• 한국균학회지
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• 제46권2호
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• pp.161-176
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• 2018

진균류 유래의 ${\beta}$-glucan은 pathogen-associated molecular patterns의 일종이기도 하며 면역촉진과 항암작용을 나타냄이 알려져 있지만 세포 내 신호전달에 관해서는 알려진 바가 많지 않다. 대식세포주인 RAW264.7 세포에 불로초에서 추출한 ${\beta}$-glucan을 처리하였을 때 세포막에서는 덱틴-1, toll-like receptor 2, 4, 6의 발현이 증가되었으며, 세포 내에서는 macrophage inflammatory protein (MIP)-1a, MIP-$1{\beta}$, MIP-$1{\gamma}$, IL-$1{\beta}$ 그리고 tumor necrosis factor (TNF)-${\alpha}$의 발현이 증가되었다. 또한 대식세포주에 불로초의 ${\beta}$-glucan과 PI3K 또는 MEK1/MEK2 억제제를 각각 처리하였을 때에 세포 내의 MIP-1a, MIP-$1{\beta}$, MIP-$1{\gamma}$, interleukin-$1{\beta}$, TNF-${\alpha}$의 발현이 감소되었다. 따라서 불로초의 ${\beta}$-glucan은 대식세포에서 MyD88의 경로인 PI3K/Akt를 경유할 뿐만 아니라 MEK 경로를 활성화시킴으로써 다양한 면역조절작용이 가능한 것으로 여겨진다.

• 대한본초학회지
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• 제28권5호
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• pp.113-119
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• 2013

Objectives : The purpose of this study was to investigate the effects of Angelicae Gigantis Radix Water Extract(AG) on the production of proinflammatory mediators in RAW 264.7 cells stimulated with lipopolysaccharide(LPS). Method : RAW 264.7 cells were cotreated with AG(50 and 100 ug/mL) and lipopolysaccharide(LPS; 1 ug/mL) for 24 hours. After 24 hour treatment, using Bead-based multiplex cytokine assay, concentrations of various cytokines such as interleukin(IL)-6, IL-$1{\beta}$, IL-10, tumor necrosis factor-alpha(TNF-${\alpha}$), granulocyte colony-stimulating factor(G-CSF), granulocyte macrophage colony-stimulating factor(GM-CSF), interferon inducible protein-10(IP-10), leukemia inhibitory factor(LIF), lipopolysaccharide-induced chemokine(LIX), monocyte chemoattractant protein-1(MCP-1), macrophage colony-stimulating factor(M-CSF), macrophage inflammatory protein(MIP)-$1{\alpha}$, MIP-$1{\beta}$, MIP-2, Regulated on Activation, Normal T cell Expressed and Secreted(RANTES) and vascular endothelial growth factor(VEGF) were measured. Result : AG significantly inhibited LPS-induced production of TNF-${\alpha}$, MIP-$1{\alpha}$, G-CSF, RANTES, IL-10, and M-CSF from LPS-stimulated RAW 264.7 cells at the concentrations of 50 and 100 ug/mL. AG significantly inhibited LPS-induced production of MIP-$1{\beta}$, MIP-2, GM-CSF, and IL-6 from LPS-stimulated RAW 264.7 cells at the concentrations of 50 ug/mL. AG significantly inhibited LPS-induced production of VEGF from LPS-stimulated RAW 264.7 cells at the concentrations of 100 ug/mL. But AG did not show any significant effect on the production of MCP-1, LIF, LIX, IP-10 and IL-$1{\beta}$ from LPS-induced RAW 264.7 cells. Conclusion : These results suggest that AG has anti-inflammatory effect related with its inhibition of proinflammatory mediators such as TNF-${\alpha}$, MIP-$1{\alpha}$, G-CSF, RANTES, IL-10, MIP-$1{\beta}$, MIP-2, GM-CSF, IL-6, VEGF and M-CSF in LPS-induced macrophages.

