• 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.

• 한국임상수의학회지
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• 제29권5호
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• pp.361-367
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• 2012

고혈당으로 야기된 염증은 당뇨병에서 합병증을 일으키는 주된 요인이다. 최근 연구에 따르면 면역세포에서 TNF-${\alpha}$ 같은 염증성 cytokine의 과도한 생성은 인슐린 저항성을 야기시킨다고 한다. Conjugated linoleic acid (CLA)는 TNF-${\alpha}$ 생산에 관여하여 면역반응을 조절하는 것으로 알려져 있다. 본 연구는 고농도의 당으로 처리한 RAW 264.7 세포에서 TNF-${\alpha}$ 생산, NF-${\kappa}B$의 활성과 $I{\kappa}B-{\alpha}$ 분해에 대한 CLA 효과를 검토하였다. 고농도의 당에 노출된 RAW세포는 저농도의 당에 노출된 RAW 세포보다 NF-${\kappa}B$의 활성과 $I{\kappa}B-{\alpha}$ 분해가 증가되었으며 RAW 세포의 배양 상층액 중에 TNF-${\alpha}$ 생산을 증가시켰다. CLA와 고농도 또는 저농도의 당을 같이 처리한 군은 당만 단일 처리한 군보다 TNF-${\alpha}$ 생산, NF-${\kappa}B$의 활성 및 $I{\kappa}B-{\alpha}$ 분해가 증가되었다. 그리고 고농도의 당과 CLA를 처리한 군에서 저농도의 당과 CLA 처리군에 비해 NF-${\kappa}B$의 활성과 $I{\kappa}B-{\alpha}$ 분해가 증가되었으며 이와 더불어 TNF-${\alpha}$의 양이 증가되었다. 이상의 결과로부터, CLA는 고농도의 당에 노출된 RAW 세포에서 NF-${\kappa}B$의 활성을 높이고 TNF-${\alpha}$ 생산을 증가시키며 이는 고혈당으로 유발되는 염증반응을 촉진하는 인자로 작용할 수 있음을 시사하였다.

• 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.

• Journal of Microbiology and Biotechnology
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• 제16권3호
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• pp.391-398
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• 2006

$NF-{\kappa}B$ is a transcription factor involved in the innate immunity against bacterial infection and inflammation. It is also known to render cells resistant to the apoptosis caused by some anticancer drugs. Such a chemoresistance of cancer cells may be related to the activation of $NF-{\kappa}B$ transcription factor; however, the mechanism of activation is not well understood. Here, we demonstrate that a chemotherapeutic agent, etoposide, independently stimulates the $I{\kappa}B{\alpha}$ degradation pathway and PI3-kinase/Akt signaling pathway: The classical $I{\kappa}B{\alpha}$ degradation pathway leads to the nuclear translocation and DNA binding of p65 subunit through $IKK{\beta}$ kinase, whereas the PI3-kinase/Akt pathway plays a distinct role in activating this transcription factor. The PI3-kinase/Akt pathway acts on the p50 subunit of the $NF-{\kappa}B$ transcription factor and enhances the DNA binding affinity of the p50 protein. It may also explain the role of the PI3-kinase/Akt pathway in the anti-apoptotic function of $NF-{\kappa}B$ during chemoresistance of cancer cells.

• Journal of Acupuncture Research
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• 제36권2호
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• pp.80-87
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• 2019

