• Molecules and Cells
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• v.39 no.10
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• pp.734-741
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• 2016

Granulocyte-macrophage colony stimulating factor (GM-CSF) has a role in inducing emergency hematopoiesis upon exposure to inflammatory stimuli. Although GM-CSF generated murine bone marrow derived cells have been widely used as macrophages or dendritic cells in research, the exact characteristics of each cell population have not yet been defined. Here we discriminated GM-CSF grown bone marrow derived macrophages (GM-BMMs) from dendritic cells (GM-BMDCs) in several criteria. After C57BL/6J mice bone marrow cell culture for 7 days with GM-CSF supplementation, two main populations were observed in the attached cells based on MHCII and F4/80 marker expressions. GM-BMMs had $MHCII^{low}F4/80^{high}$ as well as $CD11c^+CD11b^{high}CD80^-CD64^+MerTK^+$ phenotypes. In contrast, GM-BMDCs had $MHCII^{high}F4/80^{low}$ and $CD11c^{high}CD8{\alpha}^-CD11b^+CD80^+CD64^-MerTK^{low}$ phenotypes. Interestingly, the GM-BMM population increased but GM-BMDCs decreased in a GM-CSF dose-dependent manner. Functionally, GM-BMMs showed extremely high phagocytic abilities and produced higher IL-10 upon LPS stimulation. GM-BMDCs, however, could not phagocytose as well, but were efficient at producing $TNF{\alpha}$, $IL-1{\beta}$, IL-12p70 and IL-6 as well as inducing T cell proliferation. Finally, whole transcriptome analysis revealed that GM-BMMs and GM-BMDCs are overlap with in vivo resident macrophages and dendritic cells, respectively. Taken together, our study shows the heterogeneicity of GM-CSF derived cell populations, and specifically characterizes GM-CSF derived macrophages compared to dendritic cells.

• Korean Journal of Food Science and Technology
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• v.48 no.3
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• pp.289-295
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• 2016

To elucidate the new biologically active ingredient in commercial instant coffee, a crude polysaccharide (ICP-0) was isolated by ethanol precipitation, and its immunostimulatory activity was estimated. ICP-0 mainly consisted of galactose (55.5%), mannose (25.7%), arabinose (6.0%), and galacturonic acid (10.1%), suggesting the possibility of its existence as a mixture of galactomannan or pectic polysaccharide. ICP-0 showed proliferative activity in peritoneal macrophages and splenocytes. ICP-0 dose-dependently augmented the production of nitric oxide and reactive oxygen species by peritoneal macrophages. In addition, murine peritoneal macrophages stimulated by ICP-0 showed enhanced production of various cytokines (tumor necrosis factor-${\alpha}$, interleukin-6, and interleukin-12) as compared to unstimulated murine peritoneal macrophages. In an in vitro assay for assessing intestinal immunomodulation, the ICP-0-treated Peyer's patch cells showed higher bone marrow cell proliferation activity at $100{\mu}g/mL$ and higher production of granulocyte-macrophage colony-stimulating factor, compared to the untreated Peyer's patch cells. These results suggest that polysaccharides in commercial instant coffee have a potentiality for macrophage functions and the intestinal immune system.

• Proceedings of the Ginseng society Conference
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• 2006.05a
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• pp.57-58
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• 2006

Red ginseng is a classical traditional Chinese medicine. Among Chinese herbs, red ginseng has been considered as one of the tonics. Many studies indicated that red ginseng could enhance immune function of the human body. Red ginseng total saponin, ginsenoside, the most important active constituents identified in red ginseng can protect against myocardial ischaemia damage and protect endothelium against electrolysis-induced free radical injury. Macrophages play a significant role in host defense mechanisms. When activated, they inhibit the growth of a wide variety of tumor cells. The aim of this study was to determine the effects of pure ginsenoside Rb1 on immunostimulatory activity such as murine macrophage phagocytosis and proliferation of splenocytes. Furthermore, we investigated the effects of ginsenoside Rb1 on the production of nitric oxide (NO), reactive oxygen species (ROS) and proinflammatory cytokines (IL-1beta, IL-6, and TNF-alpha) in murine macrophage, RAW 264.7 cells. ROS have emerged as important signaling molecules in the regulation of various cellular processes. Ginsenoside Rb1 significantly increased production of ROS in dose dependent manner. As NO plays an important role in immune function, ginsenoside Rb1 treatment could modulate several aspects of host defense mechanisms due to stimulation. Treatment with ginsenoside Rb1 to macrophages induced the production of NO and proinflammatory cytokines and expression levels of these genes in a dose-dependent manner. Furthermore, incubation of RAW 264.7 cells with ginsenoside Rb1 showed a dose dependent increased phagocytosis activity and lymphocyte proliferation of splenocytes. Therefore, these results suggest that ginsenoside Rb1 has promising potential as a natural medicine for stimulation of the immune system.

