• Title/Summary/Keyword: MTT 검색법

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A Study of matrix metalloproteinase-9 inhibitor in root bark of ulmus davidiana planchon (유근피내의 Matrix Metalloproteinase-9 활성 억제제에 관한 연구)

  • Kong, Kwang-Hoon;Han, Kee-Jung;Lee, Kwang-Soo;Cho, Sung-Hye
    • Analytical Science and Technology
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    • v.18 no.2
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    • pp.104-111
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    • 2005
  • Several solvents were used to fractionate an extract obtained from the chapped root bark of Ulmus davidiana Planchon. The each fractionary part was condensed under reduced pressure and then examined to investigate the inhibitory effect on MMPs by modified gelatin zymography, where EA fraction showed the inhibition effect on the activity of MMPs. A compound showing inhibition effect on the MMPs was isolated and purified from EA fraction. Under IR, $^1H$- and $^{13}C$- NMR analyses it is very close to a catethin. This substance showed 48% inhibition effect on measurement of MMP-9 activity at 5 mM and 43% at 10 mM. To verify the effect of this substance on cells, human hepatoma, SK-Hep-1 cells as a cancer model, and Chang liver cells as a normal model were selected. MTT assay was performed to examine the cell viability by treatment of $1{\mu}L/mL$ of the purified substance on cells. The purified substance showed negligible toxicity on human liver cell line.

고분자량 Chitooligosaccharides의 in vitro 항종양성

  • 박헌국
    • Proceedings of the Korean Journal of Food and Nutrition Conference
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    • 2001.12a
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    • pp.127-127
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    • 2001
  • Chitosan에 Ttichoderma viride 유래의 cellulase를 처리하여 효소분해함으로써 고분자량 chitooligosaccharides를 얻었다. 생산된 올리고당을 HPLC로 분석한 결과 octamer 이상의 것이 전체 올리고당 중 49.3%에 달하는 비교적 고분자 화합물로 구성된 chitooligosaccharides의 혼합물이었다. MTT검색법에 의하여 고분자량 chitooligosaccha- rides의 정상세포주와 암세포주에 대한 세포독성을 실험하였다. 정상세포주인 아프리카 초록원숭이의 신장세포에 대한 $IC_{50}$/(50% 저해농도)값은 1,107.95$\mu$g/ml로 정상세포에 대한 독성은 거의 없었다. 폐암세포주인 A549, 방광암세포주인 J82, 대장암세포주인 SNU-C4, 위암세포인 SNU-1, 유방암세포주인 ZR75-1에 대한 $IC_{50}$/값은 각각 421.06$\mu$g/ml, 417.99$\mu$g/ml, 445.54$\mu$g/ml, 380,65$\mu$g/ml, 460.49$\mu$g/ml로 in vitro 종양세포 증식억제 활성을 나타내었다.

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Effect of glycyrrhetinic acid on the viability of human gingival fibroblasts (감초산이 인체 치은 섬유모 세포에 미치는 영향)

  • Yoo, Soo-Kyoung;Kim, Ki-Young;You, Yong-Ouk;Jang, Seon-Il;Kim, Kang-Ju;Park, Jong-Keun;Chung, Chong-Pyoung;Kurihara, Hidemi
    • Journal of Periodontal and Implant Science
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    • v.28 no.3
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    • pp.453-465
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    • 1998
  • 감초산이 인체 치은 섬유모세포에 미치는 영향을 세포의 성장과 증식, 총 교원질 합성 및 인체 치은 섬유모세포 핵내 acridine orange 결합으로 추적조사하였다. 조절이 되지 않는 성장을 해결하기 위하여 세포분화인자인 감초산이 배양 치은 섬유모세포의 활성에 미치는 효과를 검색하였다. 감초산 존재하의 배양 인체 섬유모세포의 세포성장 및 증식, 교원질 합성 및 세포 핵내 acridine orange 결합을 각각 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT)법, 4-hydroxyproline, 유식세포분석기를 이용한 acridine orange 결합으로 검색하였다. 형태학적으로 $100\;{\mu}g/ml$의 감초산으로 처리한 섬유모세포는 모양이 둥글게 되었다. 감초산은 $50\;{\mu}g/ml$ 이상의 농도에서 치은 섬유모세포의 성장과 증식을 억제하였다. 감초산 존재 시에 세포내 총 교원질 양이 감소하였고, 세포외배지내의 교원질 총 양이 증가하였다. 인체 치은 섬유모 세포를 $100\;{\mu}g/ml$의 감초산과 함께 24 시간동안 배양하였을 때, 80 채널 이상의 평균형광을 갖는 diploid 세포가 감소하였고, 80 채널 이하의 형광을 갖는 acridine orange결합이 증가하였다. 이러한 연구 결과 감초산은 인체 섬유모세포에서 세포성장 및 증식, 교원질합성 및 DNA 분절화를 유도함이 제시하였다.

