• Title/Summary/Keyword: RAW 264.7 Cell Line

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Anti-Inflammatory Effect of Peel Extracts from Citrus Fruits (감귤류 과피 추출물의 항염증 효과)

  • Lee, Sook-Hyun;Suh, Seok-Jong;Lee, Kyoung-Hae;Yang, Jong-Beom;Choi, Sung-Up;Park, Sung-Soo
    • Journal of Food Hygiene and Safety
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    • v.28 no.4
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    • pp.342-348
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    • 2013
  • The following study was presented to investigate the anti-inflammatory effect of peel extracts (PE) from three citrus fruits: Citrus unshiu, Citrus limonia Osbeck and Citrus hallabong. According to this study, cytotoxicity, NO-production and protein levels of iNOS (inducible nitric oxide synthase) in macrophage cell were analyzed, which had been incubated in murine macrophage cell line RAW 264.7 cell of PE from those three citrus fruits. According to Citrus unshiu peel extracts (CUP), Citrus limonia Osbeck peel extracts (CHP) and Citrus hallabong peel extracts (CLP) treatment, the result showed that there was no cell growth inhibited below 2 mg/mL. Comparing the NO-production of the cell with LPS (100 ng/mL) and the treatment without LPS, significant increase of NO-production was detected. However NO-production also showed decrease trend, as the concentration increased. For each treatment, at the concentration of 1 mg/mL, NO-ihibitory activity showed significant result with following order: CUP > CHP > CLP. According to the result from Western blot, the inhibitory activities of iNOS protein from CUP and CHP showed fairly similar performances. Also inhibitory activity of COX-2 showed the following order: CUP > CHP> CLP. There was no doubt that all the treatments of CUP, CHP and CLP have anti-inflammatory effect and also that the inhibitory activity of the CUP treatment was the strongest among those three.

Activation of Macrophages by Exopolysaccharide Produced by MK1 Bacterial Strain Isolated from Neungee Mushroom, Sarcodon aspratus

  • Im, Sun-A;Wang, Wenxia;Lee, Chong-Kil;Lee, Young-Nam
    • IMMUNE NETWORK
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    • v.10 no.6
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    • pp.230-238
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    • 2010
  • Background: The MK1 strain, a novel bacterial isolate from soft-rotten tissue of the Neungee mushroom, produces copious amounts of exopolysaccharide (EPS) in a dextrose minimal medium. This study examined the molecular characteristics and immunomodulatory activity of MK1 EPS. Methods: The EPS in the culture supernatant was purified by cold ethanol precipitation, and characterized by SDS- PAGE/silver staining and Bio-HPLC. The immunomodulatory activities of the EPS were examined using the mouse monocytic cell line, RAW 264.7 cells. Results: The molecular weights of the purified EPS were rather heterogeneous, ranging from 10.6 to 55 kDa. The EPS was composed of glucose, rhamnose, mannose, galactose, and glucosamine at an approximate molar ratio of 1.00 : 0.8 : 0.71 : 0.29 : 0.21. EPS activated the RAW cells to produce cytokines, such as TNF-${\alpha}$ and IL-$1{\beta}$, and nitric oxide (NO). EPS also induced the expression of co-stimulatory molecules, such as B7-1, B7-2 and ICAM-1, and increased the phagocytic activity. The macrophage-activating activity of EPS was not due to endotoxin contamination because the treatment of EPS with polymyin B did not reduce the macrophage-activating activity. Conclusion: The EPS produced from the MK1 strain exerts macrophage-activating activity.

Effects of the Extracts by Extraction Procedures from Hericium erinaceus on Activation of Macrophage (노루궁뎅이버섯(Hericium erinaceus) 추출공정별 추출물의 대식세포 활성화에 대한 효과)

