• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.23 no.1
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• pp.63-71
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• 2020

Purpose: Autoimmune hepatitis (AIH) is a chronic disease that may lead to cirrhosis. The immunopathogenesis of AIH is not fully understood and it mainly involves T-cell mediated mechanism. Macrophage migration inhibitory factor (MIF) is a pro-inflammatory cytokine that promotes T cell response and its polymorphism may serve as a severity marker of AIH. No previous study has considered investigating MIF polymorphism in children with AIH. Methods: Forty-two children with definite diagnosis of AIH were enrolled along with 100 age and sex matched controls. All participants were tested for polymorphism at -173GC (rs755622) of MIF gene. All patients received the standard protocol of steroid plus azathioprine to achieve remission. Liver biopsy was performed at time of diagnosis for all patients and only 18 of them underwent a second biopsy after treatment. Results: No statistically significant differences in the frequency of the genotypes GG and GC or in allele distribution were found in both patient and control groups (p=0.590, 0.640 respectively). Initial alanine aminotransferase (ALT) levels at the time of presentation was significantly higher in the GC group than GG group (p=0.020). GC genotype significantly correlated with disease relapse (r=0.41, p=0.007). Regression of necroinflammation and the fibrosis score in the second liver biopsy was statistically significant in the GG group (p<0.0001, p=0.010 respectively). Conclusion: MIF -173GC polymorphism is associated with clinically significant markers of pediatric AIH, including increased initial serum ALT levels, may help predict necroinflammatory/fibrosis regression effectively, following immunosuppressive treatment.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.1
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• pp.171-175
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• 2008

In the recently, increased concern has been focused on the pharmacology and clinical utility of herbal extracts and derivatives as a drug or adjunct to chemotherapy and immunotherapy. Here we investigated the effect of the water extract of Loranthi Ramulus (LR) in production of inflammatory mediators and expression of toll-like receptor (TLR)-4, CD14 from peritoneal macrophage. We assayed the effect of LR water extract in cell proliferation in vitro and Th1/Th2 cytokine level in vivo. In peritoneal macrophages, water extract of LR water extract increased the production of Nitric oxide (NO) and $TNF-{\alpha}$. Also, LR water extract increased Con A-induced cell proliferation and IgG1, IgG2a level in serum. However, i.p. injection of water extract of LR water extract did not affect the level of $TNF-{\alpha}$, $IFN-{\gamma}$, IL-2, IL-4 and IL-5 in serum of mice. These studies indicate that LR water extract induces macrophage activation and suggest the possible use of LR water extract in macrophage-based immunotherapies.

• The Journal of Korean Medicine
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• v.19 no.2
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• pp.485-501
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• 1998

Cortex Mori (Moros alba L.), the root bark of mulberry tree has been used as an autiphlogistic, diuretic and expectorant in herval medicine. Recently, a few papers reported that phenolic extract of Cortex Mori had the hypotensive, hypoglycemic, antiviral and anticancer effects, and hot water extract of Cortex Mori(CM) had inhibitory effect on the degranulation and histamine release from activated mast cells. These previous studies suggest a possibility that CM has an antidotal activity against inflammation which was mediated mainly by macrophage-secreting inflammatory factors. This study was performed to evaluate the influences of CM on carrageenan-induced edema in vivo and release of inflammatory mediators such as NO, TNF and IL-1 by macrophages stimulated with LPS or $IFN-{\gamma}$ in vitro. Subcutaneous injections of carrageenan into the mouse paw rapidly induced local edema by increasing vascular permeability, but single intraperitoneal injection of CM extract at 30 minutes before carrageenan suppressed the development of edema. NO and TNF production from macrophage stimulated by LPS or $IFN-{\gamma}$ were significantly suppressed, especially TNF secretion by up to 3-4 folds. LPS stimulated IL-1 production was also inhibited, but not significantly. Cell viability assay verified that the inhibition was not due to general cell toxicity. These results suggest that reduction of NO, TNF and IL-1 production may be one of the means by which CM prevent inflammation associated diseases.

• Journal of Microbiology and Biotechnology
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• v.27 no.2
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• pp.350-356
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• 2017

