• Molecules and Cells
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• v.20 no.3
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• pp.364-370
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• 2005

We show that sulforaphane inhibits osteoclastogenesis in the presence of macrophage colony-stimulating factor (M-CSF) and receptor for activation of nuclear factor-${\kappa}B$ ligand (RANKL) in osteoclast (OC) precursors. Sulforaphane, an aliphatic isothiocyanate, is a known cancer chemo-preventative agent with anti-oxidative properties. Nuclear factor-${\kappa}B$ (NF-${\kappa}B$) is a critical transcription factor in RANKL-induced osteoclastogenesis, and electrophoretic mobility shift assays (EMSAs) and assay of NF-${\kappa}B$-mediated secreted alkaline phosphatase (SEAP) revealed that sulforaphane selectively inhibited NF-${\kappa}B$ activation induced by RANKL. Inhibition may involve interaction of sulforaphane with thiol groups, since it was prevented by reducing agents.

• Journal of Applied Biological Chemistry
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• v.61 no.2
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• pp.205-211
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• 2018

Rebaudioside A is a natural sweetener isolated from Stevia rebaudiana Bertoni, one of the glycosides based on steviol. Recent studies have shown that rebaudioside A inhibits the inflammatory response by inhibiting cytokines secretion such as interleukin-$1{\alpha}/1{\beta}$ in activated RAW264.7 mouse macrophage cells by LPS. However, the inhibitory mechanism of inflammation by rebaudioside A in the presence of LPS has not been fully elucidated. Therefore, in this study, we tried to investigate the anti-inflammatory activity of rebaudioside A at the protein level when RAW264.7 cells were stimulated by LPS. The inducible nitric oxide synthase protein expression level was reduced in the group treated with $250{\mu}M$ rebaudioside A compared to the LPS-treated group. In addition, the mRNA expression level of $NF-{\kappa}B$, which is a representative nuclear transcription factor by inflammatory signal, was also decreased as compared with that of LPS-treated group. In addition, $NF-{\kappa}B$ and inhibitor-${\kappa}B$ ($I-{\kappa}B$) complexes that are known to be dissociated by $I-{\kappa}B$ phosphorylation and ubiquitination were less phosphorylated than LPS treated group in the presence rebaudioside A. Finally, we could find that rebaudioside A was involved in the $NF-{\kappa}B$ pathway through reducing extracellular signal-regulated kinase1/2 phosphorylation in a concentration-dependent manner. These results suggest that rebaudioside A might suppress inflammatory reaction through MAPK and $NF-{\kappa}B$ regulation in LPS-stimulated RAW264.7.

• YAKHAK HOEJI
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• v.52 no.1
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• pp.62-66
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• 2008

In this study, we examined the inhibitory effects of propenone derivatives, 1,3-diphenyl-propenone (DPhP), 3-phenyl-1-thiophen-2-yl-propenone (PhT2P), 3-phenyl-1-thiophen-3-yl-propenone (PhT3P) and 1-furan-2-yl-3-phenyl-propenone (FPhP), on $TNF-{\alpha}$-induced nuclear factor (NF)-${\kappa}B$ activity and interleukin (IL)-8-induced monocyte adhesion to colon epithelial cells. 1-Furan-2-yl-3-pyridin-2-yl-propenone (FPP-3) that is previously reported as a $NF-{\kappa}B$ inhibitor suppressed $TNF-{\alpha}$-induced monocyte-epithelial cell adhesion in a concentration-dependent manner. The propenone derivatives, DPhP, PhT2P, PhT3P, FPhP, also inhibited $TNF-{\alpha}$-induced $NF-{\kappa}B$ activation in a similar degree to FPP-3. In a DPPH radical scavenging assay, none of the compounds showed DPPH radical scavenging activity, indicating that the inhibitory actions of the propenone derivatives on redox-sensitive $NF-{\kappa}B$ activity is not due to a simple free radical scavenging activity. In addition, the propenone derivatives also suppressed the IL-8-induced monocyte adhesion to colon epithelial cells. Furthermore, the effective concentrations of the propenone derivatives on both $NF-{\kappa}B$ activation as well as IL-8 induced monocyte-epithelial cell adhesion were 1000 times lower than 5-aminosalicylic acid (5-ASA), a clinically used drug for inflammatory bowel disease. These results suggest that the propenone derivatives may be a potential lead having a strong inhibitory activity against inflammatory cytokine-induced epithelial inflammation.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.2
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• pp.385-389
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• 2008

