• Tuberculosis and Respiratory Diseases
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• v.54 no.5
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• pp.542-550
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• 2003

Background : Synthetic glucocorticoids are widely used in many chronic inflammatory diseases because of their excellent anti-inflammatory activity. Enhancing the transcription of $I{\kappa}B$ and preventing activated NF-${\kappa}B$ from binding to ${\kappa}B$ sites are thought to be the underlying mechanisms. But these data are largely derived from in vitro studies using cell lines. In this study, after administrating a steroid to volunteers, we evaluated the effect on the NF-${\kappa}B$ system. Methods : Prednisolone(0.5mg/kg/d) was orally administered to 5 healthy volunteers for 7 days. Before and after the administration, we sampled their peripheral blood monocytes, and performed western blot analysis both with stimulation, using IL-$1{\beta}$, LPS, TNF, and without stimulation(baseline). We also performed EMSA after stimulation with LPS. Results : After ingestion of the steroid, baseline expressions of $I{\kappa}B{\alpha}$ were increased in two of the subjects, while suppressed degradations of $I{\kappa}B{\alpha}$ to stimulations were observed in all five. In addition, the binding capacity of NF-${\kappa}B$ after the administration was decreased. Conclusion : Steroid plays such roles as enhancing the transcription of $I{\kappa}B{\alpha}$, suppressing the DNA binding capacity of NF-${\kappa}B$, and suppressing the degradation of $I{\kappa}B{\alpha}$.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.21 no.3
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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.

• Korean Journal of Microbiology
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• v.54 no.3
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• pp.208-213
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• 2018

Activation of nuclear factor kappa B ($NF{\kappa}B$) leads to the inflammatory process. During this $NF{\kappa}B$-dependent inflammation process, inducible nitric oxide synthase (iNOS) are expressed in the inflammatory cells. Our previous data indicated that a specific septapeptide (GVAWWMY) from the soybean extract fermented by Bacillus licheniformis B1 inhibited iNOS mRNA expression and NO production in cultured macrophage cells. Our further experiments revealed that treatment of same septapeptide resulted in inhibition of LPS-induced $NF{\kappa}B$ activation by reversing degradation of $I{\kappa}B{\alpha}$, an inhibitory protein for $NF{\kappa}B$. The molecular docking indicated that the septapeptide binds to $I{\kappa}B$ kinase ${\beta}$ ($IKK{\beta}$), and thus it can inhibit phosphorylation of $I{\kappa}B{\alpha}$. Supporting this, the binding site for the septapeptide has the highest affinity (-8.7 kcal/mol) and the site was located at the kinase domain (KD) of $IKK{\beta}$, which can significantly affect the kinase activity of $IKK{\beta}$.

• Molecules and Cells
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• v.41 no.8
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• pp.762-770
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• 2018

Adiponectin, a hormone produced by adipose tissue, is very abundant in plasma, and its anti- and pro-inflammatory effects are reported. However, the mechanisms of these pro- and anti-inflammatory effects are not fully defined. Herein, we evaluated the dual inflammatory response mechanism of adiponectin in macrophages. Short-term globular adiponectin (gAd) treatment induced $I{\kappa}B{\alpha}$ degradation, $NF-{\kappa}B$ nuclear translocation, and $TNF-{\alpha}$ production in RAW 264.7 cells. Polymyxin B pretreatment did not block gAd-induced $I{\kappa}B{\alpha}$ degradation, and heated gAd was unable to degrade $I{\kappa}B{\alpha}$, suggesting that the effects of gAd were not due to endotoxin contamination. gAd activated IKK and Akt, and inhibition of either IKK or Akt by dominant-negative $IKK{\beta}$ ($DN-IKK{\beta}$) or DN-Akt overexpression blocked gAd-induced $I{\kappa}B{\alpha}$ degradation, suggesting that short-term incubation with gAd mediates inflammatory responses by activating the $I{\kappa}B/NF-{\kappa}B$ and PI3K/Akt pathways. Contrastingly, long-term stimulation with gAd induced, upon subsequent stimulation, tolerance to gAd, lipopolysaccharide, and CpG-oligodeoxynucleotide, which is associated with gAd-induced downregulation of IL-receptor-associated kinase-1 (IRAK-1) due to IRAK-1 transcriptional repression. Conclusively, our findings demonstrate that the pro- and anti-inflammatory responses to gAd in innate immune cells are time-dependent, and mediated by the activation of the $I{\kappa}B/NF-{\kappa}B$ pathway, and IRAK-1 downregulation, respectively.

