• 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 Korean Journal of Physiology and Pharmacology
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• v.25 no.2
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• pp.111-118
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• 2021

27-Hydroxycholesterol (27OHChol) exhibits agonistic activity for liver X receptors (LXRs). To determine roles of the LXR agonistic activity in macrophage gene expression, we investigated the effects of LXR inhibition on the 27OHChol-induced genes. Treatment of human THP-1 cells with GSK 2033, a potent cell-active LXR antagonist, results in complete inhibition in the transcription of LXR target genes (such as LXRα and ABCA1) induced by 27OHChol or a synthetic LXR ligand TO 901317. Whereas expression of CCL2 and CCL4 remains unaffected by GSK 2033, TNF-α expression is further induced and 27OHChol-induced CCL3 and CXCL8 genes are suppressed at both the transcriptional and protein translation levels in the presence of GSK 2033. This LXR antagonist downregulates transcript levels and surface expression of CD163 and CD206 and suppresses the transcription of CD14, CD80, and CD86 genes without downregulating their surface levels. GSK 2033 alone had no effect on the basal expression levels of the aforementioned genes. Collectively, these results indicate that LXR inhibition leads to differential regulation of 27-hydroxycholesterol-induced genes in macrophages. We propose that 27OHChol induces gene expression and modulates macrophage functions via LXR-dependent and -independent mechanisms.

• 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.

• Journal of Microbiology and Biotechnology
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• v.33 no.7
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• pp.934-940
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• 2023

Syneilesis palmata (SP) is a traditional medicinal plant. SP has been reported to have anti-inflammatory, anticancer, and anti-human immunodeficiency virus (HIV) activities. However, there is currently no research available on the immunostimulatory activity of SP. Therefore, in this study, we report that S. palmata leaves (SPL) activate macrophages. Increased secretion of both immunostimulatory mediators and phagocytic activity was observed in SPL-treated RAW264.7 cells. However, this effect was reversed by the inhibition of TLR2/4. In addition, inhibition of p38 decreased the secretion of immunostimulatory mediators induced by SPL, and inhibition of TLR2/4 decreased the phosphorylation of p38 induced by SPL. SPL augmented p62/SQSTM1 and LC3-II expression. The increase in protein levels of p62/SQSTM1 and LC3-II induced by SPL was decreased by the inhibition of TLR2/4. The results obtained from this study suggest that SPL activates macrophages via TLR2/4-dependent p38 activation and induces autophagy in macrophages via TLR2/4 stimulation.

• The Korea Journal of Herbology
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• v.24 no.1
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• pp.151-157
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• 2009

Objectives : The purpose of this study is to investigate the effect of Water Extract from Artemisiae Argi Folium (WAAF) on mouse macrophage Raw 264.7 cells stimulated by lipopolysaccharide (LPS). Methods : Cell viabilities were measured by MTT assay. And the intracellular productions of hydrogen peroxide (H2O2) were measured by dihydrorhodamine 123 assay. TNF-$\alpha$ and IL-6 production from Raw 264.7 were measured by ELISA method. Results : The results of the experiment are as follows. 1. WAAF significantly increased the cell viability compared to the control group (treated with LPS only) at the concentrations of 10, 50, 100, 200, 400 ug/mL. 2. WAAF significantly increased the intracellular production of H2O2 compared to the control group at the concentrations of 50, 100, 200 ug/mL. 3. WAAF significantly decreased the production of TNF-$\alpha$ compared to the control group at the concentrations of 100, 200 ug/mL. 4. WAAF significantly decreased the production of IL-6 compared to the control group at the concentrations of 50, 100, 200 ug/mL. Conclusions : WAAF could be supposed to have the immune-modulating activity related with the macrophage's immunoactivity.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2022.09a
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• pp.93-93
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• 2022

Hovenia dulcis, one of the traditional medicinal plants, is currently being used as a functional ingredient for the development of health functional foods that protects the liver from alcohol damage in Korea. A variety of pharmacological effects of Hovenia dulcis have been reported so far, but studies on immune-enhancing activity are insufficient. Thus, in this study, we report that Hovenia dulcis branches (HDB) induce the activation of macrophages. HDB increased the production of immunostimulatory factors and phagocytosis in RAW264.7 cells. TLR4 inhibition blocked HDB-mediated production of immunostimulatory factors. In addition, the JNK inhibition reduced the HDB-mediated production of immunostimulatory factors, and the HDB-mediated JNK activation was blocked by the TLR4 inhibition. HDB increased the level of LC3-II and p62/SQSTM1. TLR4 inhibition blocked HDB-mediated increase in the level of LC3-II and p62/SQSTM1. These findings indicate that HDB may induce TLR4/JNK-dependent macrophage activation and TLR4-dependent macrophage autophagy.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2020.08a
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• pp.86-86
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• 2020

