• International Journal of Oral Biology
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• 제37권3호
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• pp.137-145
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• 2012

Aggregatibacter actinomycetemcomitans is the most important etiologic agent of aggressive periodontitis and can interact with endothelial cells. Monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8) are chemokines, playing important roles in periodontal pathogenesis. In our current study, the effects of A. actinomycetemcomitans on the production of MCP-1 and IL-8 by human umbilical vein endothelial cells (HUVEC) were investigated. A. actinomycetemcomitans strongly induced the gene expression and protein release of both MCP-1 and IL-8 in a dose- and time-dependent manner. Dead A. actinomycetemcomitans cells were as effective as live bacteria in this induction. Treatment of HUVEC with cytochalasin D, an inhibitor of endocytosis, did not affect the mRNA up-regulation of MCP-1 and IL-8 by A. actinomycetemcomitans. However, genistein, an inhibitor of protein tyrosine kinases, substantially inhibited the MCP-1 and IL-8 production by A. actinomycetemcomitans, whereas pharmacological inhibition of each of three members of mitogen-activated protein (MAP) kinase family had little effect. Furthermore, gel shift assays showed that A. actinomycetemcomitans induces a biphasic activation (early at 1-2 h and late at 8-16 h) of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) and an early brief activation (0.5-2 h) of activator protein-1 (AP-1). Activation of canonical NF-${\kappa}B$ pathway ($I{\kappa}B$ kinase activation and $I{\kappa}B-{\alpha}$ degradation) was also demonstrated in these experiments. Although lipopolysaccharide from A. actinomycetemcomitans also induced NF-${\kappa}B$ activation, this activation profile over time differed from that of live A. actinomycetemcomitans. These results suggest that the expression of MCP-1 and IL-8 is potently increased by A. actinomycetemcomitans in endothelial cells, and that the viability of A. actinomycetemcomitans and bacterial internalization are not required for this effect, whereas the activation of protein tyrosine kinase(s), NF-${\kappa}B$, and AP-1 appears to play important roles. The secretion of high levels of MCP-1 and IL-8 resulting from interactions of A. actinomycetemcomitans with endothelial cells may thus contribute to the pathogenesis of aggressive periodontitis.

• 한국식품영양과학회지
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• 제33권10호
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• pp.1594-1600
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• 2004

다양한 생리활성을 지닌 천연물 겨우살이와 칡뿌리 추출물의 인체피부조직세포에서 NF-$textsc{k}$B촬성에 대한조절효과와 이들 추출물의 지질 과산화물 생성 억제 및 라디칼 소거활성, reducing power와 관련된 항산화적 활성을 조사하였다. 본 실험 에 사용한 모든 시료들은 세포독성을 보이지 않는 수준에서 NF-$textsc{k}$B 활성 을 억 제 했다. 칡뿌리 추출물은 0.5 mg 농도에서 35%로 vitamin C 10 mM(8.8 mg,53%)과 함께 유의적인 NF-$textsc{k}$B활성 억제효과를 보였다(p<0.05).또한 칡뿌리 추출물은 겨우살이 추출물보다 상대적으로 더 높은 NF-$textsc{k}$B활성 억제 효과를 보였다. 겨우살이 추출물의 NF-$textsc{k}$B활성 억제 효과는 10%로 낮았지 만,지질 과산화물에 대한 항산화 효과는 5 mg 이상의 농도에서 63% 이상으로 칡뿌리 추출물보다 높았다. 겨우살이와 칡뿌리 추출물은 동일 농도 10 mg에서 vitamin C의 항산화 효과, 48%보다 각각 유의적으로 높은 56%와 75%의 항산화 활성을 보였다. 라디칼 소거활성에 대하여 칡뿌리 추출물이 겨우살이 추출물보다 동일농도에서 더 강한 소거활성을 나타냈다. 잘 알려진 항산화제 vitamin C와 비교해 볼 때, DPPH test에서 이 두 추출물은 vitamin C만큼 강한 라디칼 소거활성을 보이지는 않았다. 그러나 NO test에서 칡 뿌리 추출물의 SCn은 88 Ug으로 vitamin C($SC_{50}$/, 77 $\mu$g)만큼 강한 항산화 활성을 보였다(p<0.05). 칡뿌리 추출물의 항산화 활성은 reducing power측정 에서도 겨우살이 추출물보다 높게 나타났으나, vitamin C의 환원력보다는 매우 낮은 활성을 보였다. 본 연구의 전체 결과를 통해서, 강한 라디칼 소거활성을 지닌 칡뿌리 추출물과 상대적으로 높은 지질 과산화 억제 효과를 보인 겨우살이 추출물은 천연 항산화제로 제안될 수 있는 것으로 판단되었다. 또한 vitamin C와 칡뿌리 추출물의 결과처럼 천연 추출물의 인체피부조직세포에서 NF-$textsc{k}$B활성 조절 효과의 일부는 이들 천연 추출물이 지닌 라디칼 소거활성 또는reducing power의 항산화 활성의 역할에 의한 것으로 제안되었다.

