• Korean Journal of Clinical Laboratory Science
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• v.52 no.3
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• pp.245-252
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• 2020

Macrophage cell death contributes to the formation of plaque, leading to the development of atherosclerosis. The accumulation of triglyceride (TG) is also associated with the pathogenesis of atherosclerosis. A previous study reported that TG induces the cell death of macrophages. This study examined whether the cytoplasmic release of cathepsin B from lysosome is associated with the TG-induced cell death of macrophage. The release of cathepsin B was increased in the TG-treated THP-1 macrophages, but the TG treatment did not affect cathepsin B expression. Furthermore, the inhibition of cathepsin B by its inhibitor, CA-074 Me, partially inhibited the TG-induced cell death of macrophage. TG-triggered macrophage cell death is mediated by the activation of caspase-1, -2, and apoptotic caspases. Therefore, this study investigated whether cathepsin B is implicated in the activation of these caspases. The inhibition of cathepsin B blocked the activation of caspase-7, -8, and -1 but did not affect the activity of caspase-3, -9, and -2. Overall, these results suggest that TG-induced cytoplasmic cathepsin B causes THP-1 macrophage cell death by activating caspase-1, leading to subsequent activation of the extrinsic apoptotic pathway.

• The Journal of Pediatrics of Korean Medicine
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• v.20 no.1
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• pp.241-255
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• 2006

Objective : Allergic Inflammation is related with secretion of Cytokine. This study was performed to examine the effects of Woobangja on anti-allergic inflammation. Method : While macrophage 264.7cells was chosen as a normal group a control group was classified into three groups. One was stimulated with LPS. and another was pretreated with Woobangja for 1 hour. The third was pretreated with gydrocortisone for 1 hour. After the pretreatment, macrophage were incubated with lipopolysaccharide(LPS) 100 ng/ml for 12h and media collected and $TNF-{\alpha}$, IL-6, $IL-1{\beta}$, IL-10 concentration in supernatants were measured each by Enzyme linked immuno-sorbent assay. Woobangja were used $50\;{\mu}g/ml$, $100\;{\mu}g/ml$, $250\;{\mu}g/ml$, $500\;{\mu}g/ml$, 1 mg/ml. Hydrocortisones were used respectively $10^{-8}\;M$,$10^{-7}\;M$,$10^{-6}\;M$,$10^{-5}\;M$,$10^{-4}\;M$. Results : Woobangja showed inhibitory effect on $TNF-{\alpha}$ by LPS-stimulated macrophage 264.7. The inhibitory effect was most significant in 1mg/ml(p<0.01), and has increased according to the number of doses. Woobangja also showed inhibitory effect on IL-10 by LPS-stimulated macrophasg 264.7. The inhibitory effect was most significant in $100\;{\mu}g/ml$, and was not in a dose-dependent manner as Hydrocortisone group. Woobangja and Hydrocortison showed contrary effect on $IL-1{\beta}$ in al five concentration(p<0.01), and at the lowest concentration ($50\;{\mu}g/ml$) the level of $IL-1{\beta}$ was the lowest. On the other hand hydrocortison was observed to have inhibitory effect on $IL-1{\beta}$ in all five concentration(p<0.01). IL-6 was inhibited by hydrocortison in a roughly dose-dependent manner, but was not inhibited by Woobangja. On the contrary Woobangja obviously increased the expression of $IL-1{\beta}$ in all five concentration(p<0.01), but it was not related with concentrations. Conclusion : 1. Woobangja does significantly inhibit the expression of $TNF-{\alpha}$ by LPS-stimulated macrophage 264.7. 2. Woobangja does significantly increse the expression of IL-6 by LPS-stimulated macrophage 264.7. 3. Woobangja does significantly increse the expression of $IL-1{\beta}$ by LPS-stimulated macrophage 264.7. 4. Woobangja does significantly inhibit the expression of IL-10 by LPS-stimulated macrophage 264.7. 5. Woobangja is observer to have anti-allergic inflammatory effect through inhibiting inflammatory cytokine.

• Molecules and Cells
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• v.38 no.10
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• pp.886-894
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• 2015

