• Journal of Digital Convergence
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• v.16 no.11
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• pp.613-620
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• 2018

This study was conducted to investigate of Mori Folium Water Extract (MF) on anti-inflammation activity. MF Water extracts after 24 houres cultivation were examined to ascertain the cell viability of mouse macrophage RAW 264.7 cells. The influence of the Water extracts in RAW 264.7 macrophage cells treated with LPS was investigated. nitric oxide (NO) production, nterleukin$(IL)-1{\alpha}$ IL-6 and IL-10 increased generation of cytokines. mouse macrophage RAW 264.7 cells cell viability changes were no decreas after MTT assay of MF Water extract. The MF water extracts inhibited NO generation caused by LPS in the macrophages over $25{\mu}g/mL$. The MF water extracts increased in the control group the $IL-1{\alpha}$ and IL-6 activation generated by LPS in the macrophages over $50{\mu}g/mL$. Accordingly, it was found that different MF water extract concentrations significantly influenced certain anti-inflammation activities in RAW 264.7 macrophage cells. The results of this study are expected to be highly applicable to health - friendly functional materials. Further studies are needed to confirm the signaling pathways associated with anti-inflammation of macrophages through continuous studies.

• Journal of Life Science
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• v.33 no.5
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• pp.397-405
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• 2023

Although glutathione (GSH) has been shown to play an important role in the prevention of oxidative damage as an antioxidant, studies on immune regulation by it have not been properly conducted. In this study, we investigated whether luthione^®, a reduced GSH, has an immune enhancing effect in murine macrophage RAW 264.7 cells. The results of flow cytometry and immunofluorescence experiments indicated that luthione increased phagocytic activity, a representative function of macrophages, compared to the control cells. According to the results of the cytokine array, the expression of interleukin (IL)-5, IL-1β, and IL-27 was significantly increased in the luthione-treated cells. Luthione also enhanced the production of tumor necrosis factor-α and IL-1β through increased expression of their proteins, and increased release of the immune mediators such as nitric oxide (NO) and prostaglandin E₂ was associated with increased expression of inducible NO synthase and cyclooxygenase-2. In addition, the expression of cluster of differentiation 86, an M1 macrophage marker, was dramatically enhanced in RAW 264.7 cells treated with luthione. Furthermore, as a result of heat map analysis, we found that cytokine signaling 1/3-mediated signal transducer and activator of transcription/Janus tyrosine kinase signaling pathway was involved in the immunomodulatory effect by luthione. In conclusion, our data suggested that luthione could act as a molecular regulator in M1 macrophage polarization and enhance immune capacity by promoting macrophage phagocytic function.

• The Korea Journal of Herbology
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• v.32 no.3
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• pp.97-103
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• 2017

Objective : According to recent studies, Anemarrhenae Rhizoma has anti-inflammatory activities of DW extract, but it hasn't not yet conducted to evaluate inflammatory factors about 80% ethanol extract. Therefore, The aim of this study is to investigate the various effects of individual or combined 80% ethanol extract of Anemarrhenae Rhizoma on cell viability and various anti-inflammatory factors. Methods : Anemarrhenae Rhizoma extract was prepared with 80% ethanol. MTT assay, ELISA, and Luminex were performed in LPS-activated RAW 264.7 cell line to measure cytotoxicity, Nitric oxide (NO), cyclooxygenase-2 (COX-2), prostaglandin E2 ($PGE_2$), Leukotriene B4 ($LTB_4$), and cytokines ($IL-1{\beta}$, IL-6, and $TNF-{\alpha}$), respectively. Results : At concentration of $200{\mu}g/m{\ell}$ Anemarrhenae Rhizoma extract, cytotoxicity was observed in RAW 264.7 cells. However, at concentration less than $100{\mu}g/m{\ell}$ of Anemarrhenae Rhizoma, cytotoxicity was not observed in RAW 264.7 cells. All concentration of Anemarrhenae Rhizoma extract showed no difference of NO, and $IL-1{\beta}$level in RAW 264.7 cells compared with control group. In contrast, at concentration of $100{\mu}g/m{\ell}$ Anemarrhenae Rhizoma extract significantly inhibited LPS-induced production of COX-2, PGE2, and $LTB_4$ level in RAW 264.7 cells. In addition, the production of proinflammatory cytokines (IL-6, $TNF-{\alpha}$) in LPS-induced RAW 264.7 cells was significantly decreased at concentration of all or 10, and $100{\mu}g/m{\ell}$, respectively. Conclusion : These findings demonstrate that Anemarrhenae Rhizoma has inhibitory effect on inflammatory mediators in LPS-activated RAW 264.7 cells showing possible developed as a raw material for new therapeutics to ease the symptoms related with inflammatory.

