• Title/Summary/Keyword: RAW 264.7 Cells

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Cytokine modulation in Raw 264.7 macrophages treated with ginseng fermented by Penibacillus MBT213

  • Son, Ji Yoon;Renchinkhand, Gereltuya;Bae, Hyoung Churl;Paik, Seung-Hee;Lee, Jo Yoon;Nam, Myoung Soo
    • Korean Journal of Agricultural Science
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    • v.45 no.4
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    • pp.769-777
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    • 2018
  • The fermentation of Panax ginseng yields many compounds including ginsenosides that have various biological functions. The objective of this study was to investigate the modulation of nitric oxide (NO), Interleukin (IL)-6 and tumor necrosis factor $(TNF)-{\alpha}$ in Raw 264.7 cells treated with ginseng fermented by Penibacillus MBT213. Nitric oxide production in the Raw 264.7 cells treated for 24 hours with fermented ginseng at 3, 7, and 14 days after the treatment decreased to 74, 43, and 36%, respectively, compared with the positive control. The production of IL-6 was inhibited in all the cells treated with fermented ginseng at 3, 7, and 14 days after the treatment except for the positive control. The $TNF-{\alpha}$ production in the Raw 264.7 cells treated with fermented ginseng for 6 hours at 3, 7, and 14 days after the treatment was about 40,000, 85,000 and 65,000 pg/mL, respectively. Moreover, the $TNF-{\alpha}$ production in the Raw 264.7 cells treated with fermented ginseng for 24 hours at 7 and 14 days after the treatment was about 160,000 and 180,000 pg/mL, respectively. However, $TNF-{\alpha}$ production was inhibited in the Raw 264.7 cells at 6 and 12 hours after the treatment with fermented ginseng. herefore, it was confirmed that the immunological activity of the Raw 264.7 macrophages was affected by the treatment with fermented ginseng. It was concluded that ginseng fermented by Paenibacillus MBT213 possesses a potential anti-inflammatory activity and could be used as an ingredient in functional foods and pharmaceutical products.

Anti-inflammatory Effect of Haingkyunghonghwatang in cultured RAW264.7 cells and murine models of inflammation (RAW264.7세포주와 염증생쥐모델에서 항염증(抗炎症) 작용(作用)에 대한 행경홍화탕(行經紅花湯)의 효과(效果))

  • Min, Byung-Ho;Yoo, Dong-Youl
    • The Journal of Korean Obstetrics and Gynecology
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    • v.18 no.4
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    • pp.36-53
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    • 2005
  • Purpose : The Purpose of this research was to investigate the effects of Haingkyunghonghwatang (HKHHT) on anti-inflammatory effects. Methods : As for the parameters of inflammation, levels of several inflammatory cytokines and chemical mediators were determined in mouse lung fibroblast cells(mLFC) and RAW264.7 cells. Also, changes in pathological features by drug treatment were investigated in the in vivo edema-induced rats by carrageenin/arachidonic acid or in the colitis-induced mice by DSS treatment. Results : The cytotoxicity of HKHHT on mLFC and RAW264.7 cells wasn't observed at 100, 50, 10, and $1{\mu}g/ml$ of The treatments. $IL-1{\beta}$, IL-6 and NOS-II mRNA expression of RAW264.7 cells was inhibited by The treatments in a dose-dependent manner. HKHHT treatment of RAW264.7cells(HtRc) inhibited $TNF-{\alpha}$ and COX-2 mRNA expression. HtRc significantly inhibited IL-6 and NO production. HtRc inhibited ROS production. HKHHT inhibited rat's paw edema induced by carrageenin or arachidonate treatment in all concentrations examined. The body weight and colon length of colitis-induced mice were recovered to a normal level by DSS treatment. Clinical disease levels were significantly improved compared to the control animals. HKHHT treatment of colitis-induced mice(HtCm) significantly increased hematological values such as WBC and RBC counts, Hgb and HCT levels, but decreased PLT values. HtCm decreased IL-6 and $TNF-{\alpha}$ production significantly HtCm significantly increased CD3+(T) cell counts. In contrast, HKHHT treatment decreased CD19+ B cell counts and CD3+/CD69+ significantly, and also decreased B/T ratio (%) though not significant. Conclusion : These results indicated that HKHHT could be used for treating diverse female diseases caused by the inflammation.

