• Title/Summary/Keyword: RAW 264.7 Cell Line

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In vitro Antiinflammatory Activity of 23-Hydroxyursolic Acid Isolated from Cussonia bancoensis in Murine Macrophage RAW 264.7 Cells

  • Kim, Rung-Gyu;Shin, Kyung-Min;Park, Hee-Juhn;Park, Jong-Won;Lee, Kyung-Tae
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.320.1-320.1
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    • 2002
  • We investigated the effect of various triterpenoids isolated from the Cussonia bancoensis. such as ursolic acid. 23-hydroxyursolic acid. 3-O-${\alpha}$-L -arabinopyranosyl-23 -hydroxyursolic acid. 3-O-${\beta}$-D-glucopyranosyl-23-hydroxy-ursolic acid and 28-O-${\alpha}$-L -rhamnopyranosyl(1-4)-${\beta}$-D-glucopyranosyl( 1-6)-${\beta}$-D-glucopyranosylester of 23-hydroxyursolic acid. have been evaluated on lipopolysaccharide (LPS)-induced nitric oxide (NO) and prostaglandin E2 (PGE2) release by the macrophage cell line RAW 264.7. (omitted)

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Comparative analysis on immune response of combination with Astragali Radix and Cinnamomi Cortex (황기 육계의 배합에 따른 면역활성 비교)

  • Jung, Da-Young;Ha, Hye-Kyung;Lee, Ho-Young;Lee, Nam-Hun;Shin, Hyeun-Kyoo
    • The Korea Journal of Herbology
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    • v.26 no.4
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    • pp.187-194
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    • 2011
  • Objective : Astragali Radix (AR) and Cinnamomi Cortex (CC) are used to enhance immune response in Asian traditional medicine. Immuno-potentiation of the combination of AR and CC were evaluated on the cellular and humoral immune response using murine macrophage cell line (RAW 264.7) and OVA-immunized mice. Methods : This study was designed to investigate the immuno-potentiative effects of AR, CC, and AR with CC on nitric oxide synthesis in RAW 264.7 cells and proliferation and production levels of Intereukin-2 (IL-2) in mouse splenocytes. In addition, we evaluated the plasma-specific antibody responses and splenocyte proliferation on ovalbumin (OVA)-immunized mice treated with herbal extracts. Results : Combination treatment with AR and CC increased nitric oxide synthesis in RAW 264.7 cells and IL-2 level in splenocytes (p<0.001). Combination of AR and CC significantly enhanced the Concanavalin A- (Con A ; T cell mitogen) and lipopolysaccharide-(LPS ; B cell mitogen) induced splenocyte proliferation on the OVA-immunized mice. Combination of AR and CC also significantly enhanced plasma levels of OVA-specific IgG (p<0.01), IgG1 (p<0.05) and total IgM (p<0.01) compared with the OVA-immunized control group. Conclusion : These results suggest that combination of AR and CC could be used as therapeutic profile on activation of immune response.

Upregulation of heme oxygenase-1 by ginsenoside Ro attenuates lipopolysaccharide-induced inflammation in macrophage cells

  • Kim, Sokho;Oh, Myung-Hoon;Kim, Bum-Seok;Kim, Won-Il;Cho, Ho-Seong;Park, Byoung-Yong;Park, Chul;Shin, Gee-Wook;Kwon, Jungkee
    • Journal of Ginseng Research
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    • v.39 no.4
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    • pp.365-370
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    • 2015
  • Background: The beneficial effects of ginsenoside species have been well demonstrated in a number of studies. However, the function of ginsenoside Ro (GRo), an oleanane-type saponin, has not been sufficiently investigated. Thus, the aim of the present study was to investigate the anti-inflammatory effects of GRo in vitro using the Raw 264.7 mouse macrophage cell line treated with lipopolysaccharide (LPS), and to clarify the possible mechanism of GRo involving heme oxygenase-1 (HO-1), which itself plays a critical role in self-defense in the presence of inflammatory stress. Methods: Raw 264.7 cells were pretreated with GRo (up to $200{\mu}M$) for 1 h before treatment with 1 mg/mL LPS, and both cell viability and inflammatory markers involving HO-1 were evaluated. Results: GRo significantly increased cell viability in a dose dependent manner following treatment with LPS, and decreased levels of reactive oxygen species and nitric oxide. GRo decreased inflammatory cytokines such as nitric oxide synthase and cyclooxygenase-2 induced by LPS. Moreover, GRo increased the expression of HO-1 in a dose dependent manner. Cotreatment of GRo with tin protoporphyrin IX, a selective inhibitor of HO-1, not only inhibited upregulation of HO-1 induced by GRo, but also reversed the anti-inflammatory effect of GRo in LPS treated Raw 264.7 cells. Conclusion: GRo induces anti-inflammatory effects following treatment with LPS via upregulation of HO-1.

