• Biomolecules & Therapeutics
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• v.22 no.5
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• pp.390-399
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• 2014

Chalcones (1,3-diaryl-2-propen-1-ones), a flavonoid subfamily, are widely known for their anti-inflammatory properties. Propenone moiety in chalcones is known to play an important role in generating biological responses by chalcones. In the present study, we synthesized chalcone derivatives structurally modified in propenone moiety and examined inhibitory effect on nitric oxide (NO) production and its potential mechanisms. Among the chalcone derivatives used for this study, TI-I-174 (3-(2-Hydroxyphenyl)-1-(thiophen-3-yl)prop-2-en-1-one) most potently inhibited lipopolysaccharide (LPS)-stimulated nitrite production in RAW 264.7 macrophages. TI-I-174 treatment also markedly inhibited inducible nitric oxide synthase (iNOS) expression. However, TI-I-174 did not significantly affect production of IL-6, cyclooxygenase-2 (COX-2) and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), implying that TI-I-174 inhibits production of inflammatory mediators in a selective manner. Treatment of macrophages with TI-I-174 significantly inhibited transcriptional activity of activator protein-1 (AP-1) as determined by luciferase reporter gene assay, whereas nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activity was not affected by TI-I-1744. In addition, TI-I-174 significantly inhibited activation of c-Jun-N-Terminal kinase (JNK) without affecting ERK1/2 and p38MAPK, indicating that down-regulation of iNOS gene expression by TI-I-174 is mainly attributed by blockade of JNK/AP-1 activation. We also demonstrated that TI-I-174 treatment led to an increase in heme oxygenase-1 (HO-1) expression both at mRNA and protein level. Transfection of siRNA targeting HO-1 reversed TI-I-174-mediated inhibition of nitrite production. Taken together, these results indicate that TI-I-174 suppresses NO production in LPS-stimulated RAW 264.7 macrophages via induction of HO-1 and blockade of AP-1 activation.

• Biomolecules & Therapeutics
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• v.22 no.1
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• pp.27-34
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• 2014

Curcumin is naturally occurring polyphenolic compound found in turmeric and has many pharmacological activities. The present study was undertaken to evaluate anti-allergic inflammatory activity of curcumin, and to investigate its inhibitory mechanisms in immunoglobulin E (IgE)/Ag-induced mouse bone marrow-derived mast cells (BMMCs) and in a mouse model of IgE/Ag-mediated passive systemic anaphylaxis (PSA). Curcumin inhibited cyclooxygenase-2 (COX-2) dependent prostaglandin $D_2$ ($PGD_2$) and 5-lipoxygenase (5-LO) dependent leukotriene $C_4$ ($LTC_4$) generation dose-dependently in BMMCs. To probe the mechanism involved, we assessed the effects of curcumin on the phosphorylation of Syk and its downstream signal molecules. Curcumin inhibited intracellular $Ca^{2+}$ influx via phospholipase $C{\gamma}1$ ($PLC{\gamma}1$) activation and the phosphorylation of mitogen-activated protein kinases (MAPKs) and the nuclear factor-${\kappa}B$ (NF-${\kappa}B$) pathway. Furthermore, the oral administration of curcumin significantly attenuated IgE/Ag-induced PSA, as determined by serum $LTC_4$, $PGD_2$, and histamine levels. Taken together, this study shows that curcumin offers a basis for drug development for the treatment of allergic inflammatory diseases.

