• Title/Summary/Keyword: murine macrophage RAW 264.7 cells

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$\beta$-Glucan Suppresses LPS-stimulated NO Production Through the Down-regulation of iNOS Expression and $NF{\kappa}B$ Transactivation in RAW 264.7 Macrophages

  • Yang, Jeong-Lye;Jang, Ji-Hyun;Radhakrishnan, Vinodhkumar;Kim, Yang-Ha;Song, Young-Sun
    • Food Science and Biotechnology
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    • v.17 no.1
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    • pp.106-113
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    • 2008
  • The antioxidant and anti-inflammatory protective effects of $\beta$-glucan from barley on RAW 264.7 murine macrophage cells induced by lipopolysaccharide (LPS) were examined. The RAW 264.7 murine macrophages were preincubated with various concentrations ($0-200\;{\mu}g/mL$) of $\beta$-glucan and stimulated with LPS to induce oxidative stress and inflammation. The $\beta$-glucan treatments were found to reduce thiobarbituric acid-reactive substance (TBARS) accumulation, and enhance glutathione levels and the activities of antioxidative enzymes, including superoxide dismutase (SOD), catalase, glutathione reductase, and glutathione peroxidase (GSH-px) in the LPS-stimulated macrophages as compared to the LPS-only treated cells. Nitric oxide (NO) production was significantly suppressed in a dose-dependent manner (p<0.05) with an $IC_{50}$ of $104\;{\mu}g/mL$. Further treatment with $\beta$-glucan at $200\;{\mu}g/mL$ suppressed NO production to 2% of the LPS-control, and suppressed the levels of inducible nitric oxide synthase (iNOS) protein and mRNA in a dose-dependent manner. The specific DNA binding activity of nuclear factor ${\kappa}B\;(NF{\kappa}B)$ was significantly suppressed by $\beta$-glucan treatment with an $IC_{50}$ of $220\;{\mu}g/mL$ in a dose-dependent manner. Finally, barley $\beta$-glucan ameliorates NO production and iNOS expression through the down-regulation of $NF{\kappa}B$ activity, which may be mediated by attenuated oxidative stress in RAW 264.7 macrophages.

Anti-inflammatory Activity of Sorghum bicolor (L.) Moench var. Hwanggeumchal Grains in Lipopolysaccharide-stimulated RAW264.7 Murine Macrophage Cell Line (지질다당류-자극된 마우스 대식세포주 RAW264.7에서 황금찰수수 종자의 항염증 활성)

  • Jun, Do Youn;Woo, Hyun Joo;Ko, Jee Youn;Kim, Young Ho
    • Journal of Life Science
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    • v.32 no.12
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    • pp.929-937
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    • 2022
  • To investigate the anti-inflammatory activity of the grains of sorghum, three Sorghum bicolor (L.) Moench variants (Hwanggeumchal, Huinchal, and Chal) being cultivated in Korea, the 80% ethanol (EtOH) extracts of individual sorghum grains were compared for their inhibitory activity against nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7 murine macrophage cell line. Among them, the EtOH extract of sorghum Hwanggeumchal grains could exert the highest inhibitory effect on the LPS-induced NO production. However, under these conditions, the viability of RAW264.7 cells was not affected. When the EtOH extract of sorghum Hwanggeumchal grains was sequentially fractionated with n-hexane, methylene chloride (MC), ethyl acetate (EtOAc), and n-butanol, the anti-NO production activity was predominantly detected in both MC and EtOAc fractions. In particular, treatment with the MC fraction reduced dose-dependently the expression levels of iNOS, COX-2 and pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) in LPS-stimulated RAW264.7 cells. Simultaneously, the MC fraction could prevent LPS-induced activating phosphorylation of p38 mitogen-activated protein kinase (MAPK), c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK). HPLC analysis of the MC fraction showed gentisic acid and naringenin as the major phenolic components. Both gentisic acid and naringenin commonly exhibited a potent inhibitory activity against LPS-induced NO production in RAW264.7 cells. Together, these results provide the evidence of the inhibitory activity of Hwanggeumchal grains on LPS-induce inflammatory responses in RAW264.7 murine macrophage cells and also suggest that sorghum grains possess beneficial health effects which can be applicable in development of the grain-based functional foods.

Cilostazol Decreases Ethanol-Mediated TNFalpha Expression in RAW264.7 Murine Macrophage and in Liver from Binge Drinking Mice

