• Korean Journal of Plant Resources
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• v.25 no.3
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• pp.299-307
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• 2012

This work aimed to elucidate the anti-inflammatory effects of ethyl acetate fraction from Cnidium officinale Makino with a cellular system of LPS-stimulated RAW 264.7 and THP-1 cells. Some key pro-inflammatory cytokines and mediators including NO, iNOS, $PGE_2$, COX-2, TNF-${\alpha}$, NF-${\kappa}B$ p50 and NF-${\kappa}B$ p65 were studied by sandwich ELISA and western blot analysis. Ethyl acetate fraction could significantly inhibit the production of NO, $PGE_2$, TNF-${\alpha}$, iNOS and COX-2 in LPS-stimulated cell than that of single LPS-stimulated. And ethyl acetate fraction suppresses the activation of NF-${\kappa}B$ p50 and NF-${\kappa}B$ p65. All the results showed that ethyl acetate fraction had a good anti-inflammatory effect on LPS-stimulated RAW264.7 and THP-1 cells. Taken together, the anti-inflammatory actions of ethyl acetate fraction from Cnidium officinale Makino might be due to the down-regulation of NO, $PGE_2$, TNF-${\alpha}$, iNOS and COX-2 via the suppression of NF-${\kappa}B$ activation.

• Biomolecules & Therapeutics
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• v.17 no.1
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• pp.79-85
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• 2009

Polyphenolic compounds are reported to have various pharmacological activities such as anti-oxidative, anti-cancerous, anti-inflammatory and anti-aging effects. Although numerous papers explore their functional roles in many different cellular actions, not many studies handle their structural features in anti-inflammatory responses. In this study, therefore, we examined structural role of substituted transstilbenes in their NO inhibitory and NF-${\kappa}B$ suppressive activities. Of 10 compounds tested, 4 compounds (cinnamic acid, resveratrol, piceatannol and curcumin) displayed NO inhibitory activities in a dose-dependent manner. Similarly, these compounds blocked LPS-induced cytotoxicity of RAW264.7 cells. All NO inhibitory compounds also inhibited $I{\kappa}B{\alpha}$ phosphorylation, a hallmark for NF-${\kappa}B$ activation. However, these inhibitory compounds exhibited distinct suppressive pattern in tumor necrosis factor (TNF)-${\alpha}$- or phorbol-12-myristate-13-acetate (PMA)-induced NF-${\kappa}B$ and AP-1 activation. According to structure-activity relationship study, polarity and size of ring B seem to be important for diminishing NO production. Therefore, our data suggest that substituted trans-stilbenes can be developed as novel anti-inflammatory drug or further developed as lead compounds for another improvement.

• The Korean Journal of Physiology and Pharmacology
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• v.1 no.5
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• pp.515-521
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• 1997

Endothelial cells play a central role in the inflammatory processes, and activation of nuclear factor kappa B ($NF-_{\kappa}B$) is a key component in that inflammatory processes. Previously, we reported that tumor necrosis factor alpha($TNF{\alpha}$) had protective effect of cell death induced by serum deprivation and this protection was related to $NF-_{\kappa}B$ activation. Inducible nitric oxide synthase (iNOS) is a member of the molecules which transcription is regulated mainly by $NF-_{\kappa}B$. And the role of nitric oxide (NO) generated by iNOS on cell viability is still controversial. To elucidate the mechanism of $TNF{\alpha}$ and $NF-_{\kappa}B$ activation on cell death protection, we investigate the effect of NO on the cell death induced by serum- deprivation in bovine cerebral endothelial cells in this study. Addition of $TNF{\alpha}$, which are inducer of iNOS, prevented serum-deprivation induced cell death. Increased expression of iNOS was confirmed indirectly by nitrite measurement. When selective iNOS inhibitors were treated, the protective effect of $TNF{\alpha}$ on cell death was partially blocked, suggesting that iNOS expression was involved in controlling cell death. Exogenously added NO substrate (L-arginine) and NO donors (sodium nitroprusside and S-nitroso-N-acetylpenicillamine) also inhibited the cell death induced by serum deprivation. These results suggest that NO has protective effect on bovine cerebral endothelial cell death induced by serum-deprivation and that iNOS is one of the possible target molecules by which $NF-_{\kappa}B$ exerts its cytoprotective effect.

• Journal of Mushroom
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• v.11 no.1
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• pp.46-51
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• 2013

Sparassis crispa is a medicinal mushroom, which has been reported to have anti-cancer effect. In this study, we designed to investigate the effects of Sparassis crispa extracts on the production of nitric oxide (NO) in LPS-stimulated RAW264.7 cells. The pre-treatment of the extracts prior to add LPS in RAW264.7 cells suppressed NO production and iNOS expression at protein and mRNA levels. The phosphorylation of $I{\kappa}B{\alpha}$ was inhibited by the extracts, which was induced through suppressing the activation of $NF-{\kappa}B$. Sparassis crispa extracts showed the effect on the down-regulation of STAT-1 activation in a dose-dependent manner. In LPS-stimulated RAW264.7 cells, $NF-{\kappa}B$ was translocated into the nucleus, while the treatment of Sparassis crispa extracts induced to sequestered $NF-{\kappa}B$ in the cytosol. These experimental results determined that Sparassis crispa extracts play a inhibitory role in inflammatory reactions via regulating NO production, which suggests potential as a component of inflammatory drugs.