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Effects of Rubus coreanus Miquel Extracts on the Activity and Differentiation of MC3T3-E1 Osteoblastic Cell (복분자(Rubus coreanus Miquel) 추출물이 MC3T3-E1 조골세포의 활성과 분화에 미치는 영향)

  • Lee Ji-Won;Lee In-Seon
    • Journal of Life Science
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    • v.14 no.6 s.67
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    • pp.967-974
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    • 2004
  • The osteoblastic cell activity is important for born formation, thus, this study was performed to investigation of that the effect of edible sources, Rubus coreanus Miquel (RCM), on the proliferation and differentiation of MC3T3-E1 osteoblastic like cell. The effects of RCM extract on cell proliferation were measured by MIT assay. At 1, $10\;{\mu}g/mL$ of RCM extract treated, that were elevated of cell proliferation to 103 and $142\%$ via control, respectively. And the cell differentiation were measured as alkaline phosphatase (ALP) activity at 3, 9, 18, and 27 days. As the results, the $10\;{\mu}g/mL$ was increased ALP activity more than 2.6 times compared with control, 1.4 times via positive control at 27th day (p<0.05). The optical concentration of RC extract was rechecked by ALP staining and Alizarin Red staining for investigation of the induction of ALP activity, nodule formation by mineralization. mRNA expression analysis showed that the RCM $(10\;{\mu}g/mL)$ increased in SOX9 as well as ALP in MC3T3-E1 cells. These results suggest that RC extract was stimulates the MC3T3-E1 cell proliferation and differentiation.

Effects of ENA-A(ENA actimineral resource A) Ion Water on the Activity and Differentiation of MC3T3-E1 Osteoblastic cell (ENA-A(ENA actimineral resource A) 이온수가 MC3T3-E1 조골세포의 활성과 분화에 미치는 영향)

  • Lee, Ji-Won;Jeon, Sang-Kyung;Kim, Hyun-Jeong;Lee, In-Seon
    • Journal of Life Science
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    • v.16 no.6
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    • pp.925-931
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    • 2006
  • Culture of osteoblast is extremely valuable in analyzing biological features that are specific to bone. ENA-A, ENA actimineral resource A, is a seaweed origin alkaline water. To investigate the bioactivity of ENA which act on bone metabolism, we studied the effects of a ENA on the activity of osteoblast MC3T3-E1 cells. ENA (1, 2, 4%) dose-dependently increased survival (p<0.05) and alkaline phosphatase activity (p<0.05) on MC3T3-E1 cell. And examined histochemistry and nodule formation according to the time course. To determine the expression patterns of bone-related proteins during the MC3T3-E1 osteoblast-like cell differentiation by using RT-PCR. This study suggest that ENA may promote the function of osteoblastic cells and play an important role in bone formation.

ABTS Radical Scavenging and Anti-Tumor Effects of Tricholoma matsutake Sing. (Pine Mushroom) (송이(Tricholoma matsutake Sing.)의 ABTS Radical 소거능과 암세포 생장 억제효능의 검색)

  • Kim, Young-Eon;Yang, Ji-Won;Lee, Chang-Ho;Kwon, Eun-Kyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.5
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    • pp.555-560
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    • 2009
  • Pine mushroom (Tricholoma matsutake Sing.) is an expensive and highly prized delicacy in Korean and Japanese cuisines with its unique flavor and functional properties. The pine mushroom juice (PMJ) was investigated for its antioxidant and anti-tumor activities with ABTS radical scavenging method and MTT assay. The phenolic contents in pine mushroom juice ranged from 1.19 to 54.99 GAEs mg/100 mL at the concentrations of $1{\sim}50\;mg/mL$. The ABTS radical scavenging activities of pine mushroom juice were 7.0%, 81.7% and 91.8% at the concentrations of 1, 10 and 50 mg/mL, respectively. The $IC_{50}$ values of pine mushroom juice were 605.9, 788.4, 583.6 and $232.5{\mu}g/mL$ on the cytotoxicities against AGS, HeLa, HepG2 and HT-29, respectively, and PMJ showed the strongest growth inhibitory activity against HT-29 cell. These results suggested therapeutic potential for pine mushroom juice as an anti-oxidant and anti-tumor agent.

