• Title/Summary/Keyword: MTT 실험

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Antioxidant effect and iNOS, COX-2 Expression Inhibition on RAW 264.7 Cell of Mangifera indica L. Leafs (애플망고 잎의 추출물의 항산화 및 대식세포(RAW 264.7)에서 iNOS, COX-2 발현 저해 효과)

  • Yoo, Dan-Hee;Lee, In-Chul
    • Journal of Life Science
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    • v.30 no.9
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    • pp.783-790
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    • 2020
  • The present study investigates the antioxidant and anti-inflammatory activities of Mangifera indica L. leaf extract. The total polyphenol content was measured using the Folin-Denis method. Results showed that the M. indica L. leaf extract of water and 70% ethanol showed a content of 440.83±1.02, 475.63±1.3 mg/100 g tannic acid equivalent. To assess antioxidant activity and electron-donating ability, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging activity were measured, and all extracts were found to be highly efficacious. To assess cell viability of the extract from M. indica L. leaf on macrophage cells (RAW 264.7), a 3-[4,5-dimethyl-thiazol-2- yl]-2,5-diphenyl-tetrazolium-bromide assay was performed. The following experiments were conducted in section where cells was not shown of toxicity. In order to effectively determine anti-inflammatory activity, inhibition of lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 cells was examined using a Griess assay. The result showed that M. indica L. leaf extract concentration-dependently inhibited NO production. M. indica L. leaf extract was measured using Western blot, reverse transcription- polymerase chain reaction (RT-PCR) that to find the production of pro-inflammatory factor on stimulated RAW 264.7 cells of LPS. According to the results of this study, the M. indica L. leaf extract showed excellent effectiveness in antioxidant and anti-inflammatory activity, thus confirming its usability as a natural material and a functional raw material for cosmetics.

Anti-cancer Effect of Hot Water Extract from Mycelium in Germanium-enriched Cordyceps militaris (게르마늄 농도를 증가 시킨 동충하초 균사체 열수 추출액의 항암효과)

  • Choi, Jeong Su;Heo, Ji Hye;Kim, Dae Jin;Namkung, Su Min;Lee, Tae Bok;Lee, Min Woo;Kim, Suhng Wook
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.2
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    • pp.69-78
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    • 2017
  • Cordyceps militaris has been used in traditional Chinese medicine owing to its anticancer and immunomodulatory activities. Germanium compounds have also been shown to be associated with many pharmacological functions, such as antimicrobial, antiviral, antitumor, antimutagenic, and immunomodulating effects. In this study, we examined the biological properties of hot water extract from mycelial liquid culture of germanium-enriched C. militaris (CMGe). CMGe displayed a concentration-dependent antiproliferation activity against four human cancer cell lines. The antiproliferative activity of CMGe was 2-4-fold lower than that of hot water extract from mycelial liquid culture in C. militaris (CM). However, CM had a concentration-dependent cytotoxicity to human bone marrow-derived mesenchymal stem cells (MSCs). Contrastingly, CMGe did not cause any cellular damage to MSCs. MSCs cultured with CMGe displayed an increased proliferative activity with no cytotoxic effect. The oral administration of CMGe inhibited increased tumor volume and weight compared with the control group. CMGe has the potential to be used as an industrial product in medicinal foods as well as in pharmaceutical products.

Fermentation process increased antimutagenic and in vitro anticancer effects during Kochujang manufacturing (고추장 제조시 발효가 항돌연변이 및 in vitro 항암효과에 미치는 영향)

  • Kim, Ji-Young;Liu, Fang-Fang;Lim, Yaung-Iee;Park, Kun-Young
    • Food Science and Preservation
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    • v.21 no.6
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    • pp.878-884
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    • 2014
  • Increased antimutagenic and in vitro anticancer effects were observed by the fermentation process during Kochujang manufacturing. In order to confirm the increased functionality, wheat grain, first fermented wheat grains (FFWG), second fermented wheat grains (SFWG), final fermented wheat grains (FiFWG), red pepper powder (RPP), and commercial Kochujang (CK) were prepared. Kochujang manufactured with final fermented wheat grains and red pepper powder were further fermented for 15 days and 30 days. The antimutagenic effects were determined by counting the number of revertants in Salmonella Typhimurium TA100 against N-methyl-N'-nitro-nitrosoguanidine (MNNG, 1.0mg/mL). The final fermented wheat grains (52% inhibition) showed higher antimutagenic effects than the wheat grain (34%), and the commercial Kochujang showed the highest antimutagenic effects (55%). We tested the inhibitory effect on the growth of HT-29 human colon carcinoma cells and AGS human gastric adenocarcinoma cells by using MTT (3-(4,5-dimethyl-thiazol)-2,5-diphenyltetrazolium bromide) assay. The results showed that increased fermentation process continually increased the growth inhibitory effect on both cancer cells. The further fermentation for 15 days of the Kochujang product also increased inhibitory growth of the AGS cancer cells. In conclusion, the methanol extract from fermented wheat grains and commercial Kochujang showed sequentially increased antimutagenic and in vitro anticancer activity, and thus the final commercial Kochujang revealed the highest effect.

