygen radical absorbance capacity (ORAC) assay. SWC extracts showed strong antioxidant effect on various assay. To determine the hepatoprotective effects of SWC on t-BHP induced oxidative damage, cell viability was measured using MTT assay. Pretreatment of SWC extracts showed increasing cell viability, decreasing ROS and restoring mitochondria membrane potential on t-BHP induced oxidative stress in Chang cells. Our findings suggest that SWC extracts may be considered a potential agent for therapeutic protective effect from oxidative stress through its antioxidant activity.

Anti-stress Effects of Ethanol Extract of Ziziphus jujuba Against Corticosterone-Induced Apoptosis in PC12 Cells

  • Da Hye Song;Yu Jin Choi
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2022.09a
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    • pp.119-119
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    • 2022
  • The coronavirus disease 2019 (COVID-19) pandemic may be stressful for people. Public health actions, such as social distancing, can make people feel isolated and lonely and can increase stress and anxiety. As a result, there is a growing interest towards various materials to relieve stress. Thus, the present study aimed to investigate the anti-stress effects of ethanol extract of Ziziphus jujuba in PC12 cells treated with corticosterone and its underling mechanisms. Furthermore, the viability of the cells, the apoptosis of the cells, the level of phosphorylation of extracellular signal-regulated kinases (p-ERKs) expression were measured by MTT assay, LDH assay, Hoechst staining assay and western blotting. Our results showed that the extract of Ziziphus jujuba reversed corticosterone-induced damage in PC12 cells, which increased cell viability, decreased LDH release, and attenuated corticosterone-induced apoptosis as compared with the corticosterone-treated group. Therefore, these data suggest that the extract of Ziziphus jujuba could be a good candidate for development as a functional food supplement in the improve the anti-stress effect.

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Positive Effect of Musa paradisiaca Peel Ethanolic Extract on Antioxidant Activity and Melanin Synthesis (바나나 껍질 에탄올 추출물이 멜라닌 합성에 미치는 영향)

  • Kim, JaeRyeon;Kim, Moon-Moo
    • Journal of Life Science
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    • v.28 no.7
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    • pp.802-810
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    • 2018
  • Aging is accompanied by changes in the body, such as graying hair, wrinkles, and black spots composed of lipid peroxides and proteins. Melanin is a polymer substance produced by an oxidation polymerization reaction from tyrosine, and it determines the color of hair and skin. It has been reported that melanin is synthesized by melanocyte, and its excessive production by reactive oxygen species is associated with aging. The purpose of this study was to determine the direct effects of Musa paradisiaca peel ethanolic extract (MPEE) on antioxidative activity and melanin synthesis. It was observed that the antioxidant activity of MPEE was similar to that of vitamin C, a positive control, in both DPPH radical scavenging assay and reducing power assay. In order to examine cytotoxicity prior to cell experimentation, 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed for B16F1 cells. MPEE was not cytotoxic at $32{\mu}g/ml$ or less. In addition, MPEE increased melanin synthesis in live cells in addition to tyrosinase activity and melanin synthesis in dihydroxyphenylalanine (DOPA)-oxidation assay in vitro. Moreover, MPEE increased melanin synthesis in cells aged by pretreatment with $H_2O_2$. The expression levels of tyrosinase-related protein (TRP)-1, TRP-2, and superoxide dismutase (SOD)-2 by western blot analysis were increased in the presence of MPEE. These results suggest that MPEE could promote the melanin synthesis as an antioxidative substance.

Molecular Biological Study of Anti-cancer Effects of Bee Venom on Human Melanoma Cell (약침용봉독액(藥鍼用蜂毒液)이 흑색종세포(黑色腫細胞)에 미치는 항암효과(抗癌效果)에 대(對)한 분자생물학적(分子生物學的) 연구(硏究))

