• Title/Summary/Keyword: MTT assay

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Protective Effect of Coptidis Rhizoma on SNAP-Induced Cytotoxicity in Pancreatic RINm5F Cells (SNAP 유도성 RINm5F 세포 독성에 대한 황연 추출물의 방어효과)

  • 류도곤;권강범;양정예;김은경;김강산
    • The Journal of Korean Medicine
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    • v.24 no.2
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    • pp.159-165
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    • 2003
  • Objectives : Coptidis rhizoma (CR) is an oriental medicine that has been used in many traditional prescriptions against diabetes mellitus in Korea for centuries. Our purpose was to determine the protective effect and its action mechanism of CR on the cytotoxicity of pancreatic -cell line (RINm5F cell). Methods : In this experiment, we used methods such as MTT assay for detection of cytotoxicity, DNA fragmentation assay for detection of apoptotic cell death, LDH activity assay for detection of necrotic cell death, and measurement of $DiOC_{6}$ (3) retention for detection of mitochondrial membrane potential (MMP). Background : Nitric oxide (NO) is believed to playa key role in the process of pancreatic -cell destruction leading to insulin-dependent diabetes mellitus (IDDM). Results : Exposure of RINm5F cells to chemical NO donor such as S-nitroso-N-acetylpenicillamine (SNAP) induced cytotoxic events such as DNA fragmentation and lactate dehydrogenase (LDH) release into medium. However, pretreatment of RINm5F cells with CR extract ($10~50{\mu\textrm{g}}/ml$) for 3 hours prevented SNAP-induced DNA fragmentation and LDH release into medium through the inhibition of MMP disruption. Conclusions : These results suggest that CR may be a candidate for a therapeutic or preventing agent against IDDM.

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The Effects of Saussurea Radix and Plantaginis Herba on Cellular Viability, Proliferation, Apoptosis and Expression of Cell Cycle-related Genes in Gastric Cancer Cells (목향(木香)과 차전초(車前草)가 위암세포(胃癌細胞)의 활성(活性), 증식(增殖), 자기살해능(自己殺害能) 및 세포주기관련 유전자 발현에 미치는 영향)

  • Oh, Hee-Rah;Ko, Seong-Gyu
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.7 no.1
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    • pp.1-18
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    • 2001
  • Objective: This experimental study was carried out to evaluate the effects of Saussurea Radix and Plantaginis Herba on cellular viability, proliferation, apoptosis and expression of the cell cycle-related genes in cultured gastric cancer cells. Method :MTT assay for analysis of cellular toxicity and the effect on suppression of cellular viability, $[^{3}H]$ thymidine incorporation assay for evaluation of the effect on suppression of DNA replication, tryphan blue exclusion assay for measurement of induction of apoptosis and Quantitative RT-PCR for analysis of the effects on expression of cell cycle or apoptosis-related genes were performed. Results: Antitumor activity of Saussurea Radix associated with inhibition of cell cycle progression and promotion of apoptosis caused by transcriptional regulation of p53, p21/Wafl and the other related genes was observed.

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Antibacterial Effect of Huanggeumjakyak-tang against Methicillin-resistant Staphylococcus aureus (황금작약탕(黃芩芍藥湯)의 MRSA에 대한 항균활성에 관한 연구)

  • Kim, Esther;Choi, Chonghwan;Kim, Ilhyun;Lee, Hail;Song, Yungsun
    • Journal of Korean Medicine Rehabilitation
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    • v.23 no.3
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    • pp.15-26
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    • 2013
  • Objectives The aim of this study was to investigate antimicrobial activity of Huanggeumjakyak- tang water extract against MRSA. Methods The antibacterial activities of Huanggeumjakyak-tang were evaluated against 3 strains of Methicillin-resistant Staphylococcus aureus (MRSA) and 1 standard Methicillinsusceptible S. aureus (MSSA) strain by using the disc diffusion method, minimal inhibitory concentrations (MICs) assay, colorimetric assay using MTT test, checkerboard dilution test and time-kill assay was performed under dark. Results The MIC (minimum inhibitory concentration) of Huanggeumjakyak-tang water extract against S. aureus strains ranged from 1,000 to $2,000{\mu}g/ml$. So we confirmed that it has a strong antibacterial effect. Also the combinations of Huanggeumjakyak-tang water extract and conventional antibiotics exhibited improved inhibition of MRSA with synergy effect. Conclusions The results obtained in this study suggest that Huanggeumjakyak-tang water extract showed antibacterial effect against MRSA, and it also showed reducing effect on the side-effect problems that are the major weak points of traditional antibiotics.