  • Lee, Youn-Ju;Eun, Jong-Ryeol
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.2
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    • pp.131-138
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    • 2012
  • Alcoholic hepatitis is a leading cause of liver failure in which the increased production of tumor necrosis factor ${\alpha}$ (TNF${\alpha}$) plays a critical role in progression of alcoholic liver disease. In the present study, we investigated the effects of cilostazol, a selective inhibitor of type III phosphodiesterase on ethanol-mediated TNF${\alpha}$ production in vitro and $in$ $vivo$, and the effect of cilostazol was compared with that of pentoxifylline, which is currently used in clinical trial. RAW264.7 murine macrophages were pretreated with ethanol in the presence or absence of cilostazol then, stimulated with lipopolysacchride (LPS). Cilostazol significantly suppressed the level of LPS-stimulated TNF${\alpha}$ mRNA and protein with a similar degree to that by pentoxifylline. Cilostazol increased the basal AMP- activated protein kinase (AMPK) activity as well as normalized the decreased AMPK by LPS. AICAR, an AMPK activator and db-cAMP also significantly decreased TNF${\alpha}$ production in RAW264.7 cells, but cilostazol did not affect the levels of intracellular cAMP and reactive oxygen species (ROS) production. The $in$ $vivo$ effect of cilostazol was examined using ethanol binge drinking (6 g/kg) mice model. TNF${\alpha}$ mRNA and protein decreased in liver from ethanol gavaged mice compared to that from control mice. Pretreatment of mice with cilostazol or pentoxifylline further reduced the TNF${\alpha}$ production in liver. These results demonstrated that cilostazol effectively decrease the ethanol-mediated TNF${\alpha}$ production both in murine macrophage and in liver from binge drinking mice and AMPK may be responsible for the inhibition of TNF${\alpha}$ production by cilostazol.

Anti-inflammatory Effect of Mugi-hwan Water Extract in RAW 264.7 Cells (무기환(戊己丸)의 RAW 264.7 세포에 대한 항염증작용 연구)

  • Kim, Ilhyun;Choi, Chonghwan;Lee, Sewon;Song, Yungsun
    • Journal of Korean Medicine Rehabilitation
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    • v.23 no.3
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    • pp.27-35
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    • 2013
  • Objectives The aim of this study was to investigate anti-inflammatory activity of Mugi-hwan (MGH) Water Extract. Methods Cells were treated with 2 ug/ml of LPS 1 hour prior to the addition of MGH. Cell viability was measured by MTS assay. The production of NO was determined by reacting cultured medium with Griess reagent. The expression of COX-2, iNOS and MAPKs was investigated by Western blot, RT-PCR. The content of level of cytokines ($PGE_2$, IL-6, in media from LPS-stimulated Raw 264.7 cells was analyed by ELISA kit. Results MGH inhibited the production of NO, $PGE_2$, IL-6 as well as the expressions of iNOS, COX-2 in the murine macrophage, RAW 264.7 cells. MGH also had suppression effects of LPS induced MAPKs activation. Conclusions These results suggest that MGH has an anti-inflammatory therapeutic potential, which may result from inhibition of MAPK phosphorylation, thereby decreasing the expression of pro-inflammatory genes.

Aqueous extract of Lycii fructus suppresses inflammation through the inhibition of nuclear factor kappa B signal pathway in murine raw 264.7 macrophages

  • Kim, Beum-Seuk;Lim, Hyung-Ho;Song, Yun-Kyung;Sung, Yun-Hee;Kim, Sung-Eun;Chang, Hyun-Kyung;Shin, Mal-Soon;Kim, Chang-Ju;Lee, Hye-Jung;Kim, Dong-Hee
    • Advances in Traditional Medicine
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    • v.10 no.3
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    • pp.155-164
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    • 2010
  • Lycii fructus is the fruit of Lycium chinense Miller and is part of the Solanaceae family. Lycii fructus produces various effects such as hypotensive, hypoglycemic, anti-pyretic, and anti-stress activities. Lycii fructus is known to contain betaine, carotene, nicotinic acid, zeaxanthin, and cerebroside. In the present study, the effects of Lycii fructus aqueous extract on lipopolysaccharide (LPS)-induced inflammation in murine raw 264.7 macrophage cells were investigated. In this study we utilized the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, reverse transcriptionpolymerase chain reaction (RT-PCR), Western blotting, and nitric oxide (NO) detection. Lycii fructus aqueous extract suppressed NO production by inhibiting the LPS-induced expressions of inducible nitric oxide synthase (iNOS) and tumor necrosis factor-alpha (TNF-$\alpha$) mRNA and iNOS protein in murine raw 264.7 macrophage cells. Also, Lycii fructus aqueous extract suppressed the activation of nuclear factor-kappa B (NF-${\kappa}B$) in the nucleus. These results demonstrated that Lycii fructus aqueous extract causes an anti-inflammatory effect that was likely produced by the suppression of iNOS expression through the down-regulation of NF-$\hat{e}B$ binding activity.

Comparative Effect on Anti-Inflammatory Activity of the Phellinus linteus and Phellinus linteus Grown in Germinated Brown Rice Extracts in Murine Macrophage RAW 264.7 Cells (상황버섯과 발아현미상황버섯 열수추출물의 Murin Macrophage RAW 264.7 세포에서 항염증 반응 비교)

