• Title/Summary/Keyword: Cell-based assay

검색결과 667건 처리시간 0.028초

탈단백 우골의 골이식 대체재로서의 특성에 대한 평가;세 종류의 골 대체재의 기본 특성에 대한 비교분석 (Evaluation of deproteinized bovine bone mineral as a bone graft substitute;A comparative analysis of basic characteristics of three commercially available bone substitutes)

  • 박진우
    • Journal of Periodontal and Implant Science
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    • 제35권4호
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    • pp.863-875
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    • 2005
  • Bovine bone-derived bone substitutes are widely used for treatment of bone defects in dental and orthopedic regenerative surgery. The purpose of this study was to evaluate the basic characteristics of deproteinized bovine bone mineral as a bone graft substitute. Commercially available products from three different bovine bone minerals-Bio-Oss(GeistlichPharma, Switzerland), BBP(Oscotec. Korea), Osteograf/N-300(Dentsply Friadent Ceramed, USA) - were investigated. They were evaluated by scanning electron microscopy(SEM), energy dispersive X-ray spectrometer(EDS), surface area analysis(BET), and Kjeldahl protein analysis. Cell viability on different products was evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) assay. The results of this study indicated that each bone substitute displayed distinct surface properties. Furthermore, Kjeldahl protein analysis indicated that residual crude proteins are present in deproteinized bovine bone mineral. BBP showed relatively large amount of residual protein, which indicated that the possibility of disease transmission can not be safely ruled out. Based on the results of this study, it is suggested that active quality management is strongly needed in operations that involve processing bovine bone tissue for medical use.

Effects of N-acetylphytosphingosine on melanogenesis of B16F10 murine melanoma cells.

  • Park, M. K.;Park, C. S.;Kim, J. W.;R. M. Ahn;Y. S. Yoo;S. Y. Yi
    • 대한화장품학회:학술대회논문집
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    • 대한화장품학회 2003년도 IFSCC Conference Proceeding Book II
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    • pp.241-242
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    • 2003
  • The effects of N-acetylphytospingosine(NAPS), one of the phytospingosine derivatives, on melanogenesis of B 16F 1 0 mouse melanoma cell lines were investigated. We assessed the effect of NAPS on the depigmentation of B16F10 cells. The melanin content of cells was significantly reduced by NAPS. We examined the inhibitory effect of NAPS on tyrosinase activity using L-dopa as a substrate and the results showed that tyrosinase activity was inhibited in a does-dependent manner. The mRNA level of tyrosinase as well as that of tyrosinase related protein-l (TRP-l) and tyrosinase related protein-2 (TRP-2) genes were not affected by NAPS based on a reverse transcription-polymerase chain reaction (RT-PCR) assay. We also performed a Western blotting analysis using anti-tyrosinase antibody. It showed that there is no change in tyrosinase protein level after treatment of NAPS. These results suggest that the depigmenting mechanism of NAPS in B16F10 melanoma cells involves inhibition of melanosomal tyrosinase activity, rather than the mRNA expression or protein level of tyrosinase.

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In Vitro Cytotoxicity of a Novel Platinum(II) Coordination Complex Containing Diaminocyclohexane

  • Jung, Jee-Chang;Kim, Soon-Ae;Kim, Young-Kyu;Chang, Sung-Goo;Rho, Young-Soo
    • Biomolecules & Therapeutics
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    • 제8권3호
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    • pp.228-234
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    • 2000
  • We have synthesized a novel platinum(II) coordination complex containing trans-ι-1,2-diaminocy-clohexane (DACH) as a carrier ligand and 1,2-dichloroethane (DCE) as a leaving group. A new series of [Pt(trans-ι-DACH)(DCE)](PC) was evaluated for its cytotoxic activity on MKN-45 human gastric adenocar-cinoma cells and normal primary cultured kidney cells. The new platinum complex has demonstrated high efficacy in the cytotoxicity against MKN-45/P, MKN-45/ADM and MKN-45/CDDP cell-lines. The cytotoxicity of PC against rabbit proximal renal tubular cells, human renal cortical cells and human renal cortical tissues, determined by MTT assay, the [$^3H$]-thymidine uptake arid glucose consumption tests, was found to be quite less than those of cisplatin. Based on these results, this novel platinum(II) coordination complex appears to be better for improving antitumor activities with low nephrotoxicity and is a valuable lead in the development of new, clinically available anticancer chemotherapeutic agents.

