• BMB Reports
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• v.55 no.2
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• pp.81-86
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• 2022

Macrophages are a major cellular component of innate immunity and are mainly known to have phagocytic activity. In the tumor microenvironment (TME), they can be differentiated into tumor-associated macrophages (TAMs). As the most abundant immune cells in the TME, TAMs promote tumor progression by enhancing angiogenesis, suppressing T cells and increasing immunosuppressive cytokine production. N-myc downstream-regulated gene 2 (NDRG2) is a tumor suppressor gene, whose expression is down-regulated in various cancers. However, the effect of NDRG2 on the differentiation of macrophages into TAMs in breast cancer remains elusive. In this study, we investigated the effect of NDRG2 expression in breast cancer cells on the differentiation of macrophages into TAMs. Compared to tumor cell-conditioned medium (TCCM) from 4T1-mock cells, TCCM from NDRG2-over-expressing 4T1 mouse breast cancer cells did not significantly change the morphology of RAW 264.7 cells. However, TCCM from 4T1-NDRG2 cells reduced the mRNA levels of TAM-related genes, including MR1, IL-10, ARG1 and iNOS, in RAW 264.7 cells. In addition, TCCM from 4T1-NDRG2 cells reduced the expression of TAM-related surface markers, such as CD206, in peritoneal macrophages (PEM). The mRNA expression of TAM-related genes, including IL-10, YM1, FIZZ1, MR1, ARG1 and iNOS, was also downregulated by TCCM from 4T1-NDRG2 cells. Remarkably, TCCM from 4T1-NDRG2 cells reduced the expression of PD-L1 and Fra-1 as well as the production of GM-CSF, IL-10 and ROS, leading to the attenuation of T cell-inhibitory activity of PEM. These data showed that compared with TCCM from 4T1-mock cells, TCCM from 4T1-NDRG2 cells suppressed the TAM differentiation and activation. Collectively, these results suggest that NDRG2 expression in breast cancer may reduce the differentiation of macrophages into TAMs in the TME.

• BMB Reports
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• v.55 no.1
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• pp.48-56
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• 2022

Small extracellular vesicles (sEVs) secreted by most cells carry bioactive macromolecules including proteins, lipids, and nucleic acids for intercellular communication. Given that some immune cell-derived sEVs exhibit anti-cancer properties, these sEVs have received scientific attention for the development of novel anti-cancer immunotherapeutic agents. In this paper, we reviewed the latest advances concerning the biological roles of immune cell-derived sEVs for cancer therapy. sEVs derived from immune cells including dendritic cells (DCs), T cells, natural-killer (NK) cells, and macrophages are good candidates for sEV-based cancer therapy. Besides their role of cancer vaccines, DC-shed sEVs activated cytotoxic lymphocytes and killed tumor cells. sEVs isolated from NK cells and chimeric antigen receptor (CAR) T cells exhibited cytotoxicity against cancer cells. sEVs derived from CD8⁺ T and CD4⁺ T cells inhibited cancer-associated cells in tumor microenvironment (TME) and activated B cells, respectively. M1-macrophage-derived sEVs induced M2 to M1 repolarization and also created a pro-inflammatory environment. Hence, these sEVs, via mono or combination therapy, could be considered in the treatment of cancer patients in the future. In addition, sEVs derived from cytokine-stimulated immune cells or sEV engineering could improve their anti-tumor potency.

• Journal of Korean Medicine Rehabilitation
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• v.19 no.1
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• pp.73-89
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• 2009

Objectives : This study was carried out to investigate the effects of Polygonum cuspidatum extract on several skin functions including inflammation and wrinkle formation. Methods : To investigate in vitro anti-oxidant activity assay, ethanol extracts of medicinal plants tested by DPPH method. In the next experiment, to investigate anti-inflammatory test, the RAW 264.7 macrophage cells was cultured using DMEM including the 10% FBS. To study anti-allergic effect, we blended cultured Human Mast Cells(HMC-1), and then observe $TNF-{\alpha}$, IL-8 by ELISA Results : Polygonum Cuspidatum extract has the effects of anti-inflammation and anti-allergy, which may be due to its inhibitory potential on the macrophage activation. Furthermore, Polygonum Cuspidatum extract has the anti-wrinkle effects through the inhibitory potential on the collagnease, elastase and gelatinase activities. Conclusions : The above results suggest that Polygonum Cuspidatum extract could be applicable for improvement of several skin functions.

