• Title/Summary/Keyword: MTT reduction assay

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Inhibitory Effects of Ginsenosides on Glutamate-Induced Swelling of Cultured Astrocytes

  • Seong, Yeon-Hee;Koh, Sang-Bum;Kim, Hack-Seang
    • Journal of Ginseng Research
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    • v.24 no.3
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    • pp.138-142
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    • 2000
  • Effects of ginsenosides (Rb$_1$, Rb$_2$, Rc, Re, Rg$_1$, Rf) on L-glutamate (glutamate)-induced swelling of cultured astrocytes from rat brain cerebral cortex were studied. Following the exposure to 0.5mM glutamate for 1 hr, the intracellular water space (as measured by [$^3$H]O-methyl-D-glucose uptake) of astrocytes increased by about two-fold. Simultaneous addition of ginsenosides Rb$_2$ and Rc with glutamate reduced the astrocytic swelling in a dose-dependent manner. These ginsenosides at 0.5 mg/ml did not affect the viability of astrocytes for up to 24 hr which was determined by a colorimetric assay (MTT assay) for cellular growth and survival. These ginsenosides at 0.3 mg/ml inhibited the increase of intracellular Ca$\^$2+/ concentration ([Ca$\^$2+/]$\_$i/) induced by glutamate. These data suggest ginsenosides Rb$_2$ and Rc prevent the cell swelling of astrocytes induced by glutamate, maybe via inhibition of Ca$\^$2+/ influx.

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Effect of Celecoxib on Lung Injury Improvement by Controlling Epithelial-Mesenchymal Transition(EMT) in Chronic Obstructive Pulmonary Disease(COPD) (만성폐쇄성폐질환에서 상피중간엽이행 조절을 통한 Celecoxib의 폐 손상 개선효과)

  • Lee, Sun-Kyung
    • Journal of Convergence for Information Technology
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    • v.11 no.11
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    • pp.248-255
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    • 2021
  • This study confirmed the effects of improving lung damage of celecoxib using an animal model of chronic obstructive pulmonary disease(COPD). It was induced in models LPS + CSE and performed in vitro and in vivo. MTT assay and real-time PCR were performed in MRC5 cells as in vitro, and mRNA expression, BALF, collagen content, and protein expression were confirmed as in vivo. Celecoxib reduced the number of inflammatory cells, cytokine and soluble protein accumulation in BALF, decreased body weight and lung weight in animal models, and improved lung collagen deposition. In addition, the reduction of EMT markers was confirmed through Western blotting and real-time PCR. Consequently, celecoxib is thought to improve lung damage of COPD induced to LPS+CSE by regulating EMT.

Protective Effects of Thujae Semen against Neurotoxicity Induced by 6-hydroxydopamine in PC12 Cells (백자인의 6-하이드록시도파민으로 유도된 뇌세포독성에 대한 보호효과)

  • Kim, Hyo-Geun;Shim, Jin-Sup;Ju, Mi-Sun;Cho, Seung-Hun;Oh, Myung-Sook
    • The Korea Journal of Herbology
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    • v.23 no.3
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    • pp.19-25
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    • 2008
  • Objectives : This study was performed to evaluate the neuroprotective effect of water extracts from Thujae Semen(TSW) in PC12 cells. Methods : We performed 2,2-diphenyl-2-picrylhydrazyl(DPPH) radical scavenging assay, 2,2-azinobis- (3-ethyl-benzothiazoline-6-sulfonic acid(ABTS) cation scavenging assay, and determination of total polyphenolic content to examine the antioxidant effects of TSW. We also carried out 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay(MTT), reactve oxygen species(ROS) assay, and nitric oxide(NO) assay to examine neuroprotective effects against 6-hydroxydopamine(6-OHDA) in PC12 cells. Results : TSW showed $IC_{50}$ values of 404.3 and 219.9 ${\mu}g/mL$ in DPPH and in ABTS assays, respectively. TSW showed 9.74 ${\mu}g/mL$ of total polyphenol contents. TSW incresed cell viability in a dose dependent manner and it showed protective effect against 6-OHDA neurotoxicity at the concentration of 25-200 ${\mu}g/mL$. Moreover, it recovered 6-OHDA induced cell death at the same concentrations. The extract showed a dose dependent reduction of ROS and NO generation by 6-OHDA. Conclusions : We concluded that TSW has neuroprotective effect against 6-OHDA-induced toxicity in PC12 cells through ROS and NO inhibition.