Background: This study investigated the anti-inflammatory effects of bee venom (BV) through the inhibition of nuclear factor kappa beta ($NF-{\kappa}B$) expression in macrophages and keratinocytes. Methods: Cell viability assays were performed to investigate the cytotoxicity of BV in activated macrophages [lipopolysaccharide (LPS)] and keratinocytes [interferon-gamma/tumor necrosis factor-alpha ($IFN-{\gamma}/TNF-{\alpha}$)]. A luciferase assay was performed to investigate the cellular expression of $NF-{\kappa}B$ in relation to BV dose. The expression of $NF-{\kappa}B$ inhibitors ($p-I{\kappa}B{\alpha}$, $I{\kappa}B{\alpha}$, and p50 and p65) were determined by Western Blot analysis, and the electromobility shift assay. A nitrite quantification assay was performed to investigate the effect of BV, and $NF-{\kappa}B$ inhibitor on nitric oxide (NO) production in macrophages. In addition, Western Blot analysis was performed to investigate the effect of BV on the expression of mitogen-activated protein kinases (MAPK) in activated macrophages and keratinocytes. Results: BV was not cytotoxic to activated macrophages and keratinocytes. Transcriptional activity of $NF-{\kappa}B$, and p50, p65, and $p-I{\kappa}B{\alpha}$ expression was reduced by treatment with BV in activated macrophages and keratinocytes. Treatment with BV and an $NF-{\kappa}B$ inhibitor, reduced the production of NO by activated macrophages, and also reduced $NF-{\kappa}B$ transcriptional activity in activated keratinocytes (compared with either BV, or $NF-{\kappa}B$ inhibitor treatment). Furthermore, BV decreased p38, p-p38, JNK, and p-JNK expression in LPS-activated macrophages and $IFN-{\gamma}/TNF-{\alpha}$-activated keratinocytes. Conclusion: BV blocked the signaling pathway of $NF-{\kappa}B$, which plays an important role in the inflammatory response in macrophages and keratinocytes. These findings provided the possibility of BV in the treatment of atopic dermatitis.

• BMB Reports
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• 제50권11호
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• pp.584-589
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• 2017

Intercellular adhesion molecule-1 (ICAM-1), which is induced by tumor necrosis factor (TNF)-${\alpha}$, contributes to the entry of immune cells into the site of inflammation in the skin. Here, we show that protein tyrosine phosphatase non-receptor type 21 (PTPN21) negatively regulates ICAM-1 expression in human keratinocytes. PTPN21 expression was transiently induced after stimulation with TNF-${\alpha}$. When overexpressed, PTPN21 inhibited the expression of ICAM-1 in HaCaT cells but PTPN21 C1108S, a phosphatase activity-inactive mutant, failed to inhibit ICAM-1 expression. Nuclear factor-${\kappa}B$ (NF-${\kappa}B$), a key transcription factor of ICAM-1 gene expression, was inhibited by PTPN21, but not by PTPN21 C1108S. PTPN21 directly dephosphorylated phospho-inhibitor of ${\kappa}B$ ($I{\kappa}B$)-kinase ${\beta}$ ($IKK{\beta}$) at Ser177/181. This dephosphorylation led to the stabilization of $I{\kappa}B{\alpha}$ and inhibition of NF-${\kappa}B$ activity. Taken together, our results suggest that PTPN21 could be a valuable molecular target for regulation of inflammation in the skin by dephosphorylating p-$IKK{\beta}$ and inhibiting NF-${\kappa}B$ signaling.

• Food Science and Biotechnology
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• 제15권2호
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• pp.303-307
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• 2006

Effect of Oenanthe javanica ethanol extract (OJE) on nuclear factor-${\kappa}B$ (NF-${\kappa}B$)-mediated inflammatory reaction in RAW 264.7 macrophage cells was investigated. The OJE dose-dependently inhibited secretions of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and prostaglandins $E_2\;(PGE_2)$ from lipopolysaccharide (LPS)-stimulated RAW 264.7 cells and blocked LPS-induced expression of cyclooxygenase-2. To clarify mechanistic basis for its inhibitions of NF-${\kappa}B$ and activator protein-1 (AP-1) activations, effects of OJE on activations of NF-${\kappa}B$ and AP-1 genes by luciferase reporter activity were examined. The LPS-stimulated activations of NF-${\kappa}B$ and AP-1 were significantly blocked by 400 and $600\;{\mu$}g/mL of OJE, implicating that OJE might regulate gene expression through more than one signaling pathway. Cytosolic degradation of I-${\kappa}B{\alpha}$ was inhibited by OJE dose-dependently, indicating that the nuclear translocation of p65 was inhibited by OJE. These findings suggest that the inhibition of LPS-stimulated COX-2 expression by OJE is due to its inhibition of NF-${\kappa}B$ activation by blocking I-${\kappa}B{\alpha}$ degradation, which may be mechanistic basis of anti-inflammatory effects of OJE.