• The Korean Journal of Physiology and Pharmacology
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• v.21 no.4
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• pp.449-456
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• 2017

Beauvericin (BEA), a cyclic hexadepsipeptide produced by the fungus Beauveria bassiana, is known to have anti-cancer, anti-inflammatory, and anti-microbial actions. However, how BEA suppresses macrophage-induced inflammatory responses has not been fully elucidated. In this study, we explored the anti-inflammatory properties of BEA and the underlying molecular mechanisms using lipopolysaccharide (LPS)-treated macrophage-like RAW264.7 cells. Levels of nitric oxide (NO), mRNA levels of transcription factors and the inflammatory genes inducible NO synthase (iNOS) and interleukin (IL)-1, and protein levels of activated intracellular signaling molecules were determined by Griess assay, semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), luciferase reporter gene assay, and immunoblotting analysis. BEA dose-dependently blocked the production of NO in LPS-treated RAW264.7 cells without inducing cell cytotoxicity. BEA also prevented LPS-triggered morphological changes. This compound significantly inhibited nuclear translocation of the $NF-{\kappa}B$ subunits p65 and p50. Luciferase reporter gene assays demonstrated that BEA suppresses MyD88-dependent NF-${\kappa}B$ activation. By analyzing upstream signaling events for $NF-{\kappa}B$ activation and overexpressing Src and Syk, these two enzymes were revealed to be targets of BEA. Together, these results suggest that BEA suppresses $NF-{\kappa}B$-dependent inflammatory responses by suppressing both Src and Syk.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.33 no.2
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• pp.100-111
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• 2020

Objectives : The aim of experiment is to examine anti-inflammatory effect and anti-oxidant effect of Naesohwangryun-tang (NSHRT) in LPS-stimulated RAW264.7 macrophage cells. Methods : In the present study, The cell viability was performed by MTT assay. Nitric oxide (NO) production and prostaglandin E₂ (PGE₂) synthesis were performed by NO assay and ELISA KIT. The anti-oxidant effect was performed by DPPH and ABTS radical scavenging activity. The inhibitory effects of pro-inflammatory mediators and cytokines were confirmed by realtime PCR and western blotting. Results : NSHRT was no cytotoxicity at treated group. NO and PGE2 production were inhibited compared to the LPS treated group and also mRNA and protein expressions were significantly decreased compared to the LPS treated group. Conclusions : According to the above experiments, we confirmed that NSHRT has anti-inflammatory and anti-oxidant effects. It is suggested that NSHRT is potential ingredient of skin diseases.

• YAKHAK HOEJI
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• v.42 no.3
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• pp.302-306
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• 1998

In the previous study we described the antitumor activity of GLB, a protein-polysaccharide fraction of the growing tips of Ganoderma lucidum, against sarcoma 180 solid tu mor in ICR mice. In this study we investigated the stimulatory activity of GLB on macrophages. When analyzed using a flow cytometer, GLB ($100{\mu}g/ml$) was found to increase the phagocytic activity of the BALB/c mouse peritoneal macrophages as well as chicken macrophage BM2CL cells against FITC-labeled C.albicans by 55.2% and 21.2%, respectively. GLB also increased the spreading and the expression of MHC class II molecules of BM2CL cells as well as the mouse peritoneal macrophages. From these results, it is clear that GLB is a strong stimulator to the macrophages.

• Preventive Nutrition and Food Science
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• v.13 no.1
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• pp.1-6
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• 2008

Di-D-fructose-2,6':6,2'-dianhydride (DFA-IV) is a disaccharide consisting of two fructose residues that are prepared from levan by levan fructotransferase. Levan is a homopolysaccharide composed of D-fructofuranosyl residues joined by $\beta$-(2,6) and $\beta$-(2,1) linkages. We compared the immunomodulatory effects of levan with DFA-IV. Tumoricidal activity, phagocytosis and nitric oxide (NO) production were examined in levan- and DFA-IV-treated RAW264.7 cells. The NO production, tumoricidal and phagocytic activities were significantly increased in both treated cells. The results indicate that levan has significantly greater effects on tumoricidal activity than DFA-IV at low concentrations (1 ${\mu}g/mL$) and its effect on NO production shows a similar pattern. These results suggest that tumoricidal activity induced by both samples is mediated by NO production.