Tubeimoside-1 suppresses breast cancer metastasis through downregulation of CXCR4 chemokine receptor expression

  • Peng, Yaojin;Zhong, Yan;Li, Gao
    • BMB Reports
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    • v.49 no.9
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    • pp.502-507
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    • 2016
  • To examine the effect of TBMS1on breast cancer metastasis, and investigate the potential mechanism by which Tubeimoside-1 (TBMS1) inhibits the CXCR4 expression in breast cancer cells. The expression of CXCR4 in breast cancer cell lines was determined by immunoblotting and real-time PCR. The effect of TBMS1 on NF-κB binding activity was evaluated by EMSA assay and ChIP analysis. Cell proliferation and invasion were analyzed by MTT assay and transwell invasion assay, respectively. The effect of TBMS1 on breast cancer metastasis was further evaluated in a metastasis model of nude mice. TBMS1 suppressed the expression of CXCR4 through inhibition of NF-κB binding activity. TBMS1 inhibited CXCL12-induced invasion in breast cancer cells, while ectopic expression of CXCR4 abolished the inhibitive activity of TBMS1. TBMS1 suppressed breast cancer metastasis in the metastatic model of nude mice. TBMS1 suppressed the CXCR4-mediated metastasis of breast cancer by inhibiting NF-κB binding activity.

Anti-cancer Effects of Kamiboa-tang and some other Traditional Medical Prescriptions (가미보아탕(加味保我湯) 및 수종(數種) 한방처방의 항암효과에 대한 연구)

  • Sung, Hyun-Jea
    • The Journal of Internal Korean Medicine
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    • v.28 no.2
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    • pp.321-332
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    • 2007
  • Objectives : Anticancer and immune-modulating effects of several Korean medical prescriptions including Yukgunja-tang, Bohwa-tang, Sogam-Won, and Kamiboa-tang were investigated. Methods : In vitro anti-cancer effects were measured by cytotoxicity MTT assay using SNU-1 gastric cancer cell lines, In vivo anti-cancer effects were measured by increased life span of S-180 sarcoma-injected ICR mouse. Immune-modulating effects were analyzed by measuring hemagglutinin titer, appearance of rosette forming cells, lymphocyte proliferation, and phagocytic index in methotrexate-pretreated mice. Results : In vitro assay showed that only Sogam-won showed cytotoxic effect with $IC_{50}$ of 87.9 ${\mu}g/ml$. All other prescriptions showed no cytotoxic effects against SNU-1 gastric cancer cell line. However, in vivo assay showed that Sogam-won showed lowest anti-cancer effects in contrast to its highest cytotoxic effects, Kamiboa-tang, which showed no cytotoxic effect, showed the highest in vivo anticancer effects, with increased life span of 140%. Kamiboa-tang showed significant immune-enhancing activities by significantly increasing rosette forming cells, lymphocyte proliferation, and phagocytic index in methotrexate-pretreated mice (P<0.05). Conclusion : The anticancer effect of Kamiboa-tang is not mediated by direct inhibition of cancer cells but is mediated by improving immune reactions against cancer cells.

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Phenolic Compounds from Antioxidant Plant Materials and their Protective Effect on PC12 cells (항산화 식물의 페놀성화합물에 의한 PC12 세포보호 효과연구)

  • You, SoHyeon;Kim, Gun-Hee
    • Journal of the Korean Society of Food Culture
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    • v.33 no.1
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    • pp.86-94
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    • 2018
  • This study was designed to evaluate the antioxidant activities and protective effects on PC12 cells of the extract of Epimedium koreanum and its main constituents icariin and icariside I. After screening the seven identified flavonoid glycosides from E. koreanum through DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) assay, E. koreanum, Icariin and Icariside I exhibited significant effect on radical scavenging activity. E. koreanum, icariin and icariside I were examined using DPPH, ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) and FRAP (ferric reducing ability power) assay. In all antioxidant assays, E. koreanum, icariin and icariside I showed high radical scavenging activities in a dose-dependent manner. Protective effects against $H_2O_2-induced$ PC12 cells were assessed with MTT assay. The results indicated that cell viability and protection on PC12 cells of icariside I and icariin increased dose dependently. These study results suggest that E. koreanum, icariin and icariside showed high antioxidant capacities and cell protective effects. Icariside I, one of the metabolites of icariin, may be a new and effective flavonoid compound as a functional component.

Neuroprotective Effect according to Reactive Oxygen Species Scavenging Activity from Extracts of Cudrania tricuspidata Leaves (활성산소 소거활성에 따른 꾸지뽕잎 추출물의 신경세포 보호 효과)

