• Archives of Pharmacal Research
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• v.22 no.2
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• pp.130-136
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• 1999

Mouse guanylate-binding protein 3 (mGBP3) is a 71-kDa GTPase which belongs to GTP-binding protein family. The present study showed that the expression of mGBP3 transcript was readily induced in a dose dependent fashion in the macrophage cell line RAW264.7 treated with either $interferon-{\gamma} (IFN-\gamma)$ or lipopolysaccaride (LPS). The expression of mGBP3 protein was also apparent by 4 and 6 h after the treatment of cells with IFN-\gamma (100 U/ml) or LPS ($1{\mu}g/ml$) , and remained at palteau for at least 24 h. Cycloheximide ($10{\mu}g/ml$) had no effect on the $IFN-\gamma-$ or LPS-induced mGBP3 expression, suggesting that the mGBP3 induction did not require further protein synthesis. Interestingly, a protein kinase C (PKC) inhibitor staurosporine (50 nM) abolished the induction of mGBP3 expression by LPS, but not by $IFN-{\gamma}$. These findings suggest that mGBP3 may be involved in the macrophage activation process and both IFN-\gamma and LS induce the mGBP3 expression through distinct signal transduction pathways.

• The Korea Journal of Herbology
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• v.29 no.1
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• pp.1-6
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• 2014

Objectives : Many of currently used anti-cancer drugs were developed to target cell death mechanisms and had serious side effects by causing damage to normal cells. Hangambujeongsan or Kangai Fuzheng Powder was a mixture based on the traditional Chinese medicine. It had been used in the local Chinese hospitals to treat cancer patients for decades and had shown a certain level of beneficial effects without major toxic effects. But its mechanism of action had not been elucidated yet. Thus this study aimed to investigate the effects of Kangai Fuzheng Powder in an in vitro experiment. Methods : Cancer lines or RAW264.7 mouse macrophage cells were treated with Kangai Fuzheng Powder. Cell viability was measured by MTT assay, and morphological observation was also performed. Gene expression of cytokines in macrophages was determined by real-time polymerase chain reaction. Phagocytic function assay was also performed in macrophage cells. Results : Kangai Fuzheng Powder had no direct detrimental effect on cancer cells. When macrophages were co-cultured with cancer cells, Kangai Fuzheng Powder had toxic effect on cancer cells. After exposing macrophages to Kangai Fuzheng Powder, macrophages transformed into activated form and the mRNA level of tumor necrosis factor-alpha, interleukin-1beta, interleukin-6, interleukin-10 and monocyte chemotactic protein-1 was significantly enhanced. Phagocytic activity of macrophages was dramatically potentiated. Conclusions : We demonstrated that anti-cancer effect of Kangai Fuzheng Powder was related to activation of macrophages including enhanced cytokine production and phagocytic function.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.15 no.1
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• pp.50-62
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• 2002

In the age of puberty or 20-30th young people who are sensitive to outward appearance, acne is serious problem at beauty and has social and psychological influence on that people. So this experiment is carried out for test whether the addition temperament drugs of Yungyopaedock-san(YP) have an anti-inflammatory effect and have suppression effect on immunocyte in the state of inflammation which induced by acne. The results was as follows. 1. YP has suppress inflammatory reaction induced by carageenan. 2. YP has suppress increasing activation of abdominal cavity macrophage in the carageen and zymosan induced inflammation. 3. YP has suppress increasing activation of spleen cell in the carageenan and zymosan induced inflammation. Based on the above result, YP was improved its suppression effect to the inflammatory reaction through the suppression of spleen cell and macrophage activation. So we concluded that YP is prospected as a anti-inflammatory agent to cure inflammation induced by ance.