The leaves of Staphylea bumalda (S. bumalda) as a deciduous tree distributed in Korea, China and Japan are used to treat respiratory diseases or inflammation. However, there is no scientific research on the immune-enhancing activity of S. bumalda leaves. Thus, in this study, we investigated the effect of water extracts from S. bumalda leaves (SBL) on the macrophage activity using mouse macrophage cells, RAW264.7. SBL increased production of immunomodulators such as NO, iNOS, IL-1β, IL-6, TNF-α and MCP-1 in RAW264.7 cells and activated phagocytic activity of RAW264.7 cells. Inhibition of TLR2 and TLR4 blocked SBL-mediated production of immunomodulators in RAW264.7 cells. In addition, SBL-mediated production of immunomodulators was attenuated by JNK inhibition in RAW264.7 cells. SBL increased JNK phosphorylation, while Inhibition of TLR2 and TLR4 blocked SBL-mediated JNK phosphorylation in RAW264.7 cells. These results are thought to be evidence that SBL activates JNK through stimulation of TLR2 and TLR4 in macrophage to induce the production of immunomodulators. In LPS-stimulated RAW264.7 cells, SBL inhibited over-production of immunomodulators. Summarizing the results, SBL showed immunostimulatory activity under normal conditions and immunosuppressive activity under LPS-induced excessive immune response conditions.

• Journal of Society of Preventive Korean Medicine
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• v.27 no.3
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• pp.35-46
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• 2023

Objectives : Gumiganghwal-tang and its main components have been used for treatment of cough, headache, joint pain and fever. Using a respiratory inflammatory model, we intend to demonstrate the its anti-inflammatory effect and immune mechanism of Gumiganghwal-tang. Methods : We induced the respiratory inflammation mouse model by papain treatment. Female BALB/C mice (8 weeks old) were divided into three groups as follows: saline control group, papain treatment group (vehicle), papain and Gumiganghwal-tang (200 mg/kg) treatment group (n=4). To verify the anti-inflammatory effect of Gumiganghwal-tang extracts, we measured the infiltration of inflammatory cells in bronchoalveolar lavage fluid (BALF) and nasal lavage fluid (NALF). Additionally, the efficacy of Gumiganghwal-tang extracts on Th2 cell population and alveolar macrophage in lung were analyzed by using flow cytometry. Results : Gumiganghwal-tang extracts administration decreased inflammatory cell infiltration in BALF and NALF, especially of eosinophils. Furthermore, interleukin-5 level was reduced in lung by drug administration. Interestingly, Gumiganghwal-tang extracts treatment also decreased the Th2 cell (CD4⁺GATA3⁺) population and increased the alveolar macrophage (CD11b⁺CD11c⁺) population in lung. Conclusions : Our findings indicate that Gumiganghwal-tang extracts have anti-inflammatory effects by mediating Th2 cell and alveolar macrophage cell activation.

• Journal of Physiology & Pathology in Korean Medicine
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• v.16 no.5
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• pp.914-920
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• 2002

This experiment is carried out for test whether the addition temperament drugs of Danchisoyo-san have an anti-inflammatory effect and have suppression effect on immunocyte in the state of inflammation which induced by carageenan and zymosan. The freezing lyophilization powder which were extracted from Danchisoyo-san divided low dose group(200mg/kg-DSL) and high dose group(500mg/kg-DSH) and after melting in water, it was orally administered to the mouse. Compared with inflammation induced group which were induced by triggering-inflammation reagent carageenan and zymosan and normal contrast group, we measured the edema decrement effect, macrophage and spleen cell activation. 1. Danchisoyo-san has suppress inflammatory reaction induced by carrageenan. 2. Danchisoyo-san has suppress increasing activation of abdominal cavity macrophage cell in the carrageenan induced inflammation. 3. Danchisoyo-san has suppress increasing activation of spleen cell in the carrageenan induced inflammation. 4. Danchisoyo-san has suppress increasing activation of abdominal cavity macrophage in the zymosan induced inflammation. 5. Danchisoyo-san has suppress increasing activation of spleen cell in the zymosan induced inflammation. Based on the above result, Danchisoyo-san was improved its suppression effect to the inflammatory reaction through the suppression of spleen cell and macrophage activation. So we concluded that Danchisoyo-san is prospected as a anti-inflammatory agent to cure inflammation.

• Journal of Microbiology and Biotechnology
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• v.30 no.3
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• pp.352-358
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• 2020

In this study we investigated the immune effects of oral administration of anionic macromolecules extracted from Codium fragile (CFAM) and red ginseng extract mixture on the peritoneal macrophage cells in immune-suppressed mice. Cyclophosphamide (CY) induces the immune-suppressed condition. CY-treated mice were orally fed with different concentrations of CFAM supplemented with red ginseng extract and the peritoneal macrophages collected. CY treatment significantly decreased the immune activities of peritoneal macrophages, compared to the normal mice. The administration of CFAM mixed with red ginseng extract significantly boosted the viability of macrophage cells and nitric oxide production of peritoneal macrophages. Further, the oral administration of CFAM mixed with red ginseng extract up-regulated the expression of iNOS, COX-2, and TLR-4 as well as cytokines such as IL-1β, IL-6, TNF-α, and IFN-γ more than the red ginseng-treated group. This study showed that CFAM enhanced the immune activity of red ginseng extract in the peritoneal macrophage cells of immune-suppressed mice. Furthermore, CFAM might be used as a co-stimulant of red ginseng extract through the regulation of macrophage cells for the enhancement of human health and immunity.