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Protective Effect of Schizonepeta tenuifolia Briquet Extracts on Oxidative DNA Damage in Human Leucocytes and on Hydrogen Peroxide-induced Cytotoxicity in PC12 Cells

  • Yoon, Mi-Young;Lee, Hyun-Jin;Lee, Bo-Bae;Lee, Sang-Myeon;Kim, Ju-Young;Kim, Yong-Seong;Park, Eun-Ju;Park, Hae-Ryong
    • Food Science and Biotechnology
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    • v.16 no.5
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    • pp.858-862
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    • 2007
  • The present study was conducted to examine the antioxidant activities and neuroprotective effects of methanolic extracts from Schizonepeta tenuifolia Briquet (STE). STE ($100\;{\mu}g/mL$) showed $43.33\;{\mu}M$ of total phenolic content, 64.43% of radical scavenging activity, and 0.157 of reducing power. In addition, the effect of STE on $H_2O_2$-induced DNA damage in human leucocytes was evaluated by the comet assay, where STE was a dose dependent inhibitor of DNA damage induced by $200{\mu}M$ of $H_2O_2$. The protective effect of STE against $H_2O_2$-induced oxidative damage on PC12 cells was investigated by an 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) reduction assay and lactate dehydrogenase (LDH) release assays. After 2 hr of cell exposure to $H_2O_2\;(500\;{\mu}M)$, a marked reduction in cell survival was observed. However, this reduction was significantly prevented by $1-50\;{\mu}g/mL$ of STE. Therefore, these results suggest that STE could be a new antioxidant candidate against neuronal diseases.

Enhanced Antitumor Efficacy with Combined Administration of Astragalus and Pterostilbene for Melanoma

  • Huang, Xin-Yan;Zhang, Song-Zhao;Wang, Wen-Xi
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.3
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    • pp.1163-1169
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    • 2014
  • Astragalus, a commonly used traditional Chinese medicine, has exhibited antitumor actions in patients. In this study, in vitro and in vivo antitumor effects of astragalus and synergistic antitumor efficacy in combination with pterostilbene were investigated. Melanoma cells were treated with pterostilbene (Pt), graduated doses of astragalus injection (AI), or these in combination. Cell viability was measured using a MTT assay. Released nucleosomes and caspase activity were measured using enzyme-linked immunosorbent assay. Growth inhibition in vitro and in vivo was also assessed. Analysis of variance and t tests were used for statistical analysis. Significant reduction (p<0.05) in cellular proliferation were observed with AI and AI-Pt in a time- and concentration-dependent manner. Apoptosis and caspase-3/7 activity were significantly increased by AI and AI-Pt treatment (p<0.05). In vivo, AI inhibited melanoma tumor growth, with inhibition rates ranging from 36.5 to 62.3%, by inducing apoptosis via up-regulation Bax expression and the Bax/Bcl-2 ratio and down-regulating Bcl-2 expression. AI significantly inhibits the growth of melanoma in vitro and in vivo by inducing apoptosis. These data suggest that combined treatment of astragalus with pterostilbene enhances antitumor efficacy.

Cytotoxicity of Acetoxyscirpendiol from Paecilomyces tenuipes (Paecilomyces tenuipes로부터 분리한 Acetoxyscirpendiol의 세포사멸작용)

  • 한희창;김미정;김종수;김하원
    • YAKHAK HOEJI
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    • v.48 no.2
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    • pp.153-158
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    • 2004
  • Paecilomyces tenuipes is one of the famous Chinese medicinal entomopathogenic fungi that parasite in the lavae of silkworm. A cytotoxic compound, 4$\beta$-acetoxyscirpendiol (ASD) was isolated from a methanolic extracts of Paecilomyces tenuipes. The ASD compound belongs to scirpenol subfamily of trichothecene mycotoxin. In a continuation of the elucidation of the mechanism of ASD, we report here the evidences of induction of apoptosis by ASD in human Jurkat T cell line. In MTT reduction assay for monitoring cell viability, ASD showed strong toxicity. The 50 percent inhibitory concentrations of ASD against human T lymphoid Jurkat cell was 59.5 ng/$m\ell$. Phosphatidylserine externalization was increased by ASD at 3 and 6 hrs when compared with that of 6 hrs in the cell line showing in a time-dependent manner. When whole lysates of cells treated with ASD were subjected to western blot assay, 113 kDa poly(ADP-ribose) polymerase (PARP) was significantly cleaved to 89 kDa fragment. Time-dependent DNA fragmentation was also observed when Jurkat T cells were treated with ASD at 100 ng/$m\ell$ for 6 hrs and 18 hrs at the ratios of 8.5% and 15.0%, respectively. From these data, Jurkat T lymphocytes treated with ASD from Paecilomyces tenuipes underwent typical cascades of apoptotic cell death.

Protective Effect of Extracts from Euryale ferox against Glutamate-induced Cytotoxicity in Neuronal Cells

  • Lee, Mi-Ra;Kim, Ji-Hyun;Son, Eun-Soon;Park, Hae-Ryong
    • Natural Product Sciences
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    • v.15 no.3
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    • pp.162-166
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    • 2009
  • Oxczaasssaidative stress plays an important role in neuronal cell death, which is associated with neurodegenerative conditions such as Alzheimer's and Parkinson's disease. This study evaluated the neuroprotective effect of Euryale ferox (EF) extracts against glutamate-induced cytotoxicity in hybridoma N18-RE-105 cells. Specifically, neuroprotective effects of methanol and ethanol extracts were evaluated by the MTT reduction assay. The ethanol extracts of EF displayed dose dependent protection against neuronal cell death induced by 20 mM of glutamate. Furthermore, the ethanol extracts of EF was sequentially fractionated with hexane, diethyl ether, ethyl acetate, and water layer according to degree of polarity. The hexane fractions exhibited neuroprotective effect against glutamate-stressed N18-RE-105 cells. Overall, results suggest that EF extracts can potentially be used as chemotherapeutic agents against neuronal diseases.