• Journal of Food Hygiene and Safety
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• v.27 no.4
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• pp.406-414
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• 2012

This study was designed to investigate the effect of fucoidan on the activation of macrophage and on induction of apoptosis in AGS cell. To measure the activity of macrophages, NO and TNF-${\alpha}$ assays were performed in Raw 264.7 cell. Treatment with fucoidan significantly increased production of NO and TNF-${\alpha}$, indicating activation of macrophages. The result of MTT assay shows that cell viability was significantly decreased in a dose and time-dependent manner. Fucoidan increased to enhance mitochondrial membrane permeability, as well as the cytochrome c release from the mitochondria. Fucoidan decreased Bcl-2 and XIAP expression, whereas the expression of Bax was increased in a time-dependent manner compared to the control. In addition, the active forms of caspase-9 were increased, and the inactivation of Akt was decreased in a time-dependent manner. Caspase inhibitor, z-VAD-FMK, canceled the apoptosis of fucoidan, expression of Bax and caspase-9 were decrease. These results indicate that fucoidan induces activation of macrophage and apoptosis through activation of caspase on AGS cell.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.7
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• pp.1079-1083
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• 2015

Activating macrophage cells play an important role in the host immune defense system. In this paper, immuno-modulatory activities of polysaccharides separated from Jubak (JPS) in macrophage cells were investigated. Immuno-modulatory activities were estimated based on cell proliferation, nitric oxide (NO) and cytokine production, degree of mitogen-activated protein kinases (MAPKs), and nuclear factor (NF)-${\kappa}B$ phosphorylation in RAW264.7 macrophage cells. JPS (62.5 to $250{\mu}g/mL$) did not induce a cytotoxic event. Additionally, NO and proinflammatory cytokines (tumor necrosis factor-${\alpha}$ and interleukin-6) production significantly increased in a dose-dependent manner. Similarly, phosphorylation of MAPKs and NF-${\kappa}B$ increased upon JPS treatment. Therefore, our results suggest that polysaccharides separated from Jubak can induce macrophage activation through MAPK and NF-${\kappa}B$ signaling and induction of Th1 polarization.

• Journal of Ginseng Research
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• v.43 no.3
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• pp.394-401
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• 2019

Background: Ginsenoside Rb1, a triterpene saponin, is derived from the Panax ginseng root and has potent antiinflammatory activity. In this study, we determined if Rb1 can increase macrophage phagocytosis and elucidated the underlying mechanisms. Methods: To measure macrophage phagocytosis, mouse peritoneal macrophages or RAW 264.7 cells were cultured with fluorescein isothiocyanate-conjugated Escherichia coli, and the phagocytic index was determined by flow cytometry. Western blot analyses were performed. Results: Ginsenoside Rb1 increased macrophage phagocytosis and phosphorylation of p38 mitogenactivated protein kinase (MAPK), but inhibition of p38 MAPK activity with SB203580 decreased the phagocytic ability of macrophages. Rb1 also increased Akt phosphorylation, which was suppressed by LY294002, a phosphoinositide 3-kinase inhibitor. Rb1-induced Akt phosphorylation was inhibited by SB203580, (5Z)-7-oxozeaenol, and small-interfering RNA (siRNA)-mediated knockdown of $p38{\alpha}$ MAPK in macrophages. However, Rb1-induced p38 MAPK phosphorylation was not blocked by LY294002 or siRNA-mediated knockdown of Akt. The inhibition of Akt activation with siRNA or LY294002 also inhibited the Rb1-induced increase in phagocytosis. Rb1 increased macrophage phagocytosis of IgG-opsonized beads but not unopsonized beads. The phosphorylation of p21 activated kinase 1/2 and actin polymerization induced by IgG-opsonized beads and Rb1 were inhibited by SB203580 and LY294002. Intraperitoneal injection of Rb1 increased phosphorylation of p38 MAPK and Akt and the phagocytosis of bacteria in bronchoalveolar cells. Conclusion: These results suggest that ginsenoside Rb1 enhances the phagocytic capacity of macrophages for bacteria via activation of the p38/Akt pathway. Rb1 may be a useful pharmacological adjuvant for the treatment of bacterial infections in clinically relevant conditions.