• Molecules and Cells
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• 제27권2호
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• pp.257-261
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• 2009

Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans) is an important pathogen casuing aggressive periodontitis. The present study was designed to investigate the chemokines expression regulated by A. actinomycetemcomitans lipopolysaccharide (LPS). Chemokines genes expression profiling was performed in Raw 264.7 cells by analyses of microarray and reverse transcription-polymerase chain reaction (RT-PCR). Microarray results showed that the induction of monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-$1{\alpha}$ (MIP-$1{\alpha}$), MIP-$1{\beta}$, MIP-$1{\gamma}$, regulated upon activation, normal T-cell expressed and secreted (RANTES), macrophage inflammatory protein-2 (MIP-2), and interferon-${\gamma}$ inducible protein 10 (IP 10) by A. actinomycetemcomitans LPS was increased to 12.5, 1.53, 9.09, 17.3, 2.82, 16.1, and 18.1 folds at 18 h, respectively. To check these chemokines expression by A. actinomycetemcomitans LPS, we examined gene expressions by RT-PCR, and found that the expression of MIP-$1{\beta}$, MIP-$1{\gamma}$, RANTES, MIP-2, and IP 10 was increased 107.1, 93.6, 106.8, 86.5, and 162.0 folds at 18 h, respectively. These results indicate that A. actinomycetemcomitans LPS stimulates the several chemokines expressions (MIP-$1{\alpha}$, MIP-$1{\beta}$, MIP-$1{\gamma}$, RANTES, MIP-2, and IP 10) in Raw 264.7 cells.

• Biomolecules & Therapeutics
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• 제21권1호
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• pp.42-48
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• 2013

Nystatin, a polyene antifungal antibiotic, is a cholesterol sequestering agent. The antifungal agent alters composition of the plasma membrane of eukaryotic cells, whereas its effects on cells are poorly investigated. In the current study, we investigated the question of whether nystatin was able to induce expression of macrophage inflammatory protein-1 (MIP-1). THP-1 cells rarely express MIP-$1{\alpha}$ and MIP-$1{\beta}$, however, upon exposure to nystatin, significantly elevated expression of MIP-$1{\alpha}$ and MIP-$1{\beta}$ was observed in a dose-dependent fashion at the messenger and protein levels. Cellular factors activated by nystatin as well as involved in nystatin-induced expression of MIP-1 proteins were identified in order to understand the molecular mechanisms of action of the anti-fungal agent. Treatment with nystatin resulted in enhanced phosphorylation of Akt, ERK, p38 MAPK, and JNK. Abrogation or significant attenuation of nystatin-induced expression of MIP-$1{\alpha}$ and MIP-$1{\beta}$ was observed by treatment with Akt inhibitor IV, LY294002, and SP6001250. Inhibition of ERK or p38MAPK using U0126 and SB202190 did not lead to attenuation of MIP-1 expression. In addition, inhibitors of protein kinase C, such as GF109203X and Ro-318220, also attenuated expression of MIP-1. These results indicate that nystatin is able to activate multiple cellular kinases and, among them, Akt and JNK play primary roles in nystatin-induced expression of MIP-1 proteins.

• Journal of Periodontal and Implant Science
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• 제25권2호
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• pp.267-278
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• 1995

Human polymorphonuclear leukocytes(PMN) are the most numerous host cell in periodontal pockets and their presumed role is to form a protective barrier between the bacteria and periodontal tissues. Microbial component LPS activates macrophages to produce $IL-1{\beta}$, $MIP-1{\alpha}$, $-1{\beta}$, $TNF-{\alpha}$ and IL-6, etc. These cytokines have autocrine function to the macrophages, and paracrine function to other cell such as PMN and affect them to produce some biological functions. In the present study, human PMN were tested for the expression of $IL-1{\beta}$ and $MIP-1{\alpha}$ mRNA. Also we performed the receptor binding assay and in vitro assay for the antimicrobial action of HL-60 cell to determine whether HL-60 can replace the peripheral PMN in analyzing the biological functions. PMN were stimulated with $IL-1{\beta}$, TPA, $MIP-1{\alpha}$, LPS, IL-2 and total cytoplasmic RNA were extracted for the northern blot analysis. In order to determine the induction kinetics of $IL-1{\beta}$ or $MIP-1{\alpha}$ mRNA expression, cells were stimulated for 0,1,2,3 hours. We found peak expression of $IL-1{\beta}$ mRNA after 1hr of induction with $IL-1{\beta}$, LPS and after 2hr of induction with TPA. $MIP-l{\alpha}$ also induced but a scarce $IL-l{\beta}$ message from PMN. In contrast to the $IL-l{\beta}$ mRNA expression, $MIP-1{\alpha}$ were not induced from PMN in any culture conditions. Receptors for $MIP-1{\alpha}$ were identified on dibutyryl cyclic AMP(dbcAMP)-treated HL-60 as well as peripheral PMN. dbcAMP treatment significantly enhanced antimicrobial action of undifferentiated HL-60 cell. MIP-1 further increased enhancing effect of dbcAMP. $IL-1{\beta}$, to a lesser extent, also increased dbcAMP-induced enhancing effect of antimicrobial action of HL-60 cell.