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H460 인체 폐암세포에서 길경 물 추출물에 의해 유도된 세포사멸에서 자가포식의 역할 (The Role of Autophagy in Apoptosis Induced by Water Extract of Platycodonis Radix in H460 Human Lung Cancer Cells)

  • 홍수현;최영현
    • 대한한의학방제학회지
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    • 제29권4호
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    • pp.155-165
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    • 2021
  • Objectives : Recent studies have suggested that Platycodonis Radix has various pharmacological effects such as anti-cancer, antioxidant, anti-asthma, anti-diabetes, anti-obesity, hepatoprotective, and cardiovascular protection effects. The aim of this study was to investigate the role of water extract of Platycodonis Radix (WPR)-induced autophagy in H460 human lung cancer cells. Methods : H460 cells were treated with WPR and cell viability was calculated by an MTT assay. To evaluate changes in apoptosis- and autophagy-related genes, Western blotting was performed. Two kinds of autophagy inhibitors, 3-Methyladenine (3-MA) and bafilomycin A1, were pretreated to confirm the role of WPR-induced autophagy. Results : WPR reduced the viability of H460 cells in a treatment concentration-dependent manner, which was associated with induction of apoptosis. It was also confirmed that WPR induced autophagy based on the formation of specific intracellular vacuoles and changes in the expression of autophagy-related genes. Interestingly, pretreatment with 3-MA and bafilomycin A1 increased WPR-induced cytotoxicity and apoptosis. Conclusions : WPR induced autophagy at low concentrations and early stages of treatment, but promoted apoptosis at high concentrations and late stages. Moreover, WPR-induced autophagy had a cytoprotective role in H460 cells.

마우스 대식세포를 이용한 일당귀 물추출물의 항염효능 연구 (Anti-inflammatory Effect of Angelicae acutilobae Radix Water Extract on LPS-stimulated Mouse Macrophages)

  • 한효상
    • 대한본초학회지
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    • 제28권6호
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    • pp.129-133
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    • 2013
  • Objectives : The purpose of this study was to investigate the effects of Angelicae acutilobae Radix Water Extract (AA) on the production of cytokines in RAW 264.7 cell stimulated with lipopolysaccharide (LPS). Method : RAW 264.7 cells were cotreated with AA (50 and $100{\mu}g/mL$) and lipopolysaccharide (LPS; $1{\mu}g/mL$) for 24 hours. After 24 hours treatment, using bead-based multiplex cytokine assay, concentrations of various cytokines such as interleukin(IL)-6, tumor necrosis factor-alpha(TNF-${\alpha}$) granulocyte colony-stimulating factor(G-CSF), granulocyte macrophage colony-stimulating factor(GM-CSF), and macrophage inflammatory protein(MIP)-$1{\alpha}$ were measured. Result : AA significantly inhibited LPS-induced production of IL-6 and MIP-$1{\alpha}$ from LPS-stimulated RAW 264.7 cells at the concentration of $50{\mu}g/mL$. AA significantly inhibited LPS-induced production of TNF-${\alpha}$ from LPS-stimulated RAW 264.7 cells at the concentration of $100{\mu}g/mL$. AA significantly inhibited LPS-induced production of G-CSF and GM-CSF in RAW 264.7 cells at the concentrations of 50 and $100{\mu}g/mL$. Conclusion : These results suggest that AA has anti-inflammatory effect related with its inhibition of proinflammatory cytokines such as IL-6, TNF-${\alpha}$, G-CSF, GM-CSF, and MIP-$1{\alpha}$ in LPS-induced macrophages.

