• Journal of Life Science
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• v.18 no.4
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• pp.530-536
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• 2008

To investigate the molecular mechanism of the airway inflammation by Pseudomonas fluorescens, effects on the inflammatory mediators such as interleukin-8 (IL-8), cyclooxygenase-2 (COX-2), macophage inhibitory cytokine 1 (MIC-1) were assessed in the human alveolar epithelial cells. Exposure to P. fluorescens and its recombinant bacteria suppressed cellular viability in the A549 epithelial cells and pro-inflammatory cytokine interleukin-8 production. However, pro-inflammatory prostaglandin-producing COX-2 protein was not altered by P. fluorescens though its mRNA was slightly elevated. As the inhibitory cytokine for the pro-inflammatory mediators, MIC-1 expression was monitored in A549 cells. MIC-1 gene induction was not significantly enhanced but the protein processing was changed by exposure to P. fluorescens. Pro-protein form of MIC-1 (${\sim}40\;kD$) was cleaved into active form mature MIC-1 (${\sim}15\;kD$) and propeptide (${\sim}28\;kD$) by the bacteria exposure. MIC-1 activation can contribute to the suppression of cellular viability by P. fluorescens and can retard IL-8-induced monocyte recruitment. However, sustained activation of MIC-1 can mediate the tissue injury by P. fluorescens exposure.

• Journal of Nutrition and Health
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• v.52 no.6
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• pp.515-528
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• 2019

Purpose: This study examined the immunological activity and optimized the mixture conditions of Sargassum horneri (S. horneri) extracts in vitro and in vivo models. Methods: S. horneri was extracted using three different methods: hot water extraction (HWE), 50% ethanol extraction (EE), and supercritical fluid extraction (SFE). Splenocyte proliferation and cytokine production (Interleukin-2 and Interferon-γ) were measured using a WST-1 assay and enzyme-linked immunosorbent assay, respectively. The levels of nitric oxide and T cell activation production were measured using a Griess assay and flow cytometry, respectively. The natural killer (NK) cell activity was determined using an EZ-LDH kit. Results: Among the three different types of extracts, HWE showed the highest levels of splenocyte proliferation and cytokine production in vitro. In the animal model, three different types of extracts were administrated for 14 days (once/day) at 50 and 100 mg/kg body weight. HWE and SFE showed a high level of splenocyte proliferation and cytokine production in the with and without mitogen-treated groups, whereas EE administration did not induce the splenocyte activation. When RAW264.7 macrophage cells were treated with different mixtures (HWE with 5, 10, 15, 20% of SFE) to determine the optimal mixture ratio of HWE and SFE, the levels of nitric oxide and cytokine production increased strongly in the HWE with 5% and 10% of SFE containing group. In the animal model, HWE with 5% and 10% of SFE mixture administration increased the levels of splenocyte proliferation, cytokine production, and activated CD4⁺ cell population significantly, with the highest level observed in the HWE with 5% of SFE group. Moreover, the NK cell activity was increased significantly in the HWE with 5% of SFE mixture-treated group compared to the control group. Conclusion: The optimal mixture condition of S. horneri with immune-enhancing activity is the HWE with 5% of SFE mixture. These results confirmed that the extracts of S. horneri and its mixtures are potential candidate materials for immune enhancement.

• Journal of Life Science
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• v.29 no.11
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• pp.1218-1226
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• 2019

