• Title/Summary/Keyword: MTS Assay

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Anticancer Effects of Cisplatin in Combination with Paeonia Japonica in YD-10B Cells (YD-10B에서 Cisplatin과 백작약의 병용처리에 의한 항암 효과)

  • Kim, Eun-Jung
    • The Journal of the Korea Contents Association
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    • v.20 no.6
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    • pp.124-130
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    • 2020
  • The present study investigated the anti-proliferate and anti-invasive of Phorbol 12-myristate 13-acetate (PMA)-induced matrix metalloproteinase (MMP-2) and MMP-9 activities of combined treatment with cisplatin and ethyl acetate fractions of Paeonia japonica. Cell Proliferation was detected by the MTS assay and the activity and mRNA expression of MMP-2/-9 were examined by zymography and RT-PCR. As results, cisplatin or p. japonica treatment of YD-10B cells resulted in a dose-dependent inhibition of cell growth. Also, the viability of YD-10B cells treated with combination of 200 μM cisplatin and 50 ㎍/ml p. japonica was inhibited to 50% in compared with the cisplatin alone. In PMA-treated YD-10B cells, co-treatment of 200 μM cisplatin with 50 ㎍/ml p. japonica significantly inhibited mRNA expression and protein activation of MMP-2/-9. Therefore, This study suggest that the combination treatment of cisplatin and p. japonica potentiates a promising anti-invasive agent and has more potential anti-cancer drug for oral cancer therapy than cisplatin alone.

Primary Cultures of Drosophila melanogaster Gut Cells for Studies of Intestinal Stem Cell Regulation (장줄기세포 조절 연구를 위한 초파리 장세포의 일차배양)

  • Yoon, Young-Il;Hwang, Jae-Sam;Goo, Tae-Won;Han, Myung-Sae;Ahn, Mi-Young;Yun, Eun-Young
    • Journal of Life Science
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    • v.22 no.5
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    • pp.621-626
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    • 2012
  • $Drosophila$ $melanogaster$ has been used as a useful model to study development and disease. In this study, we established the primary culture method of $Drosophila$ in the intestine to understand how intestinal stem cells (ISCs) mediate tissue repair during infection and disease. To obtain intestinal cells, we separated intestines from adult flies and isolated single cells by enzymatic treatment. The survival of cultured cells was measured using MTS-analysis. The maximum growth rate of the cells was observed on the 9th day after seeding. In addition, the presence of ISCs and enteroendocrine cells was confirmed by delta and prospero staining. Accordingly, we supposed that $Drosophila$ $melanogaster$ gut cells established in this study are probably useful in studies about intestinal stem cell regulation and various diseases occurring in the intestine.

Antioxidant Activities of Rice Bran Extracts for Wellness Convergence (융복합적인 웰리스를 위한 미강추출물의 항산화 활성)

  • Lee, Jae-Hyeok;Park, Jeong-Suk
    • Journal of Digital Convergence
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    • v.13 no.2
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    • pp.401-406
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    • 2015
  • The aim of present study is to investigate antioxidative effect of the Rice Bran Extracts and Defatted Rice Bran Extracts. Rice Bran Extracts used Rice Bran Water Extract, Rice Bran Ethanol Extract, Defatted Rice Bran Water Extract, Defatted Rice Bran Ethanol Extract. This study was carried out to examine quenching effects of Rice bran extracts on DPPH-, Riboflavin-, and Xanthin oxidase- originated superoxide activities. In addition, in order to determine whether Rice Bran Extract can be safely applied to human skin, the cytotoxic effects of Rice Bran Extract in Human Dermal Fibroblast cells were determined using MTS Assay. These results demonstrated that RBE and DRBE had anti-oxidative properties and did not induce the cytotoxic effects in Human Dermal Fibroblast cells. Therefore, these findings suggest that anti-oxidative properties of RBE and DRBE may be considered convergence with skin care.