HepG2 간암세포에서 미토콘드리아 경로를 통한 개똥쑥 추출물의 Apoptosis 유도 효과 (Extract from Artemisia annua Linné Induces Apoptosis through the Mitochondrial Signaling Pathway in HepG2 Cells)

  • 김보민;김근태;김은지;임은경;김상용;김영민
    • 한국식품영양과학회지
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    • 제45권12호
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    • pp.1708-1716
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    • 2016
  • Akt 및 mTOR는 세포 생존에 필수적인 경로로 세포 성장과 증식 등에서 중요한 역할을 하는 것으로 알려져 있다. 본 연구에서는 항암 및 항균 효과가 있는 것으로 알려진 개똥쑥(Artemisia annua L.)에 의한 HepG2 간암세포의 apoptosis 유도 효과를 확인하였다. 본 연구 결과에 의하면 개똥쑥 추출물의 처리 농도가 증가함에 따라 HepG2 세포의 생존율은 억제되었으며, 이는 apoptosis 유도 효과에 의한 것임을 세포의 형태적 변화와 flow cytometry를 통해 확인하였다. 그리고 mitopotential assay와 caspase-3/7 activity assay, western blotting으로 Bcl-2 family 단백질을 확인함으로써 apoptosis 경로 중 내인성 경로(intrinsic pathway)에 의해 apoptosis가 일어남을 알 수 있었다. 이러한 효과는 Akt/mTOR의 활성 저해와 연관이 있었으며 Akt/mTOR의 저해제인 LY294002/rapamycin을 개똥쑥 추출물과 병행처리하였을 경우 개똥쑥 추출물에 의한 apoptosis 효과를 더욱 증대시켰다. 따라서 Akt/mTOR의 저해는 개똥쑥 추출물의 apoptosis 효과를 상승시켰으며 이에 따라 미토콘드리아의 기능 손상과 caspase 활성의 증가를 통해 이루어짐을 확인하였다.

양어 사료첨가제로서의 유산균 발효 개똥쑥의 항산화 및 항균활성 (Antioxidant and Antibacterial Activities of Lactobacillus-fermented Artemisia annua L. as a Potential Fish Feed Additive)

  • 이아란;우개민;강수경;한성구;이봉주;김수기
    • 생명과학회지
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    • 제27권6호
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    • pp.652-660
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    • 2017
  • 최근 양어에서는 전염병, 환경오염, 어분 가격 상승 등으로 어려움을 가지고 있다. 본 연구는 쑥에서 분리한 L. plantarum SK3494를 이용하여 전통 약초인 개똥쑥을 발효시켜 in vitro 생리활성을 측정하고 어류용 사료첨가제로서의 가능성을 연구하였다. 개똥쑥의 발효시 유산균수는 9.38 log10 CFU/ml이며, pH는 4.1로 나타났다. 발효개똥쑥의 항산화활성은 DPPH 측정과 MAC-T 세포 내 과산화물 감소 실험에서 뛰어난 효과를 보였다. 발효개똥쑥은 어류 병원균인Photobacterium damselae subsp. damselae와 Vibrio ichthyoenteri에 대하여 강한 항균활성을 나타내었다. 따라서 유산균을 이용한 개똥쑥 발효물은 양식용 사료첨가제로써 가능성을 가지고 있는 것으로 판단되었다.

스피루리나 첨가 샐러드 드레싱 보충 식이가 마우스 혈장 항산화 지표 및 DNA보호에 미치는 영양 (Effects of Spirulina Added Salad Dressing on the Antioxidant Index and DNA Protection in Mice)

  • 양윤형;조흔;패넬로프 펠리프;이정희;이선영;조용식;전혜경;송경빈;김미리
    • 동아시아식생활학회지
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    • 제15권4호
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    • pp.386-396
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    • 2005
  • The effects of spirulina-added salad dressing on lipid profiles and antioxidant biomarkers such as total glutathionine, TBARS value, carbonyl value, GPx, GR, SOD and paraoxonase activity in plasma or liver of mice were evaluated Sixteen male ICR mice weighing 20$\pm$2 g were divided into two groups and fed low fat ($5\%$ fat) diet (low fat control: LFC) and low fat control plus dressing diet (LFD) for eight weeks. Body weight, tissue weights of liver, heart and kidney, and the distribution of body fat deposition were not significantly different between two groups. Also, the profile of TG, TC, LDL and HDL cholesterol were similar between two groups. The DNA damage was determined using the comet assay (single cell gel assay) with alkaline electrophoresis and quantified by measuring tail length (TL). Spirulina salad dressing consumption resulted in significant decrease in lymphocyte DNA damage expressed by TL (LFC: $28.8{\mu}m$, LFD: $20.3{\mu}m$). Additionally, salad dressing consumption for 8 wks decreased the lipid peroxidation assayed by TBARS to $12.6\%$ compared with the control. The levels of antioxidant vitamins such as $\beta$-carotene were significantly higher in plasma of LFD group than those in LFC group based on HPLC method This study shows that spirulina-added salad dressing exerts degenerative disease-protective effects on oxidative DNA damage and lipid peroxidation possibly via a free radical levels.