• Journal of Applied Biological Chemistry
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• v.52 no.4
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• pp.174-179
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• 2009

Biological functionality of nineteen commercially available carbonated vinegar liquors including wood vinegar liquor (WVL), bamboo vinegar liquor (BVL) and chaff vinegar liquor (CVL) were evaluated, focusing mainly on electron donating ability to DPPH radical, reducing power against ferricyanide ($Fe^{3+}$), blockading ability to linoleic acid autoxidation and NO production from LPS-stimulated RAW264.7 cells plus cytotoxicity to RAW264.7 cells. The results showed that crude carbonated vinegar liquors, regardless of their source materials, have high capacity of antioxidation such as electron donating ability, reducing power, blockading ability to lipid peroxidation and NO production, as well as cell cytotoxicity. Refined carbonated vinegar liquors for skin care or bath showed significantly low cell cytotoxicity, however, overall antioxidant potencies were also low. Especially, these carbonated vinegar liquors revealed low levels of inhibition for NO production deeply involved in inflammation. Among nineteen carbonated vinegar liquors examined, chaff vinegar liquor was observed to be the most potent carbonated vinegar liquor with high antioxidant activities together with low cytotoxicity to mammalian cells.

• Journal of the Korean Applied Science and Technology
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• v.33 no.4
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• pp.706-716
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• 2016

In this study we applied the NAG obtained by the deacetylation of chitin extracted from the shells of crabs and shrimp as cosmetic ingredient. In order to compare NAG with GLC we identified the influence of cytotoxicity, anti-inflammatory, melanin biosynthesis formation inhibition on skin cell, and we measured the effects of the change of melanin and red spots. The results show that there was not any attentive cytotoxicity on the Raw 264.7 cell and B16F10 cell, and NO formation anti-inflammatory hindrance effect induced from Raw 264.7 by LPS was slight, and NAG suppressed the increase of melanin generation concentration-dependently after we induced the melanin generation with ${\alpha}$-MSH on B16F10 and measured the melanin biosynthesis inhibition. From this result, we identified the applicability of the cosmetics containing NAG as functional cosmetic for enhancing skin-lightening because when cream containing NAG was applied to skin the index of melanin red spots showed statistically meaningful changes.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.1
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• pp.64-70
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• 2010

Soybean is of particular interest as a food supplement of isoflavones for inhibiting bone resorption in postmenopausal woman. These beneficial effects of isoflavones are caused by functioning as partial agonists or antagonists of estrogen, of which anti-resorptive effect is mediated indirectly through paracrine factors produced by osteoblasts that act on osteoclasts. In this study, the indirect effect of soybean on osteoclastic differentiation of RAW264.7 cells were investigated. The conditioned medium was collected from MC3T3-E1 osbeoblasts treated with 0.001 mg/mL~0.1 mg/mL soybean extracts for 6 days, mixed in 1:1 ratio with osteoclast medium, and then added into RAW264.7 cells with receptor activator of nuclear factor kappa B ligand (RANKL), a differentiation inducer for 3 days. Of paracrine factors in the conditioned medium, the protein expression of osteoprotegerin (OPG) with soybean extract was specifically higher in a dose dependent manner than with $10^{-9}$ M~$10^{-6}$ M of estrogen, genistein or daidzein standards. In RAW264.7 cells, the conditioned medium with soybean inhibited RANKL induced osteoclastic differentiation as total number of multinucleated tartrateresistant alkaline phosphatase (TRAP)-positive osteoclasts and protein expression of MMP-9 were significantly decreased. Coupled with the low expression of estrogen receptor $\alpha$ and $\beta$ proteins in RANKL treated RAW264.7 cells, we demonstrate that the conditioned medium of soybean treated osteoblasts inhibits RANKL induced differentiation of osteoclasts with the selective expression of OPG in osteoblasts.