  • Kim, Sung-Phil;Choi, Yong-Hee;Kang, Mi-Young;Nam, Seok-Hyun
    • Applied Biological Chemistry
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    • v.48 no.3
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    • pp.285-291
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    • 2005
  • Effects of the aqueous or 50% ethanolic extracts prepared by various extraction procedures on macrophage activation were determined by using the mouse macrophage cell line RAW264.7 cells as a indicator cell. The results demonstrated that the fractions prepared by aqueous extraction for 2 h and by microwave extraction with 50% ethanol at 60 W for 3 min had the greatest inducing abilities for NO production, and that the greatest ROS scavenging abilities were found in the fractions prepared by hot water extraction for 2 h or 3 h, by microwave extraction with 50% ethanol at 60 W for 3 min and by 0.5% HCl extraction, respectively. Phagocytotic activities against Candida albicans were found to be highest for the 50% ethanolic extracts prepared by microwave extraction for 3 min at 60 W, 80 W and 12 W, respectively. Especially, we found that a extract prepared by microwave extraction with 50% ethanol at 60 W for 3 min enables to induce effectively overall functional activation of macrophage, such as NO production, ROS scavenging and phagocytosis of C. albicans, respectively. These results demonstrated that a 50% ethanolic extraction using microwave at 60 W for 3 min would be useful for enrichment of macrophage-activating components contained in Hericium erinaceus, implying participation of protein-bound polysaccharides as a active factor.

Studies on the Anti-inflammatory Activity of Paulownia coreana Uyeki Leaf Extract (오동나무 잎 추출물의 항염 효능에 관한 연구)

  • Kim, Nam-Kyoung;Kim, Mi-Hwa;Yoon, Chang-Soon;Choi, Shin-Wook
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.32 no.4 s.59
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    • pp.241-247
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    • 2006
  • This work was carried out to investigate the anti-inflammatory effects of Paulownia coreana Uyeki on abirritant, atopy and acne skin. Paulownia coreana Uyeki has been used as a traditional medicine having anti-febrile, anti-inflammation effect in Korea, Paulownia coreana Uyeki loaves were extracted with 70% EtOH. Its superoxide anion radical scavenging activity and inhibitory effect on LPS-induced NO production were examined. The extract inhibitied the generation of NO and $PGE_2$ induced by LPS in the macrophage cell line (Raw 264.7). Consistent with the inhibitory effects on No and $PGE_2$ generation, the extract inhibited expression of iNOS and COX-2. In further study, it was found that the extract prevented $IkB-{\alpha}$ degradation, as demonstrated by western blot analysis of $IkB-{\alpha}$ protein level. However, the extract treatment did not affect cell viability at $100{\mu}g/mL$ concentration in both human skin fibroblast and Raw 264.7 cells in vitro. Thus, the present study suggests that Paulownia coreana Uyeki leaves extract have significant anti-inflammatory activity and potential as an anti-irritation material.

Synthesis of α-cichoriin Using Deinococcus geothermalis Amylosucrase and Its Antiproliferative Effect

  • Moon, Keumok;Park, Hyun Su;Lee, Areum;Min, Jugyeong;Park, Yunjung;Cha, Jaeho
    • Microbiology and Biotechnology Letters
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    • v.50 no.2
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    • pp.218-227
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    • 2022
  • Glycosylation of aesculetin was performed using amylosucrase from the hyperthermophilic bacterium Deinococcus geothermalis DSM 11300 to improve the solubility and biological activity of aesculetin. A newly synthesized aesculetin glycoside was identified as α-cichoriin (aesculetin 7-α-D-glucoside) by nuclear magnetic resonance analysis. The solubility of α-cichoriin was 11 times higher than that of aesculetin because of the attached glucose moiety. Aesculetin and α-cichoriin had no significant effect on the proliferation of normal cells, such as RAW 264.7, but they showed a cell proliferation inhibitory effect on B16F10 melanoma cells. Unlike treatment with aesculetin and α-cichoriin, aesculin (aesculetin 6-β-D-glucoside) showed no antiproliferative activity in B16F10 cells. Based on the molecular structures of aesculin and α-cichoriin, the position where glucose binds to aesculetin and the anomeric configuration between glucose and aesculetin are thought to be important for exerting an antiproliferative effect on the B16F10 cell line. Based on these results, we propose that α-cichoriin, the α-glycosylated form of aesculetin, may serve as a model for developing phytochemical analogs with therapeutic potential for the treatment of diseases associated with tumor cell proliferation without cytotoxicity to normal cells.

