• Proceedings of the PSK Conference
• /
• 2003.10b
• /
• pp.195.3-196
• /
• 2003

In this study, we investigated the molecular pathways targeted by platycodin D, which could involve apoptosis in immortalized human keratinocytes (HaCaT). We demonstrated that platycodin D-mediated apoptosis of HaCaT cells exhibited representative features, including DNA fragmentation, caspase-3, caspase-8 activation, and upregulation of Fas and FasL expression, but not p53 activation. To investigate the events involved in activation-induced FasL upregulation, we have examined mRNA accumulation, protein expression, and NF-$\kappa$B activity to elucidate transcription level in the HaCaT cell line treated with platycodin D. (omitted)

• Archives of Pharmacal Research
• /
• v.26 no.7
• /
• pp.545-553
• /
• 2003

The expression of cyclooxygenase-2 (COX-2) is a characteristic response to inflammation, which can be inhibited with sodium salicylate. IL-1$\beta$ and TNF-$\alpha$ can induce extracellular signal-regulated kinase (ERK), IKK, IkB degradation and NF-$\kappa$B activation. Salicylate inhibited the IL-1$\beta$ and TNF-$\alpha$-induced COX-2 expressions, regulated the activation of ERK, IKK and IkB degradation, and the subsequent activation of NF-$\kappa$B, in neonatal rat ventricular cardiomyocytes. The inhibition of the ERK pathway, with a selective inhibitor, PD098059, blocked the expressions of IL-1$\beta$ and TNF-$\alpha$-induced COX-2 and $PGE_2$ release. The antioxidant, N-acetyl-cysteine, also reduced the glutathione or catalase- attenuated COX-2 expressions in IL-1$\beta$ and TNF-$\alpha$-treated cells. This antioxidant also inhibited the activation of ERK and NF-$\kappa$B in neonatal rat cardiomyocytes. In addition, IL-1$\beta$ and TNF-$\alpha$-stimulated the release of reactive oxygen species (ROS) in the cardiomyocytes. However, salicylate had no inhibitory effect on the release of ROS in the DCFDA assay. The results showed that salicylate inhibited the activation of ERK and IKK, I$\kappa$B degradation and NF-$\kappa$B activation, independently of the release of ROS, which suggested that salicylate exerts its anti-inflammatory action through the inhibition of ERK, IKK, IkB and NF-$\kappa$B, and the resultant COX-2 expression pathway in neonatal rat ventricular cardiomyocytes.

• Biomolecules & Therapeutics
• /
• v.28 no.2
• /
• pp.152-162
• /
• 2020

Cerebral ischemia exhibits a multiplicity of pathophysiological mechanisms. During ischemic stroke, the reactive oxygen species (ROS) concentration rises to a peak during reperfusion, possibly underlying neuronal death. Recombinant human erythropoietin (EPO) supplementation is one method of treating neurodegenerative disease by reducing the generation of ROS. We investigated the therapeutic effect of PEGylated EPO (P-EPO) on ischemic stroke. Mice were administered P-EPO (5,000 U/kg) via intravenous injection, and middle cerebral artery occlusion (MCAO) followed by reperfusion was performed to induce in vivo ischemic stroke. P-EPO ameliorated MCAO-induced neurological deficit and reduced behavioral disorder and the infarct area. Moreover, lipid peroxidation, expression of inflammatory proteins (cyclooxygenase-2 and inducible nitric oxide synthase), and cytokine levels in blood were reduced by the P-EPO treatment. In addition, higher activation of nuclear factor kappa B (NF-κB) was found in the brain after MCAO, but NF-κB activation was reduced in the P-EPO-injected group. Treatment with the NF-κB inhibitor PS-1145 (5 mg/kg) abolished the P-EPO-induced reduction of infarct volume, neuronal death, neuroinflammation, and oxidative stress. Moreover, P-EPO was more effective than EPO (5,000 U/kg) and similar to a tissue plasminogen activator (10 mg/kg). An in vitro study revealed that P-EPO (25, 50, and 100 U/mL) treatment protected against rotenone (100 nM)-induced neuronal loss, neuroinflammation, oxidative stress, and NF-κB activity. These results indicate that the administration of P-EPO exerted neuroprotective effects on cerebral ischemia damage through anti-oxidant and anti-inflammatory properties by inhibiting NF-κB activation.

