• Title/Summary/Keyword: MTT 실험

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Biological Activities of Dried Garlic, Red Ginseng and Their Mixture (건조 마늘, 홍삼 및 이들 혼합물의 생리활성)

  • Shin, Jung-Hye;Jung, Kang-Min;Lee, Soo-Jung;Yang, Seung-Mi;Rue, Gi-Hyun;Sung, Nak-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.12
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    • pp.1633-1639
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    • 2009
  • Water extracts were extracted on water bath for 3 hrs at 90${^{\circ}C}$ after 5 times water was added to hot-air dried garlic (DG) and red ginseng (RG), respectively. Its extracts were dried on rotary evaporator, and then the mixture samples were prepared from RG extracts added to DG extracts at 5, 10, and 15%. To test biological activities such as DPPH, nitrite scavenging, tyrosinase, $\alpha$-glucosidase, human gastric (AGS), and human colon cancer cell (HT-29) growth, dryness of DG, RG, and mixture samples were diluted 500-10,000 $\mu$g/mL adding deionized water, respectively. DPPH scavenging activities of mixture sample were slightly higher than DG and RG sample alone. Nitrite scavenging activity was the highest in DG, and the other samples were below 45%. Inhibition activity of tyrosinase was below 35.0% in tested all samples. Inhibition activity of $\alpha$-glucosidase was lower in DG or RG alone, while its activity of mixture sample was increased in proportion to concentration of RG extracts. Inhibition of AGS cell growth was more effective in mixture samples than DG and RG alone, while inhibition of HT-29 cell growth was more effective in DG or RG alone than mixture samples.

Determination of Ceramide-Induced Apoptotic Cell Death in Mouse Granulosa Cell Cultured In Vitro (생쥐 난소 과립세포의 체외배양중 세라마이드에 의한 자연세포사의 측정)

  • Kim, Jong-Hoon;Kim, Gyoung-Mi;Yoon, Yong-Dal
    • Development and Reproduction
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    • v.2 no.1
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    • pp.1-8
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    • 1998
  • In mammalian ovary, major portion(>99%) of ovarian follicles undergo atresia. Recent studies have shown that this phenomenon is mediated via GC apoptosis. Ceramide, a product of sphingomyelin hydrolysis, has been proposed as a novel lipid second messenger with specific roles in mediating antiproliferative responses including apoptosis and cell cycle arrest. In the present study, we have examined the effect of ceramide on apoptotic cell death of GC in vitro. GCs were harvested by squeezing the antral follicles from the immature mice (3-4 weeks) and cultured in MEM medium with 10% fetal bovine serum. The cells were treated with various concentrations of ceramide (0 to 50 \mu M)and cultured up to 24 h.Cell death was determined by MTT cell viability assay and apoptosis was examined by acridine orange staining, in situ 3'-end labeling(TUNEL), and flow cytometry. Ceramid treatment induced apoptotic cell death of GC in a time- and a dose-dependent manner. Results of flow cytometric analysis showed that creamide-induced cell death was mostly confined to the $G_{0}$/$G_{1}$ cells. these results provide an evidence for ceramide as a lipid second messenger of apoptosis in mouse GC.

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Experimental Studies on the Anti-tumor and the Immunomodulatory Effects of Chungsimbohyeltang (청심보혈탕(淸心補血湯)의 항암(抗癌) 및 면역증강효과(免疫增强效果)에 관(關)한 실험적(實驗的) 연구(硏究))