Mechanisms Underlying Enterococcus faecalis-Induced Tumor Necrosis Factor-$\alpha$ Production in Macrophages

  • Choi, Eun-Kyoung;Kim, Dae-Eob;Oh, Won-Mann;Paek, Yun-Woong;Kang, In-Chol
    • International Journal of Oral Biology
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    • v.35 no.2
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    • pp.43-49
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    • 2010
  • Enterococcus faecalis, a gram-positive bacterium, has been implicated in endodontic infections, particularly in chronic apical periodontitis. Proinflammatory cytokines, including tumor necrosis factor-$\alpha$ (TNF-$\alpha$), are involved in the pathogenesis of these apical lesions. E. faecalis has been reported to stimulate macrophages to produce TNF-$\alpha$. The present study investigated the mechanisms involved in TNF-$\alpha$ production by a murine macrophage cell line, RAW 264.7 in response to exposure to E. faecalis. Both live and heat-killed E. faecalis induced high levels of gene expression and protein release of TNF-$\alpha$. Treatment of RAW 264.7 cells with cytochalasin D, an inhibitor of endocytosis, prevented the mRNA up-regulation of TNF-$\alpha$ by E. faecalis. In addition, antioxidant treatment reduced TNF-$\alpha$ production to baseline levels. Inhibition of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein (MAP) kinase also significantly attenuated E. faecalis-induced TNF-$\alpha$ expression by RAW 264.7 cells. Furthermore, activation of NF-${\kappa}B$ and AP-1 in RAW 264.7 cells was also stimulated by E. faecalis. These results suggest that the phagocytic uptake of bacteria is necessary for the induction of TNF-$\alpha$ in E. faecalis-stimulated macrophages, and that the underlying intracellular signaling pathways involve reactive oxygen species, ERK, p38 MAP kinase, NF-${\kappa}B$, and AP-1.

Anti-inflammatory Effects of the Methanol Extract of Polytrichum Commune via NF-κB Inactivation in RAW 264.7 Macrophage Cells

  • Cho, Woong;Park, Seung-Jae;Shin, Ji-Sun;Noh, Young-Su;Cho, Eu-Jin;Nam, Jung-Hwan;Lee, Kyung-Tae
    • Biomolecules & Therapeutics
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    • v.16 no.4
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    • pp.385-393
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    • 2008
  • As an attempt to search for bioactive natural products exerting anti-inflammatory activity, we evaluated the effects of the methanol extract of Polytrichum commune Hedw (PCM) (Polytrichaceae) on lipopolysaccharide (LPS)-induced nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) and pro-inflammatory cytokines release in murine macrophage cell line RAW 264.7. PCM potently inhibits the production of NO, $PGE_2$, tumor necrosis factor (TNF)-$\alpha$ and interleukin (IL)-6. Consistent with these results, PCM also concentration-dependently inhibited LPS-induced inducible NO synthase (iNOS) and cyclooxygase (COX)-2 at the protein levels, and iNOS, COX-2, TNF-$\alpha$ and IL-6 at the mRNA levels without an appreciable cytotoxic effect on RAW 264.7 macrophag cells. Furthermore, PCM inhibited LPS-induced nuclear factor-kappa B (NF-$\kappa$B) activation as determined by NF-$\kappa$B reporter gene assay, and this inhibition was associated with a decrease in the nuclear translocation of p65 and p50 NF-$\kappa$B. Taken together, these results suggest that PCM may play an anti-inflammatory role in LPS-stimulated RAW 264.7 macrophages through the inhibitory regulation of iNOS, COX-2, TNF-$\alpha$ and IL-6 via NF-$\kappa$B inactivation.

