• BMB Reports
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• 제53권9호
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• pp.478-483
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• 2020

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.

• 한국응용과학기술학회지
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• 제38권1호
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• pp.217-227
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• 2021

본 연구는 국내에서 재배한 어성초를 각기 다른 조건에서 저장한 후 어성초의 추출액을 이용한 천연 화장품 소재로서의 타당성을 분석하였다. quercetin 의 함량은 건 어성초보다 생 어성초에서 더 높았으며, 총 폴리페놀 함량과 총 플라보노이드 함량을 측정한 결과 건 어성초가 생 어성초에 비해 상대적으로 높은 함량으로 분석되었으나 DPPH 및 ABTS 라디칼 소거 활성은 생 어성초가 상대적으로 높은 결과를 보였다. ROS 소거능과 산화질소 생성 억제율은 생 어성초에서 높은 억제율을 보였으나 100 ㎍/mL의 농도에서 건 어성초가 산화질소를 더 감소시키는 것으로 나타났다. 염증성 사이토카인의 감소를 측정한 결과 IL-1β는 건 어성초가 비교적 높은 감소 효과가 나타났고, IL-6와 TNF-α는 생 어성초에서 높은 감소 효과를 나타냈다. 이상의 결과로부터 생 어성초는 초기 염증 반응에 효과적이며 건 어성초는 염증이 만성 단계로 악화하는 것을 예방하는데 효과적인 것으로 예상된다.

• Journal of Ginseng Research
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• 제46권1호
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• pp.156-166
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• 2022

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 (K_D 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.

• 대한임상검사과학회지
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• 제54권3호
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• pp.209-216
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• 2022

발효된 황매화의 상업적 사용 가능성을 확인하기 위하여 항산화능 실험과 세포 독성, 항염능 실험을 실시하였다. 항산화능 실험에는 DPPH 실험, ABTS 실험, polyphenol 농도 측정, flavonoid 농도 측정을 실시하였다. polyphenol과 flavonoid 농도 측정에서는 발효군이 대조군에 비해 높은 농도를 나타내어, 발효를 통해 추출물의 유효성분 및 추출 수율을 높인다는 것을 확인하였다. DPPH, ABTS에서는 대조군에 비해 발효군의 항산화능이 높은 것을 확인하였다. 이는 S. fibuligera가 발효 과정에서 생산해 낸 효소에 의해 황매화의 세포벽 연화와 유기산과 에탄올 생성에 의한 추출 수율 증가에 의한 요인으로 생각된다. 항염능 실험에서는 세포 독성과 항염능을 알아보았다. 세포독성의 경우 대조군과 발효군 모두 낮은 세포독성을 보였으며, 염증 전달 물질인 NO 생성 억제능의 경우 발효군이 대조군에 비해 통계적으로 유의한 수준의 항염능 증가를 보였다. 염증성 cytokine인 IL-1β, IL-6의 농도를 측정한 결과 200 ㎍/mL 농도에서 각각 48.1±6.2%, 30.4±2.2%의 억제효과를 나타내었다. 이를 통해 발효 황매화가 염증 억제를 위한 물질로서 사용가능함을 보였다.

• 한국미생물·생명공학회지
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• 제50권4호
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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.

• Korean Journal of Acupuncture
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• 제39권4호
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• pp.152-171
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• 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.

• 대한임상검사과학회지
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• 제55권3호
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• pp.174-183
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• 2023

Brachythecium populeum 의 상업적 사용 가능성을 확인하기 위하여 항산화능 실험과 세포 독성, 항염능 실험을 실시하였다. 폴리페놀과 플라보노이드 농도 측정에서는 70% 에탄올 추출이 증류수 추출에 비해 높은 농도를 나타내어, 70% 에탄올이 추출물의 유효성분 및 추출 수율의 측면에서 가장 적합한 추출용매란 것을 확인하였다. DPPH, ABTS에서는 증류수 추출물에 비해 70% 에탄올 추출물의 항산화능이 높은 것을 확인하였다. 이는 B. populeum 유효성분의 극성에 따른 것으로 생각된다. 항염능 실험에서는 세포 독성과 항염능을 알아보았다. 세포독성의 경우 양 추출물 모두낮은 세포독성을 보였으며, 염증 전달 물질인 NO 생성 억제능의 경우 70% 에탄올 추출물이 증류수 추출물에 비해 통계적으로 유의한 수준의 항염능 증가를 보였다. 염증성 사이토카인인 IL-1β, IL-6, TNF-α의 농도를 측정한 결과 100 ㎍/mL 농도에서 각각 9.39%, 11.87%, 14.49%의 억제효과를 나타내었다. 이를 통해 B. populeum의 70% 에탄올이 염증 억제를 위한 물질로서 사용 가능함을 보였다.

• Radiation Oncology Journal
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• 제37권1호
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• pp.37-42
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• 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.

• Journal of Microbiology and Biotechnology
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• 제25권2호
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• pp.296-301
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• 2015

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.

• 대한의생명과학회지
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• 제10권4호
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• pp.325-332
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• 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.