  • Kang, Young-Kyoung;Lee, Eun-Ah;Park, Hae-Ryong
    • Korean journal of food and cookery science
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    • v.28 no.6
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    • pp.821-828
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    • 2012
  • In an attempt to identify the neuroprotective effect of Cudrania tricuspidata (CT) leaves against ROS (reactive oxygen species)-induced oxidative stress in neuronal cells, the extracts from CT leaves were investigated using PC12 cells and N18-RE-105 cells. The methanolic and ethanolic extracts from CT were denoted as CTM (Cudrania tricuspidata Leaves methanolic extracts) and CTE (Cudrania tricuspidata Leaves ethanolic extracts), respectively. The neuroprotective effects of the extracts were measured by DCF-DA assay, MTT reduction assay, and LDH release assay. The PC12 cells exposed to $H_2O_2$-induced oxidative stress and the N18-RE-105 cells exposed to glutamate-induced oxidative stress were treated with various concentrations of CTM and CTE. The results, CTM treatments resulted in the induction of a dose-dependent protective effect in PC12 cells and N18-RE-105 cells. Interestingly, CTE also showed neuroprotective effect in PC12 cells and N18-RE-105 cells. Therefore, these results suggest that CTM and CTE could be a new potential candidate as neuroprotective agents against ROS-induced oxidative stress in neuronal cells.

Haemocytes responses of the pearl oyster, Pincdata fucata, at different temperatures

  • Choi, Young-Joon;Hwang, Jee-Youn;Baeck, Gun-Wook;Kim, Mu-Chan;Park, Hyung-Jun;Choi, Byoung-Dae;Kang, Suk-Joong;Park, Chan-Il
    • Journal of fish pathology
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    • v.21 no.3
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    • pp.241-246
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    • 2008
  • The effects of temperatures on pearl oyster, Pincdata fucata were studied by evaluating some functional immune responses of the haemocytes. Water temperature is one of the most important factors in bivalve immune defense. Haemocytes comprise a primary line by inflammation, encapsulation and phagocytosis. These phagocytic abilities of haemocytes were observed in different temperatures. The number of the circulating haemocytes by migratory assay, phagocytic activities by MTT assay and reactive oxygen species production of haemocytes by CL assay were measured at different temperatures. Results showed that pearl oyster maintained at 20℃ and 25℃ displayed significantly higher values for all the measured immune parameters in comparison to maintained at 10, 15, and 30℃.

Effects of White Ginseng-Ejung-tang Acupuncture Solution on Nitric Oxide and Hydrogen Peroxide Production in LPS-induced Mouse Macrophages (백약(白藥)이 포함된 이중탕(理中湯)약침액의 LPS로 유발된 마우스 대식세포의 nitric oxide 및 hydrogen peroxide 생성에 미치는 영향)

  • Lee, Ji-Young;Kim, Young-Jin;Park, Wan-Su
    • Korean Journal of Acupuncture
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    • v.28 no.1
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    • pp.61-69
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    • 2011
  • Objectives : The purpose of this study is to investigate effects of White Ginseng-Ejung-tang acupuncture solution (EJ) on nitric oxide (NO) and of hydrogen peroxide production in RAW 264.7 mouse macrophages stimulated by lipopolysaccharide (LPS). Methods : Cell viability was measured by modified MTT assay. NO production was measured by Griess reagent assay. Hydrogen peroxide production was measured by dihydrorhodamine 123 (DHR) assay. Significant differences were examined by using a Student's t-test. Results : The results of the experiment are as follows. 1. EJ did not show cell toxicity against RAW 264.7 cells for 24 hr incubation at the concentrations of up to $200\;{\mu}g$/mL in RAW 264.7 cells. 2. EJ significantly inhibited NO production for 24 hr incubation in RAW 264.7 cells (p <0.05). 3. EJ significantly inhibited the LPS-induced production of NO for 24 hr incubation in RAW 264.7 cells (p <0.05). 4. EJ significantly inhibited the LPS-induced production of hydrogen peroxide for 16, 24, 40, 48, 64, and 72 hr incubation in RAW 264.7 cells (p <0.05). Conclusions : These results suggest that EJ has an anti-inflammtory property related with its inhibition of NO and hydrogen peroxide production in LPS-induced macrophages.

Effects of Anemarrhenae Rhizoma on LPS-stimulated Expression of COX-2 and iNOS in mouse BV2 microglial cells

  • Seo, Bong-Won;Kim, Ee-Hwa;Park, Se-Keun;Jang, Mi-Hyeon;Choi, Sun-Mi;Lee, Eun-Yong
    • Journal of Acupuncture Research
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    • v.22 no.2
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    • pp.155-162
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    • 2005
  • Anemarrhenae Rhizoma (AR) has been widely used for the treatment of various diseases in Oriental medicine. To investigate whether AR possesses anti-inflammatory effects against lipopolysaccharide (LPS)-induced inflammation in the BV2 microglial cells, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide) assay. reverse transcription -polymerase chain reaction (RT-PCR), and prostaglandin E2 (PGE2) assay, and nitric oxide (NO) detection assay were performed. From the present results, AR was shown to suppress PGE2 synthesis and NO production by inhibiting the LPS-stimulated enhancement of cyclooxygenase-2 and inducible nitric oxide synthase expression in BV2 microglial cells. These results suggest that AR may offer a valuable means of therapy in the treatment of inflammatory diseases by attenuating LPS-induced PGE2 synthesis and NO production.

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