• Journal of Life Science
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• v.30 no.4
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• pp.359-369
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• 2020

Although many studies on immune modulatory materials have used RAW 264.7 cells, few have used T cell-derived TK-1 cell lines. Moreover, although some studies have investigated the efficacy of plant-derived β-sitosterol, few have examined the immunomodulatory activity of its analogue, daucosterol. In this study, β-sitosterol and daucosterol were isolated from D. batatas and identified by nuclear magnetic resonance spectroscopy. To evaluate the immune-enhancing or inhibitory effects of the isolated phytosterols, the expression levels of the inflammatory response genes COX-2, TNF-α, IL-6, and iNOS were analyzed by RT-PCR. The relative expression levels of TNF-α and iNOS in RAW 264.7 cells were increased more than threefold with β-sitosterol treatment comparing to those of untreated control. In the case of TK-1 cells, the expression level of TNF-α was decreased and the expression level of iNOS was increased in a β-sitosterol concentration-dependent manner. The expression levels of COX-2, TNF-α, and IL-6 increased by approximately 0.7-1.2 times in RAW 264.7 cells treated with daucosterol compared to those of untreated control, but iNOS expression decreased by 0.8-0.18 times. In the case of daucosterol-treated TK-1 cells, the expression levels of TNF-α, IL-6, and iNOS were markedly reduced from those of TK-1 cells treated only with lipopolysaccaride. As a conclusion, β-sitosterol treatment increased TNF-α and iNOS expression levels in RAW 264.7 cells, thus exerting an immune- boosting effect. However, in TK-1 cells, iNOS expression increased while TNF-α expression decreased, indicating an immunosuppressive activity of β-sitosterol. Daucosterol appears to exert an immunosuppressive effect in both macrophages and T cell lines by inhibiting iNOS expression in RAW 264.7 cells and greatly inhibiting the expression of TNF-α, IL-6, and iNOS in TK-1 cells.

• Korean Journal of Acupuncture
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• v.25 no.3
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• pp.137-146
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• 2008

Objectives : The leaves of Artemisia argyi L. have been used for the treatment of bleeding-related diseases in traditional korean medicine. But the immunological activities with macrophage have not been sufficiently reported. This study is to investigate the immunological bioactivities of the herbal acupuncture solution obtained from leaves of Artemisia argyi L. (AAAS). Methods & Results : Against Nicotine and Acetaldehyde, AAAS increased significantly the production of hydrogen peroxide (H2O2) within mouse macrophage Raw 264.7 cells above the concentration of 10 ${\mu}g/m{\ell}$. AAAS increased significantly the production of nitric oxide (NO) in Raw 264. 7 cells above the concentration of 100 ${\mu}g/m{\ell}$ against EtOH. And AAAS increased significantly the production of nitric oxide (NO) in Raw 264. 7 cells above the concentration of 200 ${\mu}g/m{\ell}$ against Nicotine, Acetaminophen, and Acetaldehyde. Conclusions : These results suggest that AAAS could be thought to have the immunological activities related with the production of hydrogen peroxide and NO in macrophage.

• Korean Journal of Plant Resources
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• v.34 no.6
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• pp.536-541
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• 2021

In this study, we investigated whether Hibiscus manihot flowers (HMF) exhibits immunostimulatory activity in RAW264.7 cells. HMF increased the production of immunostimulatory factors such as NO, iNOS, IL-1β and TNF-α in RAW264.7 cells. TLR2 and TLR4 blocked HMF-mediated production of immunostimulatory factors in RAW264.7 cells. In addition, the inhibition of MAPK signaling pathway reduced HMF-mediated production of immunostimulatory factors. From these results, HMF is thought to promote the production of immunostimulatory factors through activating TLR2/4/MAPK signaling in macrophages. It is believed that HMF can be developed as an agent related to immune enhancement in the future.

• Microbiology and Biotechnology Letters
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• v.45 no.1
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• pp.19-26
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• 2017

Effects of resveratrol glucosylation on the immunomodulation properties of resveratrol and on the viability of macrophage cells have been studied by using murine macrophage RAW 264.7 cells. Nitric oxide (NO) and interleukin 6 (IL-6) expression in macrophages in vitro were studied after treatment with different concentrations of (E)-resveratrol, (E)-resveratrol 3-O-${\beta}$-${\small{D}}$-glucoside (R-3-G), or (E)-resveratrol 4'-O-${\beta}$-${\small{D}}$-glucoside (R-4'-G). In vitro viability of RAW 264.7 cells after treatment with the aforementioned three compounds was also studied. As demonstrated by macrophage cell viability assays, two different resveratrol monoglucosides, R-3-G and R-4'-G, exhibited 50-80% reduced cytotoxicity in comparison to (E)-resveratrol in A549 and HepG2 cells. Compared to the resveratrol aglycon, both glucosylated resveratrol derivatives positively modulated NO and IL-6 production in macrophages positively via transcriptionally up-regulating IL-6 and iNOS expression. Conjugation of a glucose moiety on resveratrol was found to enhance the immunomodulating activity of resveratrol and the viability of RAW 264.7 cells.