• 한방안이비인후피부과학회지
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• 제18권3호
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• pp.1-17
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• 2005

In many recent studies, molecular biological methods have been used to investigate the role of cytokines in pathogenesis of lung disease. This Experiment was conducted to investigate the effects of Sochungyong-tang on gene expressions in Mouse Alveolar Macrophage. Fer this purpose, we observed the cytokines ($IL-1{\beta}$, IL-6, IL-10, iNOS, $MIP-1{\alpha},\;MIP-1{\beta},\;MIP-1{\gamma},\;TGF-{\beta},\;TNF-{\alpha}$). We picked the alveolar macrophage out of mice and cultured it. We analyzed the cytokine gene expression by reverse transcription-PCR. The results obtained were as follows : 1 . Sochungyong-tang showed inhibitory effects on $IL-1{\beta}$ in time and concentration. 2. Sochungyong-tang showed inhibitory effects on IL-6 in time and concentration. 3. Sochungyong-tang showed inhibitory effects on IL-10 in concentration. 4. Sochungyong-tang showed inhibitory effects on iNOS. 5. Sochungyong-tang showed inhibitory effects on $TGF-{\beta}$ in time and concentration. 6. Sochungyong-tang showed on inhibitory effects on $MIP-1{\alpha},\;MIP-1{\beta},\;MIP-1{\gamma}$, $TCF-{\beta}$, $TNF-{\alpha}$. According to above results, it is supposed that Sochungyong-tang has the inhibitory effects on cytokine gene expression in mouse alveolar macrophage and can be usefully applied for curing inflammatory process of lung disease. Advanced studies are required to investigate the cure mechanism of Sochungyong-tang in the future.

• 동의생리병리학회지
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• 제28권3호
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• pp.330-336
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• 2014

This study aimed at examining the anti-inflammatory effects of Scutellariae Radix & Lonicerae Caulis water extract(SC). RAW 264.7 mouse macrophage cells were treated with $25{\sim}200{\mu}g/m{\ell}$ SC for 24 hours. Cell viability was then measured using MTT assays. The nitric oxide(NO) production and the creation of several cytokines in LPS-stimulated RAW 264.7 cells were investigated. SC inhibited significantly increasing the production of NO in LPS-induced RAW 264.7 cell at the density of 25, 50 and $200{\mu}g/m{\ell}$. SC inhibited significantly the TNF-${\alpha}$ of the RAW 264.7 cell induced by LPS at the density of $50{\mu}g/m{\ell}$. SC inhibited significantly the MIP-$1{\alpha}$ of the RAW 264.7 cell induced by LPS at the density of 25, 50 and $100{\mu}g/m{\ell}$. SC inhibited significantly the MIP-$1{\beta}$, MIP-2 at the density of 50, $100{\mu}g/m{\ell}$ in the RAW 264.7 cell increased by LPS, respectively. SC did not affect the production levels of VEGF in RAW 264.7 cell. As a result, SC significantly inhibited the inductions of MIP-$1{\alpha}$, MIP-$1{\beta}$, MIP-2 and NO in LPS-induced RAW 264.7 cell without causing the toxicity. These results signify that SC has anti-inflammatory effects on controlling the over inflammatory reaction on the RAW 264.7 cell.