We previously reported that the CopA3 peptide (LLCIALRKK, ${\small{D}}-form$) originally isolated from the Korean dung beetle has antimicrobial and immunosuppressive effects. However, the high cost of producing the synthetic peptide, especially the ${\small{D}}-form$, has limited the development of CopA3 for therapeutic purposes. Here, we investigated whether the CopA3 deletion derivative, CopA5, which is composed of only five amino acids (LLCIA) and has the ${\small{L}}-form$ structure, could inhibit the lipopolysaccharide (LPS)-induced activation of macrophages. Peritoneal exudate macrophages (PEM) were isolated from mice and exposed to LPS in the presence or absence of CopA5, and biomarkers of macrophage activation were measured. Our results revealed that LPS-induced nitric oxide (NO) production, tumor necrosis factor $(TNF)-{\alpha}$ secretion, and phagocytic activity of PEM were significantly inhibited by CopA5 treatment. Similar to CopA3, the structurally modified CopA5 peptide had no cell toxicity (as assessed by measurement of cell viability loss and apoptosis) in PEM. Moreover, the LPS-induced upregulation of the activating phosphorylation of signal transducer and activator of transcription 1 (STAT1) was markedly inhibited by CopA5 treatment. These results suggest that, similar to CopA3, CopA5 inhibits macrophage activation by inhibiting STAT1 phosphorylation and blocking the release of NO and $TNF-{\alpha}$. CopA5 may therefore prove therapeutically useful in the realm of immune suppression.

• Journal of Oral Medicine and Pain
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• v.32 no.2
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• pp.129-135
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• 2007

Ecdysteroids are known as insect molting hormone. At the same time, ecdysteroids and plant ecdysteroids (phytoecdysteorids) reveal beneficial effects on mammal. The present study was undertaken to determine the possible cellular mechanism of action of phytoecdysteroids in bone metabolism. The effects on the osteoblasts were determined by measuring cell proliferation, alkaline phosphatase (ALP) activity, and gelatinase activity. The effects on the osteoclasts were investigated by measuring tartrate-resistant acid phosphatase (TRAP)(+) multinucleated cells (MNCs) formation after culturing osteoclast precursors. Phytoecdysteroid treatment showed a increase in ALP activity of osteoblasts. Phytoecdysteroid increased the activity of gelatinase. In addition, phytoecdysteroid decreased the osteoclast generation induced by macrophage-colony stimulating factor (M-CSF) and receptor activator of NF-kB ligand (RANKL) in (M-CSF)-dependent bone marrow macrophage (MDBM) cell cultures. Taken these results, phytoecdysteroid may be a regulatory protein within the bone marrow microenvironment.

• BMB Reports
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• v.40 no.1
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• pp.88-94
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• 2007

Matrix metalloproteinase-9 (MMP-9) plays a pivotal role in the turnover of extracellular matrix (ECM) and in the migration of normal and tumor cells in response to normal physiologic and numerous pathologic conditions. Here, we show that the transcription of the MMP-9 gene is induced by lipopolysaccharide (LPS) stimulation in cells of a macrophage lineage (RAW 264.7 cells). We provide evidence that the NF-$\kappa$B binding site of the MMP-9 gene contributes to its expression in the LPS-signaling pathway, since mutation of NF-$\kappa$B binding site of MMP-9 promoter leads to a dramatic reduction in MMP-9 promoter activation. In addition, the degradation of l$\kappa$B$\alpha$;, and the presences of myeloid differentiation protein (MyD88) and tumor necrosis factor receptor-associated kinase 6 (TRAF6) were found to be required for LPS-activated MMP-9 expression. Chromatin immunoprecipitation (ChIP) assays showed that functional interaction between NF-$\kappa$B and the MMP-9 promoter element is necessary for LPS-activated MMP-9 induction in RAW 264.7 cells. In conclusion, our observations demonstrate that NF-$\kappa$B contributes to LPS-induced MMP-9 gene expression in a mouse macrophage cell line.

• Animal Bioscience
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• v.34 no.3_spc
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• pp.463-470
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• 2021

Objective: This experiment was conducted to find out the immunological effects of wheat phytase when long-chain inorganic polyphosphate (polyP) treated with wheat phytase was added to a macrophage cell line, Raw 264.7, when compared to intact long-chain polyP. Methods: Nitric oxide (NO) production of Raw 264.7 cells exposed to P700, a long-chain polyP with an average of 1,150 phosphate residues, treated with or without wheat phytase, was measured by Griess method. Phagocytosis assay of P700 treated with or without phytase in Raw 264.7 cells was investigated using neutral red uptake. The secretion of tumor necrosis factor α (TNF-α) by Raw 264.7 cells with wheat phytase-treated P700 compared to intact P700 was observed by using Mouse TNF-α enzyme-linked immunosorbent assay kit. Results: P700 treated with wheat phytase effectively increased NO production of Raw 264.7 cells by 172% when compared with intact P700 at 12 h exposure. At 5 mM of P700 concentration, wheat phytase promoted NO production of macrophages most strongly. P700, treated with wheat phytase, stimulated phagocytosis in macrophages at 12 h exposure by about 1.7-fold compared to intact P700. In addition, P700 treated with wheat phytase effectively increased in vitro phagocytic activity of Raw 264.7 cells at a concentration above 5 mM when compared to intact P700. P700 dephosphorylated by wheat phytase increased the release of TNF-α from Raw 264.7 cells by 143% over that from intact P700 after 6 h exposure. At the concentration of 50 μM P700, wheat phytase increased the secretion of cytokine, TNF-α, by 124% over that from intact P700. Conclusion: In animal husbandry, wheat phytase can mitigate the long-chain polyP causing damage by improving the immune capabilities of macrophages in the host. Thus, wheat phytase has potential as an immunological modulator and future feed additive for regulating immune responses caused by inflammation induced by long-chain polyP from bacterial infection.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.9
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• pp.1256-1263
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• 2015