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Anti-inflammatory effect of the water fraction from hawthorn fruit on LPS-stimulated RAW 264.7 cells

  • Li, Chunmei;Wang, Myeong-Hyeon
    • Nutrition Research and Practice
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    • v.5 no.2
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    • pp.101-106
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    • 2011
  • The hawthorn fruit (Crataegus pinnatifida Bunge var. typica Schneider) is used as a traditional medicine in Korea. The objective of this study was to understand the mechanisms of the anti-inflammatory effects of the water fractionated portion of hawthorn fruit on a lipopolysaccharide (LPS)-stimulated RAW 264.7 cellular model. The level of nitric oxide (NO) production in the water fraction and LPS-treated RAW 264.7 cells were determined with an ELISA. The cytotoxicity of the water fraction and LPS was measured with an MTT assay. Expression of nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor (TNF)-${\alpha}$, interleukin 6 (IL-6), and interleukin $1{\beta}$ (IL-$1{\beta}$) mRNA were analyzed with a reverse transcription polymerase chain reaction (RT-PCR). The water fraction of hawthorn fruit was determined to be safe and significantly inhibited NO production in LPS-stimulated RAW 264.7 cells and suppressed COX-2, (TNF)-${\alpha}$, IL-$1{\beta}$, and IL-6 expression. The observed anti-inflammatory effects of the water fraction of hawthorn fruit might be attributed to the down-regulation of COX-2, (TNF)-${\alpha}$, IL-$1{\beta}$, and IL-6 expression in LPS-stimulated RAW 264.7 cells.

Effect of Dipsaci Radix Water Extract on LPS-induced Inflammatory Response in RAW264.7 Mouse Macrophages (속단(續斷)의 RAW264.7 세포에서 LPS에 의해 유도되는 염증반응에 대한 효과)

  • Min, Ji-Young;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.189-195
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    • 2009
  • Objectives : In this study, the effect of Dipsaci Radix(DR, Dipsacus asperoides C.Y. Cheng et T. M. Ai) water extract on LPS-induced inflammatory response in RAW264.7 cells were investigated. Methods : Dried roots of DR was extracted with water for 3 h(DR-W extract). RAW264.7 cells, a mouse macrophage line, were incubated with different concentrations of DR-W extract for 30 min and then stimulated with LPS at indicated times. Cell toxicity was determined by MTT assay. The concentrations of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) were measured by Griess assay and enzyme immunoassay (EIA), respectively. The expression of inducible nitric oxide synthease (iNOS) and cyclooxyganase (COX)-2 mRNA and protein was determined by RT-PCR and Western blot, respectively. Results : DR-W extract was significantly inhibited LPS-induced productions of NO and PGE2 in RAW264.7 cells. DR-W extract was not suppressed the expressions of iNOS mRNA and protein in LPS-stimulated RAW264.7 cells. Conclusions : This study suggests that DR-W extract can attenuate inflammatory response via inhibition of the NO and PGE2 production in activated macrophages.

Anti-inflammatory Effects of Fermented Houttuyniae Herba Water Extract on LPS-induced Mouse Macrophage (발효 어성초(魚腥草) 물추출물의 마우스 대식세포 항염활성 연구)

  • Lee, Ji-Young;Lee, Young-Jong;Park, Wan-Su
    • The Korea Journal of Herbology
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    • v.25 no.3
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    • pp.27-34
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    • 2010
  • Objectives : The purpose of this study is to investigate the effect of Fermented Houttuyniae Herba Water Extract (HL) on production of proinflammatory mediators in mouse macrophage RAW 264.7 cells. Methods : Cell viabilities were measured by MTT assay. Effect of HL on nitric oxide (NO) production from RAW 264.7 cells was accessed by Griess reagent assay. Effect of HL on productions of inflammatory cytokines such as interleukine (IL)-17, Interferon $\gamma$-inducible protein (IP)-10, Eotaxin, IL-5, Monocyte Chemotactic Protein-3 (MCP-3), and IL-13 in LPS-induced RAW 264.7 cells was accessed by a multiplex bead array assay based on xMAP technology. Results : The results of the experiment are as follows. 1. Incubation with HL for 24 hours showed significant increase in cell viability of RAW 264.7 mouse macrophages (P < 0.05). 2. HL showed to inhibit NO production from RAW 264.7 cells at the concentrations of 25 and 50 ug/mL significantly (P < 0.05). 3. HL inhibited significantly NO production in LPS-induced RAW 264.7 cells at the concentrations of 25, 50, 100 and 200 ug/mL (P < 0.05). 4. HL inhibited significantly IL-17, IP-10 and Eotaxin in LPS-induced RAW 264.7 cells at the concentrations of 25, 50, 100 and 200 ug/mL (P < 0.05). Conclusions : These results suggest that HL has anti-inflammatory moiety related with its inhibition of NO, IL-17, IP-10, and Eotaxin in macrophages.