  • Park, Chan-Yol;Nam, Sang-Soo;Kim, Chang-Hwan;Lee, Jae-Dong;Kang, Sung-Keel;Lee, Yun-Ho;Ahn, Byoung-Choul
    • Journal of Acupuncture Research
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    • v.17 no.2
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    • pp.169-186
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    • 2000
  • To study anti-cancer effect and molecular biological mechanism of bee venom for aqua-acupuncture, the effects of bee venom on cell viability, apoptosis, and cell cycle were analyzed using MTT assay, tryphan blue assay, [3H]thymidine release assay, flow cytometric analysis, activity of caspase-3 protease activity assay, and immunocytometric analysis of PCNA. To explore whether anti-cancer effects of bee venom are associated with the transcriptional control of gene expression, quantitative RT-PCR analysis of apoptosis- and cell cycle-related genes was performed. The obtained results are summarized as follows: 1. The MTT assay demonstrated that cell viability was decreased by bee venom in a dose-dependant manner. 2. Significant induction of apoptosis was identified using tryphan blue assay, [$^3H$]thymidine release assay, and flow cytometric analysis of sub $G_1$ fraction. 3. In analysis of caspase-3 protease activity, the activity had increased significantly, in a dose-dependant manner. 4. Quantitative RT-PCR analysis of the apoptosis-related genes showed that Bcl-2 and $Bcl-X_L$ were down-regulated whereas Bax was up-regulated by bee venom treatment. 5. In flow cytometric analysis of cell cycle and immunocytometric analysis of PCNA expression, cell numbers of $G_1$ phase was increased by a dose-dependant manner. 6. In quantitative RT-PCR analysis of the cell cycle-related genes, p21, p27, and p57 were increased, while Cyclin D1, CDK4, c-Myc, c-Fos, and Histone H3 were decreased. In contrast, there were no remarkable changes in expression levels of CDC2 and c-Jun.

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Anti-inflammatory and Antioxidant Activities of Ulva lactuca Linnaeus Extract (참갈파래 추출물의 항염증 및 항산화 활성)

  • Min, Kyung-Cheol;Kim, Geun-Dae;Nam, Hyeon-Gyu;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.32 no.5
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    • pp.340-347
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    • 2022
  • The anti-inflammatory and antioxidant activities of the Ulva lactuca Linnaeus extracts were measured. As a result of the MTT assay of the extract, it was found that the cell viabilities were higher than 90% at all concentrations, and the cell promotion rate exceeded 100% at a certain concentration. Therefore, in all subsequent experiments, the highest concentration was set at 500 ㎍/ml. In the nitric oxide (NO) assay, one of the anti-inflammatory experiments, the Ulva lactuca Linnaeus extracts exhibited inhibitory effects of 16%, 19%, and 62% at concentrations of 5, 50, and 500 ㎍/ml, respectively, compared with the LPS-only group. In tumor necrosis factor-α (TNF-α) assay, the extracts showed inhibitory effects of 16%, 17%, and 27% at concentrations of 5, 50, and 500 ㎍/ml, respectively, compared with the LPS-only group. In the interleukin-6 (IL-6) assay, the extracts also showed inhibitory effects of 15% and 28% at concentrations of 50 and 500 ㎍/ml, respectively, compared with the LPS-only group. In the total polyphenol content assay, the extracts exhibited 8.02, 9.90, and 23.8 mg GAE/g at concentrations of 0.3, 3, and 30 mg/ml, respectively. In the ABTS assay, the extracts exhibited scavenging activities of 2.6, 12.6, and 77.3% at concentrations of 0.3, 3, and 30 mg/ml, respectively. Therefore, it is considered that Ulva lactuca Linnaeus extracts can be used as anti-inflammatory and antioxidant substances for the development of functional foods.

Antioxidative Activity of Water Extract of Different Parts of Acanthopanax divaricatus var, albeofructus (흰털오가피 부위별 물추출물의 항산화활성)

  • Lyu, Su-Yun;Kim, Ji-Young;Noh, Bin-Na;Park, Won-Bong
    • YAKHAK HOEJI
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    • v.50 no.3
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    • pp.191-198
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    • 2006
  • Acanthopanax species have traditionally been used as a tonic, a sedative as well as in the treatment of rheumatism, hypertension and diabetes. In the present study, oxidative stress was induced in Vero cells by incubating the cells with glucose and the cell viability was measured by MTT assay. The concentration of glucose which 50% of cell viability was 125 mM $(IC_{50})$ and the cell viability was increased to $87.6{\pm}8.8%$ by treatment of the extracts of Acanthopanax divaricatus var. albeofructus. The antioxidative activity of water extract of different parts of the Acanthopanax plant was investigated by DPPH (1,1-diphenyl-2-picrylhydrazyl) assay, xylenol orange assay, TBARS (thiobarbituric acid reactive substances) assay and enzyme (superoxide anion and catalase) assay. Each extract (leaves, root, stem and fruits) of the plant showed free radical and $H_2O_2$ scavenging activity. The extract also inhibited lipid peroxidation and recovered enzyme (superoxide anion dismutase and catalase) activity in Vero cells treated with glucose.