A comparison of anti-inflammatory activities of green tea and grapefruit seed extract with those of microencapsulated extracts (미세캡슐화한 녹차 및 자몽종자 추출물이 Murine RAW 264.7 대식세포주의 항염증에 미치는 영향 비교)

  • Jun, Yoon Kyung;Kim, Myung Hwan;Seong, Pil Nam;Chang, Moon-Jeong
    • Journal of Nutrition and Health
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    • v.45 no.5
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    • pp.443-451
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    • 2012
  • We compared the effects of grapefruit seed extract (GFSE), green tea extract (GT) and their microencapsulated extract on anti-inflammatory activities in murine RAW 264.7 macrophages cell line. In order to protect the bioactive compounds in the extracts, they were microencapsulated with maltodextrin and $H_2O$. Nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), inducible nitric oxide synthase (iNOS) protein expression and thiobarbiturate reactive substances (TBARS) were analyzed in LPS activated RAW 264.7 macrophages. The green tea extract at the range of $100-600{\mu}g/mL$ inhibited NO, PGE2 production and iNOS protein expression without cytotoxicity in a dose-dependent manner. Grapefruit seed extract had strong inhibitory effects on NO and PGE production and iNOS protein expression at the range of $5-20{\mu}g/mL$ without cytotoxicity. Microencapsulation of green tea extract had further inhibitory effects on NO and PGE2 production and on iNOS protein expression, whereas microencapsulated GFSE did not show any further inhibitory effects on these parameters. Taken together, our results suggest that GSFE might be a promising candidate for preventing inflammation related diseases, such as cardiovascular disease, cancer or diabetes, and the microencapsulation of green tea extract could improve its bioactivity.

Anti-Inflammatory Effects of Grasshopper Ketone from Sargassum fulvellum Ethanol Extract on Lipopolysaccharide-Induced Inflammatory Responses in RAW 264.7 Cells

  • Kim, Min-Ji;Jeong, So-Mi;Kang, Bo-Kyeong;Kim, Koth-Bong-Woo-Ri;Ahn, Dong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.29 no.5
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    • pp.820-826
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    • 2019
  • This study evaluated the anti-inflammatory potential of a grasshopper ketone (GK) isolated from the brown alga Sargassum fulvellum on lipopolysaccharide (LPS)-induced RAW 264.7 murine macrophage cell line. GK was isolated and purified from the n-hexane fraction and its structure was verified on the basis of NMR spectroscopic data. GK up to $100{\mu}g/ml$ is not cytotoxic to RAW 264.7, and is an effective inhibitor of LPS-induced NO production in RAW 264.7 cells. The production of pro-inflammatory cytokines, including IL-6, $IL-1{\beta}$, and $TNF-{\alpha}$ was found significantly reduced in $0.1-100{\mu}g/ml$ dose ranges of GK treatment (p < 0.05). We confirmed the dose-dependent and significant inhibition of iNOS and COX-2 proteins expression. In addition, it has been shown that GK induces anti-inflammatory effects by inhibiting MAPKs (ERK, JNK, and p38) and $NF-{\kappa}B$ p65 phosphorylation. Our results show that the anti-inflammatory properties of GK may be due to the inhibition of the $NF-{\kappa}B$ and MAPKs pathways, which are associated with the attenuation of cytokine secretion.

The Experimental Study on Anti-oxidant and Anti-inflammatory Effect of Sungyoutanggagambang(SYTG) (성유탕가감방(聖愈湯加減方)의 항염증(抗炎症) 및 항산화(抗酸化))

  • Kim, Eui-Il;Yoo, Dong-Youl
    • The Journal of Korean Obstetrics and Gynecology
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    • v.21 no.4
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    • pp.69-89
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    • 2008
  • Purpose: This study was performed to evaluate anti-oxidant activities and anti-inflammatory effects of Sungyoutanggagambang(SYTG). Methods: In the study of anti-oxidant activities. SYTG was investigated by DPPH radical scavenger activity. superoxide dismutase activity and superoxide anion radical scavenger activity. In the study of anti-inflammatory effects. SYTG was investigated using cultured cells and murine models. As for the parameters of inflammation. levels of several inflammatory cytokines and chemical mediators which are known to be related to inflammation were measured in mouse lung fibroblast cells(mLFCs) and RAW264.7 cells. Results: Prior to the experiment. we investigated the security of SYTG by measuring GOT and GPT in serum. 1. SYTG showed high antioxidant activity in a concentration-dependent degree by measured scavenging activity of DPPH free radical, superoxide dismutase and superoxide anion radical. 2. SYTG inhibited IL-1$\beta$, IL-6. TNF-$\alpha$, COX-2 and NOS-II mRNA expression as compared with the control group in a concentration-dependent degree in RAW264.7 cell line. 3. SYTG inhibited IL-1$\beta$, IL-6 production significantly at 100 ${\mu}g/ml$ and TNF-$\alpha$ production significantly at 50, 100 ${\mu}g/ml$ as compared with the control group in RA W264.7 cell line. 4. SYTG inhibited IL-1$\beta$, and IL-6 production significantly as compared with the control group in serum of acute inflammation-induced mice. and decreased IL-1$\beta$, IL-6 production in spleen tissue. and also decreased IL-1$\beta$, IL-6 production in liver tissue. Conclusion: These results suggest that SYTG can be useful in treating diverse female diseases caused by inflammation such as endometrosis, myoma, pelvic congestion. chronic cervicitis, chronic pelvic inflammatory disease and so on.

