• The Journal of the Society of Korean Medicine Diagnostics
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• v.17 no.3
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• pp.263-273
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• 2013

Objectives This study was designed to investigate the anti-inflammatory effect of extracts from Ligustrum obtusifolium S. fruits(LOF) in RAW 264.7 Macrophages stimulated with lipopolysaccharide(LPS). Methods We examined productions of nitric oxide(NO), reactive oxygen species(ROS), inducible isoforms of NO synthase(iNOS), cyclooxygenase-2(COX-2) to investigate the anti-inflammatory effect of LOF extracts. In addition, we measured generation of pro-inflammatory cytokines(TNF-${\alpha}$, IL-6). Results Cell viability showed that LOF extracts had no cytotoxicity in Raw 264.7 cells. The treatment with LOF extracts significantly decreased the generation of NO and pro-inflammatory cytokines(TNF-${\alpha}$, IL-6) in LPS-stimulated macrophage cells. Furthermore LOF extracts inhibited intracellular ROS generation dose dependently and reduced the expression of iNOS, COX-2 proteins. Conclusions These results showed that the LOF extracts had an anti-inflammatory effect on LPS-stimulated Raw 264.7 cells. These findings provide scientific support for the use of this Ligustrum obtusifolium S. for inflammatory-related diseases.

• The Korea Journal of Herbology
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• v.28 no.6
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• pp.129-133
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• 2013

Objectives : The purpose of this study was to investigate the effects of Angelicae acutilobae Radix Water Extract (AA) on the production of cytokines in RAW 264.7 cell stimulated with lipopolysaccharide (LPS). Method : RAW 264.7 cells were cotreated with AA (50 and $100{\mu}g/mL$) and lipopolysaccharide (LPS; $1{\mu}g/mL$) for 24 hours. After 24 hours treatment, using bead-based multiplex cytokine assay, concentrations of various cytokines such as interleukin(IL)-6, tumor necrosis factor-alpha(TNF-${\alpha}$) granulocyte colony-stimulating factor(G-CSF), granulocyte macrophage colony-stimulating factor(GM-CSF), and macrophage inflammatory protein(MIP)-$1{\alpha}$ were measured. Result : AA significantly inhibited LPS-induced production of IL-6 and MIP-$1{\alpha}$ from LPS-stimulated RAW 264.7 cells at the concentration of $50{\mu}g/mL$. AA significantly inhibited LPS-induced production of TNF-${\alpha}$ from LPS-stimulated RAW 264.7 cells at the concentration of $100{\mu}g/mL$. AA significantly inhibited LPS-induced production of G-CSF and GM-CSF in RAW 264.7 cells at the concentrations of 50 and $100{\mu}g/mL$. Conclusion : These results suggest that AA has anti-inflammatory effect related with its inhibition of proinflammatory cytokines such as IL-6, TNF-${\alpha}$, G-CSF, GM-CSF, and MIP-$1{\alpha}$ in LPS-induced macrophages.

• 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.

• The Korea Journal of Herbology
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• v.26 no.2
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• pp.75-81
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• 2011

Objectives : This study aims at examining the anti-inflammatory effects of Scutellariae Radix extract. Methods : Scutellariae Radix was hot water extracted to make the samples(SR) for the experiment. Their effects were examined on the increase of cell viability in mouse macrophage Raw 264.7 cells, the creation of nitric oxide(NO) in lipopolysaccharide(LPS)-induced Raw 264.7 cells, and the creation of cytokines of interleukin(IL)-$1{\beta}$ and others. Results : The results of the experiment are as follows. 1. The MTT assay was carried out to check the cellular toxicity of the water extract of Scutellariae Radix. The results were found no significant toxicity caused to macrophages by the water extract of Scutellariae Radix. 2. The water extract of Scutellariae Radix significantly restricted the increase of NO in the LPS-induced macrophages after 24-hour culture. 3. The water extract of Scutellariae Radix significantly restricted the creation of IL-6, IL-10, IL-12p40, IL-17, interferon-inducible protein(IP)-10, keratinocyte-derived chemokine(KC), and vascular endothelial growth factor(VEGF) in the LPS-induced macrophages at the concentration of $25{\mu}g/mL$ or higher. Conclusion : The samples(SR) of hot water extract of Scutellariae Radix caused no significant cellular toxicity to macrophages and significantly restricted the creation of NO, IL-6, IL-10, IL-12p40, IL-17, IP-10, KC, and VEGF in the LPS-induced macrophages at $25{\mu}g/mL$ or higher, thus demonstrating significant anti-inflammatory effects.