• Korean Journal of Pharmacognosy
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• v.43 no.3
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• pp.217-223
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• 2012

In the present study, we investigated anti-inflammatory activity of artemisinic acid in HaCaT cells and RAW264.7 cells. Artemisinic acid showed inhibitory activity on macrophage-derived chemokines (MDC) expression, a factor related with atopic dermatitis (AD), in interferon (IFN)-${\gamma}$ and tumor necrosis factor (TNF)-${\alpha}$-stimulated HaCaT cells. In the study on action mechanism, pretreated artemisinic acid reduced the phosphorylation of STAT1 and p38 and the degradation of $I{\kappa}B$ by IFN-${\gamma}$ and TNF-${\alpha}$ stimulations. However, artemisinic acid didn't show the inhibitory activity on LPS-induced inflammatory mediators (NO, $PGE_2$, IL-6) in RAW264.7 cell. These results indicate that artemisinic acid inhibits IFN-${\gamma}$ and TNF-${\alpha}$-induced MDC expression through inhibition of signal factors, STAT1, NF-${\kappa}B$, and p38, in HaCaT keratinocytes.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.5
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• pp.1315-1321
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• 2008

Chan-Su (Venenum bufonis) has long been for a variety of other purposes including treatment of inflammation in the folk medicine recipe. Since nitric oxide (NO) is one of the major inflammatory parameters, we first studied the effects of Chan-Su on NO production in lipopolysaccharide (LPS)-stimulated BV2 microglial cells, Chan-Su inhibited the secretion of NO in BV2 microglial cells, without affecting cell viability, The protein level of inducible nitric oxide synthase (iNOS) was decreased by Chan-Su, And Chan-Su also inhibited production of prostaglandin E2 (PGE2) and expression of cyclooxygenase (COX)-2. Proinflammatory cytokines, such as tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\beta}$ and IL-12, were inhibited by Chan-Su in a dose-dependent manner. And Chan-Su inhibited the degradation of ${IkB-\alpha}$, which was considered to be inhibitor of nuclear factor $(NF)-{\kappa}B$, one of a potential transcription factor for the expression of iNOS, COX-2 and proinflammatory cytokines. These results suggest that Chan-Su could exert its anti-inflammatory actions by suppressing the synthesis of NO through inhibition of $I{\kappa}B-{\alpha}$ degradation.

• The Journal of Pediatrics of Korean Medicine
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• v.28 no.3
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• pp.31-46
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• 2014

Objectives Forsythiae Fructus treatment has been used for inflammatory and allergic diseases in Korean Medicine. Nevertheless, the mechanism of action and the cellular targets are not understood well. The pathogenesis of allergic diseases are associated with Th2 cytokines such as IL-13, MIP-$1{\alpha}$, IL-13, IL-5, GM-CSF, IL-4, TNF-${\alpha}$ and IL-6, which are secreted by the mast cells. This study was conducted to investigate the effects of Forsythiae Fructus extracts (FF) on Th2 cytokines expression and signal transduction in MC/9 mast cells. Methods In the study, MC/9 mast cells were stimulated with DNP-IgE for 24 hours and then treated separately with CsA $10{\mu}g/m{\ell}$ and varying doses of FF for one hour. MC/9 mast cells stimulated with DNP-IgE was the control group, a treatment with CsA was the positive control group and a treatment with varying doses FF was the experimental groups. The mRNA levels of IL-13, IL-5, GM-CSF, IL-4, TNF-${\alpha}$, IL-6 were analyzed with Real-time PCR. The levels of IL-13, MIP-$1{\alpha}$ were measured using enzyme-linked immunosorbent assays(ELISA). NFAT, AP-1 and NF-${\kappa}B$ p65 were examined by Western blot analysis. Results 1. FF were observed to suppress the mRNA expression of IL-13, IL-5, GM-CSF, IL-4, TNF-${\alpha}$, IL-6 in comparison to DNP-IgE control group. 2. FF also has inhibited the IL-13, MIP-$1{\alpha}$ production significantly in comparison to DNP-IgE control group. 3. Western blot analysis of transduction factors involving Th2 cytokines expression has revealed a prominent decrease of the mast cell specific transduction factors including NFAT-1, NFAT-2, c-Jun, and NF-${\kappa}B$ p65 but c-Fos. Conclusions In conclusion, the anti-allergenic activities of FF may be strongly related to the regulation of transcription factors NFAT-1, NFAT-2, c-Jun, and NF-${\kappa}B$ p65 causing inhibition of Th2 cytokines in mast cells.