Citrus junos Sieb. ex Tanaka has been traditionally called Yuza in Korea and is used as a cuisine material or tea as well as medicinal herb. In this study, we evaluated the immune-enhancing effect of Citrus junos ethanol extract (CJE) on RAW264.7 mouse macrophage and primary immunocytes. CJE treatment showed increased macrophage activity in a dose-dependant manner. CJE also enhanced natural killer (NK) cell activity. We measured lactate dehydrogenase (LDH) level as a measurement of NK cell cytotoxicity against YAC-1 lymphoma cells. CJE treatment showed an increased LDH level in a dose-dependent manner. Finally, we evaluated the effect of CJE on mouse primary splenocyte proliferation. CJE treatment slightly increased splenocyte proliferation compared to the control. The results of this study suggest that CJE can help immune function via macrophage cytokine production, increased NK cell activity, and splenocyte proliferation.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.2
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• pp.278-286
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• 2004

The objective of the current study was to determine the effects of arabinoxylane and PSP on mouse splenocytes, T cells, B cells and macrophages in vitro. Arabinoxylane and PSP directly induced the proliferation of spleen cells in a dose-dependent manner and increased IFN-${\gamma}$ synthesis. Especially, PSP induced IL-2, IL-4 and IL-10 production, Both arabinoxylane and PSP increased PFC (plaque forming cell) and RFC (rosette forming cell) formation. Arabinoxylane was not induced the proliferation of T cells, but PSP directly induced the proliferation of T cells in a high dose. Arabinoxylane and PSP increased the proliferation of B cells and the phagocytic effects of macrophage. When arabinoxylane and PSP were used in macrophage cell line stimulation, there was a marked induction of NO synthesis in a dose-dependent and an increased TNF-$\alpha$ and IL-6 synthesis. Especially, PSP also induced IL-1$\beta$ production. When arabinoxylane and PSP treated in macrophage cell line, there was induction of MHC class II expression. These results suggest that the capacity of arabinoxylane andPSP seem to act as a potent immunomodulator causing augmentation of immune cell activity, and with the absence of notable side-effects, arabinoxylane and PSP could be used as a biological response modifier having possible therapeutic effects against immunological disorders.

• Parasites, Hosts and Diseases
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• v.28 no.2
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• pp.91-100
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• 1990

Paragonimus westermani is a tissue migrating parasite in the early stage until arriving at lung, and most of the parasites spend their life spans there. Considerable immune responses including activation of macrophages are taken place during the residence of parasites in the host. However, concerning the immunologic defense mechanisms of the host against this parasite, only a few document is available so far. In this study, the cytotoxic effect of peritoneal macrophages under the presence of antibody and/or complement against metacercariae of F. westermani was investigated in vitro. Metacercarlae were collected from the crayfish, Cambaroides similis and hatched out in Tyrode solution (pH 7.4). Plastic adherent cells from normal or infected rat (Wistar) peritoneal exudates were used as experimental macrophages. Polyclonal antibodies were obtained from infected rats and a cat. Cat IgG was fractioned with ion exchange chromatography. Fresh rabbit complement was used according to experimental scheme. Various combinations of peritoneal macrophages, normal or infected rat serum, complement and cat IgG were incubated at $36^{\circ}C$ in 5% $CO_2$ incubator for 6, 14, 24 and 48 hours. The results obtained were as follows: 1. P. westermani infection activated peritoneal macrophages non-specifically and this activation induced increases of cell adherence and cytotoxicity on metacercariae. 2. In the presence of infected rat serum the antibody.dependent cell-mediated cytotoxicity of peritoneal macrophages on metacercariae was significantly increased and showed a peak at 6-hour incubation. But the cytotoxic effect was markedly reduced after inactivation of complement and heat.labile IgE antibody by the heating of infected serum at 56$^{\circ}C$ for 30 minutes. 3. The highest cytotoxic effect (100%) of concomitant incubation with IgG and complement showed 24 hours after incubation, although cell adherence was relatively low at 6-hour incubation and 0% at 24-hour incubation. 4. Coordinative functions of complement with serum and IgG were effective in cell adherence and in cytotoxicity, but it is not clear the independent role of complement on the macrophage- mediated cytotoxicity in this study- With these results it is assumed that P. westermani infection can induce the non-specific activation of peritoneal macrophages, and strum antibodies including IgE antibody might enhance the cytotoxicity by macrophages,