• Nutrition Research and Practice
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• v.9 no.3
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• pp.219-226
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• 2015

BACKGROUND/OBJECTIVES: In this study, potential anti-inflammatory effect of enzymatic hydrolysates from Styela clava flesh tissue was assessed via nitric oxide (NO) production in lipopolysaccahride (LPS) induced RAW 264.7 macrophages and in vivo zebrafish model. MATERIALS/METHODS: We investigated the ability of enzymatic hydrolysates from Styela clava flesh tissue to inhibit LPS-induced expression of pro-inflammatory mediators in RAW 264.7 macrophages, and the molecular mechanism through which this inhibition occurred. In addition, we evaluated anti-inflammatory effect of enzymatic hydrolysates against a LPS-exposed in in vivo zebrafish model. RESULTS: Among the enzymatic hydrolysates, Protamex-proteolytic hydrolysate exhibited the highest NO inhibitory effect and was fractionated into three ranges of molecular weight by using ultrafiltration (UF) membranes (MWCO 5 kDa and 10 kDa). The above 10 kDa fraction down-regulated LPS-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), thereby reducing production of NO and prostaglandin $E_2$ ($PGE_2$) in LPS-activated RAW 264.7 macrophages. The above 10 kDa fraction suppressed LPS-induced production of pro-inflammatory cytokines, including interleukin $(IL)-1{\beta}$, IL-6, and tumor necrosis factor $(TNF)-{\alpha}$. In addition, the above 10 kDa fraction inhibited LPS-induced phosphorylation of extracellular signal-regulated kinases (ERKs), c-Jun N-terminal kinase (JNK), and p38. Furthermore, NO production in live zebrafish induced by LPS was reduced by addition of the above 10 kDa fraction from S. clava enzymatic hydrolysate. CONCLUSION: The results of this study suggested that hydrolysates derived from S. clava flesh tissue would be new anti-inflammation materials in functional resources.

• Journal of Veterinary Clinics
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• v.31 no.6
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• pp.469-476
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• 2014

The aims of this study were to explore the effects of conjugated linoleic acid (CLA) on reactive oxygen species (ROS) production in lipopolysaccharide (LPS)-naïve and LPS-stimulated RAW 264.7 macrophages and to examine whether these effects affect the regulation of tumor necrosis factor-alpha (TNF-${\alpha}$) production, and nuclear factor-kappa B (NF-${\kappa}B$) and peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) activation. Trans-10, cis-12(t10c12)-CLA increased the production of ROS, as well as TNF-${\alpha}$ in LPS-naïve RAW 264.7 cells. The CLA-induced TNF-${\alpha}$ production was suppressed by treatment of diphenyleneiodonium chloride (DPI), a NADPH oxidase inhibitor. In addition, CLA enhanced the activities of NF-${\kappa}B$ and $PPAR{\gamma}$ in LPS-naïve RAW 264.7 cells, and this effect was abolished with DPI treatment. LPS treatment increased ROS production, whereas CLA reduced LPS-induced ROS production. LPS increased both TNF-${\alpha}$ production and NF-${\kappa}B$ activity, whereas t10c12-CLA reduced TNF-${\alpha}$ production and NF-${\kappa}B$ activity in LPS-stimulated RAW 264.7 cells. DPI treatment suppressed LPS-induced ROS production and NF-${\kappa}B$ activity. Moreover, DPI enhanced the inhibitory effects of t10c12-CLA on TNF-${\alpha}$ production and NF-${\kappa}B$ activation in LPS-stimulated RAW 264.7 cells. However, neither t10c12-CLA nor DPI affected $PPAR{\gamma}$ activity in LPS-stimulated RAW 264.7 cells. Taken together, these data indicate that t10c12-CLA induces TNF-${\alpha}$ production by increasing ROS production in LPS-naïve RAW 264.7 cells, which is mediated by the enhancement of NF-${\kappa}B$ activity via $PPAR{\gamma}$ activation. By contrast, t10c12-CLA suppresses TNF-${\alpha}$ production by inhibiting ROS production and NF-${\kappa}B$ activation via a $PPAR{\gamma}$-independent pathway in LPS-stimulated RAW 264.7 cells. These results suggest that t10c12-CLA can modulate TNF-${\alpha}$ production and NF-${\kappa}B$ activation through formation of ROS in RAW 264.7 macrophages.

