• Journal of Applied Biological Chemistry
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• v.59 no.1
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• pp.37-43
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• 2016

Bioactive components of ultra-fine ground Saururus, the extraction yield increases when the leaves are ultra-fine ground. Comparison of normal-ground and ultra-fine ground Saururus chinensis leaves showed that the solid content and antiinflammatory activity of ultra-fine ground extracts was higher than that of normal-ground extracts. Lipopolysaccharide (LPS)-stimulated Raw 264.7 cells were treated with different concentrations of Saururus chinensis extract and the amount of nitric oxide (NO) was determined; LPS-treated cells produced 2 times more NO than cells that were not treated with LPS. Moreover, the NO production in cells treated with Saururus chinensis extract was inhibited in a concentration-dependent manner. Because the stimulant-induced NO production is regulated by the inducible nitric oxide synthase (iNOS), we measured the iNOS protein level to elucidate the mechanism by which the NO production was inhibited. We found that the amount of iNOS decreased dose-dependently. It was reduced by 53% at a Saururus chinensis extract concentration of $100{\mu}g/mL$. The protein expression of cyclooxygenase-2 (COX-2) in LPS-treated Raw 264.7 cells was inhibited by 31% at $100{\mu}g/mL$ of Saururus chinensis extract. Gel shift of the nuclear factor kappa B-DNA complex occurred in LPS-treated cells and the intensity of the band decreased gradually in a concentration-dependent manner. Ultra-fine ground Saururus chinensis extract had a concentration-dependent inhibitory effect on the production of prostaglandin $E_2$, tumor necrosis factor ${\alpha}$, interleukin $1{\beta}$ (IL-$1{\beta}$), IL-6, and IL-8 in LPS-treated Raw 264.7 cells, i.e., at $50{\mu}g/mL$ of Saururus chinensis extract, their levels were decreased by 53, 67, 52, 37, and 21% respectively.

• Herbal Formula Science
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• v.18 no.1
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• pp.169-179
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• 2010

The pistil of nelumbo nucifera (PNN) is used in the treatment of nocturnal pollution, hematemesis, epistaxis, metrorrhagia and diarrhoea in traditional medicine. The present study was examined to evaluate the effects of PNN on the production of pro-inflammatory mediators in vitro. After the treatment of PNN, cell viability was measured by MTT assay, nitric oxide (NO) production was monitored by measuring the nitrite content in culture medium. The protein bands were determined by immunoblot analysis and levels of cytokines were analyzed by sandwich immunoassays. In the MTT assay, the doses of PNN extract (0.03, 0.10 mg/ml) had no significant cytotoxicity. The increases of NO production and inducible nitric oxide synthase expression were detected in lipopolysaccharide(LPS)-activated Raw 264.7 cells compared with control, in contrast, these increases were significantly attenuated by pre-treatment with PNN. In cytokine assay, the massive pro-inflammatory cytokines such as tumour necrosis factor-$\alpha$, interleukin (IL)-$1{\beta}$ and IL-6 were induced in LPS-activated Raw 264.7 cells, but pre-treatment of Raw 264.7 cells with PNN caused inhibition (TNF-$\alpha$=14.17%, IL-$1{\beta}$=107.43%, IL-6=46.27%) the production of cytokines by LPS. In addition, PNN reduced prostaglandin E2 productions in a dose-dependent manner (0.03mg/ml=37.52%, 0.10 mg/ml=83.77%) as a consequence of the inhibition of cyclooxygenase-2 expression. Taken together, our data indicates that PNN can regulate the inflammatory response in macrophage cells activated by Gram-negative infection.

• The Korean Journal of Physiology and Pharmacology
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• v.21 no.3
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• pp.345-352
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• 2017

Since previous studies have reported that hydroquinone (HQ) exerted immunosuppressive and anti-inflammatory activity, various HQ derivatives have been synthesized and their biological activities investigated. In this study, we explored the anti-inflammatory activity of JS-III-49, a novel HQ derivative, in macrophage-mediated inflammatory responses. JS-III-49 suppressed the production of the inflammatory mediators nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) and down-regulated the mRNA expression of the inflammatory enzymes cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) as well as the expression of the pro-inflammatory cytokines interleukin-6 (IL-6) and IL-$1{\beta}$ without cytotoxicity in LPS-stimulated RAW264.7 cells. JS-III-49 inhibited nuclear translocation of the $NF-{\kappa}B$ transcription factors p65 and p50 by directly targeting Akt, an upstream kinase of the $NF-{\kappa}B$ pathway, in LPS-stimulated RAW264.7 cells. However, JS-III-49 did not directly inhibit the kinase activities of Src and Syk, which are upstream kinases of Akt, in LPS-stimulated RAW264.7 cells. Moreover, JS-III-49 suppressed the nuclear translocation of c-Fos, one of the components of AP-1, by specifically targeting p38, an upstream mitogen-activated protein kinase (MAPK) in the AP-1 pathway in LPS-stimulated RAW264.7 cells. These results suggest that JS-III-49 plays an anti-inflammatory role in LPS-stimulated macrophages by targeting Akt and p38 in the $NF-{\kappa}B$ and AP-1 pathways, respectively.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.46 no.4
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• pp.384-392
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• 2013