• Korean Journal of Acupuncture
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• v.24 no.3
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• pp.139-148
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• 2007

목 적 : 약침으로서 씀바귀의 염증 효과를 알아보기 위해 RAW 264.7 대식세포에서 Tnf 생성을 확인하였다. 방 법: 씀바귀를 RAW 264.7 대식세포에 미리 처치한 후 LPS로 염증 반응을 유도하였다. ELISA, Western blotting, RT-PCR, 그리고 EMSA을 통해 Tnf의 생성과 발현에 대한 효과를 평가하였다. 결 과 : LPS 유도된 세포에서 씀바귀 1, 5, 10, 50 ${\mu}g/ml$의 농도는 각각 23.7, 37.8, 66.4, 86.1% Tnf 생성을 억제한 것을 ELISA 통해 확인되었다. LPS로 유도된 대식세포에서 Tnf 생성은 농도에 따라 감소하였고, Western blotting, RT-PCR 분석에서는 씀바귀 5과 50 ${\mu}g/ml$가 LPS로 유도된 대식세포에서 Tnf의 mRNA와 단백질 발현을 저해 하는 것으로 관찰되었다. 또한 EMSA에서도 씀바귀 5과 50 ${\mu}g/ml$가 LPS로 유도된 Nf-kB를 감소시키는 것을 확인하였다. 고 찰 : 이런 결과는 씀바귀가 $Nf-{\kappa}B$를 저해하면서 LPS로 유발된 Tnf 생성을 감소시킨다는 것을 보여주었고, 이는 더 나아가 염증 질환에서 씀바귀가 약침으로써의 치료적 효과를 나타낼 수 있을 것으로 사료된다. 고 찰 : 이런 결과는 씀바귀가 $Nf-_{k}B$를 저해하면서 LPS로 유발된 Tnf 생성을 감소시킨다는 것을 보여주었고, 이는 더 나아가 염증 질환에서 씀바귀가 약침으로써의 치료적 효과를 나타낼 수 있을 것으로 사료된다.

• Korean Journal of Food Science and Technology
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• v.46 no.4
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• pp.511-515
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• 2014

In a screen for plant-derived inhibitors of nitric oxide (NO) production in lipopolysaccharide (LPS)-activated RAW 264.7 macrophage cells, a flavonol isolated from the chloroform extract of Alpinia officinarum was isolated. The structure of the flavonol was found to be 3,5,7-trihydroxy-2-phenylchromen-4-one (galangin, GLG) by using spectroscopy. GLG exhibited an inhibitory effect ($IC_{50}$ value: $26.8{\mu}M$) on NO production in LPS-stimulated RAW 264.7 murine macrophage cells. Moreover, GLG suppressed expressions of inducible nitric oxide synthase (iNOS) protein and mRNA in a dose-dependent manner.

• Korean Journal of Food Science and Technology
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• v.39 no.2
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• pp.209-216
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• 2007

The present study investigated anti-oxidative and anti-inflammatory activity of glycoprotein isolated from Morus Indica Linne (MIL glycoprotein). We found that MIL glycoprotein has a molecular weight of 32 kD and consists of carbohydrate (40.03%) and protein (59.97%), and that it has a strong scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical $({\cdot}OH)$, and superoxide anion $(O_2{\cdot}\;^-)$ radicals. In addition, MIL glycoprotein had a stable character and an optimal DPPH radical scavenging activity in the alkaline and neutral pH solution, and up to at 105. However, the results indicated that it has a minimal scavenging activity in the metal ionic solution ($Ca^{2+}$, $Mn^{2+}$, and $Mg^{2+}$) in the presence of EDTA. In addition, we further investigated whether MIL glycoprotein scavenges oxygen radicals and blocks inflammation-related signals in the bisphenol A (BPA)-stimulated Raw 264.7 cells. The results in this study showed that it has a character to scavenge the productions of reactive oxygen species (ROS) and nitric oxide (NO) dose-dependently. Also it blocked the activities of inflammation-related signals such as nuclear factor-kappa B ($NF-{\kappa}B$) and inducible nitric oxide synthase (iNOS). For example, it had an inhibitory effect on the activation of $NF-{\kappa}B$ (p50) and iNOS proteins at 200 ${\mu}g/mL$ MIL glycoprotein. Here, we speculate that MIL glycoprotein is one of natural antioxidants and of modulators of the BPA-induced inflammation.

• 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.