• YAKHAK HOEJI
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• v.42 no.3
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• pp.296-301
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• 1998

To investigate the effects of ginsenosides from Panax ginseng on mitogenic responses in macrophages and splenocytes from murine, we examined the effects of representative protopanaxadiol and protopanaxatriol ginsenosides ($Rb_1,\;Rb_2,\;Re\;and\;Rg_1$) on tumor necrosis factor-${\alpha}$ (TNF-(${\alpha}$) production in murine macrophage cell line (RAW264.7 cells) stimulated by lipopolysaccharide (LPS) and T cell proliferation in splenocytes stimulated by concanavalin A (Con A). Among the ginsenosides tested, protopanaxadiol ginsenosides ($Rb_1\;and\;Rb_2$) significantly inhibited TNF-${\alpha}$ production in a dose-dependent manner. However, protoppanaxatriol ginsenosides (Re and $Rg_1$) showed little inhibitory activity. The molar concentrations of $Rb_1\;and\;Rb_2$ producing 50% inhibition ($IC_{50}$) of TNF-${\alpha}$ production were $55.8{\mu}g/ml\;(48.0{\mu}M)\;and\;31.8{\mu}g/ml (27.9{\mu}M)$, respectively. As a positive control, prednisolone also exhibited inhibitory activity with an $IC_{50}$ value of $21.7{\mu}M$. In T cell proliferation, $Rg_1$, was not effective but $Rb_1$ and Re or $Rb_2$ significantly increased or inhibited at high concentration, 75 and $100{\mu}g/ml$. In contrast, prednisolone showed potent inhibitory activity with an $IC_{50}$ value of 6.1nM. These results suggest that ginsenosides may take part in the mitogen-induced signaling pathway for TNF-${\alpha}$ production and T cell proliferation from macrophages and splenocytes.

• Korean Journal of Pharmacognosy
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• v.30 no.4
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• pp.413-416
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• 1999

During the screening for inhibitors on nitric oxide (NO) production in LPS-activated murine macrophage, RAW 264.7 cells, two coumarins were isolated from chloroform extract of Ponciri Fructus. They were identified as imperatorin (10), phellopterin (11) on the basis of spectroscopic methods. The $IC_{50}$ values for NO formatiom were about $5.4\;{\um}M$ and $35.0\;{\um}M$, respectively, and then eleven coumarins were tested for the inhibitory effects on NO production in activated macrophages. All the test coumarins inhibited NO production in concentration-dependent manner and furanocoumarins (6-11) showed much more potent inhibitory effect than simple coumarins. Among the compounds examined xanthotoxin (8) was the most potent inhibitors of NO production $(IC_{50}=1.4\;{\mu}M)$. Analysis of the structure-activity relationship among these coumarins led to the conclusion that the substitution of C-5 position in furanocoumarins reduces greatly their inhibitory potency although the substitution of C-8 does not almost affect it.

• Biomolecules & Therapeutics
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• v.28 no.3
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• pp.250-258
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• 2020

Emphysema, a major component of chronic obstructive pulmonary disease (COPD), is a leading cause of human death worldwide. The progressive deterioration of lung function that occurs in the disease is caused by chronic inflammation of the airway and destruction of the lung parenchyma. Despite the main impact of inflammation on the pathogenesis of emphysema, current therapeutic regimens mainly offer symptomatic relief and preservation of lung function with little therapeutic impact. In the present study, we aimed to discover novel therapeutics that suppress the pathogenesis of emphysema. Here, we show that LJ-2698, a novel and highly selective antagonist of the adenosine A₃ receptor, a G protein-coupled receptor involved in various inflammatory diseases, significantly reversed the elastase-induced destructive changes in murine lungs. We found that LJ-2698 significantly prevented elastase-induced airspace enlargement, resulting in restoration of pulmonary function without causing any obvious changes in body weight in mice. LJ-2698 was found to inhibit matrix metalloproteinase activity and pulmonary cell apoptosis in the murine lung. LJ-2698 treatment induced increases in anti-inflammatory cytokines in macrophages at doses that displayed no significant cytotoxicity in normal cell lines derived from various organs. Treatment with LJ-2698 significantly increased the number of anti-inflammatory M2 macrophages in the lungs. These results implicate the adenosine A₃ receptor in the pathogenesis of emphysema. Our findings also demonstrate the potential of LJ-2698 as a novel therapeutic/preventive agent in suppressing disease development with limited toxicity.

