• Tuberculosis and Respiratory Diseases
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• 제46권2호
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• pp.185-194
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• 1999

연구배경: NF-$\kappa$B는 단백질이 생성된 후의 변형(post-translational modification)과 세포내에서의 위치 변화(subcellular localization)에 따라 그 작용이 결정되는 특성을 가진 전사 인자로서 최초에는 면역반응에 있어 중요한 역할을 하는 사실이 알려졌으나 그후 이러한 작용이외에도 급성기 염증 반응, 바이러스의 증식, 세포의 발생과 분화에 있어 중요한 작용을 한다는 사실이 알려지게 되었다. 최근의 연구들에서 NF-$\kappa$B 전사 인자가 정상 세포로부터 암세포로의 형질전환에 있어서도 어떤 기능을 할 것이라는 사실들이 알려지게 되었다. NF-$\kappa$B가 암세포로의 형질 전환, 나아가 암세포의 생성에 어떤 역할을 제공한다면 이러한 사실은 나아가 향후의 암치료에 있어서도 유용한 지식이 될 수 있다. NF-$\kappa$B 전사 인자가 인체 암세포에 있어서 세포의 형질 변환에 연관될 수 있다는 사실은 몇몇 암종에서 알려져 있으나 폐암에서의 NF-$\kappa$B 전사 인자의 종양 생성기능에 있어서는 아직 연구된 바가 없다. 방 법: 본 연구에서는 배양된 인체 폐암세포주에 있어서 NF-$\kappa$B family 전사 인자들의 발현정도를 western blot를 이용하여 관찰하고 과발현된 NF-$\kappa$B 전사 인자의 세포내 위치가 세포핵인지 세포질내에 존재하는 것인지를 각각의 단백질 분획에서 western blot를 시행하여 관찰하였고 또한 immunocy-tochemistry를 시행하여 그 발현 양상을 확인하였다. 존재하는 NF-$\kappa$B 전사 인자가 어떠한 복합체의 형태인지를 알아보기 위하여 세포주의 단백 추출물에서 NF-$\kappa$B family 전사 인자에 대한 항체를 사용하여 immunoprecipitation을 시행하였다. 세포주 단백추출물의 $\kappa$B consensus oligonucleotide에의 결합여부를 보기위하여 electrophoretic mobility shift assay를 시행하였다. 결 과: 배양된 인체 폐암세포주에서는 NF-$\kappa$B family의 p50 subunit, p65 subunit가 발현되어 있었고 p50 subunit의 발현은 세포핵내에 국한하여 위치하고 있음을 western blot와 immunocytochemistry를 통하여 관찰할 수 있었다 immunoprecipitation assay는 세포내에서 p50 subunit가 p65 subunit와 복합체를 이루는 상태로 존재하고 있음을 보여주었다. 폐암세포주의 세포핵 추출물은 NF-$\kappa$B consensus oligonucleotide와 결합할 수 있음을 electrophoretic mobility shift assay를 통하여 확인할 수 있었다. 결 론: 인체 폐암세포주에서 NF-$\kappa$B family 전사 인자의 발현이 활성화되어 있으며 NF-$\kappa$B family 전사 영자가 인체 폐암 형성에 있어 어떤 역할을 할 가능성을 시사한다.

• IMMUNE NETWORK
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• 제2권3호
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• pp.137-141
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• 2002

Among the members of the inhibitor of apoptosis (IAP) protein family, only Livin and survivin have been reported to have variant forms. We have found a variant form of c-IAP2 through the interaction with the X protein of HBV using the yeast two-hybrid system. In contrast to the wild-type c-IAP2, the variant form has two stretches of sequence in the RING domain that are repeated in the C-terminus that would disrupt the RING domain. We demonstrate that the variant form has an inhibitory effect on TNF-mediated $NF-{\kappa}B$ activation unlike the wild-type c-IAP2, which increases TNFmediated $NF-{\kappa}B$ activation. These results suggest that this variant form has different activities from the wild-type and the RING domain may be involved in the regulation of TNF-induced $NF-{\kappa}B$ activation.

• 한방안이비인후피부과학회지
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• 제21권2호
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• pp.46-53
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• 2008

Background and Objectives : The aim of this study was to investigate the anti-oxidant, anti-allergic and anti-inflammatory ability of the Taglisodog-eum(SHE) extract on the RAW 264.7 and EL4 cells Materials and Methods : Three types of experiments were implemented for this study: first, the experiment to study the anti-oxidant effect of SHE using Riboflavin; second, in vitro experiment to investigate the inhibition of Th 2 cell differentiation by SHE using EL4 cells (IL-4 mRNA expression); third, the suppression of $NF-{\kappa}B$ activation using RAW 264.7 cells (iNOS and COX-2 mRNA expression). Results : The anti-oxidant ability of SHE were dose-dependantly increased. From in vitro, the LPS-induced iNOS and COX-2 mRNA expression were dose-dependantly decreased in the RAW264.7 cells treated with SHE and the PMA-induced IL-4 mRNA expression were also dose-dependantly decreased in EL4 cells. $NF-{\kappa}B$ activation was suppressed, and iNOS & COX-2 production were inhibited by SHE Conclusion : The results suggest that SHE has dose-dependant anti-oxidant ability, and has anti-allergic and anti-inflammatory effects through the suppression of $NF-{\kappa}B$ activation and the inhibition of Th 2 cell differentiation.

