• Title/Summary/Keyword: RAW 264.7 murine macrophages

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3-(4′-hydroxyl-3′, 5′-dimethoxyphenyl) Propionic Acid Suppresses NO Production and Elevates GSH Levels in Murine Macrophages

  • Song, Young-Sun;Choi, Chun-Yeon;Suh, Hongsuk;Song, Yeong-Ok
    • Preventive Nutrition and Food Science
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    • v.9 no.3
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    • pp.270-275
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    • 2004
  • Previous studies have shown that kimchi and kimchi-derived 3-(4'-hydroxyl-3', 5'-dimethoxyphenyl) propionic acid have anti-oxidative and hypolipidemic effects in rats and rabbits. This study was designed to investigate whether chemically synthesized 3-(4'-hydroxyl-3', 5' -dimethoxyphenyl) propionic acid (HDMPPA) may ameliorate oxidative stress through the regulation of nuclear factor KB (NFkB) activation in lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophage cells. Treatment of RAW 264.7 cells with 400 uM of HDMPPA significantly reduced LPS-stimulated nitric oxide (NO) production. Treatments with HDMPPA at 100 uM to 400 uM concentrations significantly elevated glutathione (GSH) level. However, cell viability and thiobarbituric acid-reactive substances (TBARS) concentrations were not affected by the concentrations of HDMPPA used. The specific DNA binding activities of NFKB, a transcription factor which is sensitive to oxidative stress, were not down-regulated by HDMPPA treatments. These results suggest that HDMPPA may have weak anti-oxidative activity against LPS challenge by scavenging NO and stimulating GSH production.

Anti-Inflammatory Effects of Rice Bran Ethanol Extract in Murine Macrophage RAW 264.7 Cells (미강에탄올추출물의 RAW264.7 세포에서 항염증효과)

  • Park, Jeong-Suk;Kim, Mi-Hye
    • YAKHAK HOEJI
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    • v.55 no.6
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    • pp.456-461
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    • 2011
  • The aim of the present study is to investigate the anti-inflammatory effect of a Rice Bran Ethanol Extract (RBE). Inflammation, such as a bacterial infection in vivo metabolites, such as external stimuli or internal stimuli to the defense mechanisms of the biological tissue a variety of intracellular regulatory factors deulin inflammatory TNF-${\alpha}$, IL-$1{\beta}$, IL-6, IL-8, such as proinflammatory cytokines, prostagrandin, lysosomal enzyme, free radicals are involved in a variety of mediators. The present study was designed to determine the effect of the RBE on pro-inflammatory factors such as NO, iNOS expression and TNF-${\alpha}$, IL-$1{\beta}$, IL-6 in lipopolysaccharide (LPS) - stimulated RAW264.7 macrophages cells. The cell toxicity was determined by MTS assay. To evaluate of anti-inflammatory effect of RBE, amount of NO was measured using the NO detection kit and the iNOS expression was measured by reverse transcriptase polymerase chain reaction (RT-PCR). And proinflammatory cytokines were measured by ELISA kit. As a result, the RBE reduced NO, iNOS expression and TNF-${\alpha}$, IL-$1{\beta}$, IL-6 production without cytotoxicity. Our results suggest that the RBE may have an anti-inflammatory property through suppressing inflammatory mediator productions and appears to be useful as an anti-inflammatory material.

Inhibitory Effect of Galangin from Alpinia officinarum on Lipopolysaccharide-induced Nitric Oxide Synthesis in RAW 264.7 macrophages (고량강으로부터 분리된 galangin의 RAW 264.7 세포주에서 LPS로 유도된 nitric oxide 생성 저해활성)

  • Lee, Hwa Jin
    • Korean Journal of Food Science and Technology
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    • v.46 no.4
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    • pp.511-515
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    • 2014
  • In a screen for plant-derived inhibitors of nitric oxide (NO) production in lipopolysaccharide (LPS)-activated RAW 264.7 macrophage cells, a flavonol isolated from the chloroform extract of Alpinia officinarum was isolated. The structure of the flavonol was found to be 3,5,7-trihydroxy-2-phenylchromen-4-one (galangin, GLG) by using spectroscopy. GLG exhibited an inhibitory effect ($IC_{50}$ value: $26.8{\mu}M$) on NO production in LPS-stimulated RAW 264.7 murine macrophage cells. Moreover, GLG suppressed expressions of inducible nitric oxide synthase (iNOS) protein and mRNA in a dose-dependent manner.