• IMMUNE NETWORK
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• 제10권6호
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• pp.188-197
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• 2010

Background: Lichen-derived glucans have been known to stimulate the functions of immune cells. However, immunostimulatory activity of glucan obtained from edible lichen, Umbilicaria esculenta, has not been reported. Thus we evaluated the phenotype and functional maturation of dendritic cells (DCs) following treatment of extracted glucan (PUE). Methods: The phenotypic and functional maturation of PUE-treated DCs was assessed by flow cytometric analysis and cytokine production, respectively. PUE-treated DCs was also used for mixed leukocyte reaction to evaluate T cell-priming capacity. Finally we detected the activation of MAPK and NF-${\kappa}B$ by immunoblot. Results: Phenotypic maturation of DCs was shown by the elevated expressions of CD40, CD80, CD86, and MHC class I/II molecules. Functional activation of DCs was proved by increased cytokine production of IL-12, IL-$1{\beta}$, TNF-${\alpha}$, and IFN-${\alpha}/{\beta}$, decreased endocytosis, and enhanced proliferation of allogenic T cells. Polymyxin B, specific inhibitor of lipopolysaccharide (LPS), did not affect PUE activity, which suggested that PUE was free of LPS contamination. As a mechanism of action, PUE increased phosphorylation of ERK, JNK, and p38 MAPKs, and enhanced nuclear translocation of NF-${\kappa}B$ p50/p65 in DCs. Conclusion: These results indicate that PUE induced DC maturation via MAPK and NF-${\kappa}B$ signaling pathways.

• Tuberculosis and Respiratory Diseases
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• 제49권3호
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• pp.332-342
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• 2000

연구배경 : 염증매개 사이토카인은 염증성 폐질환의 중요한 매개물질이다. 폐 상피세포는 염증세포에서 분비되는 사이토카인에 의해 interleukin, chemokines, colony stimulating factors와 growth factor등을 생산 및 분비함으로써 국소 염증 부위에서의 사이토카인 network에 중요한 역할을 한다. 따라서 폐 상피세포에서 염증매개 사이토카인의 발현 기전에 대한 이해는 염증성 폐질환의 기전규명과 이에 기초한 새로운 치료법의 개발에 생각된다. 대부분의 사이토카인은 NF-${\kappa}B$전사인자에 의해 발현되는데 폐 상피세포에서 염증매개 사이토키인의 발현과 NF-${\kappa}B/I{\kappa}B$ 경로와의 관련성에 관한 연구는 부족한 실정이다. 방법 : BEAS-2B, A549, NCI-H157, NCI-H719 세포에서 IL-1$\beta$와 TNF-$\alpha$ 자극에 의한 IL-8과 TNF-$\alpha$ mRNA의 발현 양상을 평가하였고 이들의 발현과 관찰하였고 NF-${\kappa}B/I{\kappa}B$ 경로와의 관련성을 평가하기 위하여 IL-l$\beta$와 TNF-$\alpha$ 자극에 의한 NF-${\kappa}B$의 활성화 및 $I{\kappa}B{\alpha}$와 $I{\kappa}B{\beta}$의 분해 양상을 관찰하였다. 폐 상피세포의 종류에 따른 NF-${\kappa}B/I{\kappa}B$ 경로 조절의 기전을 규명하고자 IL-1$\beta$와 TNF-$\alpha$ 자극에 의한 $I{\kappa}B{\alpha}$의 인산화와 기저상태에서 IKK$\alpha$의 발현을 평가하였다. 결과 : BEAS-2B, A549, NCI-H157 세포에서는 IL-1$\beta$와 TNF-$\alpha$ 자극으로 $I{\kappa}B{\alpha}$와 $I{\kappa}B{\beta}$가 분해되었고 NF-${\kappa}B$의 활성화가 관찰되었으며 IL-8과 TNF-$\alpha$mRNA의 발현이 유도되었다. NCI-H719 세포에서는 IL-1$\beta$와 TNF-$\alpha$ 자극으로 $I{\kappa}B$ 분해에 의한 NF-${\kappa}B$의 활성화 및 염증매개 사이토카인의 발현이 관찰되지 않았다. BEAS-2B, A549, NCI-H157 세포에서는 IL-1$\beta$와 TNF-$\alpha$ 자극으로 ${\kappa}B$의 인산화가 관찰되었지만 NCI-H719 세포에서는 관찰되지 않았다. 기저상태의 IKK$\alpha$ 발현은 세포간에 차이가 관찰되지 않았다. 결론 : 폐 상피세포에서 NF-${\kappa}B/I{\kappa}B$ 경로는 염증매개 사이토카인 발현에 매우 중요한 역할을 하고, 일부 세포에서는 NF-${\kappa}B/I{\kappa}B$ 경로 조절의 차이를 보이는데 이는 IKK보다 상위 단계의 세포내 신호전달체계의 이상에 기인한 것으로 생각된다.