Effects of PALMIWON on Cell Viability of Immune Cell and ${\beta}-cell$ (취장소도세포와 면역세포에 미치는 팔미원의 영향)

  • 이인순;이인자
    • YAKHAK HOEJI
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    • v.39 no.5
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    • pp.541-547
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    • 1995
  • In order to investigate the usability of PALMIWON as antidiabetic immuno-modulating prescription for Insulin-dependent diabetes, we studies the effects of PALMIWON on immune cell and ${\beta}-cell$. U937 was used as the model of immune cell and RINm5F as the model of ${\beta}-cell$. The effects of PALMIWON was measured by cell viability in terms of MIT assay. As a result, PALMIWON and the compositional drugs showed the different effects m immune cell and ${\beta}-cell$. Cell viability of U937 was significantly decreased wheras that of RINm5F was no significantly difference between drug treated group and control, or significantly less reduction compared with U937. It implies that PALMIWON is useful as immunotherapeutic agents in the prevention and therapy of type 1 diabetes.

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The Biocompatibility Evaluation of Resin-Based Root Canal Sealers (Resin-Based Root Canal Sealer의 생체 적합성 평가)

  • Kim, Hyoung-Sun;Chon, Seong-Min;Moon, Jhong-Hyun;Lee, Kwang-Won;Yu, Mi-Kyung
    • Journal of Dental Rehabilitation and Applied Science
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    • v.23 no.1
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    • pp.95-104
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    • 2007
  • I. Objective The primary requirement of an endodontic root canal sealer is the biologic compatibility, because they remain in close contact with living periapical tissues over a long period of time. The aim of this study was the evaluation of cytotoxicity and genotoxicity of resin-based root canal sealers, AH 26 and ADSEAL. II. Material & Methods In this study, human periodontal ligament cells, human oral cancer cells (KB) and mouse osteoblasts (MC-3T3-E1) were used. Specimens of AH26, ADSEAL were eluted with culture medium for 1, 3, 5 and 7 days. Cytotoxicity was evaluated by using tetrazolium bromide reduction assay (MTT assay) for mitochondrial enzyme activity and cell viability. Genotoxicity was evaluated by using alkaline single cell gel electrophoresis assay (Comet assay). Also cell apoptosis induced by AH 26 was detected by Hoechst33258 staining. III. Results AH 26 and ADSEAL exhibited cytotoxic effects in all investigated cell groups. Genotoxicity was also noted for both sealers in mouse osteoblasts (MC-3T3-E1). But, ADSEAL presented significantly low cytotoxicity and genotoxicity compared with AH 26. Cytotoxicity and genotoxicity induced by AH 26 resulted in apopotosis. IV. Conclusion Our results clearly indicate that the recently invented ADSEAL has better biocompatibility than another resin based root canal sealer, AH 26. However ideal root canal sealer should have not only biocompatibility but also satisfactory physico-chemical properties such as sealing ability and stability. Thus continuous studies and developments should follow.

Antioxidant Activity and Protective Effects of 9-hydroxy-$\alpha$-tocopherone from Viola mandshurica Extracts (제비꽃(Viola mandshurica) 추출물로부터 분리된 9-hydroxy-$\alpha$-tocopherone의 항산화 활성 및 세포 보호효과)

  • Lee, Mi-Ra;Hwang, Ji-Hwan;Park, Jae-Hee;Kim, Hyun-Jung;Park, Eun-Ju;Park, Hae-Ryong
    • Korean Journal of Pharmacognosy
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    • v.41 no.3
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    • pp.166-173
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    • 2010
  • Oxidative stress to proteins, lipids, or DNA is higher in human autopsy tissue and in rodent models of a number of neurodegenerative conditions, including Alzheimer's and Parkinson's disease. On the basis of this information, we established a screening system using N18-RE-105 cells to identify therapeutic agents that can protect cells from glutamate toxicity. During the course of our screening program, we recently isolated the active compound 9-hydroxy-$\alpha$-tocopherone ($\alpha$-TP), which prevents glutamate-induced cell death, from Viola mandshurica. The chemical structure of $\alpha$-TP was identified using spectroscopic methods and by comparison with literature values. Antioxidant activity and protective effects of $\alpha$-TP were evaluated by DPPH radical-scavenging assay, morphological assay, MTT reduction assay, and lactate dehydrogenase (LDH) release assay. These results suggest that $\alpha$-TP could be a new potential chemotherapeutic agent against neuronal diseases.