• Korean Journal of Pharmacognosy
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• v.16 no.4
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• pp.181-190
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• 1985

To determine contents of inorganic elements of Ganoderma lucidum, the horn-shaped carpophores and the pileus of Ganoderma lucidum were incinerated and analyzed by inductively coupled plasma atomic emission spectrophotometry. The ash contents of the pileus and the horn-shaped carpophore were 1.48% and 1.40%, respectively. The pileus contained calcium, magnesium, sodium, manganese, iron, zinc and germanium in that order. The horn-shaped carpophore contained magnesium, calcium, zinc, manganese, iron, copper and germanium in that order. To examine the protein-bound polysaccharide from Ganoderma lucidum for immunopotentiating activity, its fruit bodies were extracted with hot water. Purification of the extract was carried out by acetone precipitation and dialysis. The fraction obtained during the purification procedure consisted of a polysaccharide moiety (51%) and a protein moiety (5%). When the compound was administered intraperitoneally to the mice at a dose of 50mg/kg, it enhanced the accumulation of the peritoneal exudate cells, macrophage and polymorphonuclear leucocytes, thereby indicating immunopotentiation.

• Nutrition Research and Practice
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• v.14 no.5
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• pp.453-462
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• 2020

BACKGROUND/OBJECTIVES: Platycodon grandiflorum (PG), an oriental herbal medicine, has been known to improve liver function, and has both anti-inflammatory and antimicrobial properties. However, little is known about the immune-enhancing effects of PG and its mechanism. In this study, we aimed to investigate whether fermented PG extract (FPGE), which has increased platycodin D content, activates the immune response in a murine macrophage cell line, RAW 264.7. MATERIALS/METHODS: Cell viability was determined by Cell Counting Kit-8 assay and the nitric oxide (NO) levels were measured using Griess reagent. Cytokine messenger RNA levels of were monitored by quantitative reverse transcription polymerase chain reaction. To investigate the molecular mechanisms underlying immunomodulatory actions of FPGE in RAW 264.7 cells, we have conducted luciferase reporter gene assay and western blotting. RESULTS: We found that FPGE treatment induced macrophage cell proliferation in a dose-dependent manner. FPGE also modulated the expression of NO and pro-inflammatory cytokines, such as tumor necrosis factor-α, interleukin (IL)-1β, and IL-6. The activation and phosphorylation levels of nuclear factor kappa B (NF-κB) were increased by FPGE treatment. Moreover, 5-aminoimidazole-4-carboxamide ribonucleotide, an activator of AMP-activated kinase (AMPK), significantly reduced both lipopolysaccharides- and FPGE-induced NF-κB reporter gene activity. CONCLUSIONS: Taken together, our findings suggest that FPGE may be a novel immune-enhancing agent acting via AMPK-NF-κB signaling pathway.

• BMB Reports
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• v.48 no.8
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• pp.479-484
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• 2015

In this study, we report that one of the antimicrobial peptides scolopendrasin VII, derived from Scolopendra subspinipes mutilans, stimulates actin polymerization and the subsequent chemotactic migration of macrophages through the activation of ERK and protein kinase B (Akt) activity. The scolopendrasin VII-induced chemotactic migration of macrophages is inhibited by the formyl peptide receptor 1 (FPR1) antagonist cyclosporine H. We also found that scolopendrasin VII stimulate the chemotactic migration of FPR1-transfected RBL-2H3 cells, but not that of vector-transfected cells; moreover, scolopendrasin VII directly binds to FPR1. Our findings therefore suggest that the antimicrobial peptide scolopendrasin VII, derived from Scolopendra subspinipes mutilans, stimulates macrophages, resulting in chemotactic migration via FPR1 signaling, and the peptide can be useful in the study of FPR1-related biological responses. [BMB Reports 2015; 48(8): 479-484]