• The Korean Journal of Physiology and Pharmacology
• /
• v.1 no.5
• /
• pp.515-521
• /
• 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 Acupuncture Research
• /
• v.22 no.5
• /
• pp.37-48
• /
• 2005

Objectives : The purpose of this study was to investigate the anti-caner effect of cobrotoxin on the prostatic cancer cell line(PC-3). Methods : After the treatment of PC-3 cells with cobrotoxin, we performed fluorescence microscope, MTT assay, Western blotting, Flow cytometry, PAGE electrophoresis and Surface plasmon resonance analysis to identify $NF-{\kappa}B$ the change of calcium and NO. Results : 1. The expression of $NF-{\kappa}B$ was decreased at 1nM and It·as decreased significantly at 2, 4, 8nM. 2. $I{\kappa}B,\;NF-{\kappa}B$ inhibitor, was decreased significantly at 8nM and $p-l{\kappa}Ba$, phosphrylation of $I{\kappa}B$, was decreased significantly at all concentrations of cobrotoxin. 3. The expressions of p50 and p65 were decreased significantly and dose-dependently at 1, 2, 4, 8nM. 4. The expression of p53 was increased significantly at 1, 2, 4, 8nM. 5. The calcium concentration in cell wasn't changed at 1, 2, 4, 8nM, but was increased dose-dependently at 30, 70, 130, 250nM comparing with lower dose of cobrotoxin. 6. The NO concentration in cell was increased significantly at 1, 2, 4, 8nM. 7. In immunochemical staining, we found that cobrotoxin-immunochemical complex move into intracellular space dose-dependently. Conclusion : These results indicate that cobrotoxin has anti-cancer effects by inducing apoptosis.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 2003.11a
• /
• pp.78-78
• /
• 2003

Inulin, an active component of Chicorium intybus root, has been shown to stimulate the growth of bifidobacteria, and inhibit colon carcinogenesis. NO mediates a number of the host-defense functions of activated macrophages, including antimicrobial and tumoricidal activity. We examined the effect of inulin on the synthesis of NO in RAW 264.7 cells. Inulin alone had no effect, whereas inulin with IFN-ν synergistically increased the NO production and inducible NO synthase (iNOS) expression in RAW 264.7 cells. Synergy between IFN-ν and inulin was mainly dependent on inulin-induced TNF-${\alpha}$ secretion. Also, protein kinase C (PKC)-${\alpha}$ was involved in the inulin-induced NO production. Inulin-mediated NO production was inhibited by the protein tyrosine kinase (PTK) inhibitor, tyrphostin AG126. Since iNOS gene transcriptions have been shown to be under the control of the NF -$\kappa$B/Rel family of transcription factors, we assessed the effect of inulin on NF -$\kappa$B/Rel using an EMSA. Inulin produced strong induction of NF-$\kappa$B/Rel binding, whereas AP-l binding was slightly induced in RAW 264.7 cells. Inulin stimulated phosphorylation and degradation of I$\kappa$B-${\alpha}$. These results suggest that in IFN-ν-primed RAW 264.7 cells inulin might stimulate NO synthesis via activation of PKC-${\alpha}$ and PTK, resulting in the activation of NF-$\kappa$B.

• BMB Reports
• /
• v.40 no.1
• /
• pp.88-94
• /
• 2007

Matrix metalloproteinase-9 (MMP-9) plays a pivotal role in the turnover of extracellular matrix (ECM) and in the migration of normal and tumor cells in response to normal physiologic and numerous pathologic conditions. Here, we show that the transcription of the MMP-9 gene is induced by lipopolysaccharide (LPS) stimulation in cells of a macrophage lineage (RAW 264.7 cells). We provide evidence that the NF-$\kappa$B binding site of the MMP-9 gene contributes to its expression in the LPS-signaling pathway, since mutation of NF-$\kappa$B binding site of MMP-9 promoter leads to a dramatic reduction in MMP-9 promoter activation. In addition, the degradation of l$\kappa$B$\alpha$;, and the presences of myeloid differentiation protein (MyD88) and tumor necrosis factor receptor-associated kinase 6 (TRAF6) were found to be required for LPS-activated MMP-9 expression. Chromatin immunoprecipitation (ChIP) assays showed that functional interaction between NF-$\kappa$B and the MMP-9 promoter element is necessary for LPS-activated MMP-9 induction in RAW 264.7 cells. In conclusion, our observations demonstrate that NF-$\kappa$B contributes to LPS-induced MMP-9 gene expression in a mouse macrophage cell line.

• Proceedings of the Korean Society of Toxicology Conference
• /
• 2002.05b
• /
• pp.88.2-98
• /
• 2002