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삼내자 열수추출물의 항산화 및 항염 효과 (Anti-inflammatory and Antioxidant Effects of Hot Water Extracts from Kaempferia Galanga L)

  • 진정원비너스;이지안
    • 융합정보논문지
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    • 제9권6호
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    • pp.218-226
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    • 2019
  • 본 연구의 목적은 삼내자(Kaempferia Galanga) 열수추출물의 항산화, 세포독성, 항염 효능을 조사하여 화장품 성분으로서 활용 가능성을 알아보고자 하였다. 항산화 활성 평가를 위해 DPPH, ABTS 라디컬 소거능 활성, FRAP 환원력, 총폴리페놀 함량을 측정하였다. 쥐의 대식세포인 RAW264.7 세포를 대상으로 MTT assay를 수행하여 세포독성을 확인하였다. 항염 효능을 알아보기 위해 LPS로 유도된 RAW264.7 세포에서 nitric oxide(NO), $TNF-{\alpha}$ 분비 및 iNOS, $TNF-{\alpha}$ mRNA 발현 수준을 조사하였다. 그 결과 DPPH와 ABTS 라디칼 소거능은 농도 의존적으로 라디칼 소거 활성이 증가하였다. FRAP 값은 5 mg/mL에서 24.5 uM로 가장 높게 나타났으며, 총폴리페놀 함량은 $1.28{\pm}0.064mg\;GAE/g$로 나타났다. KG 추출물 농도 0.625~2.5 mg/mL에서 세포독성은 보이지 않았다. 또한 RAW264.7 세포에서 LPS 유도에 의한 NO, $TNF-{\alpha}$ 분비 그리고 iNOS, $TNF-{\alpha}$의 mRNA 발현이 KG 추출물에 의해 유의하게 감소하였다. 이러한 결과는 삼내자 열수추출물이 안전하고 효과적인 화장품 원료로서의 활용 가능성이 있음을 보여준다.

현토단(玄兎丹)의 RAW 264.7 대식 세포에서의 항염증 효과에 관한 연구 (The study of anti-inflammatory effect of Hyeonto-dan extract in RAW 264.7 macrophage)

  • 김마룡;강옥화;공룡;서윤수;주전;김상아;김은수;신민아;이영섭;권동렬
    • 대한본초학회지
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    • 제32권2호
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    • pp.77-85
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    • 2017
  • Objectives : This study aimed to investigate the unknown mechanisms behind the anti- inflammatory activity of Hyeonto-dan(HT) 70% ethanol extract on LPS-stimulated RAW 264.7 cells. Methods : Cells were treated with Hyeonto-dan 1 h prior to addition of 200 ng/mL of LPS. Cell viability was measured by the MTS assay. Nitric oxide levels were determined by the Griess assay. $PGE_2$ were measured using EIA kit. Pro-inflammatory cytokine production was measured by the enzyme-linked immunosorbent assay (ELISA). The expression of COX-2, iNOS, and MAPKs was investigated by Western blot, qRT-PCR. $NF-{\kappa}B$/p65 localization and interaction of the TLR-4 receptor with LPS was examined by immunofluorescence assays. Results : Hyeonto-dan had no cytotoxicity at the measured concentration. Hyeonto-dan inhibited NO production and pro-inflammatory cytokines such as IL-6, $TNF-{\alpha}$, and PGE2 as well as the protein and mRNA expression of iNOS and COX-2. Moreover, Hyeonto-dan inhibited the interaction between LPS and TLR-4 in murine macrophages. It suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2), c-jun N-terminal kinase (JNK 1/2) and p38. Finally, it inhibited translocation of $NF-{\kappa}B$ in response to competitive LPS. Conclusions : Based on the results of this study, Hyeonto-dan inhibited the binding of TLR-4 receptor to LPS and inhibited the phosphorylation of extracellular signaling pathway MAPKs. These inhibitory effects are thought that the amount of $NF-{\kappa}B$ delivered to the nucleus was decreased and the inflammatory reaction was prevented by decreasing the production of LPS-induced $PGE_2$, NO, IL-6 and $TNF-{\alpha}$.