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The Effects of Bee Venom and Melittin acupuncture solution on cPLA2, TNF-α and Calcium Concentration in RAW 264.7 Cells (봉약침액(蜂藥鍼液)과 Melittin 약침액(藥鍼液)이 RAW 264.7 Cell의 cPLA2, TNF-α 및 Calcium Concentration에 미치는 영향(影響))

  • Park, Young-eun;Song, Ho-sueb
    • Journal of Acupuncture Research
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    • v.21 no.5
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    • pp.179-190
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    • 2004
  • Objectives : The purpose of this study was to investigate the effect of Bee Venom and Melittin acupuncture solution on the lipopolysaccharide and sodium nitroprusside- induced expression of cytosolic phospholipase $A_2$, tumor necrosis factor-${\alpha}$ and calcium concentration in RAW 264.7 cells, a murine macrophage cell line. Methods : The expression of cytosolic phospholipase $A_2$ and tumor necrosis factor-${\alpha}$ was determined by western blotting with corresponding antibodies, and the generation of intracellular calcium concentration was investigated by delta scan system in RAW 264.7 cells. Results : 1. Compared with control, expressions of lipopolysaccharide-induced cytosolic phospholipase $A_2$ were decreased significantly by $5{\mu}g/mL$ of bee venom and 5, $10{\mu}g/mL$ of melittin acupuncture solution and decreased by 0.5, $1{\mu}g/mL$ of bee venom. 2. Compared with control, expressions of sodium nitroprusside-induced phospholipase $A_2$ were decreased significantly by 0.5, 1, $5{\mu}g/mL$ of bee venom and by 5, $10{\mu}g/mL$ of melittin acupuncture solution. 3. Compared with control, expressions of lipopolysaccharide-induced tumor necrosis factor-${\alpha}$ were decreased significantly by $10{\mu}g/mL$ of melittin acupuncture solution and were not changed significantly by 0.5, 1, $5{\mu}g/mL$ of bee venom and $5{\mu}g/mL$ of melittin acupuncture solution. 4. Compared with control, expressions of sodium nitroprusside-induced tumor necrosis factor-${\alpha}$ were decreased significantly by 1, $5{\mu}g/mL$ of bee venom and 5, $10{\mu}g/mL$ of melittin acupuncture solution and decreased by $0.5{\mu}g/mL$ of bee venom 5. Compared with control, lipopolysaccharide-induced intracellular calcium concentrations were decreased by 0.5, 1, $5{\mu}g/mL$ of bee venom and $10{\mu}g/mL$ of melittin acupuncture solution and increased by $5{\mu}g/mL$ of melittin acupuncture solution. 6. Compared with control, sodium nitroprusside-induced intracellular calcium concentrations were decreased by 0.5, 1, $5{\mu}g/mL$ of bee venom and 5, $10{\mu}g/mL$ of melittin acupuncture solution.

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Convergence Studies of NO Homeostasis in Cellular Signalling (세포의 신호전달 과정에서 NO 항상성에 관한 융복합 연구)

  • Oh, Hee-Kyun;Do, Eun-Young;Park, Hae-Ryoung
    • Journal of Digital Convergence
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    • v.13 no.12
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    • pp.461-467
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    • 2015
  • Saussurea lappa is known for a variety of physiological activities as a component but has not known to show the effect of the cellular signaling pathway. We investigated the anti-inflammatory effects by Saussurea lappa ethanol extract on the LPS(lipopolysaccharide) induced nitric oxide (NO) production by RAW 264.7 cell line. It shows the expressions of iNOS and COX-2 at the transcriptional level (RT-PCR). The Saussurea lappa ethanol extract showed transcriptional expression levels of pro-inflammatory cytokine TNF- and IL-$1{\beta}$ induced by LPS(lipolysaccharide) in RAW264.7 cell line. Saussurea lappa ethanol extract reduced the LPS-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) at the RNA levels in a concentration-dependent manner. The finding that ethanol extract of Saussurea lappa has an influence on NO (nitric oxide) homeostasis in the study of the action mechanism on the macrophage-mediated inflammatory reaction was considered in terms of convergence. And it is to provide an important basis for the prevention and treatment of inflammatory diseases in the future.