An Experimental Study of the Anti-oxidant and the Anti-inflammatory Effects of Alum and Burnt Alum

  • Seo, Hyung-Sik
    • Journal of Pharmacopuncture
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    • v.15 no.2
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    • pp.11-14
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    • 2012
  • Objectives: The purpose of this study was to compare the antioxidant and anti-inflammatory effects of Alum (AL) and Burnt Alum (BAL), which are commonly used as external ointments. Methods: Extracts of AL and BAL were classified into three groups: 20, 50, and $100mg/{\mu}{\ell}$. The cytotoxicity was measured by using MTT assays in human keratinocyte cell line (HaCaT). The anti-oxidant effect was measured by using the DPPH (1, 1-diphenyl-2-picryl-hydrazyl-hydrate) radical scavenger. The anti-inflammatory effect was measured by using the inhibitory efficacy for the amount of nitric-oxide (NO) produced in mouse macrophage cell line (RAW 264.7). Results: BAL showed a higher level of cytotoxicity than AL. The AL groups showed a concentration-dependent scavenging effect on DPPH radicals, but no significant relevance was found. The BAL groups showed a concentration-dependent scavenging effect on DPPH radicals. The scavenging effects of the BAL groups were almost insignificant, but the values for the 20, 50, and $100{\mu}g/m{\ell}$ trials were different. The BAL groups showed significant concentration-dependent inhibitory effects on NO production, but the AL groups did not. Conclusions: AL showed an anti-oxidant effect more efficiently than BAL did, which demonstrated a superior anti-inflammatory effect. Therefore, for external usage, AL must be distinguished from BAL.

Anti-inflammatory Effects of Abeliophyllum distichurn Flower Extract

  • Lee, Jin Wook;Kang, Yoon Joong
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.89-89
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    • 2018
  • Abeliophyllum distichum is a medicinal plant used in regional traditional medicine to relieve pain in inflammatory processes. In this study, anti-inflammatory effects of Abeliophyllum distichum stem (ADS) ethyl acetate extract were examined. Furthermore, possible molecular mechanisms of the anti-inflammatory effects were dissected. The anti-inflammatory activity was investigated by inhibition of lipopolysaccharide (LPS) induced pro-inflammatory cytokine production in murine macrophage-like cell line Raw264.7 cells and human microglial cell line BV2 cells. The measurement of the induced pro-inflammatory cytokine levels were carried out by ELISA. The phosphorylation of ERK1/2, JNK, and MAPK, and the nuclear expression of nuclear factor $NF-{\kappa}B$ p65 were investigated by Western blot analysis. The extract of ADS significantly decreased the production of pro-inflammatory cytokines. In addition, the extract suppressed the phosphorylation of ERK1/2, JNK, and p38 MAPK, and the nuclear translocation of $NF-{\kappa}B$ p65 in activated cells. Our findings provide evidence for the popular use of Abeliophylli distichum in inflammation around Goesan region and also suggest that the stem extract has potential therapeutic benefits against several inflammatory diseases.

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A Novel Synthetic Compound, YH-1118, Inhibited LPS-Induced Inflammatory Response by Suppressing IκB Kinase/NF-κB Pathway in Raw 264.7 Cells

  • Yun, Chang Hyun;Jang, Eun Jung;Kwon, Soon Cheon;Lee, Mee-Young;Lee, Sangku;Oh, Sei-Ryang;Lee, Hyeong-Kyu;Ahn, Kyung-Seop;Lee, Ho-Jae
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.1047-1055
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    • 2015
  • For the search of a potent first-in-class compound to inactivate macrophages responsible for inflammatory responses, in the present study, we investigated the anti-nflammatory effects of YH-1118, a novel synthetic compound, in a lipopolysaccharide (LPS)-stimulated mouse macrophage cell line, Raw 264.7. YH-1118 inhibited LPS-induced nitric oxide (NO) production and inducible NO synthase (iNOS) expression at both the protein and mRNA levels. The suppression of LPS-induced iNOS expression by YH-1118 was mediated via nuclear factor kappa B (NF-κB), but not activator protein-1 (AP-1) transcription factor. This was supported by the finding that YH-1118 attenuated the phosphorylation of inhibitor of κBα (IκBα) and nuclear translocation and DNA binding activity of NF-κB. Through the mechanisms that YH-1118 inhibited the activation of IκB kinases (IKKs), upstream activators of NF-κB, or p38 MAPK, YH-1118 significantly suppressed LPS-induced production of pro-inflammatory cytokines, tumor necrosis factor-α, interleukin-1β (IL-1β), and IL-6 (p < 0.05). In conclusion, our results suggest that YH-1118 inhibits LPS-induced inflammatory responses by blocking IKK and NF-κB activation in macrophages, and may be a therapeutic candidate for the treatment of various inflammatory diseases.