• Applied Chemistry for Engineering
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• v.30 no.4
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• pp.479-486
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• 2019

This study investigated the anti-inflammatory effects of mixed extracts of Achyranthes japonica Nakai (Aj) and Aralia continentalis Kitagawa (Ac) (ratios of 1 : 2, 1 : 3, 1 : 5, 2 : 1, 3 : 1 and 5 : 1) on RAW264.7 macrophages. Cell toxicity was determined using a cell counting kit (CCK) assay. We evaluated anti-inflammatory effects of the mixed extracts of Aj and Ac by measuring interleukin $(IL)-1{\beta}$, IL-6, and tumor necrosis factor $(TNF){\alpha}$ using an enzyme-linked immunosorbent assay (ELISA) kit assay. The mixed extracts of Aj and Ac inhibited lipopolysaccharide (LPS)-induced $IL-1{\beta}$ and $TNF{\alpha}$ in LPS-stimulated macrophages. Comparing different ratios of the mixed extracts, the 2 : 1 ratio of Aj and Ac has much more potency and inhibited the production of $TNF{\alpha}$ in LPS-induced RAW264.7 cells. The results of the present study showed that the mixed extracts of Aj and Ac have potential anti-inflammatory effects on RAW264.7 macrophages. Therefore, these extracts may be used as a good source of functional foods for the protection against inflammatory diseases.

• Journal of Digital Convergence
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• v.16 no.7
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• pp.309-316
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• 2018

The purpose of this study was to investigate the effects of Violae Herba Water Extract (VH) on the proinflammatory factors of lipopolysaccharide (LPS)-induced on the production of inflammatory mediators in RAW 264.7 mouse macrophages cells. We examined effect of Violae Herba Water Extract on the cell viability of RAW 264.7 mouse macrophages cells. Futhermore, After 24 hours treatment we investigated anti-inflammatory effect of Violae Herba Water Extract by the production of Bio-Plex cytokine assay, concentrations of various cytokines such NO, $interleukin(IL)-1{\beta}$, tumor necrosis factor ${\alpha}(TNF-{\alpha})$ and IL-6. The water extract of Violae Herba significantly inhibited the production of NO, $IL-1{\beta}$, $TNF-{\alpha}$ and IL-6 at the concentration of 25, 50, 100 and $200{\mu}g/mL$ in the LPS-induced RAW 264.7 mouse macrophages cells with no changes in the cell viability of them. These results suggest that water extract of Violae Herba has anti-inflammatory effect related with its inhibition of proinflammatory cytokines such as $IL-1{\beta}$, $TNF-{\alpha}$ and IL-6 in the LPS-induced RAW 264.7 mouse macrophages cells. Further research is needed to develop therapeutic agents for inflammatory diseases using Violae Herba.

• 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.

• Korean Journal of Pharmacognosy
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• v.47 no.4
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• pp.295-300
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• 2016

Flavonoids are widely reported to be beneficial to human health. Among flavonoids, in general, flavonoid aglycons have better biological activities than flavonoid glycosides, in that aglycons can easily penetrate through cell membrane because of their low polarity. Therefore, kaempferol, quercetin and various their glycosides were evaluated for their abilities to inhibit NO and $PGE_2$ productions in LPS-induced RAW 264.7 cells. Of these flavonoids and flavonoid glycosides, kaempferol-7-O-${\beta}$-D-glucoside(kp-7-glu) which possesses a glycoside at C-7 position of the A ring in kaempferol, potently inhibited NO, $PGE_2$ and $TNF-{\alpha}$, $IL-1{\beta}$, IL-6 productions in LPS-induced RAW 264.7 macrophages.