• Korean Journal of Oriental Medicine
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• v.16 no.3
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• pp.155-166
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• 2010

Objetives : The purpose of this study was to examine the pathway of anti-allergic effects of Aconiti Ciliare Tuber (ACT). Methods : We examined cell viability, ${\beta}$-hexosaminidase release, pro-inflammatory cytokines secretion and mRNA expressions, nuclear factor-kappa B (NF-${\kappa}B$) (p65) activation, inhibitor kappa B-alpha ($I{\kappa}B-{\alpha}$) degradation, and MAPKs activation from RBL-2H3 cells pre-treatment by ACT of 1.0 mg/ml, 2.0 mg/ml separately. Results : We observed that ACT reduced the secretion of ${\beta}$-hexosaminidase, TNF-${\alpha}$, IL-4 and the expression of COX-2 mRNA in RBL-2H3 cells. Futhermore, ACT inhibited the levels of activation of NF-${\kappa}B$ (p65) protein, ERK MAPK, and degradation of $I{\kappa}B-{\alpha}$ in RBL-2H3 cells. Conclusions : These results show that ACT has an anti-histamine effect and inhibitory effect of NF-${\kappa}B$ (p65) through regulation of $I{\kappa}B-{\alpha}$ degradation. This improves that ACT could be used as an anti-allergic medicine.

• The Journal of Pediatrics of Korean Medicine
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• v.24 no.1
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• pp.93-103
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• 2010

Objectives The purpose of this study was to examine the pathway of anti-allergic effects of Cheonmaec-tang (CMT). Methods We examined the cell viability, $\beta$-hexosaminidase release, pro-inflammatory cytokines secretion and mRNA expressions, nuclear factor-kappa B (NF-${\kappa}B$) (p65) activation, inbibitor kappa B-alpha ($I{\kappa}B-{\alpha}$) degradation, and MAPKs activation in RBL-2H3 cells pre-treated by CMT of 2.0 mg/ml, 4.0 mg/ml separately. Results We observed that CMT reduced the secretion of $\beta$-hexosaminidase, TNF-$\alpha$, IL-4 and the expression of COX-2 mRNA in RBL-2H3 cells. Furthermore, CMT inhibited the levels of activation of NF-${\kappa}B$ (p65) protein, ERK MAPK, and degradation of $I{\kappa}B-{\alpha}$ in RBL-2H3 cells. Conclusions These results show that CMT has an anti-histamine effect and inhibitory effect of NF-${\kappa}B$ (p65) through regulation of $I{\kappa}B-{\alpha}$ degradation. These suggest that CMT could be used as an anti-allergic medicine.

• Journal of Physiology & Pathology in Korean Medicine
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• v.26 no.4
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• pp.483-490
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• 2012

This study was designed to identify the effects of Polygoni cuspidati Radix(PCR) on the generation of superoxide anion radicals (${\cdot}O_2{^-}$), nitric oxide (NO), peroxynitrite ($ONOO^-$) in the renal epithelial cells of mouse(LLC-$PK_1$). The effects of PCR on the expression of inflammation-related proteins, IKK-${\alpha}$, phospho-$I{\kappa}B-{\alpha}$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, IL-$1{\beta}$, VCAM-1, were examined by western blotting. For this study, the fluorescent probes, namely dihydrorhodamine 123 (DHR 123), 2',7'-dichloro dihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) were used. Protein expression levels of IKK-${\alpha}$, phospho-$I{\kappa}B-{\alpha}$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, IL-$1{\beta}$, VCAM-1 were assayed by western blot. PCR reduced $H_2O_2$-induced cell death dose-dependently. It inhibited the generation of ${\cdot}O_2{^-}$, NO, $ONOO^-$ and $PGE^2$ in the $H_2O_2$-treated LLC-PK1 cells in vitro. PCR inhibited the espression of IKK-${\alpha}$, phospho-$I{\kappa}B-{\alpha}$, COX-2, iNOS, IL-$1{\beta}$ and VCAM-1 genes by means of decreasing the NF-${\kappa}B$ activation. These results suggest that PCR is an effective NO, ${\cdot}O_2{^-}$, $ONOO^-$ scavenger, and this substance recommended to be applied in treatment for the inflammatory process and inflammation-related disease.