Effects of Solidago virga-aurea var. gigantea Miq. Root Extracts on the Activity and Differentiation of MC3T3- E1 Osteoblastic Cell (미역취(Solidago virga-aurea var. gigantea Miq.) 뿌리 추출물이 MC3T3-E1 조골세포의 활성과 분화에 미치는 영향)

  • Park, Jung-Hyun;Lee, Ji-Won;Kim, Hyun-Jeong;Lee, In-Seon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.7
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    • pp.929-936
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    • 2005
  • The purpose of this study was to examine the ability of alkaline phosphatase (ALP) synthesis of MC3T3-E1 cells when above edible sources, Solidago virga-aurea var. gigantea Miq. root (SVR) extracts, were supplimented. MC3T3-E1 cells were cultured with $\alpha-MEM$(vehicle control), dexamethasone and genestein (positive control), and SVR extracts for 27 days. The effects of SVR MeOH extracts and its fractions on cell proliferation were measured by MTT assay. At 10, 100${\mu}g/mL$ of SVR methanol extract treated, that were elevated of cell proliferation to 140 and $120\%$ via vehicle control, respectively. And then ALP synthesis was measured by spectrophotometer for enzyme activity and by naphthol AS-BI staining for morphometry at 3, 9, 18, and 27th day. As the results, every extracts and fractions were promoted ALP activity by time course at 1, 10, 100${\mu}g/mL$, except n-hexane and chloroform fractions. Remarkably, the MeOH extracts were increased ALP activity more than 4.4 times compared with vehicle control, 2.2 times via positive control at 27th day (p<0.05). The SVR MeOH extracts treated cells, especially at a concentration of 10${\mu}g/mL$, showed remarkably higher than vehicle-treated control cells of mineralization which were checked by Alizarin red staining. These results indicate that SVR methanol extract have an induction ability of proliferation and differentiation on osteoblast.

Skin Regeneration, Anti-wrinkle, Whitening and Moisturizing Effects of Cheongsangbangpung-tang Aqueous Extracts with Cytotoxicity (청상방풍탕 열수 추출물의 피부재생, 주름개선, 미백, 보습 효과 및 세포독성 평가)

  • Woo, Chang-Yoon;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.30 no.2
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    • pp.49-70
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    • 2017
  • Objectives: The present study is to observe the skin-regeneration, anti-wrinkle, whitening and skin moisturizing effects of Cheongsangbangpung-tang (CSBPT) with cytotoxicity. Methods: In the present study, cytotoxicity of CSBPT lyophilized aqueous extracts (yield=18.71%) was experimented against human normal fibroblast cells and B16F10 murine melanoma cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide (MTT) assay, and skin regeneration and anti-wrinkle effects were also showed through the assay of collagen type I synthesis by an enzyme immunoassay (EIA) kit as comparing with transforming growth factor (TGF)-${\beta}1$, hyaluronidase, collagenase and matrix metalloproteinase (MMP)-1 inhibitory assays as comparing with oleanolic acid (OA), and elastase inhibitory effects as comparing with phosphoramidon disodium salt (PP). In addition, whitening effects of CSBPT were observed by tyrosinase inhibitory assay and melanin formation test in B16/F10 melanoma cells as comparing with arbutin, and skin moisturizing effects were measured through mouse skin water contents test, respectively. Results: No CSBPT treatment related cytotoxic effects were demonstrated against human normal fibroblast cells and B16/F10 murine melanoma cells. CSBPT concentration-dependent increased collagen type I synthesis at human normal fibroblast cells. It also effectively suspreessed hyaluronidase, collagenase, elastase and MMP-1 activities, which were enzymes that related to declining of ECM and formation of wrinkle. CSBPT supressed B16/F10 melanoma cells's melanin productions with tyrosinase activity, which was an enzyme connected with melanin formation, and dose-dependent and significant increases of skin water contents were detected in CSBPT treated mouse skin as compared with vehicle control skins. Conclusions: CSBPT showed favorable and enough skin regeneration, anti-wrinkle, whitening and skin moisturizing effects at least in a condition of this experiment. However, more detail mechanism and in vivo skin protective efficacy studies should be conducted in future with the screening of the biological active compounds in individual herbs of Cheongsangbangpung-tang.