• 생약학회지
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• 제35권1호통권136호
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• pp.92-97
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• 2004

Effect of AGI $({\alpha}-Glucosidase\;Inhibitor)-1120$, pine (Pinus densiflora) bark extract and Chaga mushroom (Inonotus obliquus) - and Chaga mushroom mycelium extracts on cellular $NF-{\kappa}B$ activation in malignant human keratinocytes (SCC-13) were evaluated to elucidate the possible correlation of $NF-{\kappa}B$ with antioxidant activity. The antioxidant activities of these natural products were examined in three different evaluation methods, i.e., lipid peroxidation value (POV) evaluation test, and 1,1diphenyl-2-picrylhydrazyl radical (DPPH) and nitric oxide (NO) scavenging test. In a cell-based $NF-{\kappa}B$ monitoring assay systern, all samples revealed the downregulatory profiles on the cellular $NF-{\kappa}B$ activity. AGI -1120 (1, 2 mg) and Chaga mushroom extract (0.05, 0.1 mg) downregulated the $NF-{\kappa}B$ activity in a dose-dependent manner. Chaga mushroom mycelium extract (5 mg) significantly inhibited the $NF-{\kappa}B$ activity (p<0.05). Although AGI-1120 and Chaga mushroom mycelium extract exhibited no antioxidant activities evaluated in pay, Chaga mushroom extract showed antioxidant in a dose-dependent manner at concentrations of $0.05{\sim}1$ mg. While AGI-1120 and Chaga mushroom extract possessed a relatively potential DPPH radical scavenging activity, the NO scavenging activity of Chaga mushroom extract $(SC_{50}:47\;{mu}g)$ was higher than the known antioxidant, vitamin C $(SC_{50}:77\;{mu}g)$. These results suggest that AGI-1120 and Chaga mushroom- and Chaga mushroom mycelium extracts may serve as an useful radical scavenging antioxidant agents with $NF-{\kappa}B$ inhibitory effect in human skin.

• Bulletin of the Korean Chemical Society
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• 제35권8호
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• pp.2361-2366
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• 2014

A new aliphatic acid amide, ZP-amide F (1), and eight known compounds, including bungeanumamide A (2), tumuramide C (3), ZP-amide A (4), ZP-amide B (5), ZP-amide D (6), hyperin (7), quercitrin (8), and (-)-sesamin (9), were isolated from pericarps of Zanthoxylum piperitum. The effects of these compounds on $TNF{\alpha}$-induced NF-${\kappa}B$ activation and transactivational activity of PPARs, including $PPAR{\alpha}$, $PPAR{\beta}({\delta})$ and $PPAR{\gamma}$ subtypes, were evaluated. Compounds 7 and 9 exhibited potent inhibitory effects on $TNF{\alpha}$-induced NF-${\kappa}B$ activation with $IC_{50}$ values of 5.50 and $8.10{\mu}M$, respectively. Aliphatic acid amide compounds 3, 4 and 6 displayed enhanced effects on PPAR transactivational activity with $EC_{50}$ values of 47.12, 19.13 and $12.02{\mu}M$, respectively. Among them, compound 4 demonstrated an increase in $PPAR{\alpha}$ transactivational activity, compound 3 showed a moderate increase on all PPAR subtypes, whereas compound 6 displayed weak PPAR transactivational activity.

