• BMB Reports
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• v.46 no.11
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• pp.544-549
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• 2013

High mobility group box 1 (HMGB1) is involved in the pathogenesis of vascular diseases. Unlike activated protein C (APC), the activation of PAR-1 by thrombin is known to elicit proinflammatory responses. To determine whether the occupancy of EPCR by the Gla-domain of APC is responsible for the PAR-1-dependent antiinflammatory activity of the protease, we pretreated HUVECs with the PC zymogen and then activated PAR-1 with thrombin. It was found that thrombin downregulates the HMGB1-mediated induction of both TNF-${\alpha}$ and IL-6 and inhibits the activation of both p38 MAPK and NF-${\kappa}B$ in HUVECs pretreated with PC. Furthermore, thrombin inhibited HMGB1-mediated hyperpermeability and leukocyte adhesion/migration by inhibiting the expression of cell adhesion molecules in HUVECs if EPCR was occupied. Collectively, these results suggest the concept that thrombin can initiate proinflammatory responses in vascular endothelial cells through the activation of PAR-1 may not hold true for normal vessels expressing EPCR under in vivo conditions.

• Journal of Applied Biological Chemistry
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• v.63 no.2
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• pp.117-123
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• 2020

We examined the anti-inflammatory effect of the peel extract of the newly bred Korean apple (Malus domestica Borkh.) cultivar Green ball. To test its possible use as anti-inflammatory functional material, Raw 264.7 macrophages were treated with pro-inflammatory lipopolysaccharide (LPS) in the presence or absence of Green ball apple peel ethanol extract (GBE). Notably, up to 500 ㎍/mL of GBE did not result in any signs of inhibition on cellular metabolic activity or cytotoxicity in Raw 264.7 macrophages. Supplementation with GBE to LPS-treated Raw 264.7 macrophage significantly suppressed various pro-inflammatory responses in a dose-dependent manner, including i) nitric oxide (NO) production, ii) accumulation of inducible NO synthase and cyclooxygenase-2, iii) phosphorylation of nuclear factor-kappa B (NF-κB) subunit p65, and iv) expression of pro-inflammatory biomarker genes, including tumor necrosis factor alpha, interleukin 1 beta, interleukin 6, monocyte chemoattractant protein-1, and prostaglandin E synthase 2.

• Journal of Microbiology and Biotechnology
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• v.29 no.8
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• pp.1248-1254
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• 2019

Identification of novel probiotic strains is of great interest in the field of functional foods. Specific strains of heat-killed bacteria have been reported to exert immunomodulatory effects. Herein, we investigated the immune-stimulatory function of heat-killed Lactobacillus plantarum KCTC 13314BP (LBP). Treatment with LBP significantly increased the production of $TNF-{\alpha}$ and IL-6 by macrophages. More importantly, LBP was able to enhance the phagocytic activity of macrophages against bacterial particles. Activation of p38, JNK, ERK, $NF-{\kappa}B$, and STAT3 was involved in the immunomodulatory function of LBP. LBP treatment significantly increased production of $TNF-{\alpha}$ by bone marrow-derived macrophages and splenocytes, further confirming the immunostimulatory effect of LBP in primary immune cells. Interestingly, the immunomodulatory effects of LBP were much stronger than those of Lactobacillus rhamnosus GG, a well-known probiotic strain. These results indicate that LBP can be a promising immune-enhancing functional food agent.

• Journal of Applied Biological Chemistry
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• v.57 no.4
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• pp.373-377
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• 2014

In this study, we demonstrated whether the extract of Fomes fomentarius (FFE; FF extract) could be used to stimulate macrophages (RAW 264.7 cells). All four doses of FFE (5, 10, 20, and $40{\mu}g/mL$) had no significant cytotoxicity during the entire experimental period. FFE potently increased the production of nitric oxide (NO). Consistent with these observations, inducible NO synthase levels were increased by FFE in a dose-dependent manner. Moreover, FFE increased the production of tumor necrosis factor-${\alpha}$, interleukin (IL)-$1{\beta}$, and IL-6 in the same cells. These stimulating effects of FFE were found to be caused by the stimulation of phosphorylation of $I{\kappa}B{\alpha}$ and MAP kinases (p38, ERK, and JNK). These results suggest that FFE may be used as new agents for wide application in the immune study of mushroom.

• Korean Journal of Medicinal Crop Science
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• v.24 no.3
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• pp.228-236
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• 2016

Background: Sedum takesimense Nakai has been used as folk medicine in Korea. The present study aimed to determine the biological activity of S. takesimense by investigating the anti-inflammatory effects of S. takesimense water extract (SKLC) on the lipopolysaccharide-induced inflammatory response in RAW 264.7 cells. Methods and Results: Cytotoxicity of SKLC on RAW 264.7 cells was determinded by performing MTS assay was found to have no cytotoxic effect on RAW 264.7 cells at a concentration range of $62-500{\mu}g/m{\ell}$. Further, pretreatment of SKLC inhibited lipopolysaccharide-induced nitric oxide (NO) production in a dose-dependent manner. To determined the inhibitory mechanisms of SKLC on inflammatory mediators, we assessed the inducible nitric oxide synthase (iNOS) and cyclooxygnease-2 (COX-2) pathways. The activities of these pathways were decreased in a dose-dependent manner by SKLC. The production of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin $(IL)-1{\beta}$, and IL-6 were also reduced. Conclusions: These results suggest that the down regulation of iNOS, COX-2, TNF-${\alpha}$, IL-$1{\beta}$, and IL-6 expression by SKLC are mediated by the down regulation of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activity, a transcription factor necessary for pro-inflammatory mediators. This might be the mechanism underlying the anti-inflammatory effects of SKLC.

