• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.3
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• pp.385-392
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• 2011

Although monoculture methods have been remarkably useful due to their simplicity, they have serious limitation because of the different types of cells communication with each other in many physiological situations. We demonstrated levels of markers of endothelial dysfunction such as tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and interleukin-1$\beta$ (IL-1$\beta$) as well as stimulation of receptor of advanced glycation endproducts (AGEs) on monoand co-culture system such as only monocyte (THP-1) cultivation system, only endothelial cell (HUVEC) cultivation system, and co-cultivation system of THP-1 and HUVEC. The mRNA levels of TNF-$\alpha$ and IL-1$\beta$ on HUVEC increased by the co-culture with monocyte after 4 hr at 100 ${\mu}g/mL$ glyceraldehyde-AGE. The secreted protein contents into medium of TNF-$\alpha$ and IL-1$\beta$ increased after 8 hr approximately 2~2.5 times compared to mono-cultivation. In contrast, the mRNA level of receptor of AGE (RAGE) was relatively insensitive on the co-culture system. The mediators by which monocytes activate endothelial cell have not been fully elucidated. In this study we confirmed production of soluble cytokines such as TNF-$\alpha$ and IL-1$\beta$ by monocytes. Use of monocyte conditioned medium, which contains both cytokines, can activate endothelial cell.

• Molecules and Cells
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• v.22 no.3
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• pp.308-313
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• 2006

Stromal cell-derived factor (SDF-1) is a CXC chemokine that selectively activates the CXCR4 chemokine receptor. Fibronectin is an intracellular matrix component that binds integrin and mediates cell-matrix adhesion. Activation of the integrin receptor can occur in two ways: by ligand binding (outside-in signaling), and in response to intracellular events (inside-out signaling). In the current study we showed that SDF-$1{\alpha}$ inhibited adhesion of T lymphocyte Jurkat cells resulting from binding high concentrations of fibronectin as well as that of THP-1 monocytes. The effect of SDF-$1{\alpha}$ on fibronectin-mediated adhesion was partly reversed by the CXCR4 receptor antagonist T140. Our results suggest that an SDF-1/CXCR4 signal pathway modulates fibronectin-mediated lymphocytes adhesion.

• International Journal of Oral Biology
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• v.43 no.4
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• pp.217-222
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• 2018

Phagocytosis is a fundamental process in which phagocytes capture and ingest foreign particles including pathogenic bacteria. Several oral pathogens have anti-phagocytic strategies, which allow them to escape from and survive in phagocytes. Impaired bacteria phagocytosis increases inflammation and contributes to inflammatory diseases. The purpose of this study is to investigate the influences of various agents on oral pathogenic phagocytosis. To determine phagocytosis, Streptococcus mutans, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis were stained with 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE), and was measured using flowcytometery and confocal microscopy. The influencing factors on phagocytosis were evaluated through the pretreatment of ROS inhibitor (N-acetyl-L-cysteine (NAC)), lysozyme, potassium chloride (KCI) and adenosine triphosphate (ATP) in THP-1 cells. Expression of pro-inflammatory cytokines was determined by enzyme-linked immunosorbent assay (ELISA). The phagocytosis of various bacteria increased in a MOI-dependent manner. Among the tested bacteria, phagocytosis of P. gingivalis showed the highest fluorescent intensity at same infection time. Among the tested inhibitors, the NAC treatment significantly inhibited phagocytosis in all tested bacteria. In addition, NAC treatment indicated a similar pattern under the confocal microscopy. Moreover, NAC treatment significantly increased the bacteria-induced secretion of $IL-1{\beta}$ among the tested inhibitors. Taken together, we conclude that the phagocytosis occurs differently depending on each bacterium. Down-regulation by ROS production inhibited phagocytosis and lead increased of oral pathogens-associated inflammation.

• The Journal of Internal Korean Medicine
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• v.29 no.1
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• pp.12-24
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• 2008