  • Park, Se-Ki;Ryu, Bong-Ha;Park, Dong-Won;Ryu, Ki-Won
    • The Journal of Internal Korean Medicine
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    • v.19 no.1
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    • pp.221-246
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    • 1998
  • The aim of this experimental study was to evaluate the anti-tumor effects of Chungsimbohyeltang through investigation about viability of tumor cell by MTT assay, survival period of mice transplanted with L-1210 cells, growth inhibition on the tumor cell, body weight variation in mice transplanted with sarcoma-180 cells and its immunomodulatory effects through the investigation on delayed type hyper-sensitivity, the hemagglutinin and hemolysin titers for humoral immune response, the appearance of rosette forming cells for cell-mediated immune response, the natural killer cell activity, the transformation of lymphocyte, the productivity of Interleukin-2 and phagocytic index K was performed in immune-depressed ICR mice induced by methotrexate treatments. The results were as follows ; 1. $IC_{50}$ of Chungsimbohyeltang treated group was 5.85mg/ml in SNU-C4 cell, 1.38mg/ml in SNU-396 cell, 0.21mg/ml in SNU-1 cell, so it had low anti-tumor activity. 2. The both groups of Chungsimbohyeltang extract 10mg/kg and Chungsimbohyeltang extract 20mg/kg had no toxicity and the group of Chungsimbohyeltang 20mg/kg which was shown 120% in ILS had the effect of life prolongation in mice transplanted with L-1210 cells. 3. In the growth inhibition on the tumor cells, only the group of Chungsimbohyeltang extract 20mg/kg was noted and in the weight variation in mice transplanted with sarcoma-180 cells, both groups of Chungsimbohyeltang extract had a significant effect. 4. In the delayed type hypersensitivity and appearance of rosette forming cells, both groups of Chungsimbohyeltang extract didn't have any significant effect. 5. The hemagglutinin titers was slightly increased with no significance, and the hemolysin titers was significantly increased in the only group of Chungsimbohyeltang extract 20mg/kg. 6. The natural killer cell activity of the Chungsimbohyeltang extract groups was significantly increased in the ratio of 100:1 of effector and target cells, but it was not significantly increased in the ratio of 50:1, 10:1. 7. The transformation of lymphocyte and the productivity of Interleukin-2 were increased significantly and in dose-dependent manner in both group of Chungsimbohyeltang extract. 8. The phagocytic effect of macropage was significantly increased in both groups of Chungsimbohyeltang extract. Considering the results above, we could conclude that Chungsimbohyeltang have an indirect anti-tumor effect through the modulation of immunme response, although it had not toxicity on the tumor cell it self.

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Free Redical Scavenging and Cytotoxicity Activitives of Soybean Germ Saponin (대두배아 사포닌의 유리기 생성 억제 및 세포독성)

  • 류병호;이홍수;김현대
    • The Korean Journal of Food And Nutrition
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    • v.15 no.2
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    • pp.151-157
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    • 2002
  • This study was carried out to investigate functional activities of the free radical scavenging and germ of Glycin max. Merrill fur cytotoxicity toward P338 and L1210 cells derived from mouse. Effect of crude saponin were examined to oxygen radicals and their scavenger enzymes in liver fractions of Spragae-Dawley(SD) rats. Male rats were fed basic diets of control and experiment diets of 0.5∼1.0% crude saponin. There were no significant differences in hydroxy radical($.$OH) formation of liver mitochondria and microsomes in 1.0% group, while $.$OH formations were significantly decrease in 0.5% and 1.0% saponin compared with control group. Their oxygen radical(O$_2$$\^$$.$/) scavenging activities were significantly decrease in liver cytosol of 0.5% and 1.0% saponin group compared with control group. Soybean germ saponin was isolated purified by the method of HPLC to investigate the cytotoxicity of mouse cells by using the MTT assay. SA-1 saponin fraction of soybean germ showed to inhibit toward growth cell of P338 and L1210 cells and its showed less than 50% cytotoxicity These results suggest that the saponin may play a effective role in attenuating a oxygen radical formations and increasing a scavenger enzyme activities.

Antioxidant and Anti-inflammatory Effect of Extracts from Flammulina velutipes (Curtis) Singer (팽이버섯 추출물의 항산화 및 항염증 활성)

  • Kang, Hyun-Woo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.8
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    • pp.1072-1078
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    • 2012
  • The potential antioxidant and anti-inflammatory effect of water and ethanol extracts from Flammulina velutipes (Curtis) Singer (FVS) on hydrogen peroxide ($H_2O_2$) and LPS-induced oxidative damage in PC-12 and RAW264.7 cells were investigated. The DPPH radical scavenging activities of the water extract from FVS was the highest, and the 50% inhibitory concentration value was 0.388 mg/mL. Also, the antioxidant activities of water and ethanol extracts were determined by ferric reducing antioxidant power, 2,2'-azino-bis-(3-ethybenzothiazoline-6-sulphonic acid) radical scavenging activity. In addition, water extract from FVS showed a strongly inhibitory effect on lipid peroxidation by measuring ferric thiocyanate values. The water extract decreased cell apoptosis in PC-12 cells against $H_2O_2$-induced oxidative damage. In addition, FVS extracts exhibited the strongest nitric oxide (NO) inhibition activity. It was also found that FVS extract inhibited LPS-induced iNOS and COX-2 expression in RAW264.7 cells. The findings of the present study suggest that extracts of FVS exhibit anti-oxidative and anti-inflammatory activity against oxidative stress and/or stimulated cells.