• Journal of Acupuncture Research
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• v.36 no.2
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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.

• Journal of Life Science
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• v.10 no.2
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• pp.55-60
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• 2000

We examined the effects of Kamgil-Tang on the process of lipopolysaccharide (LPS)-induced unclear factor (NF)-${\kappa}$ Bp65 and inhibitory (I)-${\kappa}$ B${\alpha}$ alteration in RAW 264.7 cell and acute lung injury in rats. Immunoblot analysis showed that LPS-induced degradation of I-${\kappa}$ B${\alpha}$ in RAW 264.7 was inhibited by pretreatment of Kamgil-Tang. The total cells of bronchoalveolar lavage fluid by LPS challenge markedly decreased in the Kamgil-Tang pretreatment rats. Kamgil-Tang pretreatment caused also 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 Kamgil-Tang pretreated rats compared with LPS alone treated ones. It may be concluded that Kamgil-Tang attenuates the development of LPS-induced inflammation by reduction of NF-${\kappa}$ Bp65 activation and neutrophil-mediated acute lung injury. Kamgil-Tang would be useful as a therapeutic agent for endotoxin-induced lung disease.

• Tuberculosis and Respiratory Diseases
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• v.54 no.5
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• pp.551-562
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• 2003

Background : NF-${\kappa}B$ is a characteristic transcriptional factor which has been shown to regulate production of acute inflammatory mediators and to be involved in the pathogenesis of many inflammatory lung diseases. There has been some evidence that PI3K/Akt pathway could activate NF-${\kappa}B$ in human cell lines. However, the effect of PI3K/Akt pathway on the activation of NF-${\kappa}B$ varied depending on the cell lines used in the experiments. In this study we evaluated the effect of PI3K/Akt pathway on the activation of NF-${\kappa}B$ in human respiratory epithelial cell lines. Methods : BEAS-2B, A549 and NCI-H157 cell lines were used in this experiment. To evaluate the activation of Akt activation and I${\kappa}B$ degradation, cells were analysed by western blot assay using phospho-specific Akt Ab and $I{\kappa}B$ Ab. To block PI3K/Akt pathway, cells were pretreated with wortmannin or LY294002 and transfected with dominant negative Akt (DN-Akt). For IKK activity, immune complex kinase assay was performed. To evaluate the DNA binding affinity and transcriptional activity of NF-${\kappa}B$, electrophoretic mobility shift assay (EMSA) and luciferase assay were performed, respectively. Results : In BEAS-2B, A549 and NCI-H157 cell lines, Akt was activated by TNF-$\alpha$ and insulin. Activation of Akt by insulin did not induce $I{\kappa}B{\alpha}$ degradation. Blocking of PI3K/Akt pathway via wortmannin/LY294002 or DN-Akt did not inhibit TNF-$\alpha$-induced $I{\kappa}B{\alpha}$ degradation or IKK activation. Inhibition of PI3K/Akt did not affect TNF-$\alpha$-induced NF-${\kappa}B$ activation. Overexpression of DN-Akt did not block TNF-$\alpha$-induced transcriptional activation of NF-${\kappa}B$, but wortmannin enhanced TNF-$\alpha$-induced in NF-${\kappa}B$ transcriptional activity. Conclusion : PI3K/Akt was not involved in TNF-$\alpha$-induced $I{\kappa}B{\alpha}$ degradation or transcriptional activity of NF-${\kappa}B$ in human respiratory epithelial cell lines.