Induction of Apoptosis in AGS Human Gastric Cancer Cells by Platycarya strobilacea Leaf Extract (굴피나무 잎 추출물의 위암세포에 대한 세포사멸 유도 효과)

  • Lee, Hyeong-Seon
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.283-288
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    • 2021
  • This study investigated the anticancer activity of methanol extract from Platycarya strobilacea leaf in AGS human gastric cancer cells. We determined the cell viability effect of P. strobilacea using MTS assay. Apoptosis induction and cell cycle arrest were confirmed by fluorescein isothiocyanate and propidium iodide staining using cellometer K2. The mRNA expression levels of the Bcl-2 family were confirmed by reverse transcription-polymerase chain reaction. The cell viability was decreased in a dose-dependent manner treated with different concentrations of P. strobilacea. Total, early, and late apoptotic cells were dramatically increased, and the cell cycle was arrested at the sub-G1 phase. The mRNA expressions of Bcl-2 and Bcl-xL were reduced, whereas pro-apoptotic factors, Bax and Bak, were increased in a dose-dependent manner. These results suggested that P. strobilacea leaf extract induced significant apoptotic activity through an intrinsic mitochondria pathway.

Antibacterial and anti-obesity effects of Abeliophyllum distichum Nakai: an in vitro study

  • Song, Dong Cheol;Lee, Ji Hwan;Oh, Han Jin;Kim, Yong Ju;An, Jae Woo;Chang, Se Yeon;Go, Young Bin;Cho, Hyun Ah;Cho, Jin Ho
    • Korean Journal of Agricultural Science
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    • v.48 no.3
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    • pp.557-565
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    • 2021
  • Interest in research on various medicinal plants has increased globally over the last few decades, possibly due to their possible antibacterial and antioxidant activities. The present study was conducted to verify the antioxidant effects, antibacterial activity, and collagen synthesis and cell viability outcomes of adipocytes upon exposure to Abeliophyllum distichum Nakai (AdN). Antibacterial activity was measured through the Disc diffusion method to compare the growth ability of pathogenic microorganisms (E.coli, Salmonella). The absorbance was measured at 560 nm to calculate the active oxygen scavenging ability. Fibroblasts were dispensed in a 96-well plate at a density of 1 × 105 cells·well-1. The amount of procollagen was measured in each case using a procollagen type 1 C-peptide EIA KIT. The cytotoxicity of the Abeliophyllum distichum Nakai extract against animal adipocytes (Hanwoo backfat cells) was determined using a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay, a method that measures the conversion of MTS to Formazan by means of mitochondrial dehydrogenases. The concentrations of the samples were made to be 0.0125, 0025, 0.05, 0.1, and 0.5% and all were -completely absorbed into the disc in an incubator at 37℃ for 24 to 36 hours. For the 0.125 mg·disc-1, effects of Abeliophyllum distichum Nakai on the antioxidant effect, antibacterial activity, and cell viability of adipocytes were found. However, Abeliophyllum distichum Nakai had no effect on collagen synthesis, thus suggesting that AdN extracts may be useful for the prevention and/or treatment of obesity.

Screening Assay for Identification of Endocrine Disruptors with Androgen Activities using LNCaP Cells (LNCaP 세포주를 이용한 내분비계장애물질중 안드로겐성 확인시험을 위한 검색법)