Antimicrobial Effect of Carvacrol against Cariogenic and Periodontopathic Bacteria (치아우식증 및 치주질환 원인균에 대한 Carvacrol의 항균효과)

  • Park, Soon-Nang;Lee, Dong-Kyun;Lim, Yun-Kyong;Kim, Hwa-Sook;Cho, Eu-Gene;Jin, Dongchun;Kim, Saeng-Gon;Kook, Joong-Ki
    • Korean Journal of Microbiology
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    • v.48 no.1
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    • pp.52-56
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    • 2012
  • The aim of this study was to evaluate the antimicrobial effect of carvacrol against periodontopathic and cariogenic bacteria and its cytotoxicity in human oral tissue cells. We tested their antibacterial properties against mutans streptococci and five major periodontopathic bacterial species involved in periodontal disease. The antimicrobial activity was evaluated by the minimal inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The cell viability of carvacrol on normal human gingival fibroblast (NHGF) cells was tested by metyl thiazolyl tetrazolium assay. The data showed that carvacrol had remarkable antimicrobial effect on tested bacteria with a MIC and MBC values ranged from 16 to $128{\mu}g/ml$ and from 32 to $128{\mu}g/ml$, respectively. In cell toxicity studies, carvacrol had significantly decreased cell viability when NHGF cells were treated at $128{\mu}g/ml$. These findings suggest that carvacrol has a strong antimicrobial activity against periodontopathic and cariogenic bacteria. However, in order to use it as a component of gargling solution or toothpaste, its concentration should be below $64{\mu}g/ml$ and other compounds having an antimicrobial activity against periodontopathic and cariogenic bacteria should be used together.

Antibacterial, antioxidant and antitumor activities of mushroom mycelium mixed culture extracts (버섯 균사체 혼합 배양 추출물의 항균, 항산화 및 항암 활성)

  • Kim, Man-Cheol;Kim, Ju-Sang;Heo, Moon-Soo
    • KSBB Journal
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    • v.23 no.2
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    • pp.158-163
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    • 2008
  • In this study, we have investigated the antibacterial, antioxidant, and antitumor activities of mycelium cultural extract from mushroom. Mushroom mycelium was grown in a synthetic liquid media such as PD broth, YM broth or citrus extracts. In antibacterial activity test, the best result was achieved when mycelium cultural extracts from Phellinus linteus and Coriolus versicolor were incubated together on YM broth. On the other hand, mushroom mycelium cultured on citrus extracts showed better activity than that on PD broth. We have also tested the antioxidant activity at concentration up to 10 mg of mycelium cultural extract/mL. The more it is in higher concentration, the more the activity increases. The higher antioxidant activity was observed both on PD broth containing the Phellinus linteus and Coriolus versicolor mycelium and citrus extract containing the same. The complex culture extracts obtained from the synthetic medium and citrus extract medium showed 10-89% of the 1,1-diphenyl-2-picrylhydrazyl radical scavenger activity. The antitumor activity of mycelium cultural extract was examined by using MTT assay on A549 cells. Mushroom mycelium cultured on citrus extracts showed interestingly higher antitumor activity than that on synthetic liquid media.

Antioxidant and Cell Proliferation Effects of Acanthopanax senticosus for Extract in Human Osteoblast-like MG-63 Cell Line (가시오가피 추출물의 항산화활성 및 MG-63 조골세포 증식과 alkaline phosphatase 활성에 미치는 효과)

