Kudoa septempunctata is a myxozoan parasite that causes food poisoning in individuals consuming olive flounder. The present study aimed to investigate the currently insufficiently elucidated early molecular mechanisms of inflammatory responses in the intestine owing to parasite ingestion. After Kudoa spores were isolated from olive flounder, HT29 cells were exposed to spores identified to be alive using SYTO-9 and propidium iodide staining or to antigens of Kudoa spores (KsAg). IL-1β, IL-8, TNF-α and NFKB1 expression and NF-κB activation were assessed using real-time PCR, cytokine array and western blotting. The immunofluorescence of FITC-conjugated lectins, results of ligand binding assays using Mincle-Fc and IgG-Fc, CLEC4E expressions in response to KsAg stimulation, and Mincle-dependent NF-κB activation were assessed to clarify the early immune-triggering mechanism. Inflammatory cytokines (IL-1β, GM-CSF and TNF-α), chemokines (IL-8, CCL2, CCL5 and CXCL1) and NF-κB activation (pNF-κB/NF-κB) in HT29 cells increased following stimulation by KsAg. The immunofluorescence results of spores and lectins (concanavalin A and wheat germ agglutinin) suggested the importance of Mincle in molecular recognition between Kudoa spores and intestinal cells. Practically, data for Mincle-Fc and KsAg binding affinity, CLEC4E mRNA expression, Mincle immunofluorescence staining and hMincle-dependent NF-κB activation demonstrated the involvement of Mincle in the early immune-triggering mechanism. The present study newly elucidated that the molecular recognition and immune-triggering mechanism of K. septempunctata are associated with Mincle on human intestinal epithelial cells.
Journal of the Korean Applied Science and Technology
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v.38
no.1
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pp.217-227
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2021
In this study, Korean-cultivated Houttuynia cordata Thunb. (HC) was extracted anti-inflammatory effects of extracts were compared in order to confirm the possibility of natural cosmetics as a raw material. HPLC pattern analysis, cell viability assay, total phenolic contents, ABTS/DPPH radical scavenging analysis, measurement of ROS production, griess reagent assay, and luminex technique. The content of quercetin was higher in fresh Houttuynia cordata (HC-F) than in dried Houttuynia cordata (HC-D). Also, content of HC extracts shows a difference in the content of the two representative compounds (polyphenol and flavonoid) according to the stored differently. ROS production showed higher inhibitory rates at HC-F. The anti-inflammatory activity of HC-D was shown higher than that of HC-F. But, in the measurement of the reduction in inflammatory cytokines, IL-1β showed that HC-D had a relatively high reduction effect, and IL-6 and TNF-α showed a high reduction effect. And these results will be a basis for the development of cosmetology in skin care and improvement of skin problem.
Background: Panax ginseng Meyer (P. ginseng), a herb distributed in Korea, China and Japan, exerts benefits on diverse inflammatory conditions. However, the underlying mechanism and active ingredients remains largely unclear. Herein, we aimed to explore the active ingredients of P. ginseng against inflammation and elucidate underlying mechanisms. Methods: Inflammation model was constructed by lipopolysaccharide (LPS) in C57BL/6 mice and RAW264.7 macrophages. Molecular docking, molecular dynamics, surface plasmon resonance imaging (SPRi) and immunofluorescence were utilized to predict active component. Results: P. ginseng significantly inhibited LPS-induced lung injury and the expression of proinflammatory factors, including TNF-α, IL-6 and IL-1β. Additionally, P. ginseng blocked fluorescencelabeled LPS (LPS488) binding to the membranes of RAW264.7 macrophages, the phosphorylation of nuclear factor-κB (NF-κB) and mitogen-activated protein kinases (MAPKs). Furthermore, molecular docking demonstrated that ginsenoside Ro (GRo) docked into the LPS binding site of toll like receptor 4 (TLR4)/myeloid differentiation factor 2 (MD2) complex. Molecular dynamic simulations showed that the MD2-GRo binding conformation was stable. SPRi demonstrated an excellent interaction between TLR4/ MD2 complex and GRo (KD value of 1.16 × 10-9 M). GRo significantly inhibited LPS488 binding to cell membranes. Further studies showed that GRo markedly suppressed LPS-triggered lung injury, the transcription and secretion levels of TNF-α, IL-6 and IL-1β. Moreover, the phosphorylation of NF-κB and MAPKs as well as the p65 subunit nuclear translocation were inhibited by GRo dose-dependently. Conclusion: Our results suggest that GRo exerts anti-inflammation actions by direct inhibition of TLR4 signaling pathway.