• Biomolecules & Therapeutics
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• v.21 no.6
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• pp.481-486
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• 2013

Macrophages play a role in innate immune responses to various foreign antigens. Many products from primary tumors influence the activation and transmigration of macrophages. Here, we investigated a migration of macrophages stimulated with cancer cell culture-conditioned medium (CM). Macrophage activation by treatment with CM of B16F10 cells were judged by the increase in protein levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX2). The location where macrophages were at 4 h-incubation with control medium or CM was different from where they were at 5 h-incubation in culture dish. Percentage of superimposed macrophages at every 1 h interval was gradually increased by CM treatment as compared to control. Total coverage of migrated track expressed in coordinates was smaller and total distance of migration was shorter in CM-treated macrophages than that in control. Rac1 activity in CM-treated macrophages was also decreased as compared to that in control. When macrophages were treated with CM in the presence of dexamethasone (Dex), an increase in COX2 protein levels, and a decrease in Rac1 activity and total coverage of migration were reversed. In the meanwhile, biphasic changes were detected by Dex treatment in section distance of migration at each time interval, which was more decreased at early time and then increased at later time. Taken together, data demonstrate that macrophage motility could be reduced in accordance with activation in response to cancer cell products. It suggests that macrophage motility could be a novel marker to monitor cancer-associated inflammatory diseases and the efficacy of anti-inflammatory agents.

• The Korean Journal of Physiology and Pharmacology
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• v.15 no.2
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• pp.101-106
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• 2011

We demonstrate that glycoprotein isolated from Dioscorea batatas (GDB) has immunostimulatory effects including macrophage activation. Analysis of infiltration of inflammatory cells into peritoneal cavity showed GDB treatment significantly increased the recruitment of macrophages, lymphocytes, neutrophils, and monocytes into the peritoneal cavity. Treatment of spleen cells isolated from C57BL/6 mice with GDB significantly increased the proliferation of B cells and T cells induced by LPS and ConA, respectively. Treatment with GDB significantly increased the cytolytic capacity of NK cells and macrophages against YAC-1 and B16 cells, respectively. In order to further confirm and investigate the mechanism of GDB on macrophage activation, we analyzed the effects of GDB on the cytokine expression including iNOS, IL-1${\beta}$, and TNF-${\alpha}$ in mouse macrophage cell line, RAW 264.7 cells. RT-PCR and ELISA showed that GDB increased the expression of IL-1${\beta}$, and TNF-${\alpha}$, whereas iNOS was not induced by GDB. Collectively, this series of experiments indicates that GDB stimulates immune system including macrophage activation.

• Journal of Animal Science and Technology
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• v.47 no.6
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• pp.947-954
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• 2005

This study was conducted to investigate the effects of hydrolyzed Lactobacillus supplementation with digestive enzymes treatment on the macrophage activation, the induction of nitric oxide(NO), interleukin (IL)-6 and tumor necrosis factor(TNF)-$\alpha$. The RAW 264.7 murine macrophage was exposed to porcine Lactobacillus strains which were digested with both pepsin and pancreatin. The production of NO, TNF-$\alpha$ and IL-6 in the macrophage were strain and dose-dependent, respectively. The induction of NO and cytokines were higher in both 3149 and 3146 strains compared with other Lactobacillus strains. Overall, the level of NO was observed at the lower range between 10 and 150 μg hydrolysates per ml, whereas IL-6 and TNF-$\alpha$ were observed at relatively higher concentration between 50 and 300 μg hydrolysates per ml. Lactobacillus strains which produced a high level of NO also showed a high induction of TNF-$\alpha$ and IL-6. Therefore, the present results suggest that hydrolysates of Lactobacillus strains are related to induction of several macrophage mediators, and then it could be beneficially used to modulate gastrointestinal immune function of the host. Also, the methodogly employed in this study might be useful to investigate the effects of lactic acid bacteria on gastrointestinal immunity.

• Biomedical Science Letters
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• v.4 no.1
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• pp.35-42
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• 1998

The effects of CRP purified from human ascites fluid on phagocytic activity of the human macrophage were investigated. CRP was purified using affinity chromatography including absorption on p-diazonium phosphocholine or C-polysaccharide coupled sepharose 4B and gel filtration on hydroxylapatite column chromatography. Macrophage was separated ficoll hypaque gradient density and absorption method, and then was confirmed phagocytic uptake test using latex method. CRP was able either to inhibit or to enhance phagocytic activity of human macrophage against bacteria in vitro. The effects of CRP on phagocytic activity of human macrophage were in time and dose-dependent manners. The additional sequence of reaction mixture against bacteria in vitro shows a threshold stimulus on the activation of phagocytic response upon the CRP.