Comparative Analysis on the Cytotoxicity of Naegleria fowleri and N. gruberi to Macrophages by the Addition of Saccharides

  • Jung, Suk-Yul
    • Biomedical Science Letters
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    • v.16 no.4
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    • pp.221-227
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    • 2010
  • To elucidate the invasion mechanism of pathogenic Naegleria fowleri, especially a receptor-ligand recognition, we investigated the in vitro cytotoxicity of pathogenic N. fowleri and nonpathogenic N. gruberi to murine macrophages, RAW 264.7, by adding four kinds of saccharides, ${\alpha}$-fucose, ${\beta}$-galactose, ${\alpha}$-D-mannopyranoside (${\alpha}$-mannose) and xylose. There was not enough of a difference in the cytotoxicity of N. fowleri treated with 10 mM of each saccharide. In particular, the cytotoxicity of N. fowleri was highly inhibited by 100 mM ${\alpha}$-mannose, which was 62.3% inhibition calculated by the analysis of lactate dehydrogenase (LDH) release assay. Although murine macrophages were not significantly destroyed by nonpathogenic N. gruberi under hematoxylin staining, the cytotoxicity of N. gruberi was inhibited from 31.5% to 14.5% (P<0.01) by 100 mM ${\alpha}$-mannose treatment. The binding of N. fowleri to macrophages was inhibited from 33% to 50% by 100 mM ${\alpha}$-mannose. Furthermore, as results of the adhesion assays which were performed to determine whether binding of Naegleria is mediated by saccharides-binding protein, the binding ability of N. fowleri as well as N. gruberi was inhibited by 100 mM ${\alpha}$-mannose.

Anti-inflammatory Activity of Dichloromethane Extract of Auricularia auricula-judae in RAW264.7 Cells

  • Damte, Dereje;Reza, Md. Ahsanur;Lee, Seung-Jin;Jo, Woo-Sik;Park, Seung-Chun
    • Toxicological Research
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    • v.27 no.1
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    • pp.11-14
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    • 2011
  • The present study investigated the anti-inflammatory effects of dichloromethane extract of Auricularia auricula-judae. Dichloromethane extract of Auricularia auricula-judae inhibited Lipopolysaccharide (LPS)-induced nitric oxide (NO) production significantly in a dose-dependent manner in the concentration ${\geq}\;10\;{\mu}g/ml$ (p < 0.05). Furthermore, RT-PCR results of this study indicated that the extract markedly reduced the expressions of inflammatory cytokines (IL-6, TNF-$\alpha$ and IL-$1{\beta}$) mRNA in LPS-treated murine RAW 264.7 macrophages, which could possibly ameliorate the inflammation. Nevertheless, dichloromethane extract of Auricularia auricula-judae did not show complete inhibition of IL-6 mRNA expression. The inhibition of IL-$1{\beta}$ cytokine at protein level was also observed in a dose dependent manner. In conclusion, the current study revealed the previously unknown effect of dichloromethane ethyl extract of Auricularia auricula-judae inhibitions of the production of NO, IL-6, TNF-$\alpha$ and IL-$1{\beta}$ in LPS-stimulated macrophages.

