• Journal of Applied Biological Chemistry
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• v.66
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• pp.447-454
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• 2023

In the environment in which humans live, there are various antigens that invade the human body and interfere with humans leading a healthy life, so the immune system recognizes the antigen then removes them through a complex mechanism. Macrophages are widely distributed immune cells involved in the innate immune system, and produce various immune modulators such as inducible nitric oxide synthase-induced nitric oxide, cyclooxygenase-2 induced prostaglandin E2 and proinflammatory cytokines such as tumor necrosis factor-alpha. On the other hand, Protaetia brevitarsis seulensis larvae are a type of edible insect that have emerged as an alternative to the future food supply problem. The immuno-modulatory effect through the activation of murine macrophage RAW264.7 cell via mitogen-activated protein kinases (MAPKs)/nuclear factor-kappa B (NF-κB) signaling pathways has been reported. Based on this report, in this study, we confirmed how the expression of immune modulators induced by Protaetia brevitarsis seulensis larvae extracts in RAW264.7 cells was changed by treatment with pharmacological inhibitors of toll-like receptor 4 (TLR4), MAPKs and NF-κB signaling pathways. As a result, reduction of immune modulators was confirmed in the c-Jun N-terminal kinase (JNK) inhibitor treatment group and NF-κB inhibitor treatment group among the Protaetia brevitarsis seulensis larvae-treated RAW264.7 cell. Furthermore, in the TLR4 inhibitor-treated group, decreases in phosphorylation of JNK and NF-κB factors were confirmed in Protaetia brevitarsis seulensis larvae-treated RAW264.7 cell, as well as decreases in immune modulators. This results suggest that Protaetia brevitarsis seulensis larvae activates RAW264.7 cells by the engagement of TLR4-JNK/NF-κB signaling pathway.

• Journal of the Korean Applied Science and Technology
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• v.35 no.3
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• pp.797-806
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• 2018

This study attempted to investigate the effects of Paeonia Lactiflora Pallas (P. lactiflora) on the inhibition of oxygen free radical, anti-inflammation and MMP-1 inhibitory activity and examine its possibility as a functional cosmetic material. For test methods, the inhibition of oxygen free radical after measuring reactive oxygen species (ROS) in the cell, cytotoxicity assessment and anti-inflammation were measured, and MMP-1 inhibitory effects in the HDF cell were measured. According to the test, the inhibition of ROS was confirmed in RAW 264.7 and HDF cells. In terms of cytotoxicity assessment, 90% or higher cell viability was detected at $5/10{\mu}g/mL$ Paeonia Lactiflora Pallas extract while it was 80% or higher at other concentration levels in both RAW 264.7 and HDF cells. In addition, NO production was inhibited in the RAW 264.7 cell while MMP-1 was significantly inhibited in the HDF cell. The above results reveal a possibility of Paeonia Lactiflora Pallas extract as a functional cosmetic material after confirming the inhibition of ROS synthesis in the cell, antioxidant and anti-inflammatory effects by inhibiting NO synthesis, low toxicity on skin cells and anti-aging effect through MMP-1 inhibition.

• The Korean Journal of Mycology
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• v.45 no.4
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• pp.336-344
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• 2017

The anti-inflammatory effects of cell-free extracts from wild yeasts, Meyerozyma guilliermondii YJ34-2 and Rhodotorula graminis YJ36-1, caused by the inhibition of nitric oxide (NO) activity and cytotoxic effects were determined. Cell-free extracts from these two yeast strains had dose-dependent inhibitory effects on the production of lipopolysaccharide-induced NO and there were no cytotoxic effects on the treated cells or negative effects on their proliferation. Their cell-free extracts were also shown to have inhibitory effects on pro-inflammatory cytokines, such as tumor necrosis factor $(TNF)-{\alpha}$ and $prostaglandin-E_2$, in a dose-dependent manner. Maximal inhibitory activity on NO production occurred in cell-free extracts of Meyerozyma guilliermondii YJ34-2 cultivated at $30^{\circ}C$ for 24 hr and Rhodotorula graminis YJ36-1 cultivated at $25^{\circ}C$ for 24 hr in the yeast extract-peptone-dextrose (YPD) media.