An ethanolic extract of Undaria pinnatifida was fractionated using several solvents. Of the fractions, the ethyl acetate fraction had the greatest inhibitory effect on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 macrophage cells. Using this fraction (U. pinnatifida ethyl acetate extract, UPE), we investigated the molecular mechanism underlying its inhibitory effect on LPS-stimulated RAW 264.7 cells. Pretreatment of the cells with up to $100{\mu}g/mL$ UPE significantly inhibited NO production and inducible nitric oxide synthase (iNOS) expression, in a dose-dependent manner. Similarly, UPE treatment markedly reduced the production of pro-inflammatory cytokines, such as interleukin (IL)-1, IL-6 and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), while it strongly suppressed the nuclear translocation of nuclear factor-kappa B (NF-${\kappa}B$) by preventing proteolytic degradation of inhibitor of nuclear factor ${\kappa}B$ $(I{\kappa}B)-{\alpha}$. Moreover, UPE treatment significantly reduced the phosphorylation of phosphatidylinositol 3-kinase (PI3K)/Akt and mitogen-activated protein kinase (MAPK) in LPS-stimulated cells. These results indicate that UPE contains anti-inflammatory compounds and suggest that it might be used as a functional food material that assists in prevention of inflammatory diseases.

• Journal of Periodontal and Implant Science
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• v.48 no.2
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• pp.70-83
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• 2018

Purpose: The aim of this study was to evaluate the capacity of single and combined applications of the bark of the stems and roots of Magnolia officinalis Rehd. et Wils. (Magnoliae Cortex) and Zea mays L. (maize) to modulate inflammation in RAW 264.7 cells stimulated with Porphyromonas gingivalis. Methods: RAW 264.7 cells were stimulated with P. gingivalis, and Magnoliae Cortex and/or maize was added. Cytotoxicity and the capacity to modulate inflammation were determined with a methylthiazol tetrazolium (MTT) assay, nitrite production, enzyme-linked immunosorbent assay (ELISA), and western blotting. Results: Treatment with Magnoliae Cortex and/or maize inhibited nuclear transcription factor ${\kappa}B$ ($NF-{\kappa}B$) pathway activation and nuclear p44/42 mitogen-activated protein kinase (MAPK) and inducible nitric oxide synthase (iNOS) protein expression in P. gingivalis-stimulated RAW 264.7 cells. Moreover, the treatments suppressed cytokines (prostaglandin $E_2$ [$PGE_2$], interleukin $[IL]-1{\beta}$, and IL-6) and nitrite production. Conclusions: Both Magnoliae Cortex and maize exerted an anti-inflammatory effect on P. gingivalis-stimulated RAW 264.7 cells, and this effect was more pronounced when the extracts were combined. These findings show that these extracts may be beneficial for slowing the progression of periodontal disease.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.1
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• pp.14-19
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• 2011

In this study, we extracted a whole polysaccharide fraction from the sea hare, Aplysia kurodai, and screened its functional properties using cell lines. The functionalities of polysaccharide and glycosaminoglycan (GAG) were investigated with RAW 264.7 cell lines. The crude polysaccharides and GAG purified DEAE-Sepharose chromatography did not show the toxicity on RAW 264.7 cell line in the range of $10\sim200{\mu}g$/mL, whereas they increased the cell growth rate. The crude polysaccharides and purified GAG also increased the production of NO, interleukin-6 and tumor necrosis factor-$\alpha$ on RAW 264.7 cell. Particularly, the purified GAG inhibited the proliferation of stomach cancer cell line, AGS, up to 40% for 72 hr incubation, but not the intestinal epithelial IEC-6 cell lines.