Background and Objective : Chungsangboha-tang (CSBHT) has analgesic, sedative, anti-convulsive and anti-histamine effects, so it alleviates the symptoms of asthma. For the comparison of anti-inflammatory effect(s) on CSBHT, PD098059 was used as a negative control. Materials and Methods : This study emphasized THP-1 cells, which had been well characterized as a human monocytic leukemic cell line. The cells resemble monocytes with respect to several criteria and can be differentiated into macrophage-like cells by treatment with PMA. By using the MTS assay, it was possible to prove the safety of CSBHT. Results : Results shows that the CSBHT did not affected cell survival within $10^{1}$ ng/ml to $10^{5}$ ng/ml. Especially, $10^{5}$ ng/ml CSBHT treated cells show 70% deduction of $TNF-{\alpha}$ gene expression against that of LPS treated group. Furthermore, $IL-1{\beta}$, IL-6, IL-8, IL-10 and $TNF-{\alpha}$ levels are down-regulated when treated with CSBHT with concentrations up to 100 ug/ml on monocyte-derived macrophages. Interestingly, CSBHT-treated samples showed that overall transcriptional activities were down-regulated to 20% of that of PD098059 ($TNF-{\alpha}$ inhibitor). At protein level, the expression of $TNF-{\alpha}$ showed similar results as that of transcriptional activity. Results show that the protein level decreased more in the CSBHT-treated group (487 ${\pm}$ 87 pg/ml) than in the LPS-treated group (703 ${\pm}$ 103 pg/ml). In addition, the protein level of IL-8 in the CSBHT treated-group (9.84 ${\pm}$ 3.28 ng/ml) decreased similar as the expression of the control and PD098059-treated groups. Conclusion : CSBHT affects immune response, especially allergic responses and suppression of inflammatory reaction. The results provide us an alternative way to care for clinical inflammatory diseases, not only asthma but also the other possible general inflammatory and allergic diseases.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.1
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• pp.1-7
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• 2017

Zerumbone is a major component of the essential oil from Zingiber zerumbet Smith, which is a kind of wild ginger. In addition, various biological functions, such as liver protection, pain relief, atherosclerosis, and antimicrobial activity have been reported. It is also known to be effective in the proliferation of immune cells and the expression of cytokines. In this study, we investigated the effects of zerumbone on monocyte activation. First, it was confirmed that the proliferation of THP-1 cells was increased by zerumbone. The strongest increase in THP-1 proliferation after lipopolysaccharide treatment was observed at $5{\mu}M$ zerumbone treatment, and the increase of cell proliferation without lipopolysaccharide was the highest at $10{\mu}M$. Conversely, when treated with $50{\mu}M$ zerumbone, a rapid decrease of proliferation was observed regardless of the presence of lipopolysaccharide (LPS). The phosphorylation of signaling protein, Erk, induced by LPS was also increased by zerumbone. The strongest increase in phosphorylation was observed when treated with $50{\mu}M$ of zerumbone with reduced proliferation. The activity of transcription factor $NF-{\kappa}B$ was not significantly altered by zerumbone alone, but increased when treated with lipopolysaccharide. Furthermore, the transcription of the inflammatory cytokines $TNF-{\alpha}$ and IL-8, which are regulated by $NF-{\kappa}B$, is also increased by zerumbone. These results suggest that zerumbone can enhance the proliferation and activity of monocytes. Furthermore, it is believed that zerumbone can enhance rthe immune responses through increased monocyte activity in bacterial infections with LPS, thereby helping to treat effective bacteria.

• IMMUNE NETWORK
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• v.16 no.5
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• pp.296-304
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• 2016

It has been reported that fatty acid binding proteins (FABPs) do not act only as intracellular mediators of lipid responses but also have extracellular functions. This study aimed to investigate whether extracellular liver type (L)-FABP has a biological activity and to determined serum L-FABP levels in patients with end-stage renal disease (ESRD). We isolated L-FABP complementary deoxyribonucleic acid (cDNA) from the Huh7 human hepatocarcinoma cell line and expressed the recombinant L-FABP protein in Escherichia coli. A549 lung carcinoma and THP-1 monocytic cells were stimulated with the human recombinant L-FABP. Human whole blood cells were also treated with the human recombinant L-FABP or interleukin (IL)-1α. IL-6 levels were measured in cell culture supernatants using IL-6 enzyme-linked immunosorbent assay (ELISA). Human recombinant L-FABP induced IL-6 in a dose-dependent manner in A549, THP-1 cells, and whole blood cells. The blood samples of healthy volunteers and patients with ESRD were taken after an overnight fast. The serum levels of L-FABP in healthy volunteers and ESRD patients were quantified with L-FABP ELISA. The values of L-FABP in patients with ESRD were significantly lower than those in the control group. Our results demonstrated the biological activity of L-FABP in human cells suggesting L-FABP can be a mediator of inflammation.