Raloxifene, a Selective Estrogen Receptor Modulator, Inhibits Lipopolysaccharide-induced Nitric Oxide Production by Inhibiting the Phosphatidylinositol 3-Kinase/Akt/Nuclear Factor-kappa B Pathway in RAW264.7 Macrophage Cells

  • Lee, Sin-Ae;Park, Seok Hee;Kim, Byung-Chul
    • Molecules and Cells
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    • v.26 no.1
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    • pp.48-52
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    • 2008
  • We here demonstrate an anti-inflammatory action of raloxifene, a selective estrogen receptor modulator, in lipopolysaccharide (LPS)-induced murine macrophage RAW264.7 cells. Treatment with raloxifene at micromolar concentrations suppressed the production of nitric oxide (NO) by down-regulating expression of the inducible nitric oxide synthase (iNOS) gene in LPS-activated cells. The decreased expression of iNOS and subsequent reduction of NO were due to inhibition of nuclear translocation of transcription factor NF-${\kappa}B$. These effects were significantly inhibited by exposure to the phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor, LY294002, or by expression of a dominant negative mutant of PI 3-kinase. In addition, pretreatment with raloxifene reduced LPS-induced Akt phosphorylation as well as NF-${\kappa}B$ DNA binding activity and NF-${\kappa}B$-dependent reporter gene activity. Thus our findings indicate that raloxifene exerts its anti-inflammatory action in LPS-stimulated macrophages by blocking the PI 3-kinase-Akt-NF-${\kappa}B$ signaling cascade, and eventually reduces expression of pro-inflammatory genes such as iNOS.

Downregulation of inducible nitric oxide synthase expression by a ceramide analogue in RAW 264.7 murine macrophages

  • Park, Sung-Sik;Chulbu Yim;Kim, Mie-Young;Chun, Young-Jin
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2003.05a
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    • pp.50-50
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    • 2003
  • Nitric oxide (NO) has been studied and found to be an important intracellular modulator. The excess NO produced by the inducible nitric-oxide synthase (iNOS) is implicated in various inflammatory diseases and cellular injury. Inflammatory cytokines such as TNF- or IL-6 increase intracellular ceramide and ceramide may induce NO production and inflammation. (omitted)

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Anti-inflammatory properties of chloroform extracts from GW10-45, a new cultivar derived from Pleurotus ferulae, in RAW264.7 murine macrophages. (아위느타리 신품종 GW10-45 클로로포름 추출물의 항염증 효과)

  • Choi, Hyung-Wook;Kim, Eun-Joo;Kim, Keun-Ki;Shin, Pyung-Gyun;Kim, Gun-Do
    • Journal of Mushroom
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    • v.14 no.4
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    • pp.220-224
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    • 2016
  • Chronic inflammation, which results from continuous exposure to antigens, is one of major reasons for tissue damage and diseases such as rheumatoid arthritis and type 2 diabetes. In this study, we investigated the anti-inflammatory effects of extracts (hexane, $CHCl_3$, MeOH, $MeOH/H_2O$, and $H_2O$) from GW10-45, which is our new cultivar of an edible mushroom Pleurotus ferulae (ASI 2803 and ASI 2778), in RAW264.7 murine macrophages. None of the extracts showed cytotoxicity in RAW264.7 cells and the hexane, CHCl and H extracts reduced nitric oxide (NO) production, an important inflammatory marker, in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Particularly, the extract (CG45) inhibited NO production more than the other extracts did. To elucidate the effects of CG45 on molecular targets involved in pro-inflammatory responses, we performed western blot analysis. Expression of inducible nitric oxide (iNOS) significantly decreased in LPS and CG45 co-incubated cells compared to that in LPS only-treated cells. Additionally, another protein thatplays a critical role in inflammation, was down-regulated in cells treated with both LPS and CG45. In the nuclear factor $(NF)-{\kappa}B$ pathway, phosphorylation of $I{\kappa}B{\alpha}$ decreased in RAW264.7 cells treated with both LPS and CG45. Furthermore, CG45 inhibited the phosphorylation of $NF-{\kappa}B$ in LPS-stimulated RAW264.7 cells. Conclusively, CG45 could suppress pro-inflammatory responses in LPS-stimulated RAW264.7 cells by down-regulating not only the phosphorylation of $NF-{\kappa}B$ and $I{\kappa}B{\alpha}$ but also the expression of iNOS and COX-2 without any cytotoxicity.