  • Jeoung, Young-Jun;Choi, Se-Young;An, Chi-Sun;Jeon, Yun-Hee;Park, Dong-Ki;Lim, Beoung-Ou
    • Korean Journal of Medicinal Crop Science
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    • v.17 no.2
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    • pp.97-101
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    • 2009
  • The present study describes the preliminary evaluation of the anti-inflammatory activities of Phellinus linteus (PL) and Phellinus linteus Grow in Germinated Brown Rice (BRPL). In order to effectively screen for anti-inflammatory agents, we first examined the extracts' inhibitory effects on the expression of pro-inflammatory cytokines activated with lipopolysaccharide. Moreover, we examined the inhibitory effects of the PL and BRPL extracts on pro-inflammatory factors such as NO, iNOS, $TNF-{\alpha}$ and $IFN-{\gamma}$ in murine macrophage RAW 264.7 cells. NO production and iNOS expression was significantly augmented in LPS treated cell, the production of NO and iNOS was greater in the BRPL than in the PL group. In addition, protein and mRNA levels of $TNF-{\alpha}$ and $IFN-{\gamma}$ in BRPL showed relatively more potent pro-inflammatory-activity inhibition compared to that of PL. These results suggest that BRPL may have significant effects on inflammatory factors, and may be a potential anti-inflammatory therapeutic materials.

In vitro Anti-inflammatory Activity of the Artemisia fukudo Extracts in Murine Macrophage RAW 264.7 Cells (큰비쑥(Artemisia fukudo) 추출물의 murine macrophage RAW 264.7 세포에서 in vitro 항염효과)

  • Yoon, Weon-Jong;Lee, Jung-A;Kim, Kil-Nam;Kim, Ji-Young;Park, Soo-Yeong
    • Korean Journal of Food Science and Technology
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    • v.39 no.4
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    • pp.464-469
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    • 2007
  • The present study describes the preliminary evaluation of the anti-inflammatory activities of Artemisia fukudo extracts. The 80% ethanol extract of A. fukudo was sequentially fractionated with n-hexane, dichloromethane, ethylacetate, and butanol. In order to effectively screen for anti-inflammatory agents, we first examined the extracts’ inhibitory effects on the production of pro-inflammatory cytokines activated with lipopolysaccharide. Moreover, we examined the inhibitory effects of the A. fukudo extracts on pro-inflammatory factors (NO, iNOS, COX-2, and $PGE_{2}$) in murine macrophage RAW 264.7 cells. The protein levels were determined by immunoblotting. Of the sequential solvent fractions, the n-hexane and dichloromethane fractions inhibited the mRNA expression of pro-inflammatory cytokines (TNF-${\alpha}$, IL-$1{\beta}$, and IL-6), production of NO and $PGE_{2}$, and the protein levels of iNOS and COX-2. These results suggest that A. fukudo may have signifIcant effects on inflammatory factors, and may be a potential anti-inflammatory therapeutic plant.

Investigation of the IL-1β, TNF-α and iNOS gene differential expression in Raw 264.7 cells by the water extract of Angelicae Radix from Korea, China and Japan (참당귀, 중국당귀, 일당귀 열수 추출물의 RAW 264.7 대식세포에서 IL-1β, TNF-α, iNOS 유전자 차등 발현 연구)

  • Han, Hyo-Sang;Hong, Seong-Gyun
    • Journal of Digital Convergence
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    • v.15 no.11
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    • pp.513-522
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    • 2017
  • We tried to analyze the inflammation reactions by treatment of AG, AS and AA in murine RAW 264.7 cells. To investigate the effect of AG, AS and AA on cell viability of RAW 264.7 cells, AG, AS and AA were treated for 24 h and MTS assay was performed. Cell viabilities were increased in $1,600{\mu}g/ml$ concentration by AS, AA and AG treatments, respectively. The mRNA expression levels of $IL-1{\beta}$, $TNF-{\alpha}$ and iNOS were increased by AG and AA treatment at a concentration of $200{\mu}g/ml$ in RAW 264.7 cells without Lipopolysaccharide (LPS) treatment. The mRNA expression levels of $IL-1{\beta}$, $TNF-{\alpha}$ and iNOS were increased by AG and AA 6 h treatment at a concentration of $200{\mu}g/ml$ with LPS treatment. In this study, we observed that AG, AS and AA show various activities on inflammation reaction depend on their treatment time. In the future, studies should be conducted to investigate the effects of AG, AS and AA on the various inflammatory responses of macrophages.

In vitro Antiinflammatory Activity of the Essential oil Extracted from Chrysanthemum sibiricum in Murine Macrophage RAW 264.7 Cells

  • Lee, Kyung-Tae;Kim, Ryung-Kyu;Ji, Sa-Young;Shin, Kyoung-Min;Choi, Jong-Won;Jung, Hyun-Ju;Park, Hee-Juhn
    • Natural Product Sciences
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    • v.9 no.2
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    • pp.93-96
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    • 2003
  • This research was undertaken to find the in vitro inflammatory action of the essenetial oil (CS-oil) extracted from Chrysanthemum sibiricum (Compositae) herbs. We investigated the effects of the CS-oil not only on the formation NO, $PGE_2$, and $TNF-{\alpha}$ but also on inducible nitric oxide synthase and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-induced murine macrophage RAW 264.7 cells. The data obtained were consistent with the modulation of iNOS enzyme expression. A similar fashion was also observed when LPS-induced $PGE_2$ release and COX-2 expression were tested. The significant inhibitory effects were shown in concentration-dependent manners. In addition, CS-oil also mildly but significantly reduced the formation of TNF-a. These findings support the application of CS-oil as an antiinflammatory essential oil.