The effect of Saccharomycopsis fibuligera fermentation on the antioxidant and anti-inflammatory activity of Kerria japonica (K. japonica) extracts was studied. First, the antioxidant activity of the fermented extract was measured using the 2,2-diphenyl1-picrylhydrazyl (DPPH) and 2,2'-azinobis (3-ethylbenzthiazoline 6-sulfonic acid (ABTS) methods. Also, the quantification of polyphenols and flavonoids, which are representative components with antioxidant activity, was performed. The results of the DPPH and ABTS assays showed an increase in the antioxidant activity by 14.39% and 21.74%, respectively, due to fermentation. The polyphenol concentration increased by 10.5%, and the flavonoid concentration increased by 100.0%. In the cell experiment, a cytotoxicity test and a nitric oxide (NO) production inhibitory test were performed using RAW 264.7 cells. Both the control group and the fermentation group showed no cytotoxicity. In the NO production inhibition experiment, the fermentation group showed a 6.85% higher inhibition of NO production compared to the control group. When the inhibitory effects of the extracts on inflammatory cytokine production were assessed, the fermentation group showed 12.4% and 23.5% higher inhibition of interleukin (IL)-1β and IL-6 production, respectively, compared to the control group. In conclusion, due to its potential for inhibiting NO and inflammatory cytokine production, fermented K. japonica extracts could be considered a source of anti-inflammatory compounds.
Ji-Won, Park;Jin-Mi, Park;Sangmi, Eum;Jung Hee, Kim;Jae Hoon, Oh;Jinseon, Choi;Tran The, Bach;Nguyen, Van Sinh;Sangho, Choi;Kyung-Seop, Ahn;Jae-Won, Lee
Microbiology and Biotechnology Letters
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v.50
no.4
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pp.574-583
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2022
Ficus vasculosa Wall. ex Miq. (FV) has been used as a herbal medicine in Southeast Asia and its antioxidant activity has been shown in previous studies. However, it has not yet been elucidated whether FV exerts anti-inflammatory effects on activated-macrophages. Thus, we aimed to evaluate the ameliorative property of FV methanol extract (FM) on lipopolysaccharide (LPS)-induced inflammatory responses and the underlying molecular mechanisms in RAW264.7 macrophages. The experimental results indicated that FM decreased the production of inflammatory mediators (NO/PGE2) and the mRNA/protein expression of iNOS and COX-2 in LPS-stimulated RAW264.7 cells. FM also reduced the secretion of interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α and monocyte chemoattractant protein (MCP)-1 in LPS-stimulated RAW264.7 cells. Results also demonstrated that FM improved inflammatory response in LPS-stimulated A549 airway epithelial cells by inhibiting the production of cytokines, such as IL-1β, IL-6 and TNF-α. In addition, FM suppressed MAPK activation and NF-κB nuclear translocation induced by LPS. FM also upregulated the mRNA/protein expression levels of heme oxygenase-1 and the nuclear translocation of nuclear factor erythroid 2-related factor 2 in RAW264.7 cells. In an experimental animal model of LPS-induced acute lung injury, the increased levels of molecules in bronchoalveolar lavage (BAL) fluid were suppressed by FM administration. Collectively, it was founded that FM has anti-inflammatory properties on activated-macrophages by suppressing inflammatory molecules and regulating the activation of MAPK/NF-κB signaling.