  • Lim, So-Young;Leem, Jae-Yoon;Lee, Choong-Soo;Jang, Yu-Jung;Park, Jeong-Woo;Yoon, Sun
    • Korean Journal of Food Science and Technology
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    • v.39 no.6
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    • pp.694-700
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    • 2007
  • Acanthopanax senticosus is a common Asian herb also known as "Siberian Ginseng". It is often used as a traditional herbal medicine for reducing damage in the liver, kidney, bone and muscle. In the present study we investigated the ferric reducing/antioxidant power and total phenolic contents of the ethanol-/water-extracts obtained from the stems and leaves of Acanthopanax senticosus. Osteoblast cellular proliferation was evaluated using the MTT and alkaline phosphatase activity assays in the human osteoblast-like MG-63 cell line. Acanthopanax senticosus extracts exerted remarkable ferric reducing/antioxidant power and contained high amount of phenolics. Among the extracts the stem-/ethanol-extract showed the highest antioxidant activity and total phenol content. Interestingly a highly positive correlation was found between antioxidant activity and total phenol content (p < 0.01). Proliferation of MG-63 osteoblast cells was highest in the stem-/ethanol-extract and alkaline phosphatase activity significantly increased in the water-extract of the stems (p < 0.05). These findings suggest that Acanthopanax senticosus extracts have antioxidant activity for preventing oxidative stress-related diseases and may have beneficial effects on bone health through the proliferation of osteoblast cells.

Growth Inhibitory and Quinone Reductase Activity Stimulating Effects of Internal Organs of Aplysia kurodai Fractions on Cancer Cell Lines In vitro (군소내장 분획물의 in vitro에서의 암세포 성장억제 및 quinone reductase 유도 활성 증가 효과)

  • Shin, Mi-Ok
    • Journal of Life Science
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    • v.20 no.6
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    • pp.877-884
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    • 2010
  • We investigated the growth inhibitory effect of internal organs of Aplysia kurodai (AK) on proliferation in cancer cell lines in vitro. The internal organs of AK were extracted with methanol (AKM), which were then further fractionated into four subfractions by using solvent partition method, resulting in hexane (AKMH), methanol (AKMM), butanol (AKMB), and aqueous (AKMA) soluble fractions. We determined the cytotoxic effect of these four fractions in four kinds of cancer cell lines - HepG2, MCF-7, HT29 and B16-F10 - by MTT assay. Among the four subfractions of AKM, AKMM showed the strongest cytotoxic effects on all cancer cell lines which were used. Morphological changes such as membrane shrinking and blebbing of cells were also observed in AKMM treatment in HepG2 cells. In addition, we also observed quinone reductase (QR) induced effect in the methanol layer (AKMM) of HepG2 cells. AKMM showed the highest induction activity of quinone reductase on HepG2 cells among the partition layers. The QR induced effect of AKMM was determined to be 2.4 at $100\;{\mu}g/ml$ level with a control value of 1.0. Although further studies are needed, the present work suggests that internal organs of Aplysia kurodai (AK) may be a chemopreventive agent for the treatment of human cells.

Studies on Skin Whitening Effects of Resorcinol Dipentyl Ether (레조시놀다이펜틸에터의 피부 미백 효능 평가)

  • Cha, Young Kwon;Cho, Hyun Dae;Cho, Wan Goo;Byun, Sang Yo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.43 no.2
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    • pp.115-124
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    • 2017
  • The objective of this study was to investigate skin whitening effect of Resorcinol dipentyl ether [1,3-di(pentyloxyl)benzene] by in vitro experiments. Resorcinol dipentyl ether was prepared by alkylation of resorcinol with 1-bromopentane. The reaction products were confirmed by NMR, MS and other analytical equipments. In order to evaluate the skin safety of resorcinol dipentyl ether, the cytotoxicity of the cells constituting the skin (keratinocyte, melanocyte, fibroblast) was analyzed and similar cell viability was observed in all cell lines as compared with the control group. Inhibition of extracellular melanin synthesis effect of resorcinol dipentyl ether was approximately 65.75% at $20{\mu}g/mL$ and inhibition of intracellular melanin synthesis effect of resorcinol dipentyl ether was approximately 53.89% at $20{\mu}g/mL$. The real-time PCR and western blot analysis of mRNA expression and protein expression of tyrosinase, TRP-1, and TRP-2 related to melanogenesis revealed that melanin inhibitory effect of resorcinol dipentyl ether was inhibited from the transcription stage respectively. Finally, this study suggested applicability of Resorcinol dipentyl ether [1,3-di(pentyloxyl)benzene] as a whitening functional cosmetic new material.