Macrophages are divided into two subpopulations: classically activated macrophages (M1) and alternatively activated macrophages (M2). BCG (Bacilli Calmette-$Gu{\acute{e}}rin$) activates disabled $na{\ddot{i}}ve$ macrophages to M1 macrophages, which act as inflammatory, microbicidal and tumoricidal cells through cell-cell contact and/or the release of soluble factors. Various transcription factors and signaling pathways are involved in the regulation of macrophage activation and polarization. We discovered that BCG-activated macrophages (BAM) expressed a new molecule, and we named it Novel Macrophage Activated Associated Protein 1 (NMAAP1). 1 The current study found that the overexpression of NMAAP1 in macrophages results in M1 polarization with increased expression levels of M1 genes, such as inducible nitric oxide synthase (iNOS), tumor necrosis factor alpha (TNF-${\alpha}$), Interleukin 6 (IL-6), Interleukin 12 (IL-12), Monocyte chemoattractant protein-1 (MCP-1) and Interleukin-1 beta (IL-$1{\beta}$), and decreased expression of some M2 genes, such as Kruppel-like factor 4 (KLF4) and suppressor of cytokine signaling 1 (SOCS1), but not other M2 genes, including arginase-1 (Arg-1), Interleukin (IL-10), transforming growth factor beta (TGF-${\beta}$) and found in inflammatory zone 1 (Fizz1). Moreover, NMAAP1 overexpression in the RAW264.7 cell line increased cytotoxicity against MCA207 tumor cells, which depends on increased inflammatory cytokines rather than cell-cell contact. NMAAP1 also substantially enhanced the phagocytic ability of macrophages, which implies that NMAAP1 promoted macrophage adhesive and clearance activities. Our results indicate that NMAAP1 is an essential molecule that modulates macrophages phenotype and plays an important role in macrophage tumoricidal functions.

• Journal of Digital Convergence
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• v.16 no.11
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• pp.613-620
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• 2018

This study was conducted to investigate of Mori Folium Water Extract (MF) on anti-inflammation activity. MF Water extracts after 24 houres cultivation were examined to ascertain the cell viability of mouse macrophage RAW 264.7 cells. The influence of the Water extracts in RAW 264.7 macrophage cells treated with LPS was investigated. nitric oxide (NO) production, nterleukin$(IL)-1{\alpha}$ IL-6 and IL-10 increased generation of cytokines. mouse macrophage RAW 264.7 cells cell viability changes were no decreas after MTT assay of MF Water extract. The MF water extracts inhibited NO generation caused by LPS in the macrophages over $25{\mu}g/mL$. The MF water extracts increased in the control group the $IL-1{\alpha}$ and IL-6 activation generated by LPS in the macrophages over $50{\mu}g/mL$. Accordingly, it was found that different MF water extract concentrations significantly influenced certain anti-inflammation activities in RAW 264.7 macrophage cells. The results of this study are expected to be highly applicable to health - friendly functional materials. Further studies are needed to confirm the signaling pathways associated with anti-inflammation of macrophages through continuous studies.

• Journal of Society of Preventive Korean Medicine
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• v.16 no.2
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• pp.113-124
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• 2012

The oxidative modification of low density lipoprotein(LDL) has been implicated in the development of atherosclerosis. Oxidized LDL(oxLDL) is captured into macrophage and stimulates to form macrophage foam cell. And it can induce an inflammation and smooth muscle proliferation in atherosclerotic plaque. Objective : In this study, we aimed to investigate the effect of Bupleuri radix(SH) on the foam cell formation, a critical initiation stage of atherosclerosis. Methods : To achieve the goal, we examined the effect of SH on LDL oxidation, nitric oxide production in RAW264.7, and the effect of SH on cupuric sulfate-induced cytotoxicity, LDH release, and macrophage activity. Results : SH inhibited the formation of oxidized LDL from native LDL in RAW264.7 cell culture, and decreased the release of LDH from cupric sulfate-stimulated RAW264.7 cell. In other experiments, SH activated RAW264.7 cell, and prolonged the survival time, and inhibited foam cell formation induced by oxLDL in Raw 264.7 cells. Conclusion : These results showed that SH might prevent atherosclerosis by controlling the early stages of foam cell formation.

• Journal of Acupuncture Research
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• v.27 no.4
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• pp.223-231
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• 2010

Objectives : The objective of this study is to study the effects of Euonymi Lignum Suberatatum pharmacopuncture solution on NO and IL-6 production in macrophage. Methods : At first, the RAW 264.7 macrophage was subclutured. In order to evaluate cytotoxicity, MTT assay performed. Then, the cell was induced by LPS, INF-$\gamma$ and Experimental groups were divided into five(Normal, Control, Euonymi Lignum Suberatatum 100, 200, $300{\mu}g/m{\ell}$). Then Euonymi Lignum Suberatatum pharmacopuncture solution was put into cell. We measured IL-6, iNOS, NO. Results : The cytotoxic effect of Euonymi Lignum Suberatatum pharmacopuncture solution in RAW 264.7 macrophage was not appeared. $300{\mu}g/m\ell$ Euonymi Lignum Suberatatum pharmacopuncture solution inhibited IL-6 production in LPS, INF-$\gamma$-stimulated RAW 264.7 macrophages significantly. Euonymi Lignum Suberatatum pharmacopuncture solution inhibited iNOS revelation in LPS, INF-$\gamma$-stimulated RAW 264.7 macrophages. All group of Euonymi Lignum Suberatatum pharmacopuncture solution inhibited NO production in LPS, INF-$\gamma$-stimulated RAW 264.7 macrophages significantly. Conclusions : Our study demonstrated that Euonymi Lignum Suberatatum pharmacopuncture solution had an inhibition effect on NO production, iNOS revelation, IL-6 production. So Euonymi Lignum Suberatatum pharmaco puncture solution may have an Anti-inflammation effect.