A wide arry of naturally occurring substances particularly those present in dietary and medicinal plants, have been reported to possess substantial cancer chemopreventive properties. Certain phytochemicals retain strong antioxidative and anti-inflammatory properties which appear to contribute to their chemopreventive or chemoprotective activities. Inducible cyclooxygenase(COX-2) and nitric oxide synthase (iNOS) are important enzymes that mediate inflammatory processes. There is some evidence that expression of both COX-2 and iNOS is co-regulated by the eukaryotic transcription factor NF-$textsc{k}$B. Increased expression of COX-2 and/or iNOS has been associated with pathophysiology of certain types of human cancers as well as inflammatory diseases. Since inflammation is closely linked to tumor promotion, substances with potent anti-inflammatory activies are anticipated to exert chemopreventive effects on carcinogenesis, particularly in the promotion stage. An example is curcumin, a yellow pigment of turmeric (Curcuma longa L., Zingiberaceae), that strongly occurring diaryl heptanoids structurally related to curcumin have substantial anti-tumor promotional activities in two-stage mouse skin carcinogenesis. Thus, yakuchinone A [1-(4'-hydroxy-3'-methoxyphenyl)-7-phenyl-3heptanone] and yakuchinone B [1-(4'-hydroxy-3'methoxyphenyl)-7-phenylhept-1-en-3-one] present in Alpinia oxyphylla Miquel (Zingiberacease) attenuate phorbol ester-induced inflammation and papilloma formation in female ICR mice. These diarylheptanoids also suppressed phorbol ester-induced activation of epdermal ornithine decarboxylase and its mRNA expression when applied onto shaven backs of mice. Yakuchinone A and B as well as curcumin inhibited phorbol ester-induced expression of COX-2 and iNOS and their mRNA in mouse skin via inactivation of NF-$textsc{k}$B. Capsaicin, a major pungent ingredient of red pepper also attenuated phorbol ester-induced NF-$textsc{k}$B activation. Similar suppression of COX-2 and iNOS and down-regulation of NF-$textsc{k}$B activation for its DNA binding were observed with the ginsenosied Rg3 and the ethanol extract of Artemisia asiatica. We have also found that certain anti-inflammatory phytochemicals exert inhibitory effects on phorbol ester-induced COX-2 expression and NF-$textsc{k}$B activation in immortalized human breast epithelial (MCF-10A) cells in culture. One of the plausible mechanisms undelying inhibition by aforementioned phytochemicals of phorbol ester-induced NF-$textsc{k}$B activation involves interference with degragation of the inhibitory unit, I$textsc{k}$Ba, which blocks subsequent nuclear translocation of the functionally active p65 subunit of NF-$textsc{k}$B. the activation of epidermal NF-$textsc{k}$B by phorbol ester and subsequent induction of COX-2 hence appear to play an important role in intracellular signaling pathwasy leading to tumor promotion and targeted inhibition of NF-$textsc{k}$B may provide a new promising cancer chemopreventive strategy.

• Membrane and Water Treatment
• /
• v.6 no.3
• /
• pp.173-187
• /
• 2015

The goal of present work is the preparation of a novel positively charged nanofiltration (NF) membrane and its development for the cation removal of aqueous solutions. This NF membrane was fabricated by the surface modification of polysulfone (PSf) ultrafiltration support. The active top-layer was formed by interfacial cross-linking polymerization of poly(ethyleneimine) (PEI) with p-xylylene dichloride (XDC) and then quaternized with methyl iodide to form a perpetually positively charged layer. In order to improve the efficiency of nanofiltration membrane, the concentration of PEI, XDC and methyl iodide solutions, PEI coating and cross-linking time have been optimized. As a result, a high water flux and high $CaCl_2$ rejection (1,000 ppm) was obtained for the composite membrane with values of $18.29L/m^2.h$ and 93.62% at 4 bar and $25^{\circ}C$, respectively. The rejections of NF membrane for different salt solutions followed the order of $Na_2SO_4$ < $MgSO_4$ < NaCl < $CaCl_2$. Molecular weight of cut off (MWCO) was calculated via retaining of PEG solutions with different molecular weights that finally, it revealed the Stokes and hydrodynamic radius of 1.457 and 2.507 nm on the membrane selective layer, respectively. The most efficient positively charged nanofiltration membrane exhibited a $Ni^{2+}$ rejection of 96.26% for industrial wastewater from Shamse Hadaf Co. (Kashan, Iran).

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
• /
• v.22 no.1
• /
• pp.33-45
• /
• 2009

Objective : Lacca Sinic: Exsiccate (LSE) extracted from Rhus vemicitlus Stokes (RVS) has been used traditionally as a remedy for inflammation in Korea, China, and Japan. However, as yet there is no clear explanation of how LSE affects the production of inflammatory cytokines. This study was to determine the effects of LSE on the mast cell-mediated inflammatory responses. Method : We measured the amount of inflammatory cytokine production induced by the phorbol myristate acetate (PMA) plus calcium ionophore(A231S7) in the human mast cell line (HMC-l) incubated with various concentrations of Laces Sinica Exsiccate (LSE). The $TNF-\alpha$, IL-6 and IL-8 secreted protein levels were measured by the ELISA assay. The $TNF-\alpha$, IL-6 and IL-8 mRNA levels were measured by the RT-PCR analysis. Nuclear and cytoplasmic proteins were examined by Western blot analysis. The NF-${\kappa}B$ promoter activity was examined by a luciferase assay. Result : LSE inhibited the PMA + A231S7-induced $TNF-\alpha$, IL-6, and IL-8 expression and suppressed NF-${\kappa}B$ activation in the stimulated-HMC-1. In addition, LSE inhibited induction of NF-${\kappa}B$ promoter-mediated luciferase activity. Conclusion : In this study, we have found that LSE is an inhibitor of NF-${\kappa}B$ and cytokines on the mast cell-mediated inflammatory responses.