• The Korea Journal of Herbology
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• v.35 no.4
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• pp.37-43
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• 2020

Objectives : Allium hookeri is a well-known traditional herbal remedy and its root used for treatment of inflammation and tumor. However, the mechanism of anti-inflammatory effect of Allium hookeri is still unknown. This study aims to examine the mechanism of anti-inflammatory effect of Allium hookeri on mouse macrophage cell line, RAW 264.7 cells. Methods : Anti-oxidant effect of water extract of Allium hookeri (WEAH) was measured by 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay. 3- (4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was performed to determine the effect of WEAH on cell viability in RAW 264.7 cells. In addition, anti-inflammatory effect of WEAH was investigated in RAW 264.7 cells. Inflammation of RAW 264.7 cells induced by lipopolysarccharide (LPS) treatment and expression levels of inflammatory cytokine interleukin 1 β (IL-1β) and interleukin 6 (IL-6) gene were analyzed using quantitative reverse transcription PCR (qRT-PCR) analysis. Furthermore, the phosphorylation of inhibitor of nuclear factor kappa B (IκBα) after LPS treatment with WEAH-treated RAW 264.7 cells was confirmed by immunoblot analysis. Results : WEAH showed a strong anti-oxidant effect and no cytotoxicity to RAW 264.7 cells up to 2 mg/㎖ concentration. The LPS-induced mRNA expression levels of IL-1β and IL-6 were decreased by WEAH treatment. Furthermore, the LPS-induced phosphorylation of IκBα is attenuated by WEAH treatment. Conclusions : Through experimental demonstration of anti-oxidant and anti-inflammatory effects of WEAH, we suggest that Allium hookeri is a valuable material for prevention and treatment of various inflammatory diseases.

• The Journal of Korean Obstetrics and Gynecology
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• v.18 no.3
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• pp.92-109
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• 2005

Purpose : The Purpose of this research was to investigate the effects of Cheongyeolhawlhyeoltanggagyehyeoldeung (CYHHT) 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 CYHHT on mLFC and RAW264.7 cells was not observed at 100, 50, 10, and $1{\mu}g/ml$ of CYHHT treatments. $IL-1{\beta}$, IL-6 and NOS-IImRNA expression of RAW264.7 cells was inhibited by CYHHT treatments in a dose-dependent manner. CYHHT treatment of RAW264.7 cells inhibited $TNF-{\alpha}$ and COX-2 mRNA expression. CYHHT treatment of RAW264.7 cells significantly inhibited IL-6 and NO production. CYHHT treatment of RAW264.7 cells inhibited ROS production. CYHHT 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. CYHHT treatment of colitis-induced mice significantly increased hematological values such as WBC and RBC counts, Hgb and HCT levels, but decreased PLT values. CYHHT treatment of colitis-induced mice decreased IL-6 and $TNF-{\alpha}$ production significantly CYHHT treatment of colitis-induced mice significantly increased CD3+(T) cell counts. In contrast, CYHHT treatment decreased CD19+ B cell counts and CD3+/CD69+ significantly, and also decreased B/T ratio (%) though not significant. Conclusion : These results indicated that CYHHT could be used for treating diverse female diseases caused by the inflammation.