• Korean Journal of Pharmacognosy
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• v.50 no.1
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• pp.11-17
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• 2019

Cassia tora L. have been used as a folk medicine in Korea. This study investigated anti-inflammatory effect of aurantio-obtusin isolated from C. tora. We isolated aurantio-obtusin from 50% ethanol extracts of C. tora L. We investigated the anti-inflammatory effects of aurantio-obtusin on the lipopolysaccharide (LPS)-stimulated inflammatory response in murine macrophage cell line (Raw 264.7). To investigate the cytotoxicity of aurantio-obtusin on RAW 264.7 cells, MTS assay was performed. RAW 264.7 cells were treated with aurantio-obtusin at different concentrations (12.5, 25, 50, $100{\mu}M$) for 30 h. The result showed that aurantio-obtusin had no cytotoxic effect in a concentration range of $12.5-100{\mu}M$. To determine the effect of aurantio-obtusin on LPS-induced NO production, the NO concentration measurement was performed. RAW 264.7 cells were treated with aurantio-obtusin at 12.5, 25, 50 and $100{\mu}M$ for 24 h, and the results showed that the NO production of aurantio-obtusin-treated cells compared to LPS alone treated group was significantly decreased in a dose-dependent manner. Pretreatment of aurantio-obtusin inhibited LPS-induced NO production in a dose-dependent manner. To find out inhibitory mechanisms of aurantio-obtusin on inflammatory mediators, we examined the $PGE_2$ pathways. As a result, $PGE_2$ were decreased in a dose-dependent manner by aurantio-obtusin. The release of interleukin-$1{\beta}$ (IL-$1{\beta}$) and IL-6 were also reduced. Moreover, aurantio-obtusin suppressed LPL-induced $I{\kappa}B-{\alpha}$ degradation. These results suggest that the down regulation of NO, $PGE_2$, IL-$1{\beta}$ and IL-6 expression by aurantio-obtusin are achieved by the downregulation of NF-${\kappa}B$ activity.

• The Journal of Korean Medicine
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• v.44 no.2
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• pp.119-131
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• 2023

Objectives: Betula Platyphylla(BP) has been used as a analgesic, anti-microbial, anti-oxidant drug in Eastern Asia. However, it is still unknown whether BP ethanol extract could exhibit the inhibitory activities against ultraviolet B(UVB)-induced skin injury on human keratinocytes, HaCaT cells. This study was aimed to investigate the protective activity of BP ethanol extract on UVB-irradiated skin injury in HaCaT cells. Methods: The skin injury model of HaCaT cells was established under UVB stimulation. HaCaT keratinocyte cells were pre-treated with BP ethanol extract for 1 h, and then stimulated with UVB. Then, the cells were harvested to measure the cell viability, production of reactive oxygen species(ROS), pro-inflammatory cytokines such as interleukin(IL) 1-beta, IL-6, and tumor necrosis factor(TNF)-𝛼, hyaluronidase, type 1 collagen, matrix metalloproteinase(MMP)s. In addition, we examined the mitogen activated protein kinases(MAPKs) and inhibitory kappa B alpha(I𝜅;-B𝛼) as inhibitory mechanisms of BP ethanol extract. Results: The treatment of BP ethanol extract inhibited the UVBinduced cell death and ROS production in HaCaT cells. BP ethanol extract treatment inhibited the UVB-induced increase of IL-1beta, IL-6, and TNF-𝛼. BP ethanol extract treatment inhibited the increase of hyaluronidase, MMP and decrease of collagen. BP ethanol extract treatment inhibited the activation of MAPKs and the degradation of I𝜅-B𝛼. Conclusions: Our result suggest that treatment of BP ethanol extract could inhibit the UVB-induced skin injury via deactivation of MAPKs and nuclear factor kappa B(NF-𝜅B) in HaCaT cells. This study could suggest that BP ethanol extract could be a beneficial agent to prevent skin damage or inflammation.