Activation of Murine Macrophage Cell Line RAW 264.7 by Korean Propolis

  • Han, Shin-Ha;Sung, Ki-Hyun;Yim, Dong-Sool;Lee, Sook-Yeon;Cho, Kyung-Hae;Lee, Chong-Kil;Ha, Nam-Joo;Kim, Kyung-Jae
    • Archives of Pharmacal Research
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    • v.25 no.6
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    • pp.895-902
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    • 2002
  • Monocytes and macrophages playa major role in defense mechanism of the host response to tumor, in part through the secretion of several potent products and macrophage cytokines. Monocytes and tissue macro phages produce at least two groups of protein mediators of inflammation, interleukin 1 (IL-1) and tumor necrosis factor (TNF). Recent studies emphasizes that TNF and IL-1 modulate the inflammatory function of endothelial cells, leukocytes, and fibroblasts. In this study, our work is directed toward studying the in vitro effects of Korean propolis on the ability to induce cellular and secretory responses in murine macrophage cell line, RAW 264.7. It was found that Water Extract of Korean Propolis (WEP) could activate macro phages by producing cytokines. The production of the macrophage cytokines, IL-1 and TNF-$\alpha$, by RAW 264.7 treated with WEP was examined from 2.5 $\mu\textrm{g}$/ml up to 25 $\mu\textrm{g}$/ml with dose dependent manner. Nitric oxide (NO) production was also increased when cells were exposed to combination of LPS and WEP from 2.5 $\mu\textrm{g}$/ml up to 25 $\mu\textrm{g}$/ml. At high dose of WEP (50 to 100 $\mu\textrm{g}$/ml) used to prescribe for anti-inflammatory and analgesic medicine showed inhibition of NO production in LPS-stimulated macrophage. Besides cytokine production, NO release, surface molecule expression and cell morphologic antigen expression were increased in response to the stimulation by WEP. These results suggested WEP may function through macrophage activation.

Effects of Semen jugrandis on the iNOS Expression and Superoxide Formation in the RAW264.7 Cells (호도(胡挑) 추출물이 마우스 대식세포주인 RAW264.7 세포주의 iNOS 발현 및 Superoxide 형성에 미치는 영향)

  • Moon, Goo;Ko, Su-Mi
    • The Journal of Korean Medicine
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    • v.20 no.1 s.37
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    • pp.151-160
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    • 1999
  • Nitric oxide(NO) is synthesized via the oxidation of L-arginine by a family of nitric oxide synthases(NOS), which are either constitutive(cNOS) or inducible(iNOS). The induction of iNOS in tissues can lead to the sustained production of high concentrations of NO which may exert pro-inflammatory effects including vasodilation. edema, cyototoxicity, and its activity can be mediated by various pro-inflammatory cytokine, including interferon ${\gamma}(INF-{\gamma})$. tumor necrosis factor, IL- 1 and IL-6. The enzyme, iNOS, became a new target for pharmacologcal research with the aim to find new substances for the treatment of chronic inflammatory disorders. Murine macrophages produce large amounts of NO when activated with $TFN-{\gamma}$ plus LPS. The murine macrophage-like cell line, RAW 264.7, is a suitable cell model on which to perform vitro studies regarding the iNOS system. Semen jugrandis is a fatty walnut seed found in Korea. The walnut have been used in foik medicine to improve virility, to relieved asthma, and to relieve constipation. Sesquiterpenelactones were isolated from this plant. In the course of screening for NO inhibitory activity from medicnial plants, the aqueous extract of this plant was found to have a significant activity. The result are summarized as followings. 1. The viability of cells incubated in the presence of semen jugrandis increased mare than non incubated cells. 2. Semen jugrandis suppressed the production of NO in tissues dependent on density. 3. Semen jugrandis suppressed the induction of iNOS in tissues dependent on density can lead to reduced production of NO. 4. Semen jugrandis suppressed the production of superoxide in tissue depend on density. According to the above mentioned results, semen jugrandis could be applied production of NO and superoxide can lead to reduction of chronic inflammatary. And as a depence matter come into a virus of microbe and tumor cells.

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