• The Journal of Pediatrics of Korean Medicine
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• v.32 no.4
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• pp.71-86
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• 2018

Objectives The purpose of this study is to investigate the effects of Gardenia jasminoides for. grandiflora extracts' (GAJ) anti-inflammatory effect on RBL-2H3 mast cells and OVA/alum-sensitized mice. Methods In this study, IL-4 and IL-13 production was measured via ELISA analysis, and mRNA expressions of GM-CSF, IL-4, IL-5, $TNF-{\alpha}$, IL-6 were analyzed by real-time PCR. In addition, MAPKs and $NF-{\kappa}B$ p65 transcription factors were examined using western blotting, and ELISA was used to understand IgE, IL-4, and IL-13 production in ovalbumin-allergic mice in in vitro study. Results As a result of this study, 1. GAJ were observed to suppress the mRNA expression of GM-CSF, IL-4, IL-13, IL-5, $TNF-{\alpha}$, IL-6 in comparison to PMA 50 ng/ml, ionomycin $0.5{\mu}M$ (PI) control group. 2. GAJ also inhibited the IL-4, IL-13 production in comparison to PI control group. 3. Western blot analysis showed decrease on the expression of mast-cell-specific transcription factors, including MAKPs (ERK, JNK, p38) and $NF-{\kappa}B$ p65. 4. Orally-administered GAJ group in OVA/alum induced Balb/c mice showed decreased level of OVA-specific IgE in the serum. This group also has shown decreased the level of IL-4, IL-13 in the splenocyte culture supernatant. Conclusions Obtained results suggest that GAJ may regulate the allergic inflammation by transcription factors MAKPs (ERK, JNK, p38) and $NF-{\kappa}B$ p65 causing inhibition of Th2 cytokines in mast cells and OVA/alum-sensitized mice.

• Journal of the Korean Applied Science and Technology
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• v.36 no.2
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• pp.468-478
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• 2019

The aim of this study is to evaluate the efficacy of cordyceps militaris extracts for the improvement of cognitive dysfunction in PC12 and BV2 cells. Cordyceps militaris extracts was prepared by extracting with distilled water. Cell viability was assessed by MTT assay using PC12 cells and BV2 cells. Confirmed effects of L-glutamate induced cytotoxicity test, Acetylcoline (ACh) concentration, and Acetylcolinestase (AChE) activity in PC12 cells. Anti-inflammatory activities of cordyceps militaris extracts was measured through changes in the levels of nitric oxide (NO), and prostaglandin E2 ($PGE_2$) on lipopolysaccharide(LPS)-induced BV2 cell. In addition, we measured the expression of $NF-{\kappa}B$, p38, JNK, and caspase-3 in western blot analysis. Cordyceps militaris extracts showed no cytotoxicity at the concentrations of 1, 10, and $100{\mu}g/m{\ell}$ except for the concentration of $200{\mu}g/m{\ell}$. Cordyceps militaris extracts protected the cell and exhibited significant increases in the ACh concentration and a significant decrease in the AChE activity in L-glutamate induced PC12 cells. Moreover, cordyceps militaris extracts inhibited the productions NO, and PGE2 level and the protein expression of $NF-{\kappa}B$, p38, JNK, caspase-3 in LPS-induced BV2 cells. These results indicate that cordyceps militaris extracts possible prevented and improved cognitive dysfuction symptoms. Thus, cordyceps militaris extracts may be a novel natural material option for the improvement of cognitive dysfunction.