Modulatory Effects of 21 kinds of Medicinal Herbs Including Herba Pogostemi (Agastache rugosa) on Nitric Oxide Production in Macrophage Cell line RAW 264.7 cells (곽향(Agastache rugosa)을 포함한 21종의 한약재가 대식세포주 RAW 264.7 세포의 nitric oxide(NO) 생산 조절에 미치는 효과)

  • Kim, Seung-Hyun;Kang, Mi-Young;Nam, Seok-Hyun
    • Applied Biological Chemistry
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    • v.48 no.4
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    • pp.411-417
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    • 2005
  • Aqueous extracts were prepared from 21 medicinal herbs including Herba Pogostemi (Agastache rugosa) to examine their modulatory effects on NO production in mouse macrophage cell line RAW264.7 cells. While almost all medicinal herb extracts failed to show marked scavenging activities to NO produced by LPS stimulation, only Herba Pogostemi showed a rather strong induction of NO production in RAW264.7 cells without stimulation with LPS. When we treated the cell with $200{\mu}M\;of\;N^G-monomethyl-L-arginine\;(N^GMMA)$, a NOS2 inhibitor, a significant reduction in NO production could be observed. Moreover, a treatment of $100{\mu}M$ pyrrolidine dithiocarbamate (PDTC) led to about a 79% reduction of NO production. These results demonstrated that the aqueous extract of Herba Pogostemi might provide a second signal for the expression of NOS2 in RAW264.7 cells, and suggested that Herba Pogostemi induces NO production through L-argininedependent pathway.

Anti-inflammatory Effects of Fermented Laminaria japonica and Hizikia fusiforme Water Extracts with Probiotics in LPS-stimulated RAW264.7 Macrophage Cell Line (RAW 264.7 대식세포에서 유산균으로 발효한 다시마와 톳의 항염증 효과)

  • Hwang, Yeon-ji;Chae, Insook;Lee, Yunkyoung
    • Journal of the East Asian Society of Dietary Life
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    • v.27 no.1
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    • pp.1-8
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    • 2017
  • This study was conducted to investigate alterations of seaweed composition upon Lactobacillus rhamnosus GG (LGG) fermentation as well as potential anti-inflammatory effects and mechanism (s) of water extracts and fermented water extracts of Laminaria japonica (LJ) and Hizikia fusiforme (HF) in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. Total polyphenol, total sugar, and reducing sugar contents were measured in LJ and HF water extracts before and after fermentation by LGG. Alterations of inflammatory cytokine levels in cell culture media were measured by ELISA, and levels of phosphorylation of c-jun NH2-terminalkinase (JNK) and extra cellular signal regulated kinase (ERK) were examined by Western blot analysis. LGG fermentation of LJ and HF altered total polyphenol and sugar contents in water extracts of LJ and HF. LPS-induced production of pro-inflammatory cytokines such as IL-6 and $TNF-{\alpha}$ was significantly reduced by HF-f compared to control in RAW264.7 cells. Consistent with reduction of anti-inflammatory cytokine, interleukin (IL)-6, and tumor necrosis factor $(TNF)-{\alpha}$ levels by HF-f, HF-f also significantly reduced phosphorylation of ERK and JNK in LPS-stimulated RAW264.7 cells. In addition, LJ-f and HF also significantly reduced phosphorylation of JNK and ERK induced by LPS in RAW264.7 cells. Overall, our result suggests that HF-f among the four tested seaweed extracts is the most potent anti-inflammatory agent, and its mechanism of action is partially mediated by reduction of JNK and ERK phosphorylation as well as IL-6 and $TNF-{\alpha}$ production in LPS-stimulated RAW264.7 cells.