• Journal of Physiology & Pathology in Korean Medicine
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• v.36 no.2
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• pp.66-72
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• 2022

Flos Lonicerae Japonicae (the flower buds of Lonicera japonica Thunberg) has been used as an antibacterial and antiviral drug in Korean Medicine. The aim of this study is to evaluate the effect of Flos Lonicerae Japonicae water extract (FL) on the production of cytokines in RAW 264.7 mouse macrophages stimulated by lipopolysaccharide (LPS). After 24 h treatment, the production of various cytokines from RAW 264.7 was measured with multiplex cytokine assay using Bio-Plex 200 suspension array system. FL at concentrations of 50, 100, and 200 ㎍/mL significantly inhibited productions of tumor necrosis factor-α, macrophage inflammatory protein (MIP)-1β, and MIP-2 in LPS-stimulated RAW 264.7 cells; FL at concentrations of 100 and 200 ㎍/mL significantly inhibited productions of leukemia inhibitory factor, LIX (CXCL5), and RANTES in LPS-stimulated RAW 264.7 cells; FL at concentrations of 200 ㎍/mL significantly inhibited productions of granulocyte-macrophage colony-stimulating factor and macrophage colony-stimulating factor in LPS-stimulated RAW 264.7 cells; FL at concentrations of 50 and 100 ㎍/mL significantly increased productions of interleukin (IL)-10 in LPS-stimulated RAW 264.7 cells; FL at concentrations of 50, 100, and 200 ㎍/mL significantly increased productions of IL-6 and interferon gamma-induced protein-10 in LPS-stimulated RAW 264.7 cells; FL at concentrations of 100 and 200 ㎍/mL significantly increased productions of monocyte chemoattractant protein-1 in LPS-stimulated RAW 264.7 cells. Taken together, these data mean that FL might modulate productions of cytokines, chemokines, and growth factor in LPS-stimulated macrophages. Further study needs to verify the exact mechanism for modulatory activities of FL with macrophages.

• The Korea Journal of Herbology
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• v.37 no.6
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• pp.19-28
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• 2022

Objectives : To develop natural ingredients that help prevent or treat anti-inflammatory-related diseases and use themas basic data, we investigated anti-inflammatory activity of Cynanchi Atrati Radix Et Rhizoma water extracts(CWE) in lipopolysaccharide(LPS)-induced murine macrophage cell line, RAW 264.7 cells. Methods : The cell viabilities were evaluated with RAW 264.7 cells. The production of nitric oxide(NO), prostaglandin E2(PGE2), pro-inflammatory cytokines such tumor necrotic factor(TNF)-α and interleukin(IL)-6 were assessed in LPS-induced RAW 264.7 cell treated with CWE. Furthermore, the protein expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2(COX-2), and mitogen-activated protein kinase(MAPK) were assessed by western blotting. Results : In RAW 264.7 cell, the cell viability by CWE treatment was more than 98.4% at a concentration of 100-400 ㎍/mL. At a concentration of 800 ug/ml of CWE, the cell viability was as low as 86%. At doses of 100, 200 and 400 ㎍/mL, CWE inhibited the production of NO, PGE2, TNF-𝛼 and IL-6 in a dose-dependent manner and also decreased the expression of iNOS and COX-2 from LPS-induced RAW 264.7 cells. In addition, CWE significantly inhibited the MAPK pathway including decreased the phosphorylation of the p38, c-Jun N-terminal kinase(JNK) and extracellular signal-regulated kinase(ERK1/2). Conclusions : Our study provides evidence that CWE inhibits the production of main pro-inflammatory molecules in LPS-induced RAW 264.7 cells via expression of p38, JNK, and ERK1/2 MAPK signaling pathways. Therefore, CWE is expected to be widely used as a natural ingredient for anti-inflammatory functional foods or pharmaceuticals in the future.