Nitric oxide (NO) has been suggested to play an important role in endotoxin-mediated shock and inflammation. In this study, we investigated the effect of Rosa laevigata Michx. (Rosaceae) on the production of NO and the molecular mechanism of its action. Rosa laevigata inhibited NO generation and iNOS expression in LPS-stimulated murine macrophages. Activity of nuclear $factor{-\kappa}B\;(NF{-\kappa}B)$ and the degradation of $I{\kappa}B-{\alpha}$ were suppressed by Rosa laevigata. Furthermore, extracellular signal-stimulated kinase (ERK), which is known to be involved in $NF{-\kappa}B$ activation, is inhibited by Rosa laevigata. These results suggest that Rosa laevigata could exert its anti-inflammatory actions by suppressing the synthesis of NO through inhibition of $NF{-\kappa}B$ activity.

• Molecules and Cells
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• v.20 no.1
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• pp.105-111
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• 2005

Mannasantin B, a dilignan structurally related to manssantin A, is an inhibitor of NF-${\kappa}B$ transactivation. In the present study, we found that it inhibited PMA-induced expression of IL-$1{\beta}$, IL-$1{\beta}$ mRNA, and IL-$1{\beta}$ promoter activity in U937 cells with $IC_{50}$ values of about 50 nM. It also inhibited NF-IL6- and NF-${\kappa}B$-induced activation of IL-$1{\beta}$, with $IC_{50}$ values of 78 nM and $1.6{\mu}M$, respectively, revealing a potent inhibitory effect on NF-IL6. Electrophoretic mobility shift assays showed that manassantin B had an inhibitory effect on DNA binding by NF-IL6, but not by NF-${\kappa}B$. Further analysis revealed that transactivation by NF-IL6 was also inhibited. Our results indicate that manassantin B suppresses expression of IL-$1{\beta}$ in promonocytic cells by inhibiting not only NF-${\kappa}B$ but also NF-IL6 activity. Furthermore, our observations suggest that manassantin B may be clinically useful as a potent inhibitor of NF-IL6 activity.

• 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.

• IMMUNE NETWORK
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• v.3 no.1
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• pp.38-46
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• 2003

Background: Platelet-activating factor (PAF) induces nuclear factor $(NF)-{\kappa}B$ activation and angiogenesis and increases tumor growth and pulmonary tumor metastasis in vivo. The role of $NF-{\kappa}B$ activation in PAF-induced angiogenesis in a mouse model of Matrigel implantation, and in PAF-mediated pulmonary tumor metastasis were investigated. Methods: Angiogenesis using Matrigel and experimental pulmonary tumor metastasis were tested in a mouse model. Electrophoretic mobility shift assay was done for the assessment of $NF-{\kappa}B$ translocation to the nucleus. Expression of angiogenic factors, such as tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\alpha}$, basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) were tested by RT-PCR and ELISA. Results: PAF induced a dose- and time-dependent angiogenic response. PAF-induced angiogenesis was significantly blocked by PAF antagonist, CV6209, and inhibitors of $NF-{\kappa}B$ expression or action, including antisense oligonucleotides to p65 subunit of $NF-{\kappa}B$ (p65 AS) and antioxidants such as ${\alpha}$-tocopherol and N-acetyl-L-cysteine. In vitro, PAF activated the transcription factor, $NF-{\kappa}B$ and induced mRNA expression of $TNF-{\alpha}$, $IL-1{\alpha}$, bFGF, VEGF, and its receptor, KDR. The PAF-induced expression of the above mentioned factors was inhibited by p65 AS or antioxidants. Also, protein synthesis of VEGF was increased by PAF and inhibited by p65 AS or antioxidants. The angiogenic effect of PAF was blocked when anti-VEGF antibodies was treated or antibodies against $TNF-{\alpha}$, $IL-1{\alpha}$, and bFGF was co-administrated, but not by antibodies against $TNF-{\alpha}$, $IL-1{\alpha}$, and bFGF each alone. PAF-augmented pulmonary tumor metastasis was inhibited by p65 AS or antioxidants. Conclusion: These data indicate that PAF increases angiogenesis and pulmonary tumor metastasis through $NF-{\kappa}B$ activation and expression of $NF-{\kappa}B$-dependent angiogenic factors.