• Journal of Microbiology and Biotechnology
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• v.25 no.5
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• pp.738-744
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• 2015

Tuberculosis is one of the most threatening infectious diseases to public health all over the world, for which Mycobacterium tuberculosis (MTB) is the etiological agent of pathogenesis. Ursolic acid (UA) has immunomodulatory function and exhibits antimycobacterial activity. However, the intracellular killing effect of UA has yet to be elucidated. The aim of this study was to evaluate the intracellular killing effect of UA during mycobacterial infection. The intracellular killing activity of UA was evaluated in the macrophage cell line THP-1 by the MGIT 960 system as well as by CFU count. The production of reactive oxygen species (ROS) and the level of nitric oxide (NO) were measured using DCF-DA and Griess reagent, respectively. Phagocytosis was observed by a fluorescence-based staining method, and the colony forming units were enumerated on 7H11 agar medium following infection. In addition, MRP8 mRNA expression was measured by qRT-PCR. UA significantly decreased the number of intracellular Mycobacterium through generation of ROS and NO. In addition, it profoundly activated the phagocytosis process of THP-1 cells during MTB-infection. Furthermore, our data demonstrated that UA activated the phagocytosis process in human monocyte cells through MRP8 induction. These data suggest that UA firmly contributes to the intracellular killing effect of macrophages during mycobacterial infection.

• Journal of the Korea Convergence Society
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• v.13 no.5
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• pp.113-117
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• 2022

In this study, the antibacterial and phagocytosis regulation effects of Hordeum vulgare extract and pine needle extract on S. mutans, the causative bacteria of dental caries, were investigated. Ethanol extracts of domestic Hordeum vulgare powder and pine needle powder were used, and the antibacterial and phagocytic ability against S. mutans was confirmed according to the concentration of the extracts. As a result, S. mutans colony formation did not show a significant difference in the Hordeum vulgare extract but was significantly decreased in the pine needle extract. As a result of confirming the phagocytic ability of THP-1 cells for S. mutans, there was no significant difference in the Hordeum vulgare extract, but the phagocytic ability of immune cells was improved in the pine needle extract. Therefore, it suggests that pine needle extract can be used as a material for preventing dental caries.

• BMB Reports
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• v.46 no.7
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• pp.352-357
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• 2013

Atherosclerosis, which manifests as acute coronary syndrome, stroke, and peripheral arterial diseases, is a chronic inflammatory disease of the arterial wall. Prunella vulgaris, a perennial herb with a worldwide distribution, has been used as a traditional medicine in inflammatory disease. Here, we investigated the effects of P. vulgaris ethanol extract on TNF-${\alpha}$-induced inflammatory responses in human aortic smooth muscle cells (HASMCs). We found that P. vulgaris ethanol extract inhibited adhesion of monocyte/macrophage-like THP-1 cells to activated HASMCs. It also decreased expression of intercellular adhesion molecule-1, vascular cell adhesion molecule-1, E-selectin and ROS, No production in TNF-${\alpha}$-induced HASMCs and reduced NF-${\kappa}B$ activation. Furthermore, P. vulgaris extract suppressed TNF-${\alpha}$-induced phosphorylation of p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinase (ERK). These results demonstrate that P. vulgaris possesses anti-inflammatory properties and can regulate TNF-${\alpha}$-induced expression of adhesion molecules by inhibiting the p38 MAPK/ERK signaling pathway.

• Journal of Korean Neurosurgical Society
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• v.55 no.5
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• pp.237-243
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• 2014

Objective : Symptomatic disc degeneration develops from inflammatory reactions in the annulus fibrosus (AF). Although inflammatory mediators during annular inflammation have been studied, the roles of matrix metalloproteinases (MMPs) and their inhibitors have not been fully elucidated. In this study, we evaluated the production of MMPs and tissue inhibitors of metalloproteinase (TIMPs) during annular inflammation using an in vitro co-culture system. We also examined the effect of notochordal cells on annular inflammation. Methods : Human AF (hAF) pellet was co-cultured for 48 hours with phorbol myristate acetate-stimulated macrophage-like THP-1 cells. hAF pellet and conditioned media (CM) from co-cultured cells were assayed for MMPs, TIMPs, and insulin-like growth factor (IGF)-1 levels using real-time reverse-transcriptase polymerase chain reaction and enzyem-linked immunosorbent assay. To evaluate whether notochordal cells affected MMPs or TIMPs production on annular inflammation, hAF co-cultured with notochordal cells from adult New Zealand White rabbits, were assayed. Results : MMP-1, -3, -9; and TIMP-1 levels were significantly increased in CM of hAF co-cultured with macrophage-like cells compared with hAF alone, whereas TIMP-2 and IGF-1 levels were significantly decreased (p<0.05). After macrophage exposure, hAF produced significantly more MMP-1 and -3 and less TIMP-1 and -2. Interleukin-$1{\beta}$ stimulation enhanced MMP-1 and -3 levels, and significantly diminished TIMP-2 levels. Co-culturing with rabbit notochordal cells did not significantly influence MMPs and TIMPs production or COL1A2 gene expression. Conclusion : Our results indicate that macrophage-like cells evoke annular degeneration through the regulation of major degradative enzymes and their inhibitors, produced by hAF, suggesting that the selective regulation of these enzymes provides future targets for symptomatic disc degeneration therapy.