The white-spotted flower chafer Protaetia brevitarsis seulensis is a medicinally beneficial and important edible insect species. We previously performed an in silico analysis of the Protaetia brevitarsis seulensis transcriptome to identify putative antimicrobial peptides and then tested their antimicrobial and hemolytic activities. These peptides had potent antimicrobial activities against bacteria and yeast without inducing hemolysis. In the present study, the cationic antimicrobial peptide, protaetiamycine 2, was selected for further assessment of its anti-inflammatory properties in mouse macrophage Raw264.7 cells. Protaetiamycine 2 treatment of Raw264.7 cells suppressed LPS-induced nitric oxide production and reduced the expression of inducible nitric oxide synthase and cyclooxygenase-2, as determined by real-time PCR and western blotting. The expression of proinflammatory cytokines ($TNF-{\alpha}$, IL-6, and $IL-1{\beta}$) was also attenuated through the MAPKs and $NF-{\kappa}B$ signaling. We also confirmed that protaetiamycine 2 bound to bacterial cell membranes by a specific interaction with LPS. Collectively, these data obtained from LPS-induced Raw264.7 cells indicated that protaetiamycine 2 could have both antimicrobial and anti-inflammatory properties.

• Journal of Life Science
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• v.19 no.4
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• pp.479-485
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• 2009

Bulnesia sarmienti (BS), a traditional South American herbal medicine native to Gran Chaco, has been used to treat various human ailments. We investigated the cytotoxic activities and the inhibitory effects of BS bark extract(0, 50, 100 and $200\;{\mu}g/\;mL$) on the production of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), cyclooxygenase (COX) and proinflammatory cytokines ($IL-1{\beta}$, IL-6 and $TNF-{\alpha}$) in the lipopolysaccharide (LPS) (100 ng/ml)-stimulated murine macrophage cell line RAW264.7. The levels of NO, COX, PGE2 production and proinflammatory cytokines ($IL-1{\beta}$, IL-6 and $TNF-{\alpha}$) were measured by ELISA kit. Cell viability, as measured by the MTT assay, showed that BS extract had no significant cytotoxicity in RAW264.7 cells. BS extract significantly inhibited the LPS-induced NO, $PGE_2$ and COX production accompanied by an attenuation of $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ formation in macrophages. These results suggest that BS extract has potential as an herbal medicine for the treatment of inflammatory diseases.

• Journal of Life Science
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• v.25 no.9
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• pp.993-999
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• 2015

The beetle Popillia flavosellata has been no reported its functional effects. In this study, we investigated the anti-inflammatory effect of P. flavosellata ethanol extract (PFE) on RAW 264.7 mouse macrophage cells treated with lipopolysaccharide (LPS) for the induction of inflammation. First, we examined the cytotoxicity of PFE in the RAW 264.7 cells at a concentration of 2,000 μg/ml or less. To evaluate the anti-inflammatory effects of PFE, we investigated the expression levels of proinflammatory cytokines, such as tumor necrosis factor (TNF)-α and interleukin (IL)-6, and proinflammatory enzymes, such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in LPS-induced RAW 264.7 cells. In addition, we examined whether PFE inhibited the translocation of nuclear factor kappa B (NF-κB) p65 into the nucleus in the LPS-induced RAW 264.7 cells. We found that the protein levels of TNF-α and IL-6 were decreased in the LPS-induced RAW 264.7 cells after the treatment with PFE in a dose-dependent manner. In addition, we confirmed that PFE inhibited the translocation of NF-κB p65 into the nucleus, as well as the protein expression levels of iNOS and COX-2. Accordingly, we propose that PFE exerts an anti-inflammatory effect through the down-regulation of NF-κB p65, TNF-α, IL-6, iNOS, and COX-2 via the toll like receptor (TLR)-4 inflammatory signaling pathway.