• BMB Reports
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• 제43권4호
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• pp.257-262
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• 2010

The aim of work was to investigate changes of 7-ketocholesterol synthesis in alveolar macrophages in the dynamic of lung mechanical ventilation with injurious parameters. The goal of in vitro part of work was to observe influence of 7-ketocholesterol on iNOS and MIP1 $\beta$ production in bronchoalveolar lavage fluid (BALF) cells. We used 17 healthy domestic pigs randomly assigned into two treatment groups: group I with mechanical ventilation with physiological parameters; group II underwent injurious ventilation with high volume tidal (VT) and low positive end expiratory pressure (PEEP). Cells were analyzed for CYP27A1 protein and gene expression levels, 7-ketocholesterol production. In alveolar macrophages of group II, we obtained increase of production of CYP27A1 protein and 7-ketocholesterol, as well as the expression of the CYP27A1 gene at the 2nd hour of ventilation. In the in vitro experiments we show dose-dependent increase of MIP1 $\beta$ and decrease of CYP27A1, iNOS protein production after 7-ketocholesterol treatment.

• 대한한방내과학회지
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• 제25권4호
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• pp.289-299
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• 2004

Background : Although the pathophysiology of asthma has been reported, its mechanism has not been fully elucidated. The mast cell is an effector cells in allergic inflammation and secretes a number of chemokines. Chemokines are important for the recruitment of leukocytes to sites of infection, which is essential in host defense. Chemokines also contribute to the pathogenesis of several disorders such as asthma, chronic bronchitis, atopic dermatitis, allergic rhinitis, and rheumatoid arthritis. Objective : In this study, the aim was to identify the effect of Baekryunchihyo-tang(白蓮治哮湯) on expression of chemokines. This was examined by RT-PCR using the human mast cell line (HMC-l) Materials and Methods : HMC-l cells were used, which is known to secrete and express chemokines. In order to investigate the protective effect of Baekryunchihyo-tang(白蓮治哮湯), HMC-l cells were incubated with pretreatment of Baekryunchihyo-tang(白蓮治哮湯) for 24 hrs. RT-PCR analyses of chemokine genes of cells pretreated with Baekryunchihyo-tang(白蓮治哮湯) showed that expressions of IL-8, $MIP-l{\beta}$, and RANTES genes in these cells were lower and $MIP-l{\alpha}$ showed a similar pattern compared to the calcium ionophore-treated group. In addition, cell cytotoxicity concentration measurements were performed by MTT assay method. Results : After stimulation with 1 uM calcium ionophore A23178 for 2 hrs, IL-8, major one of CXC chemokines, was highly expressed, and expression of $MIP-l{\beta}$ and RANTES (CC chemokines) increased, while expression of $MIP-l{\alpha}$ did not change. The cell cytotoxicity of Baekryunchihyo-tang(白蓮治哮湯) with treatments at various concentrations and times was not observed, respectively. Conclusion : This study suggests that Baekryunchihyo-tang(白蓮治哮湯) has dose-dependent effects on mRNA expression of IL-8(CXC chemokines), $MIP-l{\beta}$ and RANTES(CC chemokines) in human mast cellline(HMC-l). So these herbal medicines may inhibit the inflammatory process of asthma. Advanced studies are required to investigate the mechanism of inhibition by herbal medicine in the asthma model. This study provides basic data on the possibility of the clinical treatment of Baekryunchihyo-tang(白蓮治哮湯) for allergic disorders.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제18권2호
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• pp.286-297
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• 1996

The proliferation of bone marrow stem cell compartment is thought to be under both positive and negative controls by cytokines and colony stimulation factors. Macrophage inflammatory $protein-1{\alpha}(MIP-1{\alpha})$ has been assessed for its potential to protect hematopoietic stem cells from cytotoxic effects of a cycle-specific antineoplastic agents. We have tested the ability of $MIP-1{\alpha}$ to suppress the proliferation of stem cell line Du.528.101 in variety of active status by using $[^{3}H]-thymidine$ incorporation test. The results were as follows. 1. The effect of $MIP-1{\alpha}$ on steady-state Du.528.101 cell represented the cell growth suppression at the concentration of 10, 50, 100nM of $MIP-1{\alpha}$(P<0.001). 2. $MIP-1{\alpha}$ stimulated the proliferation of Du.528.101 cells previously treated with IL-1 at the concentration of 5, 50nM of $MIP-1{\alpha}$(P<0.01). 3. The suppression effect of MIP-1 on Du.528.101 cells at the concentration of 5, 50nM was shown when cells were treated with $MIP-1{\alpha}$ before activation with $IL-1{\beta}(P<0.01)$. 4. The growth rate of synchronized cells were slower than that of non-synchronized ones, and $MIP-1{\alpha}$ represented the similar suppression effect on both synchronized and non-synchronized cells.