• Journal of Microbiology and Biotechnology
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• v.23 no.1
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• pp.64-68
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• 2013

Previous studies have shown that lactic acid bacteria can inhibit inflammatory responses, but the mechanisms are very little known. In this study, transaction and expression of three proinflammatory factors, iNOS, PTGS-2, and IL8, which are closely related to the inflammatory response, were investigated by luciferase reporter assay and RTPCR in HT-29 cells treated by Lactobacillus acidophilus. The results showed that the live L. acidophilus sharply down-regulated the transcription of these three genes. Because there was a NF-${\kappa}B$ binding site located at -265 bp, -225 bp, and -95 bp upstream of the iNOS, PTGS-2, and IL8 promoters, respectively, we further addressed the effects of NF-${\kappa}B$ on transaction of the three promoters by cotransfection. As was expected, NF-${\kappa}Bs$ remarkably upregulated the activity of the reporter gene and, no effect of NF-${\kappa}B$s on IL-8 promoter transaction was found after NF-${\kappa}B$ binding site mutation of the IL8 promoter in HT-29 cells. In conclusion, the live L. acidophilus decreased the transcriptional activity of NF-${\kappa}B$ and, in turn, inhibited the transaction of NF-${\kappa}B$ on the three proinflammatory factors mentioned above.

• Tuberculosis and Respiratory Diseases
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• v.52 no.6
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• pp.616-626
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• 2002

Background : The present study was performed to further improve our understanding of molecular mechanisms involved in the activation of NFkB, a major transcriptional factor involved in the inflammatory response in the lung, by particulate matter in lung epithelial cells with an aerodynamic diameter of less than $2.5{\mu}m$(PM2.5). Materials and Methods : Immediate production of reactive oxygen species (ROS) and nitrogen species (RNS), with the PM2.5 induced expression of inducible nitric oxide synthase (iNOS), $I{\kappa}B$ degradation and $NF{\kappa}B$-dependent transcriptional activity, in 549 cells, were monitored. Addition, we also examined the effect of the iNOS inhibitor, L-N6-(1-iminoethyl) lysine hydrochloride (L-NIL), on the PM2.5-induced $NF{\kappa}B$ activation in A549 cells. Results : The rapid degradation of $I{\kappa}B$ and the increase of transcriptional activity of the $NF{\kappa}B$-dependent promotor were observed in A549 cells exposed to PM2.5. The immediate production of ROS in response to PM2.5 in A549 cells was not clearly detected, although immediate responses were observed in RAW264.7 cells. A 549 cells, cultured in the presence of PM2.5, produced an increase in NO, which was noticeably significant after 15 min of exposure with the expression of iNOS mRNA. The addition of L-NIL, an iNOS inhibitor, significantly inhibited the PM2.5-induced $I{\kappa}B$ degradation and the increase of the $NF{\kappa}B$-dependent transcriptional activity. Conclusion : These results suggest that PM2.5 stimulates the immediate production of RNS, leading to the activation of $NF{\kappa}B$ in the pulmonary epithelium.

• Journal of Veterinary Clinics
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• v.32 no.2
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• pp.135-140
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• 2015

Trans-10, cis-12-conjugated linoleic acid (t10c12-CLA) has been shown to participate in the regulation of anti-inflammatory effects. The objectives of this study were to examine the effects of t10c12-CLA on reactive oxygen species (ROS) and nitric oxide (NO) production and nuclear factor-kappaB (NF-${\kappa}B$) activation in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells and to determine whether these effects were associated with change of intracellular calcium ion ($Ca^{2+}$). ROS production was increased in LPS-stimulated RAW 264.7 cells, and this effect was suppressed by 1,2-bis-(o-aminophenoxy) ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA/AM), a calcium chelator. t10c12-CLA suppressed ROS production in LPS-stimulated RAW 264.7 cells, which was further more decreased by treatment with BAPTA/AM. These indicated that t10c12-CLA decreases $Ca^{2+}$-dependent ROS production in LPS-stimulated RAW 264.7 cells. Similarly, NF-${\kappa}B$ p65 DNA binding activity and NO production were decreased by treatment with either t10c12-CLA, BAPTA/AM, or t10c12-CLA and BAPTA/AM combination. However, there were no differences between t10c12-CLA and BAPTA/AM treatment in NO production of LPS-stimulated RAW 264.7 cells. These data indicate that t10c12-CLA inhibits the increases in ROS and NO production and the NF-${\kappa}B$ activation in LPS-stimulated condition. These results suggested that CLA exerts potent anti-inflammatory effects by suppression of LPS-induced ROS and NO production, and NF-${\kappa}B$ activationn via $Ca^{2+}$-dependent pathway.