• Microbiology and Biotechnology Letters
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• v.44 no.4
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• pp.571-576
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• 2016

In this study, the anti-inflammatory activities of the 80% ethanol extract of Dystaenia takeshimana (DT) were investigated using Raw 264.7 cells treated with lipopolysaccharide (LPS). The effect of DT extract on the production of pro-inflammatory factors (iNOS, COX-2) in LPS-stimulated Raw 264.7 macrophages was examined. The cytotoxic effect of DT extract on macrophage cells (Raw 264.7) was examined by the 3-[4, 5-dimethyl-thiazol-2-yl]-2, 5-diphenyl-tetrazoliumbromide (MTT) assay. Treatment with DT extract showed 100% or more cell viability at the concentration $1,000{\mu}g/ml$. The inhibitory effect of DT extract on protein expression of inducible NOS (iNOS) and cyclooxygenase-2 (COX-2) was measured by western blotting using the concentrations 50, 100, and $500{\mu}g/ml$, with ${\beta}-actin$ used as the positive control. Consequently, the protein expression of iNOS, and COX-2 as observed by western blotting, was decreased by 56%, 61.6%, respectively with $500{\mu}g/ml$ DT extract. Inhibition of iNOS and COX-2 mRNA expression was measured by reverse transcription- polymerase chain reaction (PCR) using DT extract concentrations 50, 100, and $500{\mu}g/ml$, with GAPDH used as a positive control. Consequently, the mRNA expression of iNOS and COX-2 as observed by reverse-transcription-PCR was decreased by 77.9% and 83.3%, respectively at $500{\mu}g/ml$ concentration of DT extract. In conclusion, DT extract may affect inflammatory factors as a potential anti-inflammatory agent.

• Korean Journal of Plant Resources
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• v.31 no.5
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• pp.466-477
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• 2018

In this study, the anti-inflammatory activities of the extracts of different parts of Hovenia dulcis such as leaves, stems, and roots were investigated. Among them, the roots extract (RE) showed the most potent suppressive effect against pro-inflammatory mediators in LPS-stimulated mouse macrophage cells. RE induced dose-dependent reduction of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) and concomitantly reduced the production of NO and $PGE_2$. Additionally, pre-treatment with RE significantly suppressed the production of inflammatory cytokines, such as tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), interleukin $(IL)-1{\beta}$, and IL-6, as well as mRNA levels. Moreover, phosphorylation of mitogen-activated protein kinases (MAPKs) and nuclear translocation of nuclear factor-kappa B (NF-kB) were also strongly attenuated by RE in RAW264.7 cell. Furthermore, RE induced HO-1 expression through nuclear translocation of nuclear factor E2-related factor 2 (Nrf2) and increase HO-1 activity in RAW264.7 macrophages. Therefore, these results indicate that RE strongly inhibits LPS-induced inflammatory responses by blocking NF-kB activation, inhibiting MAPKs phosphorylation, and enhancing HO-1 expression in macrophages, suggesting that RE of H. dulicis and a major component, 27-O-protocatechuoylbetulinic acid could be applied as a valuable natural anti-inflammatory material.

• Korean Journal of Food Science and Technology
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• v.40 no.6
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• pp.707-711
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• 2008

The principal objective of the current study was to isolate a purpurogallin derivative as an oxidation product from gallic acid, in an effort to assess the anti-inflammatory effects of this compound. Purpurogallin derivative is known to be the one of the oxidation products of gallic acid. This compound has been identified as a minor chemical component in fermented tea products. It has been previously demonstrated that theaflavins, the oxidation products of catechins found in fermented tea products, exert profound antioxidant and anti-inflammatory effects. However, the biological activities of a minor chemical component in fermented teas have yet to be evaluated. Purpurogallin carboxylic acid (PCA) was identified as a major oxidation product of gallic acid from a peroxidase/hydrogen peroxide oxidation model system. The identity of the PCA was verified by $^{1}H$ NMR, $^{13}C$ NMR and MS techniques. PCA treatment significantly suppressed the generation of pro-inflammatory mediators including nitric oxide and IL-6 in lipopolysaccharide (LPS)-stimulated RAW264.7 murine macrophages. According to the nitrite assay, PCA 100, 75, and $50{\mu}g/mL$ treatment dose-dependently inhibited NO production by 57.6, 41.5, and 21.8%, respectively, in LPS-stimulated RAW264.7 murine macrophage cells. Moreover, IL-6 production was inhibited to a significant degree with PCA treatment of 100 and $75{\mu}g/mL$ at 43.1 and 23.9%, respectively. PCA treatment also significantly suppressed $PGE_2$ production at levels of 100 and $75{\mu}g/mL$. These results showed that PCA exerts inhibitory effects on the production of inflammatory mediators.