Antimutagenic and Cytotoxic Effects of Hovenia dulcis Thumb Leaves Extracts (헛개나무 잎 추출물의 항돌연변이원성 및 암세포 성장억제효과)

  • Park, Sun-Hee;Chang, Eun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.11
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    • pp.1371-1376
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    • 2007
  • The effect of Hovenia dulcis Thumb leaves on the mutagenicity in salmonella assay and inhibitory effects on the growth of cancer cells were studied. On antimutagenicity as evaluated by Ames test, the extract and fractions of Hovenia dulcis Thumb leaves had no effect on the mutagenicity by themselves. However, methanol extract and fractions from Hovenia dulcis Thumb showed strong inhibitory effect on the mutagenesis induced by N-methyl-N#-nitro-N-nitroso-guanidine (MNNG) and benzo(a)pyrene (B(a)P). Among the solvent fractions of methanol extract, the hexane, chloroform and butanol fraction exhibited stronger inhibitory activity against MNNG and B(a)P induced mutagenesis than water fraction. For anticancer effects, Hovenia dulcis Thumb loaves extract and fractions against cancer cell lines including HepG2 and HT29 were investigated. The methanol extract, the hexane fraction and the chloroform fraction of Hovenia dulcis Thumb leaves inhibited growth of cancer cells but they had no effect on the cytotoxicity of normal human liver cells under the same conditions.

Growth-inhibitory Effect of the Extract of Porphyran-Chungkookjang on Cancer Cell (Porphyran-청국장 추출물의 암세포 성장 억제효과)

  • Min, Hyun-Kyeng;Kim, Hyo-Ju;Chang, Hae-Choon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.7
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    • pp.826-833
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    • 2008
  • The effects of porphyran-chungkookjang on cytotoxicity of human normal cell line (BJ) and human cancer cell lines (AGS and HT-29) were examined. Porphyran, which was prepared from laver (Porphyra yezoensis), decreased the viability of the cancer cells, however, it did not affect the viability of normal cells. Porphyranchungkookjang was prepared by the addition of 5% (w/w) porphyran into chungkookjang which was fermented by starter, Bacillus subtilis DJI. The cytotoxicity effects of the chungkookjang and porphyran-chungkookjang were evaluated with MTT assay. The methanol and the water extract of porphyran-chungkookjang at 1.0 mg/mL showed $23{\sim}38%$ decreases in proliferation of cancer cells (AGS and HT-29). However, the methanol and the water extracts of porphyran-chungkookjang did not inhibit the growth of normal cell. Moreover, the methanol extract of porphryan-chungkookjang at 1.0 mg/mL showed $1.2{\sim}1.5$ fold higher anticancer effects than that of the chungkookjang.

Hepatoprotective Effects of Sumaeyaksuk (Artemisia argyi H.) Extract on LPS-mediated Inflammatory Response (LPS에 의해 유도된 염증반응에서 섬애약쑥 추출물의 간보호 효과)

  • Kim, Dong-Gyu;Kang, Min-Jung;Shin, Jung-Hye
    • Journal of Life Science
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    • v.26 no.11
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    • pp.1282-1288
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    • 2016
  • Artemisia, a plant widely used as traditional herbal medicine in many countries, has drawn attention of the researchers. And its extracts or compounds are known to have an efficacy of antioxidant, anti-diabete, anti-cancer, anti-inflammation and neuroprotection. Sumaeyaksuk is a variant of the Artemisia argyi and major constituents are eupatilin and jaceosidin. This study was performed to investigate the effects of the sumaeyaksuk aqueous extract on inflammatory response induced by lipopolysaccharide (LPS) in human hepatoma HepG2 cells. To examine the potential hepatoprotective properties of sumaeyaksuk extract, cell viability, as well as nitric oxide (NO), reactive oxygen species (ROS), macrophage colony-stimulating factor (M-CSF), interleukin-8 (IL-8) levels, alanine transaminase (ALT), and aspartate transaminase (AST) activities, were measured. Cytotoxic activity of extracts on HepG2 cells was measured by MTT assay. Sumaeyaksuk extract did not induce cytotoxicity at concentrations of $0{\sim}400{\mu}g/mL$. NO and ROS levels significantly decreased with increasing concentration of the extract. The secretion levels of M-CSF and IL-8 were suppressed by sumaeyaksuk extract in a dose-dependent manner. Moreover, ALT (75.4%) and AST (61.6%) levels significantly decreased in sumaeyaksuk extract-treated cells at $400{\mu}g/mL$. These results suggested that the sumaeyaksuk extract attenuates the LPS-induced hepatotoxicity resulting from regulation of inflammatory factors and could potentially be used as a hepatitis therapeutic agent.