• Toxicological Research
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• v.22 no.3
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• pp.293-299
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• 2006

We demonstrate that magnolol, a hydroxylated biphenyl compound isolated from Magnolia officinalis, inhibits LPS-induced expression of iNOS gene in RAW 264.7 cells(murine macrophage cell line). Treatment of RAW 264.7 cells with magnolol inhibited LPS-stimulated nitric oxide production in a dose-related manner. RT-PCR analysis showed that the decrease of NO was due to the inhibition of iNOS gene expression. Western immunoblot analysis of phosphorylate p38 kinase showed magnolol significantly inhibited the phosphorylation of p38 kinase which is important in the regulation of iNOS gene expression. The specific p38 inhibiter SB203580 abrogated the LPS-induced NO generation and iNOS expression, whereas the selective MEK-1 inhibitor PD98059 did not affect the NO induction. Immunostaining of p65 and reporter gene assay showed that magnolol inhibited NF-${\kappa}/Rel$ nuclear translocation and transcriptional activation, respectively. Collectively, this series of experiments indicates that magnolol inhibits iNOS gene expression by blocking NF-k/Rel and p38 kinase signaling. Due to the critical role that NO release plays in mediating inflammatory responses, the inhibitory effects of magnolol or iNOS suggest that magnolol may represent a useful anti-inflammatory agent.

• Korean Journal of Food Science and Technology
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• v.48 no.4
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• pp.392-397
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• 2016

Zaluzanin C is a sesquiterpene lactone isolated from Ainsliaea acerifolia, a Korean medicinal plant. In the present study, the anti-inflammatory effects of zaluzanin C were demonstrated in lipopolysaccharide (LPS)-stimulated murine macrophages (RAW264.7 cells). Zaluzanin C inhibited the release of nitric oxide (NO) by alleviating the protein expression of inducible NO synthase in LPS-treated RAW264.7 cells. Furthermore, it suppressed the release of interleukin-6 induced by LPS. Zaluzanin C was also found to block the translocation of the p65 subunit of nuclear factor-kB from the cytosol to the nucleus, which is one of the underlying mechanisms of the anti-inflammatory action of zaluzanin C. These data suggest the potential of zaluzanin C in the treatment of inflammatory diseases.

• Journal of Microbiology and Biotechnology
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• v.26 no.3
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• pp.469-476
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• 2016

This study was undertaken to evaluate the immunomodulatory effect of dead nano-sized Lactobacillus plantarum (nLp) in RAW 264.7 cells and murine primary splenocytes. nLp is a dead, shrunken, processed form of L. plantarum nF1 isolated from kimchi (a traditional Korean fermented cabbage) and is less than 1 μm in size. It was found that nLp treatment stimulated nitric oxide (NO) production more in RAW 264.7 macrophages than pure live L. plantarum (pLp), and that the stimulatory properties were probably largely derived from its cell wall. In addition, nLp induced murine splenocyte proliferation more so than pLp; in particular, a high dose of nLp (1.0 × 10¹¹ CFU/ml) stimulated proliferation as much as lipopolysaccharide at 2 μg/ml. Moreover, according to our cytokine profile results in splenocytes, nLp treatment promoted Th1 (TNF-α, IL-12 p70) responses rather than Th2 (IL-4, IL-5) responses and also increased Th17 (IL-6, IL-17A) responses. Thus, nLp stimulated NO release in RAW 264.7 cells and induced splenocyte proliferation more so than pLp and stimulated Th1 and Th17 cytokine production. These findings suggested that dead nLp has potential use as a functional food ingredient to improve the immune response, and especially as a means of inducing Th1/Th17 immune responses.

• The Korean Journal of Physiology and Pharmacology
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• v.17 no.1
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• pp.73-79
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• 2013

The fruit of the black raspberry (Rubus coreanus Miquel) has been employed in traditional medicine, and recent studies have demonstrated its measureable biological activities. However, the root of the black raspberry has not been studied. Therefore, in this study, we evaluated the anti-inflammatory and antibacterial properties of the root and unripe fruit polyphenols of the black raspberry. Both polyphenols proved to have anti-inflammatory activity as evidenced by the decreased nitric oxide (NO), cytokines (IL-$1{\beta}$, IL-6, and IL-10) and prostaglandin E2 ($PGE_2$) levels in lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophages. However, root polyphenols showed stronger anti-inflammatory activity than fruit polyphenols. LPS-induced mRNA and protein expressions of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 levels were also decreased, confirming the anti-inflammatory activity. Root polyphenols showed lethal activity against methicillin-resistant Staphy-lococcus aureus (MRSA), carbapenem-resistant Acinetobacter baumannii (CRAB), and Bacillus anthracis. In contrast, the black raspberry fruit did not demonstrate these properties. These data provide the first demonstration that black raspberry root has potential anti-inflammatory and anti-superbacterial properties that can be exploited as alternatives for use in the food and cosmetic industries and/or as pharmaceuticals.