• Journal of Life Science
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• v.24 no.4
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• pp.370-376
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• 2014

Although the grasshopper Oxya chinensis sinuosa has long been used as food in Korea, there is little data on its functional effects. In this study, we investigated the anti-inflammatory effect of O. c. sinuosa ethanol extract (OCE) in RAW 264.7 mouse macrophage cells treated with lipopolysaccharide (LPS) for induction of inflammation. First, we determined that there is no cytotoxicity at $2,000{\mu}g/ml$ or less of OCE in RAW 264.7 cells. To evaluate the anti-inflammatory effects of OCE, we investigated expression levels of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-6, and pro-inflammatory enzymes such as inducible nitric oxide synthase (iNOS) and cyclo-oxygenase-2 (COX-2) in LPS-induced RAW 264.7 cells. In addition, we examined whether OCE could inhibit translocation of NF-${\kappa}B$ p65 into the nucleus in LPS induced RAW 264.7 cells. As a result, we found that the mRNA and protein levels of TNF-${\alpha}$ and IL-6 decreased in LPS-induced RAW 264.7 cells after treatment with OCE in a dose-dependent manner. In addition, we confirmed a $2,000{\mu}g/ml$ concentration of OCE inhibited translocation of NF-${\kappa}B$ p65 by immunnostaining and Western blot analysis, and a decrease in the protein expression levels of iNOS and COX-2. Accordingly, we suppose that OCE has an anti-inflammatory effect through down-regulation of TNF-${\alpha}$, IL-6, iNOS, and COX-2 related to ${\kappa}B$ p65 inflammatory signaling pathways.

• Tuberculosis and Respiratory Diseases
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• v.46 no.6
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• pp.775-785
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• 1999

Background : The herbicide paraquat can cause severe lung injury and fibrosis in experimental animals. In this study we have investigated the changes in lung endothelin-1(Et-1) levels and immunohistochemical localization in relation to treatment with cyclophosphamide and methylprednisolone in paraquat induced pulmonary fibrosis in guinea pigs. Material and methods : 29 male Hartley guinea pigs were divided into 4 groups. Group I was normal control. Paraquat was instilled into the lung of guinea pig of group II, III and IV unilaterally. Group II was treated with cyclophosphamide and methylprednisolone. Group III was treated with methlprednisolone. Group IV was not treated. The degree of fibrosis was evaluated by H-E stains and Masson's trichrome stains and cell activity was assessed by Et-1 immunohistochemical stains. Statistical evaluation was performed using the Kruskawallis oneway analysis. Results : Paraquat induced an increase in numbers of fibroblasts and total amount of lung collagen in Group IV compared to the normal controls. There was no significant difference in total numbers of fibroblasts between any of paraquat instilled groups, but there was significant increase in total amount of collagen in Group IV compared to group II and III (p<0.05). The treatment of cyclophosphamide and methyprednisolone suppressed the growths of both fibroblasts and collagen, but this suppression was stastically significant only in the case of collagen Et-1 immunoreactivities of bronchial epithelium, type II pneumocytes, endothelial cells and fibroblast in group II and III were decreased compared to those in group IV. Conclusion : These results demonstrate that Et-1 is an important contributing factor in the pathogenesis of pulmonary fibrosis. Et-1 is synthesized and released by bronchial epithelium, Type II pneumocyte, endothelial cells, alveolar macrophages and fibroblasts. Especially they are associated with alveolar macrophage and fibroblasts. We conclude that combined therapy of cyclophosphamide and methylprednisolone are more effective in the control of Et-1 expression and collagen deposition.