Inhibitory effects of Sam-Myo-San on the LPS-induced production of nitric oxide and $TNF-{\alpha}$ in RAW 264.7 cells and BV-2 Microglia cells (삼묘환(三妙丸)의 LPS에 의해 활성화된 RAW 264.7 cells과 BV-2 Microglia cells로부터 생성되는 nitric oxide 및 $TNF-{\alpha}$의 생성억제효과)

  • Lee, Jae-Hyun;Jung, Hyo-Won;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.21 no.4
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    • pp.59-67
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    • 2006
  • Objectives : Sam-Myo-Whan(SMW) has been known traditional prescription with anti- anthritis activities. We investigated inhibitory effects of SMW on lipopolysaccharide (LPS)-induced nitric oxide(NO), $TNF-{\alpha}$ and inducible nitric oxide synthase(iNOS) production from RAW264.7 cells and BV-2 Microglia cells. Methods : SMW, which had been extracted with 70% MeOH, concentrated and freeze-dried was used for this experiment. After BV2 mouse brain macrophages and RAW264.7 mouse peritoneal macrophages were pretreated with increasing concentrations of SMW extract for 30min, and then activated with LPS. To investigate cytotoxicity of SMW extract, cell viability was measured by MTT assay. NO production was measured in each culture supernatant by Griess reaction. mRNA expression of iNOS in two type cells was investigated by RT-PCR. $TNF-{\alpha}$ production was measured in each culture supernatant by ELISA. Results : SMW extract significantly inhibited LPS-induced NO and $TNF-{\alpha}$ production in BV2 cells and RAW264.7 cells dose-dependently. SMW extract also greatly suppressed mRNA expression of iNOS in both type cells activated with LPS. Conclusion : These data suggests that SMW extract may have an anti-inflammatory effect through the inhibition of iNOS expression.

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Anti-inflammatory effect of a mixture of Astragalus membranaceus and Lithospermum erythrorhizon extracts by inhibition of MAPK and NF-κB signaling pathways in RAW264.7 cells (RAW264.7 대식세포에서 MAPK 및 NF-κB 신호전달 경로 억제를 통한 황기 및 지치 복합물의 항염증 효과)

  • Choi, Doo Jin;Kim, Geum Soog;Choi, Bo-Ram;Lee, Young-Seob;Han, Kyung Sook;Lee, Dong-Sung;Lee, Dae Young
    • Journal of Applied Biological Chemistry
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    • v.63 no.4
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    • pp.421-428
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    • 2020
  • This study investigated a mixture of Astragalus membranaceus (AM) and Lithospermum erythrorhizon (LE) extracts (ALM16), exerts anti-inflammatory effects in lipopolysaccharide (LPS)-induced RAW264.7 macrophage cells, and its underlying mechanism. ALM16 was prepared by mixing AM and LE extracts in a ratio of 7:3 (w/w). Cytotoxicity of ALM16 in RAW264.7 cells was not shown up to 200 ㎍/mL of ALM16. The results of this study showed that ALM16 does-dependently inhibits the production of nitric oxide, prostaglandin E2 and pro-inflammatory cytokines (interleukin-1β, interleukin-6, and tumor necrosis factor-α) in LPS-induced RAW264.7 cells. ALM16 not only markedly reduced the protein expression levels of inducible nitric oxide synthase and cyclooxygenase-2 (COX-2) in LPS-stimulated RAW264.7 cells, but also inhibited the nuclear translocation and DNA-binding activity of nuclear factor-kappa B (NF-κB). In addition, ALM16 specifically inhibited the phosphorylation of c-Jun N-terminal kinase and extracellular signal-regulated kinases in LPS-stimulated RAW264.7 cells. In conclusion, these results suggest that ALM16 may exert anti-inflammatory effect by modulating mitogen-activated protein kinase and NF-κB signaling pathways.

The Study on the Physiological Activities of Beta vulgaris such as Antioxidant and Anti-inflammatory in RAW 264.7 cells (RAW 264.7 세포에서 레드비트의 항산화 및 항염증 등의 생리활성 연구)

  • Ji, Joong-Gu
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.1
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    • pp.309-317
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    • 2021
  • This study was designed to examine the cell cytotoxic, anti-inflammatory, and antioxidant activity for raw material of Beta vulagaris tea. Antioxidative ability was evaluated by bioassays using DPPH and ABTS radical scavenging activity. Cell viability was assessed by MTT assay using RAW 264.7 cells, and investigated production levels of reactive oxide speies, and inflammatory meditors(i.e., nitric oxide, tumor necrosis factor-α, and interleukin-6) in LPS-induced RAW 264.7 cells. As a results, DPPH and ABTS raidcal scavenging activity were increased in a dose-dependent manner, and confirmed no cytotoxicity in all concentration. Also, it was significantly decreased level of ROS, NO, IL-6, and TNF-a in LPS-induced RAW 264.7 cells. Therefore these results suggest Beta vulagaris has considerable potential as a raw material of leached tea with safe anti-oxidative and anti-inflammatory effects.