  • 김진호;정혜주;김영옥;정승태;박재현;조대현;김동섭
    • Toxicological Research
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    • v.18 no.1
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    • pp.59-64
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    • 2002
  • Substantial evidences have been accumulated about the hormone-like effects of exogenous substances such as pesticides and industrial chemicals during past years. The effects of these substances on the endocrine system are believed to be either enhancing or reducing of various endocrine action. It is necessary to identify putative causal agents by the batter system and to assess their ability to disrupt the endocrine system. A variety of in vitro and In vivo approaches have been used to determine the androgenic effects of environmental chemicals. To establish the method for assessment of the putative endocrine disruptors with androgenic activity, we carried out the cell proliferation assay by MTS method after treatment with the various concentration of testosterone in LNCaP cells (human prostatic cancer cell line) and also observed the expression of androgen-related genes by quantitative RT-PCR. In the cell proliferation assay, the results showed that the grouth of LNCaP cells increased within level of at least 10pM testosterone. We measured by quantitative RT-PCR method on the effects of testosterone on mRNA expression of androgen receptor (AR), prostate-specific antigen (PSA), bone morphogenetic protein (BMP) and BMP receptor (BMPR) In LNCaP cells. The results demonstrated that mRNA expression of PSA and BMPR-IB was observed differently within level of at least 0.01 pM testosterone compared with non-treated control. These observations suggest that the detection of PSA and BMPR-IB mRNA by the quantitative RT-PCR in LNCaP cells is very sensitive method to identify the endocrine disruptors to have the androgenic effects.

Fibrinolytic, thrombin inhibitory, anti-oxidative and anti-inflammatory activities of Pleurotus ferulea (아위버섯 추출물의 혈전용해, 트롬빈저해, 항산화 및 항염증 활성)

  • Kim, Eun-Jung;Kim, Jun-Ho
    • Journal of Mushroom
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    • v.13 no.1
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    • pp.30-36
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    • 2015
  • Our study investigated the fibrinolytic, thrombin inhibitory, anti-oxidative and anti-inflammatory activities of the water extract and solvent fractions isolated from Pleurotus ferulea. Fibrinolytic activity was investigated using the fibrin plate method. Thrombin inhibitory activity was used to analyze thrombin inhibitor assay. The DPPH assay was used to estimate anti-oxidative activity. Inhibition of NO production was measured for anti-inflammatory activity in LPS-activated murine RAW 264.7 macrophage cells. An MTS assay was used to evaluate the effects of the water extract and solvent fractions isolated from Pleurotus ferulea on cell viability. Our results showed the fibrinolytic activity to be strong in the ethyl acetate fraction at 1.33 plasmin units. The ethyl acetate fraction also showed high thrombin inhibitory activity at 94.45%. The anti-oxidative activity of the water extract was 37.01% and the anti-inflammatory activity of the chloroform fraction was 98.13%. These findings suggest that Pleurotus ferulea's extract and fractions could be applicable in the development of functional foods for the treatment and prevention of cardiovascular diseases.

The effects of Bee Venom on NO, H2O2 in Raw 264.7 cells and IL-1 in D10S cells (봉약침액(蜂藥鍼液)이 NO, H2O2, IL-1에 미치는 영향(影響)에 관(關)한 연구(硏究))

  • Song, Jeong-Yeol;Lee, Seong-No;Jo, Hyun-Chul;Kim, Kee-Hyun
    • Journal of Pharmacopuncture
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    • v.5 no.2
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    • pp.52-62
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    • 2002
  • Objectives : The purpose of this study was to investigate the effects of Bee Venom on NO, $H_2O_2$ expression induced by LPS in Raw 264.7 cells as a murine marcrophage cell line and on IL-1 expression induced by LPS in D10S cells. Methods : The expression of NO was measured by MTT Assay and IL-1 by MTS Assay. The expression of $H_2O_2$ was measured as ROS level within the cell using by FACS analysis. The non-toxic concentration(from $0.1\;{\mu}g/ml\;to\;5\;{\mu}g/ml$) of Bee Venom was determined by MTT Assay. Results : 1. Bee Venom inhibited the NO expression. The effective concentration of Bee Venom was $5\;{\mu}g/ml$ after 3 hours, 1 and $5\;{\mu}g/ml$ after 1 day and 2 days. The all concentration of Bee Venom inhibited the NO expression after 6, 12 hours and 3 days. 2. Bee Venom inhibited the $H_2O_2$ expression in a dose-dependent manner compared to the control. 3. Bee Venom could not significantly inhibit the IL-1 expression.