• Biomolecules & Therapeutics
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• 제18권3호
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• pp.308-315
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• 2010

Essential oil-excluded Artemisia princeps Pamp var Ssajuarissuk (AP) was fermented with Lactobacillus brevis K-1, which was isolated from cabbage Kimchi, and the anti-inflammatory effects of AP and fermented AP (FAP) on lipopolysaccharide (LPS)-induced inflammatory response in peritoneal macrophages were investigated. AP and FAP inhibited LPS-induced TNF-$\alpha$, IL-$1{\beta}$, COX-2, iNOS and COX-2 expression, as well as NF-${\kappa}B$ activation. AP and FAP also reduced ear thickness, inflammatory cytokine (TNF-$\alpha$, IL-$1{\beta}$ and IL-6) expression and NF-${\kappa}B$ activation with 12-O-tetradecanoylphorbol-13-acetate (TPA) induced dermatitis in mice. Furthermore, AP and FAP also reduced exudate volume, cell number, protein amount, inflammatory cytokines (TNF-$\alpha$, IL-$1{\beta}$ and IL-6) expression and NF-${\kappa}B$ activation in carrageenan-induced air pouch inflammation in mice. The inhibitory effects of FAP were more potent than those of non-fermented AP. Based on these findings, we propose that FAP can improve inflammatory diseases, such as dermatitis, by inhibiting the NF-${\kappa}B$ pathway.

• 대한의생명과학회지
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• 제20권1호
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• pp.32-38
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• 2014

Potassium cyanate (KOCN) that is known as an inducer of the protein carbamylation is an inorganic compound and is the conjugate based of cyanic acid (HOCN). Based on these studies, we confirmed that KOCN induces the apoptosis of the human colorectal cancer cell line, HCT 116 cells, by various mitochondrial pathways. To investigate other mechanisms of KOCN-mediated apoptosis, in the present study, we examined KOCN-induced cytokines production in HCT 116 cells and identified the intracellular signaling pathway in these processes. We first demonstrated that KOCN considerably increased the cell apoptosis via intracellular $Ca^{2+}$ signaling, mitochondrial dysfunction and ROS production. And then we examined TNF-${\alpha}$ and IL-$1{\beta}$ levels mediated by KOCN in HCT 116 cells. Although IL-$1{\beta}$ was not involved in KOCN-mediated HCT 116 cell apoptosis, the release of TNF-${\alpha}$ was mediated by KOCN in HCT 116 cells via NF-${\kappa}B$ activation. Apoptosis was also enhanced by incubation with supernatants from HCT 116 cells after KOCN treatment and this effect was partially reduced by BAY 11-7085 pre-treated supernatant. Taken together, our results indicate that KOCN-induced apoptosis in HCT 116 cells is dependent on the releases of TNF-${\alpha}$ and the increased factors and that the mechanism involves the activation of NF-${\kappa}B$.

• 대한의생명과학회지
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• 제15권3호
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• pp.257-260
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• 2009

Toll-like receptors (TLRs) recognize molecular structures derived from microbes including bacteria, viruses, yeast, and fungi. TLRs have emerged as a major signaling component of the mammalian host defense. TLR4 is a member of the Toll family that senses lipopolysaccharide (LPS), a cell wall component of gram negative bacteria. LPS recognition by TLR4 requires an additional accessory molecule, MD-2. LPS induces the activation of NF-${\kappa}B$ and IRF3 through MyD88 or TRIF-dependent pathways. The activation of NF-${\kappa}B$ leads to the induction of inflammatory gene products including cytokines and cyclooxygenase-2 (COX-2). This study was carried out to investigate the anti-inflammatory effects of oak wood vinegar. Oak wood vinegar inhibits the NF-${\kappa}B$ activation and COX-2 expression induced by LPS. These results provide new ideas to understand the mechanism of oak wood vinegar for its anti-bacterial and anti-inflammatory activities.

• 동의생리병리학회지
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• 제21권4호
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• pp.916-920
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• 2007

Flowers of Magnolia denudata has been reported to possess a variety of pharmacological activities. In this study, we investigated the anti-inflammatory effects and mechanism of the water extract of Flowers of Magnolia denudata(MD) in lipopolysacchride (LPS)-mediated inflammatory mediators in murine peritoneal macrophages. MD itself does not have any toxic effects in murine peritoneal macrophages. MD inhibits LPS-induced nitric oxide (NO), tumor necrosis factor $(TNF)-{\alpha}$, IL-6 and IL-12 production in murine peritoneal macrophages. Furthermore, we have found that MD inhibited LPS-induced $NF-{\kappa}B$ but not c-Jun N-terminal kinase (JNK), p38 and extracellular signal-ragulated kinase (ERK) activation. These results suggested that MD inhibit LPS-induced production of $TNF-{\alpha}$, IL-6 and IL-12 via suppression of the $NF-{\kappa}B$ activation.