• 미생물학회지
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• 제54권3호
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• pp.208-213
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• 2018

Nuclear factor kappa B ($NF{\kappa}B$)의 활성화는 염증을 일으킨다. 이 때, inducible nitric oxide synthase (iNOS)가 발현된다. 우리 선행 연구에 의하면 Bacillus licheniformis B1에 의해 제조된 발효대두에 있는 septapeptide (GVAWWMY)가 대식세포에서 iNOS mRNA 발현과 NO 생산을 억제하였다. 본 연구에서 septapeptide의 처리가 $I{\kappa}B{\alpha}$ ($NF{\kappa}B$ 억제 단백질)의 분해를 억제하여 LPS에 의해 유도되는 $NF{\kappa}B$의 활성화를 억제함을 확인했다. 분자 docking에 의해 septapeptide가 ${\kappa}B$ kinase ${\beta}$ ($IKK{\beta}$)에 부착하여 $I{\kappa}B{\alpha}$의 인산화를 방해할 가능성이 있다. Septapeptide는 높은 친화도로(-8.7 kcal/Mol) $IKK{\beta}$의 kinase domain에 부착해서 kinase 활성에 크게 영향을 미칠 수 있다.

• 동의생리병리학회지
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• 제21권4호
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• pp.1017-1024
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• 2007

Thymus and activation-regulated chemokine (TARC/CCL-17), produced by keratinocytes, is a CC chemokine known to selectively Th2 type T cells via $CCR4^+$ and is implicated in the development of atopic dermatitis (AD). TARC/CCL17 expression was induced by cytokines such as tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and interferon-${\gamma}$ (IFN-${\gamma}$). We recently found that the wogonin, a flavone isolated from Scutellaria baicalensis, suppressed TARC expression via heme oxygenase 1 (HO1) in human keratinocytes induced with mite antigen. However, little is known about the inhibitory mechanism of wogonin on TARC/CCL-17 expression stimulated with cytokines. To investigate the inhibitory mechanism, I determined the inhibitory effects of wogonin on the activation of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) and $I{\kappa}B{\alpha}$ phosphorylation, and also examined the activation of p38 MAP kainase in HaCaT keratinocytes stimulated with TNF-${\alpha}$ and IFN-${\gamma}$. Wogonin inhibited NF-${\kappa}B$-DNA complex, NF-${\kappa}B$ binding activity, and the phosphorylation of $I{\kappa}B{\alpha}$ in a dose dependent manner. Wogonin also inhibited the translocation of NF-${\kappa}B$ from cytosol to nucleus. Moreover, the phosphorylation of of p38 MAP kinase in the TNF-${\alpha}$ and IFN-${\gamma}$-stimulated HaCaT keratinocytes were suppressed by wogonin in a dose dependent manner. These results suggest that wogonin may inhibit cytokine-induced NF-${\kappa}B$ activation by $I{\kappa}B{\alpha}$ degradation via suppression of p38 MAP kinase signaling pathway in keratinocytes and modulation of wogonin signaling pathway may be beneficial for the treatment of AD.

• 한국임상수의학회지
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• 제28권2호
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• pp.190-195
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• 2011

본 연구에서 돼지 PBMC에 t10c12-CLA 처리는 TNF-${\alpha}$생산을 증가시켰으나, LPS 자극 PBMC에서는 TNF-${\alpha}$생산을 감소시켰다. t10c12-CLA 처리는 PBMC의 inhibitory ${\kappa}B$ ($I{\kappa}B$)-${\alpha}$ 단백질 분해를 증가시키고 NF-${\kappa}B$ p65 활성 수준을 증가시켰다. 그러나 LPS 자극 PBMC에서는 상반되는 효과가 관찰되었다. 특히, LPS 비자극 PBMC에서 t10c12-CLA는 NF-${\kappa}B$ 저해제인 caffeic acid phenethyl ester (CAPE)를 처리한 경우 NF-${\kappa}B$ p65 활성 수준을 증가시켰으나 반대로 LPS로 자극한 CAPE 처리 PBMC에서는 NF-${\kappa}B$ p65 활성 수준을 억제시켰다. 이상의 결과는 t10c12-CLA가 돼지 PBMC에 있어 LPS 자극 유무에 따라 다른 효과를 가질 수 있으며, 이는 NF-${\kappa}B$ p65 활성도의 변화와 관련성이 있음을 보여주고 있다.