Investigation of the Effect of Sappan Lignum and Brazilin on Expression of Tight Junction Related-genes in Human Keratinocyte (소목(蘇木)과 그 지표물질인 brazilin이 인간 유래 각질 형성 세포의 tight junction 유전자 발현에 미치는 영향)

  • Cheon, Seong Hye;Choi, Sun Kyung;Cho, Nam Joon;Kim, Kee Kwang;Lee, Woong Hee;Hwang, Hyung Seo;Kim, Kyoon Eon;Han, Hyosang
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.32 no.2
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    • pp.106-112
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    • 2018
  • The aim of this research was to determine the diverse effects of Sappan Lignum extract and brazilin on human keratinocyte HaCaT cells. We confirmed the antioxidant effect of Sappan Lignum extract and brazilin was analyzed by using an ABTS assay, confirming the efficacy of water extraction method. Also, we examined effect of Sappan Lignum extract and brazilin on the cell viability, using the MTS assay in HaCaT cells. mRNA expression levels of tight junction-related genes associated with skin barrier in HaCaT cells were analyzed using quantitative real-time PCR analysis. Sappan Lignum extract increased the cellular activity of HaCaT cells and the expression of the tight junction-related genes claudin 3, claudin 6, and ZO-2. Brazilin displayed the same effects as that of the extract on HaCaT cells activity and tight junction-related genes expression. Furthermore, dispase assay demonstrated altered cell-cell adhesion strength of Sappan Lignum extract or brazilin treated HaCaT cells. Sappan Lignum extract or brazilin might be an useful ingredient in skin-mosturizinng and anti-wrinkle cosmetics, given its effects of altering mRNA expression of tight junction-related genes and enhancing cell-cell adhesion strength of HaCaT cells.

Anti-atherosclerosis Effect of Imyosan Extract in Human Aortic Smooth Muscle Cells (사람 대동맥 평활근 세포에서 이묘산(二妙散)의 항동맥경화 활성)

  • Yun, Hyun-Jeong;Heo, Sook-Kyoung;Yi, Hyo-Seung;Kim, Tae-Hoon;Kim, Dong-Wan;Kim, Sun-Mo;Park, Sun-Dong
    • The Korea Journal of Herbology
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    • v.23 no.4
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    • pp.113-120
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    • 2008
  • Objectives: This study was evaluated to elucidate the inhibitory potential of Imyosan(IMS) and its components, Phellodendri Cortex(PC: Phellodendron amurense Rupr., Hwangbaek in Korean) and Atractylodis Rhizoma(AR: Atratylodes lancea D.C., Changchool in Korean), on human aortic smooth muscle cells(HASMC) migration and production of matrix metalloproteinase (MMP)-2 and MMP-9 by TNF-${\alpha}$ treatment. Methods: Cytotoxic activity of IMS and its components on HASMC was using 5-(3-caroboxy meth-oxyphenyl)-2H-tetra-zolium inner salt(MTS) assay. Effect of IMS, PC and AR on TNF-${\alpha}$-induced HASMC migration underside of matrigel filter was stained with hematoxylin-eosin. And total number of cells that migrated to the underside of the filter was counted. MMP-2 and MMP-9 activity was evaluated by gelatin zymography assay. Results: The matrigel migration assay showed that IMS effectively inhibited the TNF-${\alpha}$-induced migration of HASMC. Moreover, IMS significantly inhibited MMP-9 activity. Our present study demonstrates that IMS and its components inhibit TNF-${\alpha}$-induced HASMC migration and MMP-9 activity. The inhibitory effect of IMS extract is more potent than that of its component herb extracts. Conclusions: These results provide evidence that IMS has multiple effects in the inhibition of HASMC migration and may offer a therapeutic approach to block HASMC migration.

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