Whitening Effect and Skin Regeneration Effect of Red Sea Cucumber Extract (홍해삼 추출물의 멜라닌 형성 억제를 통한 미백효과 및 피부 재생효과에 관한 연구)

  • Jeon, Mi Ji;Kim, Eun Ji;Kim, Geun Tae;Kim, Ga Yeon;Lee, Seung Jae;Jung, In Cheol;Kim, Sang-Yong;Kim, Young Min
    • Journal of Life Science
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    • v.28 no.6
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    • pp.681-687
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    • 2018
  • Recently, several researchers have been developing cosmetics from natural ingredients for skin whitening and anti-aging products. The red sea cucumber (RSC), Apostichopus japonicas, is a species of sea cucumber in the family stichopodiae, which is widely distributed in China, Japan, and Korea. To use Red Sea Cucumber as a cosmetic ingredient, its inhibitory effects on melanogenesis and the anti-aging effects of RSC extracts were investigated. First, a tyrosinase activity assay was performed, which showed that RSC inhibited tyrosinase activity at a concentration of $200{\mu}g/ml$. An MTT assay was carried out to evaluate cell toxicity, and the results showed that RSC extract has no cytotoxicity in HaCaT cells. Furthermore, the mRNA expression levels of tyrosinase, tyrosinase related protein 1 (TRP-1), tyrosinase related protein 2 (TRP-2), microphthalmia-associated transcription factor (MITF), and matrix metalloproteinase (MMPs) genes treated with RSC extract in B16F10 and HaCaT cells decreased. Moreover, a wound-healing assay was performed to identify the cell regeneration effect of RSC extracts. Also, a skin turnover effect was confirmed by creating a three-dimensional cell culture with HaCaT and human fibroblasts. Altogether, the results suggested that Red Sea Cucumber may possess a high ability to induce whitening and anti-wrinkle effects as a cosmeceutical ingredient.

Effects of Plant Water Extract Codonopsis Lanceolatae on Mouse Immune Cell Activation Ex Vivo (더덕 물 추출물의 경구 투여가 마우스 면역 세포 활성에 미치는 효과)

  • Ryu, Hye-Sook;Kim, Kyoung-Ok;Kim, Hyun-Sook
    • Journal of Nutrition and Health
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    • v.42 no.3
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    • pp.207-212
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    • 2009
  • Codonopsis lanceolatae has been used as one of the traditional remedies as well as food source. However, few studies on their immunomodulating effects have been reported. We previously reported that ex vivo supplementation of Codonopsis lanceolatae water extracts enhanced splenocyte proliferation compared to the control group. In order to elucidate its ex vivo effect, six to seven week old balb/c mice were fed ad libitum on a chow diet and water extracts of Codonopsis lanceolatae were orally administrated every other day for four weeks at two different concentrations (50 and 500 mg/kg B.W). After preparing the single cell suspension, the proliferation of splenocytes was determined by MTT (3- [4,5-dimethylthiazol-2-y] -2,5-diphenyl terazolium bromide) assay. The production of cytokine (IL-1${\beta}$, IL-6, TNF-${\alpha}$), secreted by macrophages stimulated with LPS or not, was detected by ELISA assay using a cytokine kit. After 48 hrs of incubation with the mitogen (ConA or LPS) stimulation, the mice splenocyte proliferation in experimental group was statistically increased at two different concentrations than that in control group. The cytokines production was more significantly enhanced at the lower supplementation (500 mg/kg B.W.) group rather than higher concentration (500 mg/kg B.W.) compared to the control group. The results of this study may suggest that the supplementation of water extract of plant mixture could regulate the immune function by increasing the splenocyte proliferation and enhance the immune function through regulating cytokine production capacity by activated macrophages in mice.

Studies on Constituents of Higher Fungi of Korea (LXXIII) Antitumor Components of the Cultured Mycelia of Paxillus atrotomentosus (한국산 고등 균류의 성분 연구(제73보) -좀우단버섯 배양 균사체의 항암 성분-)

  • Kwag, Sang-Deok;Bok, Jin-Woo;Hyun, Jin-Won;Choi, Eung-Chil;Kim, Byong-Kak
    • The Korean Journal of Mycology
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    • v.20 no.3
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    • pp.240-251
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    • 1992
  • To find antitumor components from higher fungi, the cultured mycelia of Paxillus atrotomentosus were extracted with hot water. The water soluble fraction was purified and separated by DEAE-cellulose ion exchange chromatography and Sepharose CL-4B gel filtration method. The separated fractions(Fr.) were designated CR A, B, C and D. Fr. A showed the highest inhibition ratio of 68.51% among the five tractions at a dose of 20 mg/kg/day. When Fr. A was examined for immunopotentiation activity, it increased the amount of the superoxide anion from activated macrophages to 1.1 fold and the number of plaques in hemolytic plaque assay to 2.3 fold, respectively. Otherwise, it did not show direct cytotoxity in sarcoma 180. Delayed type hypersensitiyity reaction showed that the decreased footpad swelling of tumor-hearing was restored to the normal. These results indicate that antitumor activity was exerted through immunopotentiation. Its chemical analysis showed 86.36% polysaccharide, 1.52% protein and 1.64% hexosamine. The polysaccharide consisted of fucose, galactose, glucose, mannose and xylose. This component was named paxillan.

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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.