In Vitro Cytotoxicity, Skin Regeneration, Anti-wrinkle, Whitening and In Vivo Skin Moisturizing Effects of Oncheongeum (온청음 물 추출물의 세포독성, 피부재생, 주름개선, 미백 및 보습 효과)

  • An, Tteul-E-Bom;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.29 no.1
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    • pp.14-34
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    • 2016
  • Objectives: The objective of this study was to evaluate the effects of cytotoxicity, skin regeneration, anti-wrinkle, whitening and skin moisturizing of Oncheongeum (OCE).Methods: The cytotoxicity of OCE lyophilized aqueous extracts (yield=13.82%) was observed against human normal fibroblast cells and B16/F10 murine melanoma cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium Bromide (MTT) assay, and skin regeneration and anti-wrinkle effects were also evaluated through the assay of collagen type I synthesis compared to the transformation of the growth factor (TGF)-β1, hyaluronidase, collagenase and matrix metalloproteinase (MMP)-1 inhibitory assays compared to oleanolic acid (OA), and elastase inhibitory effects compared to phosphoramidon disodium salt (PP). In addition, OCE’s whitening effects were measured by a tyrosinase inhibitory assay and melanin formation test in B16/F10 murine melanoma cells compared to arbutin, and skin moisturizing effects were observed through a mouse skin water content test, respectively. Results: No OCE treatment-related cytotoxic effects appeared on human normal fibroblasts and B16/F10 murine melanoma cells. OCE concentration-dependently increased the collagen Type I synthesis on human normal fibroblast cells, and also effectively inhibited hyaluronidase, elastase, collagenase and MMP-1 activities. In addition, OCE inhibited melanin production of B16/F10 murine melanoma cells and activity of tyrosinase. And significant and dose-dependent increases of skin water content were detected in OCE-treated mouse skin compared to vehicle control skins. Conclusions: OCE showed favorable and sufficient effects in skin regeneration, anti-wrinkle, whitening and skin moisturizing in this experiment. But more detail mechanisms and studies on the skin protective efficiency of in vivo are needed with the screening of active biological compounds in individual OCE herbs.

The Antioxidant and Antitumor Effects of the Extract of Bulnesia sarmientia (Bulnesia sarmienti 추출물의 항산화 및 항암효과)

  • Jo, Dae-Hyoun;Min, Kyung-Jin;Cha, Chun-Geun
    • Journal of Food Hygiene and Safety
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    • v.22 no.2
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    • pp.120-126
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    • 2007
  • Present study have been performed to develop Bulnesia sarmienti as a functional food. Methanol, n-hexane, chloroform, ethyl acetate and butanol extracts of Bulnesia sarmienti contained total phenol by 5.81 to 7.47%. It is high content than fruits which were known as high contests of total phenol. The electron donating ability of the extract of Bulnesia sarmienti were increased along with increasing concentrations of extracts. At $500{\mu}g/mL\;and\;1000{\mu}g/mL$, the all extracts showde more than 80% of scavenging abilities, which means the equal effect of the antioxidant, BHT. Nitrite scavenging abilities were measured as follows: methanol, butanol, 5.53, 5.77% at $100{\mu}g/mL$, respectively. The ethyl acetate extract was 73.29% at $1000{\mu}g/mL$ which showed the highest activity and methanol, butanol, n-hexane, chloroform and water extract were 65.65, 65.02, 47.49, 52.51, 45.54% which also showed relatively high activities. The growth inhibitory effects of each solvent extract on tumor cell were as follows: test against SUN-1, the gastric carcinoma cell, exhibited the highest inhibitory effects at $100{\mu}g/mL$ where the n-hexane extract was 61.6%. The ethyl acetate and water extracts did not revealed any inhibitory effects. Hela, the uterine carcinoma cell, exhibited the highest inhibitory effects at $100{\mu}g/mL$ where the n-hexane extract was 75.1%. The water extracts did not revealed any inhibitory effects. HT-29, the colon carcinoma cell, also exhibited the highest inhibitory effects at $100{\mu}g/mL$ where n-hexane extract was 57.4%. In conclusion, Bulnesia sarmienti have been shown the antioxidant and antitumor effects, and that it is expected to be developed as functional foods.