• Journal of Applied Biological Chemistry
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• v.66
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• pp.282-288
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• 2023

Due to the COVID-19 pandemic, the immuneenhancing health functional food market that protects our bodies from pathogens such as viruses continues to grow. In this study, we aimed to prove the Cheonggukjang, a high-nutrient food with high protein, fat, and dietary fiber content, as an immuneenhancing nutraceutical. Cheonggukjang water extract (CWE) increased the production of nitric oxide, reactive oxygen species, and cytokines such interleukin (IL)-6, IL-1β, and tumor necrosis factor-α without affecting viability in RAW 264.7 cells. Furthermore, CWE significantly upregulated the expression of inducible nitric oxide synthase and cyclooxygenase-2 in RAW 264.7 cells. CWE enhanced the phosphorylation of I kappa B kinase α/β and I kappa B (IκB)α, as well as the degradation of IκBα. CWE also induced increased phosphorylation of nuclear factor-kappa B p65 and facilitated the redistribution of p65 from the cytoplasm to the nucleus in RAW 264.7 cells. These findings suggest that CWE has potential as a health functional food material that can enhance the innate immune response.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.04a
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• pp.13-13
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• 2018

The anti-inflammatory (INF) compounds (1-15) were isolated from Agrimonia pilosa Ledeb. (APL) by activity-guided isolation technique. The isolated compounds (1-15) were identified as quercetin-7-O-rhanmoside (1), apigenin-7-O-glycoside (2), kaempferol-7-O-glycoside (3), apigenin-7-O-[6"-(butyl)-glycoside] (4), querceitn (5), kaempferol (6), apigenin (7), apigenin-7-O-[6"-(pentyl)-glycoside] (8), agrimonolide (9), agrimonolide-6-O-glucoside (10), desmethylagrimonolide (11), desmethylagrimonolide-6-O-glucoside (12), luteolin (13), vitexin (14) and isovitexin (15). Flavonoids, compound 2, 3, 11, and 14-15 have been found in APL for the first time. Furthermore, two novel flavone derivatives, compound 4 and 8, have been isolated inceptively in plant. In the no cytotoxicity concentration ranges of $0-20{\mu}M$, nitric oxide (NO) production level of 1-15 was estimated in LPS-treated Raw 264.7 macrophage cells. The flavone aglycones, 7 (apigenin, $IC_{50}=3.69{\pm}0.34{\mu}M$), 13 (luteolin, $IC_{50}=4.62{\pm}0.43{\mu}M$), 6 (kaempferol, $IC_{50}=14.43{\pm}0.23{\mu}M$) and 5 (quercetin, $IC_{50}=19.50{\pm}1.71{\mu}M$), exhibited excellent NO inhibitory (NOI) activity in dose-dependent manner. In the structure activity relationship (SAR) study of apigenin-derivatives (APD), apigenin; Api, apigenin-7-O-glucoside; Api-G, apignenin-7-O-[6"-(butyl)-glycoside]; Api-BG and apignenin-7-O-[6"-(pentyl)-glycoside]; Api-P, from APL on INF activity was investigated. The INF mediators level such as NO, INF-cytokines, NF-KB proteins, iNOS and COX-2 were sharply increased in Raw 264.7 cells by LPS. When pretreatment with APD in INF induced macrophages, NOI activity of Api was most effective than other APD with $IC_{50}$ values of $3.69{\pm}0.77{\mu}M$. And the NOI activity was declined in the following order: Api-BG ($IC_{50}=8.91{\pm}1.18{\mu}M$), Api-PG ($IC_{50}=13.52{\pm}0.85{\mu}M$) and API-G ($IC_{50}=17.30{\pm}0.66{\mu}M$). The NOI activity of two novel compounds, Api-PG and Api-BG were lower than their aglycone; Api, but more effective than Api-G (NOI: Api-PG and Api-BG). And their suppression ability on INF cytokines such as $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6 mRNA showed the similar tendency. Therefore, the anti-INF mechanism study of Api-PG and Api-BG on nuclear factor-kappa B ($NF-{\kappa}B$) pathway, representative INF mechanism, was investigated and Api was used as positive control. Api-BF was more effectively prevent the than phosphorylation of $pI{\kappa}B$ kinase (p-IKK) and p65 than Api-PG in Raw 264.7 cells. In contrast, Api-PG and Api-BG were not reduced the phosphorylation of inhibitor of kappa B alpha ($I{\kappa}B{\alpha}$). Moreover, pretreatment with Api-PG and Api-BG, dose-dependently inhibited LPS-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) mRNAs and proteins in macrophage cells, and their expression were correlated with their NOI activity. Therefore, APL can be utilized to health promote agent associated with their AIN metabolites.

• Food Science and Preservation
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• v.22 no.5
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• pp.735-743
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• 2015

This study investigated the anti-inflammatory activity of barley leaf extract in lipopolysaccharide (LPS)-stimulated RAW264.7 cells and hairless mice. Pre-treatment with barley leaf extract significantly inhibited the protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-II (COX-II) in a dose-dependent manner in LPS-stimulated RAW264.7 cells. Barley leaf extract also significantly inhibited the secretion of inflammatory cytokines, such as tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), and interleukin-6 (IL-6). Moreover, phosphorylation of mitogen-activated protein kinases (MAPKs) and nuclear translocation of nuclear factor-kappa B (NF-${\kappa}B$) were strongly suppressed by barley leaf extract in LPS-stimulated cells. In hairless mice, barley extract significantly decreased the pathological phenotypes of contact dermatitis, such as erythema, edema, and scabs. These results indicate that barley leaf extract has an anti-inflammatory effect and therefore a possible role in the treatment of inflammatory diseases or in functional cosmetics.