You Bin, Shin;Han Byeol, Park;Jae Su, Kim;Hyun Jong, Lee;Sung Chul, Lim;Yun Kyu, Lee
Korean Journal of Acupuncture
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v.39
no.4
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pp.152-171
/
2022
Objectives : This study was designed to investigate the effects of Sunbanghwalmyung-eum gamibang on Monosodium iodoacetate-induced osteoarthritis rats. Methods : Forty Sprague-Dawley (SD) rats were divided into 5 groups of 8 rats each. Osteoarthritis (OA) was induced by injecting MIA (2 mg/50 µl) into the joint cavity of the left knee of SD rats belonging to the experimental group, and normal saline was injected into the joint cavity of the left knee instead of MIA in the normal group. To the normal group and the controlled group (OA group), 2 ml of distilled water was orally administered. To the positive control group (Indomethacin group), indomethacin 2 ml at a concentration of 2 mg/kg, to the low concentration group of SHG (Low group), 2 ml of SHG at a concentration of 2 mg/kg, and to the high concentration group of SHG (High group), 2 ml of SHG at a concentration of 4 mg/kg ml was orally administered. The drug was administered for a total of 4 weeks, and histological changes were analyzed by Hematoxylin-Eosin staining and Safranin-O staining. In addition, inflammatory cytokines such as TNF-α, IL-1β, and IL-6, and MMP-13, TIMP-1, and GAGs were immunohistochemically analyzed. Finally, hematological examination, blood biochemical examination, and liver and kidney biopsy were performed. Results : SHG groups (Low and High) inhibited the matrix destruction and damage of the knee joint cartilage in SD rat model, and significantly prevented the reduction in cartilage thickness. In SHG groups, the expressions of TNF-α, IL-1β, IL-6 and MMP-13 were significantly decreased, and the expressions of TIMP-1, GAGs were significantly increased compared with OA group. The safety indicators had no significant differences among five groups. Conclusions : These results show that SHG has cartilage protection capacity, anti-inflammatory effect.
Antioxidant, cytotoxic, and anti-inflammatory assays were conducted to determine the commercial viability of Brachythecium populeum. The antioxidant activity was assessed by performing the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays. This was followed by the quantification of polyphenols and flavonoids. Results of the DPPH and ABTS assay showed that antioxidant activities of the ethanol extract of B. populeum were 3.7 and 3.6 times higher than water extract, respectively. The polyphenol concentration was also determined to be 4.1 times higher and the flavonoid concentration was 5.3 times higher than the water extract. The cell-based experiments, 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay and nitric oxide assay, were performed using RAW 264.7. Results of the MTT assay revealed that both extracts exerted no cytotoxicity on the cells (based on 80% viability). In the nitric oxide (NO) production inhibition experiment, inhibition of NO production was determined to be 15.42% more when exposed to ethanol extract as compared to water extract. Furthermore, the ethanol extract exerted greater inhibition of inflammatory cytokines interleukin (IL)-1β, IL-6, and tumor necrosis factor-α production (9.39%, 11.87%, and 14.49% more, respectively) when compared to the water extract. Due to the good antioxidant activity and potential for inhibiting NO and inflammatory cytokine production, B. populeum ethanol extracts are prospective sources of anti-inflammatory compounds.