• Journal of Life Science
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• v.21 no.10
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• pp.1377-1384
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• 2011

Astaxanthin (ATX) is a red-orange carotenoid pigment that occurs naturally in a wide variety of living organisms. In this study we investigated the inhibitory effects of ATX on the induction of inducible nitric oxide synthase (iNOS), nitric oxide (NO), proinflammatory cytokines, nuclear factor-kappa B(NF-${\kappa}B$) and reactive oxygen species (ROS) in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. In addition, we tested the superoxide radical scavenging activity of ATX by scavenging assay. iNOS and NF-${\kappa}B$ expressions were determined by immunoblot analysis. Interleukin (IL)-6 and tumour necrosis factor-${\alpha}$ (TNF-${\alpha}$) were assayed by ELISA. NO production was monitored by measuring the amount of nitrite. ROS was examined by using the 2', 7'-Dichlorodihydrofluorescin diacetate (DCFH-DA) method. At a concentration of 100 ${\mu}M$, ATX inhibited the expression level of LPS-induced NF-${\kappa}B$, as well as the production of LPS-induced NO and proinflammatory cytokines (IL-6 and TNF-${\alpha}$), by suppressing iNOS expression. In particular, the maximal inhibition rate of IL-6 and TNF-${\alpha}$ production by ATX (100 ${\mu}M$) was 65.2----- and 21.2-----, respectively. In addition, ATX inhibited the LPS-induced transcriptional activity of NF-${\kappa}B$, and this was associated with suppressing the translocations of NF-${\kappa}B$ from the cytosol to the nucleus. Moreover, at various concentrations (25-100 ${\mu}M$), ATX inhibited the intracellular level of ROS. At a concentration of 5 mg/ml, the superoxide radical scavenging activity of ATX was 1.33 times higher than ${\alpha}$-tocopherol of the same concentration. These results showed that ATX inhibited the expression of iNOS and the production of NO and proinflammatory cytokines resulting from ROS production and NF-${\kappa}B$ activation in macrophages. Furthermore, ATX was found to be more effective in superoxide radical scavenging activities compared to ${\alpha}$-tocopherol. These findings are expected to strengthen the position of ATX as anti-inflammatory medicine and antioxidant.

• Food Science and Biotechnology
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• v.16 no.5
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• pp.728-733
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• 2007

Bamboo grass, Sasa quelpaertensis, is a native plant to Jeju Island, Korea. The leaves of Sasa plants are widely used in traditional Korean medicine to treat inflammation-related diseases. We investigated the effect of hot water extract from Sasa quelpaertensis leaves (HWE-SQ) on nitric oxide (NO) production and nuclear $factor-{\kappa}B\;(NF-{\kappa}B)$ activation in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. HWE-SQ inhibited LPS-induced NO production and inducible NO synthase (iNOS) protein expression in a dose-dependent manner. Reporter gene assays indicated that HWE-SQ decreases LPS-induced $NF-{\kappa}B$ transcriptional activation. However, HWE-SQ did not affect the phosphorylation and degradation of inhibitory ${\kappa}B{\alpha}\;(1{\kappa}B{\alpha})$. HWE-SQ also directly inhibited iNOS enzyme activity in a dose-dependent manner. These results suggest that HWE-SQ suppresses NO synthesis in macrophages by attenuating $NF-{\kappa}B-mediated$ iNOS protein expression and inhibiting iNOS enzymatic activity, thereby implicating a mechanism by which HWE-SQ is able to ameliorate inflammation-related diseases by limiting excessive or prolonged NO production in pathological events.

• 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.

• Biomolecules & Therapeutics
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• v.23 no.2
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• pp.110-118
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• 2015

According to the expansion of lifespan, neuronal disorder based on inflammation has been social problem. Therefore, we isolated shikonin from Lithospermum erythrorhizon and evaluated anti-inflammatory effects of shikonin in lipopolysaccharide (LSP)-stimulated BV2 microglial cells. Shikonin dose-dependently inhibits the expression of the proinflammatory mediators, nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), and tumor necrosis factor-${\kappa}B$ (TNF-${\alpha}$) as well as their main regulatory genes and products such as inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), and TNF-${\alpha}$ in LPS-stimulated BV2 microglial cells. Additionally, shikonin suppressed the LPS-induced DNA-binding activity of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) to regulate the key regulatory genes of the proinflammatory mediators, such as iNOS, COX-2, and TNF-${\alpha}$, accompanied with downregulation of reactive oxygen species (ROS) generation. The results indicate that shikonin may downregulate the expression of proinflammatory genes involved in the synthesis of NO, $PGE_2$, and TNF-${\alpha}$ in LPS-treated BV2 microglial cells by suppressing ROS and NF-${\kappa}B$. Taken together, our results revealed that shikonin exerts downregulation of proinflammatory mediators by interference the ROS and NF-${\kappa}B$ signaling pathway.

• Natural Product Sciences
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• v.20 no.4
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• pp.227-231
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• 2014

Cimicifuga heracleifolia extract (CHE) was investigated for its effects on the release of nitric oxide (NO) and at the level of inducible nitric oxide synthase (iNOS) gene expression in mouse macrophages. We found that C. heracleifolia elicited a dose-dependent increase in NO production and the level of iNOS mRNA. Since, iNOS transcription has been shown to be under the control of the transcription factor $NF-{\kappa}B$, the effects of CHE on $NF-{\kappa}B$ activation were examined. Transient expression assays with $NF-{\kappa}B$ binding sites linked to the luciferase gene revealed that the increased level of iNOS mRNA, induced by CHE, was mediated by the $NF-{\kappa}B$ transcription factor complex. By using DNA fragments containing the $NF-{\kappa}B$ binding sequence, CHE was shown to activate the protein/DNA binding of $NF-{\kappa}B$ to its cognate site, as measured by electrophoretic mobility shift assay. These results demonstrate that C. heracleifolia stimulates NO production and is able to up-regulate iNOS expression through $NF-{\kappa}B$ transactivation.

• 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.