• Journal of the Korean Society of Visualization
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• v.15 no.3
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• pp.41-46
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• 2017

The apoptosis of macrophages occurs throughout all stages of atherosclerosis. It is known to constitute atheromatous plaque, increase plaque instability, and thus contribute to the development of atherosclerosis. However, there still remains much to be elucidated on how the apoptotic macrophages affect the endothelial cells and also how they contribute to the development of atherosclerosis. Here we present a microfluidic system, which enables co-culture of apoptotic macrophages and endothelial cells in fibrin gel that mimics in vivo extracellular matrix. With the system, we can investigate the effect of macrophage apoptosis on vascular endothelial cells by quantitatively analyzing the level of reactive oxygen species of HUVECs, integrity of VE-cadherin and cell proliferation. We expect that this system could be utilized further for understanding different mechanisms of apoptotic macrophage on the development of atherosclerosis.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.4
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• pp.815-820
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• 2008

Macrophage is the important cell for the immune system. Many of herbal drugs were searched about their immune-modulating activity. The purpose of this study is to investigate the biological effect of water extract from ARTEMISIAE ARGI FOLIUM (WAAF) on mouse macrophage Raw 264.7 cells. ARTEMISIAE ARGI FOLIUM was known to have the antibacterial, immune-enhancing, and anticoagulative properties. Cytotoxicity of WAAF was verified by MTT assay. The intracellular production of hydro peroxide ($H_2O_2$) by WAAF was examined. The productions of nitric oxide (NO) and $TNF-{\alpha}$ from Raw 264.7 cell by WAAF were also examined. WAAF showed no cytotoxicity on RAW 264.7 cells for 3 hours. WAAF increased the production of $H_2O_2$ in Raw 264.7 cells. WAAF decrease the production of NO from the cells at low concentrations but increased at high concentrations. WAAF increased the production of $TNF-{\alpha}$ from the cells. Therefore, It could be suggested that WAAF has the immune-modulating effect.

• Journal of Microbiology and Biotechnology
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• v.4 no.3
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• pp.195-199
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• 1994

Immunostimulation effects of the cell wall components isolated from Lactobacillus plantarum were investigated by studying the macrophage s tumorcidal activity, splenocyte proliferation, anticomplementary activity and the inhibition of peritoneal tumor cell growth measured with ICR mice inoculated with sarcoma 180. The immunopotentiating cell wall components were a complex of peptidoglycan and exopolysaccharides. The tumorcidal activity of macrophage against Yacl and B16 tumor cells was enhanced when the cell wall components were added into the macrophage s culture medium. They also stimulated splenocytes to proliferate up to the same level as when the concanavalin A was added into the splenocyte's culture medium. The complementary activity was inhibited by 50% when the cell wall components were incubated with the sheep red blood cells treated with hemolysin and guinea pig complement. This result confirmed that the cell wall components had an antitumor effect, because the anticomplementary activity is usually accompanied by an antitumor activity at the same time. This fact was confirmed again by the inhibition of the growth of sarcoma 180 when the cell wall components were injected intraperitoneally into ICR mice inoculated with sarcoma 180. As a result, it is concluded that the cell wall components isolated from Lactobacillus plantarum had multifunctional immunostimulation effects in vitro and in vivo.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.8
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• pp.1145-1149
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• 2004

Immunomodulatory feed additives might offer alternatives to antimicrobial growth promoters in poultry production. This experiment was carried out to test the effect of $\beta$-glucan supplementation on the growth performance and immune response in broilers. Total of 160 day-old broilers were randomly assigned to 4 treatment groups fed corn-soybean diets containing 0, 0.012, 0.025 or 0.05% of $\beta$-glucan supplement in a 6 week feeding experiment. Growth performance, antibody titer against New Castle vaccine, lymphocyte blastogensis, and peritoneal macrophage chemotaxis activity of broilers were evaluated. Results showed that there were no significant differences in weight gain and feed efficiency among the treatments, and no differences in antibody titer was observed. Supplementation of $\beta$-glucan did not elevate the lymphocyte blastogensis among treatments, following stimulation with different mitogens. However, supplementation with 0.025 and 0.05% $\beta$-glucan enhanced the macrophage chemotaxis activity of broilers. These results suggest that $\beta$-glucan may enhance some cell-mediated immune responses of chickens by modulate macrophages ability.