• Biomedical Science Letters
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• v.23 no.3
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• pp.272-276
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• 2017

HOX genes are transcription factors that play important roles in body patterning and cell fate specification during normal development. In previous study, we found aberrant overexpression of HOXB5 in breast cancer tissues and cell lines, and demonstrated that HOXB5 is important in regulation of cell proliferation, tamoxifen resistance, and invasiveness through the epithelial-mesenchymal transition (EMT). Although the relationship between HOXB5 and phenotypic changes in MCF7 breast cancer cells has been studied, the molecular function of HOXB5 as a transcription factor remains unclear. IL-6 has been reported to be involved in not only inflammation but also cancer progression, which is characterized by the increase of growth speed and invasiveness of tumor cells. In this study, we selected Interleukin-6 (IL-6) as HOXB5 putative downstream target gene and discovered that HOXB5 transcriptionally up-regulated the expression of IL-6 in HOXB5 overexpressing MCF7 cells. The upstream region (~1.2 kb) of IL-6 promoter turned out to contain several putative HOX consensus binding sites. Chromatin immunoprecipitation assay confirmed that HOXB5 directly binds to the promoter region of IL-6 and positively regulated the expression of IL-6. These data all together, indicate that HOXB5 promotes IL-6 transcription by actively binding to the putative binding sites located in the upstream region of IL-6, which enable to increase its promoter activity in MCF7 breast cancer cells.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.2
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• pp.368-373
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• 2009

The purpose of this study was to investigate the anti-inflammatory effects of water extract from Euonymus alatus (Thunb.) Sieb. (EAS) on the RAW 264.7 cells. To evaluate of anti-inflammatory of EAS, we examined the cytokine productions on lipopolysaccharide (LPS)-induced RAW 264.7 cells and also inhibitory mechanisms using Western blot. EAS reduced LPS-induced production of nitric oxide (NO), interleukin (IL)-1b, IL-6, IL-10 and tumor necrosis factor-a (TNF-a) in RAW 264.7 cells. EAS inhibited the activation of mitogen-activated protein kinases (MAPKs) such as p38, extracelluar signal-regulated kinase (ERK 1/2) and c-Jun NH2-terminal kinase (JNK) but not of inhibitory kappa B a (Ik-Ba) degradation in the LPS-stimulated RAW 264.7 cells. In conclusion, EAS down-regulated LPS-induced NO and cytokines production, which could provide a clinical basis.

• Korean Journal of Plant Resources
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• v.31 no.5
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• pp.478-488
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• 2018

Mushrooms have been widely cultivated and consumed as foods and herbal medicines owing to their various biological properties. However, few studies have evaluated the anti-inflammatory effects of mushrooms. Here, we investigated the effects of mushroom extracts (MEs) on lipopolysaccharide (LPS)-induced inflammation in macrophages (RAW264.7 cells). First, we extracted MEs with either water or ethanol. Using LPS-treated RAW264.7 cells, we measured cell proliferation and NO production. Gene expression of tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), interleukin (IL)-6 (IL-6), and $IL-1{\beta}$ was assessed by RT-PCR, and protein abundance of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) and phosphorylation of p65 were determined by immunoblotting. MEs prepared using both water and ethanol inhibited LPS-induced inflammation in RAW264.7 cells. Nitric oxide (NO) levels induced by LPS were reduced by treatment with MEs. Isaria japonica Yasuda water extracts and Umbilicaria esculenta (Miyoshi) Minks ethanol extracts significantly decreased the mRNA expression of inflammation-related cytokine genes including $TNF-{\alpha}$, IL-6, and $IL-1{\beta}$. Similarly, the protein abundance of iNOS and COX-2 was also decreased. The phosphorylation of p65, a subunit of nuclear $factor-{\kappa}B$ was at least partly suppressed by MEs. This study suggests that mushrooms could be included in the diet to prevent and treat macrophage-related chronic immune diseases.

• Korean Journal of Pharmacognosy
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• v.47 no.2
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• pp.150-157
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• 2016

Earth worm (Eisenia andrei) and clam worm (Perinereis linea) have been used as anti-bacterial and anti-inflammatory agent. However, it is unclear how they exerted their physiological effects in macrophages. In this experiment, the anti-inflammatory and antioxidant effects of clam worm extract (CWE) and earth worm extract (EWE) in RAW264.7 cells were examined by measuring MDA, catalase, SOD, GSH-Px and inflammatory cytokines (nitric oxide, iNOS, interleukin-$1{\beta}$ and tumor necrosis factor-${\alpha}$). Treatment with CWE significantly increased the activities of catalase, SOD and GSH-Px in RAW264.7 cells and decreased the level of MDA. Interestingly, treatment with CWE induced more activities of SOD than EWE. In addition, CWE decreased NO production, iNOS, COX-2, TNF-${\alpha}$ and IL-$1{\beta}$ in RAW264.7. The EWE also decreased NO production and iNOS, but increased COX-2 and IL-$1{\beta}$ suggesting that CWE could be better resources for anti-inflammatory and antioxidant agent than EWE. Taken together, these results indicate that CWE has the potential as a natural antioxidant and a therapeutic for inflammation-related diseases.