• Journal of Life Science
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• v.29 no.8
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• pp.904-915
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• 2019

Prostate cancer (PCa) is one of the most metastatic tumor. Although hormone therapy or surgical castration is mostly conducted to treat PCa, it has a lot of side effects. Recently, many researchers have been exploring the tumor microenvironment to remedy these circumstances. Immune cells, especially macrophages, are an important composition of the tumor microenvironment. Under normal conditions, macrophages exhibit mild tumoricidal activity against tumors. However, once activated by interferon gamma or lipopolysaccharides, macrophages can kill cancer cells directly or indirectly by secreting cytokines and chemokines. In this study, murine macrophage RAW 264.7 cells were treated with Phellinus linteus extract. To analyze their pro-inflammatory phenotype, we were used several assays such as a real-time polymerase chain reaction, an enzyme-linked immunosorbent and nitric oxide assay. Prostate cancer cells were treated with the RAW 264.7-conditioned media, which was identified as a pro-inflammatory nature, for 48 h, and the expression of epithelial-mesenchymal transition (EMT)-related genes was determined. Not only N-cadherin, Snail, Twist, Slug, and Cadherin 11, which are mechenchymal-related proteins, were decrease, but epithelial marker of E-cadherin was increased. In addition, the mRNA level of vimentin, ccl2, and vegfa were decreased, as the EMT is closely related to the migration and invasion of cancer cells. In conclusion, the RAW 264.7-conditioned media stimulated with P. linteus extract inhibited migration and invasion and regulated the EMT pathway in human prostate cancer cells.

• Journal of Life Science
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• v.33 no.6
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• pp.463-470
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• 2023

Desmarestia tabacoides Okamura is a brown macroalgae that is found worldwide. Although several genera of Desmarestia have been reported as having anti-tumorigenic, anti-melanogenic, and photoprotective properties, the anti-inflammatory activity of D. tabacoides Okamura has not yet been evaluated. In this study, we analyzed the anti-inflammatory mechanisms of D. tabacoides Okamura ethanol extract (DTEE) via the inhibition of nitric oxide (NO) and prostaglandin (PG) E₂ production and the expression of their corresponding enzymes, inducible NO synthase (iNOS), and cyclooxygenase (COX)-2. In addition, their upstream signaling molecules were evaluated by Western blot analysis, such as nuclear factor (NF)-κB, mitogen-activated protein kinase (MAPK), and phosphoinositide-3-kinase (PI3K)/Akt, in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. The DTEE treatment significantly inhibited LPS-induced NO and PGE₂ production as well as the expression of their corresponding enzymes, iNOS, and COX-2 without cytotoxicity. The stimulated transcription factor NF-κB and upstream signaling molecules extracellular signal-regulated kinase (ERK), c-Jun NH₂-terminal kinase (JNK), and p38 were attenuated by the DTEE treatment, which was statistically significant, while Akt did not provide any inhibitory effect. Moreover, the DTEE treatment significantly mitigated the LPS-activated adaptor molecules, toll-like receptor 4 (TLR4), and myeloid differentiation primary response 88 (MyD88) in the RAW 264.7 cells. These results suggest that DTEE attenuates TLR4-mediated inflammatory responses by inhibiting NF-κB activation and suppressing MAPK phosphorylation in LPS-stimulated RAW 264.7 cells.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.4
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• pp.815-820
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• 2008

Macrophage is the important cell for the immune system. Many of herbal drugs were searched about their immune-modulating activity. The purpose of this study is to investigate the biological effect of water extract from ARTEMISIAE ARGI FOLIUM (WAAF) on mouse macrophage Raw 264.7 cells. ARTEMISIAE ARGI FOLIUM was known to have the antibacterial, immune-enhancing, and anticoagulative properties. Cytotoxicity of WAAF was verified by MTT assay. The intracellular production of hydro peroxide ($H_2O_2$) by WAAF was examined. The productions of nitric oxide (NO) and $TNF-{\alpha}$ from Raw 264.7 cell by WAAF were also examined. WAAF showed no cytotoxicity on RAW 264.7 cells for 3 hours. WAAF increased the production of $H_2O_2$ in Raw 264.7 cells. WAAF decrease the production of NO from the cells at low concentrations but increased at high concentrations. WAAF increased the production of $TNF-{\alpha}$ from the cells. Therefore, It could be suggested that WAAF has the immune-modulating effect.