Kim, Jae Sik;Kim, Hak Jae;Lee, Me-Yeon;Moon, Kyung Chul;Song, Seung Geun;Kim, Han-Soo;Han, Ilkyu;Kim, Il Han
Radiation Oncology Journal
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v.37
no.1
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pp.37-42
/
2019
Purpose: To identify prognostic factors influencing progression-free survival (PFS) of aggressive fibromatosis (AF) after postoperative radiotherapy (PORT) and assess correlations between immunohistochemistry (IHC) features of β-catenin/smooth muscle actin (SMA) and PFS. Materials and Methods: Records of 37 patients with AF treated by PORT from 1984 to 2015 were retrospectively reviewed. Fifteen patients underwent wide excision for AF and 22 patients received debulking operation. The median total dose of PORT was 59.4 Gy. IHC staining results of β-catenin and SMA were available for 11 and 12 patients, respectively. Results: The median follow-up duration was 105.9 months. Five-year PFS rate was 70.9%. Tumor size or margin status was not related to PFS in univariate analysis (p = 0.197 and p = 0.716, respectively). Multivariate analysis showed that increased interval from surgery to PORT (>5.7 weeks) was a marginal risk factor for PFS (p = 0.054). Administration of PORT at the initial diagnosis resulted in significantly improved PFS compared to deferring PORT after recurrence (p = 0.045). Patient with both risk factors of deferring PORT after recurrence and interval from surgery to PORT >5.7 weeks had significantly lower 5-year PFS than patients without risk factor (34.1% vs. 100.0%; p = 0.012). Nuclear β-catenin intensity tended to inversely correlate with 5-year PFS, although it did not reach statistical significance (62.5% at low vs. 100.0% at high; p = 0.260). SMA intensity was not related to PFS (p = 0.700). Conclusion: PORT should be performed immediately after surgery irrespective of margin status or tumor size especially in recurrent case. Nuclear β-catenin staining intensity of IHC might correlate with local recurrence.
Chronic kidney diseases are based on uncontrolled immunological and inflammatory responses to pathophysiological renal circumstances such as glomerulonephritis, which is caused by immunological mechanisms of glomerular inflammation with increased production of renal pro-inflammatory cytokines. Pentoxifylline (PTX) exhibits anti-inflammatory properties by inhibiting cytokine and chemokine production through aggregation of erythrocytes and thrombocytes. We synthesized a series of 1,7-substituted methylxanthine derivatives by the Traube purine reaction, and the formation of purine ring was completed through nitrosation, a reduction of the nitroso to the amine by catalytic hydrogenation as derivatives of PTX. Then we studied biological activities such as renal anti-inflammatory effects of the synthesized compounds in the production of cytokines such as nitric oxide (NO), interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) and of chemokines such as monocyte chemoattractant protein-1 and IL-8 in Raw 264.7 and HK-2 cells. Renal antiinflammatory activities of this novel series of N-1 and N-7-substituted methylxanthine showed that the N-7 methyl-group-substituted analogs (S7b) showed selective 61% and 77% inhibition of the production of NO and IL-8. The other replacement of the N-1-(CH2)4COCH3 roup, as in the case of compound S6c, also showed an effective 50% and 77% inhibition of TNF-α and IL-8 production in LPS-stimulated Raw 264.7 and HK-2 cells.
Sun Hu-Nan;Fang Wan;Jin Mei-Hua;Han Ying-Hao;Kim Sun-Uk;Lee Sang-Han;Kim Nam-Soon;Kim Cheol-Hee;Lee Dong-Seok
Biomedical Science Letters
/
v.10
no.4
/
pp.325-332
/
2004
Inflammatory factor such as Interleukin-1 play important roles in determining the fate of both acute and chronic neurological disorders. We investigated whether inhibitors of PKC or PTK can serve as pharmacological agents to reduce IL-I production and the mechanisms underlying their pharmacological effects in a mixed population of glia. Inhibitors of PKC such as H7, Go6976 and Ro31-8220 significantly reduced both the mRNA and protein levels of IL-1α and IL-β in lipopolysaccharide-activated primary glial cells. While the PTK inhibitor genistein also significantly reduced the production of these cytokines, it did not affect the expression of their mRNA. Taken together, inhibitors of PKC and PTK could serve as pharmacological agents to reduce IL-1 production. However, the mechanisms underlying their pharmacological effects are different. Our results provide evidence that inhibitors of protein kinases can serve as pharmacological agents to modulate IL-1 production in glial cell, and in turn, alleviate neuronal injury.
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