• 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.

• Journal of Life Science
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• v.32 no.7
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• pp.491-500
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• 2022

HK shiitake mushroom mycelium (HKSMM), containing 14% β-glucan, is a health functional food ingredient individually approved by the Korea Ministry of Food and Drug Safety for liver health. The anti-inflammatory effect of a 50% aqueous ethanol extract of HKSMM (designated HKSMM50) was studied in RAW 264.7 macrophage cells treated with lipopolysaccharide (LPS). An active hexose correlated compound (AHCC) was used as a positive control. LPS-activated RAW 264.7 cells were treated with HKSMM50 and AHCC (0, 20, 100, 500 ㎍/ml) and cultured for 24 hr. Inflammation-related elements in the supernatant were measured using enzyme-linked immunosorbent assay (ELISA) kits, and the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins in the cells was analyzed by Western blotting. The HKSMM50 lowered iNOS and COX-2 protein expressions, and nuclear factor-kappa B (NF-κB), nitric oxide (NO) and prostaglandin E₂ (PGE₂) contents in a concentration-dependent manner as compared to LPS treatment. Similarly, the HKSMM50 lowered the content of pro-inflammatory cytokines interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4) and interleukin-6 (IL-6) contents and increased the activity of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT). The efficacy of the AHCC treatment was similar to that of the HKSSM50 treatments. These results indicate that HKSMM50 showed an anti-inflammatory effect in LPS-treated RAW 264.7 cells by down-regulation of NF-κB signaling and suggest that HKSMM could be used as a health functional food ingredient to help improve immune function.

• International Journal of Oral Biology
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• v.34 no.1
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• pp.37-42
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• 2009

The attachment and adhesion of RAW 264.7 and MC3T3-E1 cells to titanium (Ti) discs with various degrees of roughness was investigated. The attachment, adhesion, and proliferation of these cells were evaluated after 4 hr, 24 hr and 7 day incubations. Both RAW 264.7 and MC3T3-E1 cells showed a time-dependant correlation between attachment and adhesion on the surface of the titanium discs. Both types of cells tended to have higher survival rate on these discs as the surface roughness increased. The percentage of adherent inflammatory RAW 264.7 cells was greater than MC3T3-E1 cells at 24 hr, but this was reversed at 7 days in culture. The morphology of osteoblastic MC3T3-E1 cells at 24 hr, determined using a surface emission microscope (SEM), appeared flattened and spread out while inflammatory RAW 264.7 cells were predominantly spherical in shape. The adhesion of both cell types on the titanium discs was dependant on the levels of fibronectin adsorbed on the disc surface, indicating that serum constituents modulate the efficient adhesion of these cells. Our data indicate that the cellular response to the titanium surface is dependent on the types of cells, surface roughness and serum constituents.

• Toxicological Research
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• v.13 no.3
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• pp.247-250
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• 1997

We have studied cytotoxicity of S-nitroso-N-acetyl- N-DL-penicillamine(SNAP), a Nitric oxide (NO)-releasing compound, in RAW264.7 macrophages. SNAP is cytotoxic to RAW264.7 cells in a concentration-dependent manner. PMA(200 nM) stimulated cells to produce superoxide anton radical($O_2^{-\cdot}$) and caused a little loss of RAW264.7 cell viability for 12 hr and diminished the cytotoxicity of SNAP. The mechanism by which PMA can protect cells against NO-mediated cytotoxicity was studied by peroxynitrite-enhanced chemiluminescence method. Observed results suggested that $O_2^{-\cdot}$ produced by PMAstimulated RAW264.7 cells may quench NO released by SNAP and reduce NO, thus attenuating NO-related damages.

• The Korea Journal of Herbology
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• v.23 no.3
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• pp.163-173
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• 2008

Objectives : Imyosan (IMS), a drug preparation comprised of Phellodendri Cortex (PC) and Atractylodis Rhizoma (AR), is commonly used as a traditional herbal medicine in Korea and China for the treatment of various inflammatory diseases. However, little is known about the effect of IMS and its component herbs on inflammatory mediators in RAW 264.7 cells. Therefore, in this study, methanol extracts of IMS and its component herbs were examined to determine if they inhibited inflammatory effects in RAW 264.7 cells. Methods : Cytotoxic activity of IJHT and its components on RAW 264.7 cells was using 5-(3-carboxymethoxyphenyl)-2H-tetrazolium inner salt (MTS) assay. The nitric oxide (NO) production was measured by Griess reagent system. And proinflammatory cytokines were measured by ELISA kit. The levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression were detected by western blot. Results : Methanol extract of IMS and its component herbs were significantly reduced iNOS and COX-2 expression as well as NO, PGE2, $IL-1{\beta}$ and IL-6 production in RAW 264.7 cells. Conclusions : The results of this study indicated that the anti-inflammatory effect of Imyosan extract is more potent than that of extracts of its component herbs in macrophages.

• 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.