The Protective Effect of Chlorophyll a Against Oxidative Stress and Inflammatory Processes in LPS-stimulated Macrophages

  • Park, Ji-Young;Park, Chung-Mu;Kim, Jin-Ju;Noh, Kyung-Hee;Cho, Chung-Won;Song, Young-Sun
    • Food Science and Biotechnology
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    • v.16 no.2
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    • pp.205-211
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    • 2007
  • This study was designed to investigate the suppressive effect of chlorophyll a on nitric oxide (NO) production and intracellular oxidative stress. In addition, chlorophyll a regulation of nuclear factor (NF) ${\kappa}B$ activation and inducible NO synthase (iNOS) expression were explored as potential mechanisms of NO suppression in a lipopolysaccharide (LPS)-stimulated macrophage cell line. RAW 264.7 murine macrophages were preincubated with various concentrations ($0-10\;{\mu}g/ mL$) of chlorophyll a and stimulated with LPS to induce oxidative stress and inflammatory response. Treatment with chlorophyll a reduced the accumulation of thiobarbituric acid-reactive substances (TBARS), enhancing glutathione level and the activities of antioxidative enzymes including superoxide dismutase, catalase, glutathione peroxidase (GSH-px), and glutathione reductase in LPS-stimulated macrophages compared to LPS-only treated cells. NO production was significantly suppressed in a dose-dependent manner (p<0.05) with an $IC_{50}$ of $12.8\;{\mu}g/mL$. Treatment with chlorophyll a suppressed the levels of iNOS protein and its mRNA expression. The specific DNA binding activities of NFkB on nuclear extracts from chlorophyll a treated cells were significantly suppressed in a dose-dependent manner with an $IC_{50}$ of $10.7\;{\mu}g/mL$. Chlorophyll a ameliorates NO production and iNOS expression through the down-regulation of NFkB activity, which may be mediated by attenuated oxidative stress in RAW 264.7 macrophages.

Expression of Neurotensin/Neuromedin N Precursor in Murine Mast Cells

  • Ahn, Hyun-Jong;Cho, Jeong-Je
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.6
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    • pp.495-501
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    • 2001
  • We have cloned the mouse neurotensin/neuromedin N (NT/N) gene from the murine mast cell line Cl.MC/C57.1 for the first time. The murine NT/N cDNA clone consisted of 765 nucleotides and coded for 169 peptide residues with an N-terminal signal peptide, and the C-terminal region contained of one copy of neurotensin (NT) and one copy of neuromedin N (NN). Total of four Lys-Arg dibasic motifs were present; one each at the middle of the open reading frame, at the N-terminal of NN, at the C-terminal of NT, and between NN and NT. Amino acid sequence analysis of the mouse NT/N revealed 90% homology to that of the rat NT/N gene. NT/N is expressed in murine mast cell lines (Cl.MC/C57.1 and P815), but not in murine bone marrow-derived mast cells (BMMCs), murine macrophage cell line (RAW 264.7), nor in murine T cell line (EL-4). NT/N mRNA in C1.MC/C57.1 is highly inducible by IgE cross-linking, phorbol myristate acetate, neurotensin, and substance P. Following the treatment of demethylating agent, 5-azacytidine (5-azaC), the NT/N gene was induced in BMMCs in response to IgE cross-linking. 5-azaC-treated BMMCs did not express the NT/N gene without additional stimuli. These findings suggested that the regulation of NT/N gene expression was dependent on the effects of not only gene methylation but also enhancer and/or repressor proteins acting on the NT/N promoter.

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