• Korean Journal of Plant Resources
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• v.25 no.6
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• pp.682-692
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• 2012

Sea Buckthorn (Hippophae rhamnoides L.) has been used in traditional medicine for the treatment of cough, indigestion, circulatory problems and pain. The associated anti-inflammatory effect of this agent is achieved via the inhibition of Nf-${\kappa}B$ signaling, a property that has been demonstrated to effectively control the symptoms of various skin disorders, including atopic dermatitis. Accordingly, the purpose of this study was to assess the efficacy of Sea Buckthorn in reducing the production of lipopolysaccharide (LPS) activated nitric oxide (NO) by inhibiting the Nf-${\kappa}B$ pathway, as measured by the symptoms of atopic dermatitis (AD) occurring secondarily to inflammation and immune dysregulation. Our data demonstrate that Sea Buckthorn significantly decreased the LPS-induced production of NO (p<0.001). Atopic dermatitis was induced by repeated application of 2,4-dinitrochlorobenzene to the dorsal skin of mice. Topical application of 5% Sea Buckthorn extract improved the symptoms of AD, specifically reducing disease severity scores, scratching behaviors and epidermal thickness. When compared to the control group, animals treated with Sea Buckthorn exhibited increased serum IL-12 levels and decreased serum TNF-${\alpha}$, IL-4 and IL-5 levels. Such a modulation of biphasic T-helper (Th)1/Th2 cytokines may result in a reduction in serum IgE levels. Our findings suggest that mechanism of action of Sea Buckthorn in the treatment of AD is associated with a marked anti-inflammatory effect as well as an inhibition of Th2-mediated IgE overproduction via the modulation of biphasic Th1/Th2 cytokines. Such results suggest that topical Sea Buckthorn extract may prove to be a novel therapy for AD symptoms with few side effects.

• Korean Journal of Food Science and Technology
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• v.39 no.2
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• pp.175-180
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• 2007

Toll-like receptors induce innate immune responses recognizing conserved microbial structural molecules that are known as pathogen-associated molecular patterns (PAMPs). Ligand-induced homotypic oligomerization was found to proceed in LPS-induced activation of TLR4 signaling pathways. TLR2 is known to heterodimerize with TLR1 or TLR6 and recognize diacyl- or triacyl-lipopeptide, respectively. These results suggest that ligand-induced receptor dimerization of TLR4 and TLR2 is required for the activation of downstream signaling pathways. Therefore, receptor dimerization may be one of the first lines of regulation in the activation of TLR-mediated signaling pathways and induction of subsequent innate and adaptive immune responses. Here, we report biochemical evidence that curcumin from the plant Curcuma longa inhibits activation of $NF-{\kappa}B$, expression of COX-2, and dimerization of TLRs induced by TLR2, TLR3 and TLR4 agonists. These results imply that curcumin can modulate the activation of TLRs and subsequent immune/inflammatory responses induced by microbial pathogens.

• Biomolecules & Therapeutics
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• v.23 no.2
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• pp.119-127
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• 2015

Chalcones (1,3-diaryl-2-propen-1-ones), a subfamily of flavonoid, are widely known to possess potent anti-inflammatory and anti-oxidant properties. In this study, we investigated the effect of 3-(4-Hydroxyphenyl)-1-(thio3-(4-Hydroxyphenyl phen-2-yl)prop-2-en-1-one (TI-I-175), a synthetic chalcone derivative, on endotoxin-induced expression of monocyte chemoattractant protein-1 (MCP-1), one of the key chemokines that regulates migration and infiltration of immune cells, and its potential mechanisms. TI-I-175 potently inhibited MCP-1 mRNA expression stimulated by lipopolysaccharide (LPS) in RAW 264.7 macrophages without significant effect on cell viability. Treatment of cells with TI-I-175 markedly prevented LPS-induced transcriptional activation of activator protein-1 (AP-1) as measured by luciferase reporter assay, while nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activity was not inhibited by TI-I-175, implying that TI-I-175 suppressed MCP-1 expression probably via regulation of AP-1. In addition, TI-I-175 treatment significantly inhibited LPS-induced Akt phosphorylation and led to a significant decrease in reactive oxygen species (ROS) production by LPS, which act as up-stream signaling events required for AP-1 activation in RAW 264.7 macrophages. Taken together, these results indicate that TI-I-175 suppresses MCP-1 gene expression in LPS-stimulated RAW 264.7 macrophages via suppression of ROS production and Akt activation.