• 대한의생명과학회지
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• 제19권2호
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• pp.112-117
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• 2013

Toll-like receptors (TLRs) recognize pathogen-associated molecular patterns (PAMPs) and regulate the activation of innate immunity. All TLR signaling pathways culminate in the activation of NF-${\kappa}B$, leading to the induction of inflammatory gene products such as cyclooxygenase-2 (COX-2). Triptolide (TP), a natural component of Tripterygium wilfordii Hook. F, has been used as folk remedies to treat many chronic diseases for many years. In the present report, we present biochemical evidence that TP inhibits the NF-${\kappa}B$ activation induced by polyriboinosinic polyribocytidylic acid (Poly[I:C], TLR3 agonist) and lipopolysaccharide (LPS, TLR4 agonist). TP also inhibits COX-2 expression induced by Poly[I:C] and LPS. These results suggest that TP can modulate the immune responses regulated by TLR3 and TLR4 signaling pathways.

• 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.

• Biomolecules & Therapeutics
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• 제17권4호
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• pp.403-411
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• 2009

Thiacremonone, the main component isolated from heated garlic (Allium sativum L.), is interested for using as a cancer preventive or therapeutic agent since garlic has been known to be useful plant in the treatment of cancers. Nuclear factor kappaB (NF-${\kappa}B$) is constitutively activated in the prostate cancer and activation of NF-${\kappa}B$ is implicated in drug resistance in cancer cells. Docetaxel, a semisynthetic analog of paclitaxel, is an antineoplastic drug widely used for advanced various cancer. In previous studies, we found that thiacremonone inhibited activation of NF-${\kappa}B$ in cancer cells and marcrophages. In the present study, we investigated whether thiacremonone could increase susceptibility of prostate cancer cells (PC-3 and DU145) to docetaxel via inactivation of NF-${\kappa}B$. We found that the combination treatment of thiacremonone (50 ${\mu}g$/ml) with docetaxel (5 nM) was more effective in the inhibition of prostate cancer cell growth and induction of apoptosis accompanied with the significant inhibition of NF-${\kappa}B$ activity than those by the treatment of thiacremonone or docetaxel alone. It was also found that NF-${\kappa}B$ target gene expression of Bax, caspase-3 and caspase-9 was much more significantly enhanced, but the expression of Bcl-2 was also much more significantly inhibited by the combination treatment. These results indicate that thiacremonone inhibits NF-${\kappa}B$, and enhances the susceptibility of prostate cancer cells to docetaxel. Thus, thiacremonone could be useful as an adjuvant anti-cancer agent.

• 대한의생명과학회지
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• 제10권2호
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• pp.143-151
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• 2004

Matrix metalloproteinases (MMPs) are capable of degrading extracellular matrix, a process that is necessary for angiogenesis, tumor invasion and metastasis. Platelet-activating factor (PAP) increases angiogenesis, tumor growth and metastasis through nuclear factor (NF)-$\kappa$B activation. Based on these facts, the involvement of MMPs in PAF-induced pulmonary metastasis was investigated in murine sarcoma cells, MMSV-BALB/3T3. Messenger RNA expression and enzymatic activity of MMP-9 were assessed by RT-PCR and zymography, and cell migration and metastasis were done for the detection of MMP-9 functional activity. PAP induced mRNA expression and enzymatic activity of MMP-9, and its effects were either inhibited by the PAP antagonist, WEB 2170 or by the NF-$\kappa$B inhibitor, parthenolide, or p65 antisense oligonucleotide in a dose-dependent manner. In addition, PAF induced promoter activity of MMP-9, which was inhibited by WEB 2170, phenanthroline, NAC, PDTC. These results indicate that PAF induces mRNA expression and enzymatic activity of MMP-9 in NF-$\kappa$B dependent manner. Cell migration assay showed that PAF induced MMSV-BALB/3T3 migration, and its effect was significantly inhibited by treatment with phenanthroline. PAF enhanced pulmonary metastasis of murine sarcoma cells, MMSV-BALB/3T3 was also reduced by phenanthroline. These results suggest that PAF-enhanced cell migration and pulmonary metastasis is mediated through the expression of MMP. In conclusion, It is suggested that PAF enhances pulmonary metastasis by inducing MMP-9 expression via the activation of NF-$\kappa$B.