• 대한한의학회지
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• 제26권1호
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• pp.18-25
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• 2005

Objective & Methods: We examined the effects of Platycodi radix on the process of lipopolysaccharide (LPS)-induced nuclear factor $NF-{\kappa}Bp65$ and inhibitory $(I)-{\kappa}B{\alpha}$ alteration in RAW 264.7 cells and acute lung injury in rats. Results: Immunoblot analysis showed that LPS-induced degradation of $I-{\kappa}B{\alpha}$ in RAW 264.7 was inhibited by pretreatment of Platycodi radix. The total cells of bronchoalveolar lavage fluid by LPS challenge markedly decreased in the Platycodi radix pretreatment rats. Platycodi radix pretreatment also caused a decline in neutrophils infiltration into interstitium of the lung. In the alveolar macrophages and neutrophils, decreased $NF-{\kappa}Bp65$ and inducible nitric oxide synthase and increased $I-{\kappa}B{\alpha}$ immunoreaction were detected in Platycodi radix pretreated rats compared with LPS alone treated ones. Conclusion : It may be concluded that Platycodi radix attenuates the development of LPS-induced inflammation by reduction of $NF-{\kappa}Bp65$ activation and neutrophil-mediated acute lung injury. Platycodi radix would be useful as a therapeutic agent for endotoxin-induced lung disease.

• 한방안이비인후피부과학회지
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• 제21권3호
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• pp.82-93
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• 2008

Objective: Previously, the methanol extracts of the semen of Xanthium strumsrium could involved anti-inflammatory effects in lipopolysaccharide (LPS)-stimulated Raw 264,7 cells, We evaluated the anti-allergic effects of X. strumarium on rat basophilic leukemia (RBL-2H3) cells, Methodes : To investigate the effect of X. strumarium on the phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187-induced RBL-2H3 cells. The effects of X. strumarium on the degranulation and the pro-inflammatory cytokines secretion and expression from RBL-2H3 cells were evaluated with $\beta$-hexosaminidase assay, ELISA, and RT-PCR analysis, In addition, we examined the effects of X. strumarium on nuclear factor (NF)-${\kappa}B$ activation and $I{\kappa}B-\alpha$ degradation using Western blot analysis. Results : X. strumarium inhibited degranulation and secretions and expressions of pro-inflammatory cytokines, such as tumor necrosis factor-alpha ($TNF-\alpha$), interleukin (IL)-4 and cyclooxygenase (COX)-2, on stimulated RBL-2H3 cells, however, X. strumarium not affect cell viability. In stimulated RBL-2H3 cells, the protein expression level of nuclear factor-kappa B (NF-${\kappa}B$) was decreased in the nucleus by X. strumarium. In addition, X. strumarium suppressed the degradation of inhibitory protein $I{\kappa}B-{\alpha}$ protein in RBL-2H3 cells. Conclusion : These results suggest that X. strumarium inhibits the degranulation and secretion of pro-inflammatory cytokines through blockade of NF-${\kappa}B$ activation and I $I{\kappa}B-{\alpha}$ degradation.

• 생명과학회지
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• 제19권4호
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• pp.514-519
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• 2009

혈관벽에 단핵구, 대식세포 등의 세포와 지질 등의 축적은 중요한 동맥경화 발병 요인이다. 이들 세포의 혈관벽으로의 이동에 있어서 chemokine인 MCP1이 중요한 역할을 한다는 것이 많이 알려져 있다. 본 연구에서는 사람 평활근세포에서 $IL-1{\beta}$의 처리에 의하여 MCP1의 발현이 증가되는 기전을 알아보고자 실험을 진행하였다. $IL-1{\beta}$의 처리는 전사인자 $NF-{\kappa}B$의 활성화를 통해 MCP1 발현을 전사단계에서 증가시켰다. 이러한 $IL-1{\beta}$에 의해 증가된 MCP1 발현을 억제하는 물질을 찾기 위해 여러 항염증작용을 하는 물질들을 전처리하여 확인해 본 결과 luteolin이 선택적으로 $IL-1{\beta}$에 의해 증가된 MCP1의 발현을 전사단계에서 저해하는 것을 확인하였고 이는 전사인자 $NF-{\kappa}B$가 핵으로 이동하는 것을 감소시킴으로써 나타나는 현상임을 확인하였다. Luteolin이 염증작용을 조절하는데 있어서 중요한 전사인자인 $NF-{\kappa}B$의 활성을 조절한다는 것을 본 실험을 통해 알 수 있었고 이는 식용식물에서 일반적으로 발견되는 luteolin이 어떠한 기전으로 항 염증작용을 하는지에 대한 이해를 높여줄 것이다.