• Environmental Mutagens and Carcinogens
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• v.22 no.4
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• pp.312-318
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• 2002

Dioxin represents a group of halogenated aromatic hydrocarbons of which 2,3,7,8-tetrachlorod-ibenzo-p-dioxin (TCDD) is well known for its extremely toxic properties as well as ubiquitous presence in our environment and ecosystems. In order to better assess the carcinogenic mechanism of dioxin, we should utilize the reliable biomarkers that can precisely and correctly reflect multi-stage carcinogenesis. When MCF10A cells were exposed to TCDD (10 nM), expression of both CYP1A1 and CYP1B1 was induced in a time-related manner. The expression as well as activity of ornithine decarboxylase was transiently induced by TCDD treatment. In contrast, the induction of COX-2 that is implicated in carcinogenesis as well as inflammation, was not induced by TCDD. In another study, gap-junctional intercellular communication (GJIC) was attenuated by TCDD treatment as revealed by the dye-transfer assay. Based on these findings, TCDD has both tumor initiating and promoting potential in human breast epithelial cells in culture. Also, treatment of MCF10A cells with the carcinogen 7,12-dimethylbenz[a]anthracene plus TCDD resulted in malignant cell transformation as revealed by increased anchorage-independent growth of exposed cells. Additional studies may be necessary to assess the effects of TCDD on multi-stage carcinogenesis in vivo.

• Korean Journal of Exercise Nutrition
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• v.25 no.4
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• pp.45-53
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• 2021

[Purpose] In this study, we investigated whether a 70% ethanolic (EtOH) extract of Sargassum horneri had antioxidant and anti-inflammatory effects in lipopolysaccharide (LPS)-stimulated macrophage-like RAW 264.7 cells. [Methods] The proximate composition, fatty acids, amino acids, and dietary fiber of S. horneri, various biologically active compounds, and antioxidant activity were analyzed. [Results] The DPPH and ABTS free radical scavenging activities, as well as the reduction power, of the S. horneri extract used here were significantly increased in a concentration-dependent manner. This indicates that S. horneri contains bioactive compounds, such as phenols and flavonoids, that have excellent antioxidant activity. The cellular viability and metabolic activity results confirmed that the extract had no discernible toxicity at concentrations up to 100 ㎍/mL. The levels of nitrites and cytokines (PGE₂, TNF-α and IL-6), which mediate pro-inflammatory effect, were significantly inhibited by treatment with either 50 or 100 ㎍/mL S. horneri extract, whereas that of IL-1β was significantly inhibited by treatment with 100 ㎍/mL of the extract. Similarly, the expression of iNOS and COX-2 proteins also decreased according to 50 or 100 ㎍/mL extract concentrations. NF-κB binding to DNA was also significantly inhibited by treatment with 100 ㎍/mL of extract. [Conclusion] These results suggest that 70% EtOH extracts of S. horneri can relieve inflammation caused by disease or high intensity exercise.

• The Korea Journal of Herbology
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• v.30 no.5
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• pp.67-74
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• 2015

Objectives : TheHaedokgumhwa-sanwater extract (HDKHS) is used in Korea, Japan and China as a traditional therapeutic agent to cure an infectious disease. But its study is not enough. Therefore, the present study focused on the elucidation of HDKHS to investigate the anti-inflammatory effects and to established the possible mechanisms involved in its action on LPS-stimulated immune response in murine macrophages.Methods : Inflammatory status was induced by LPS and measured by increasement of inflammatory mediators. LPS induced secretions of NO and PGE₂in RAW 264.7 cells were measured using griess reagent and enzyme-linked immunosorbent assay (ELISA) kit respectively. production of IL-6 was examined using ELISA kit and expression of IL-6 mRNA was measured by RT-PCR method. To investigate the effects of HDKHS on inflammatory mediators, such as iNOS, COX-2 and MAPKs, western blot and RT-PCR were performed.Results : HDKHS significantly reduced production of NO and PGE2 which were induced by LPS. Also, activation of IL-6 was reduced both protein and mRNA levels. The expressions of inflammatory mediator include iNOS and COX-2 were decreased by pretreatment with HDKHS. futhermore The result showed HDKHS down-regulate the LPS induced phosphorylation of ERK 1/2, one of the MAPK family, which is considered as a main regulator of transmission from pathogens to nucleus of immune cells.Conclusions : Our results suggest that the anti-inflammatory properties of HDKHS may stem from the inhibition of pro-inflammatory mediators via suppression of initiation of inflammatory response by inhibiting MAPKs signaling pathways.

• Animal cells and systems
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• v.9 no.2
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• pp.101-105
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• 2005

[ ${\beta}ig-h3$ ] is a secretory protein that is induced by $TGF-{\beta}$ and implicated in various disease conditions including fibrosis. We have previously reported that ${\beta}ig-h3$ expression is implicated in astrocyte response to brain injury. In this study, we further investigated potential roles of ${\beta}ig-h3$ protein in the injured central nervous system (CNS). We specifically assessed whether the treatment of microglial cells with ${\beta}ig-h3$ can regulate microglial activity. Microglial cells are the prime effector cells in CNS immune and inflammatory responses. When activated, they produce a number of inflammatory mediators, which can promote neuronal injury. We prepared conditioned medium from the stable CHO cell line transfected with human ${\beta}ig-h3$ cDNA. We then examined the effects of the conditioned medium on the LPS- or $IFN-{\gamma}-mediated$ induction of proinflammatory molecules in microglial cells. Preincubation with the conditioned medium significantly attenuated LPS-mediated upregulation of $TNF-{\alpha},\;IL-1{\beta}$, iNOS and COX-2 mRNA expression in BV2 murine microglial cells. It also reduced $IFN-{\gamma}-mediated$ upregulation of $TNF-{\alpha}$ and COX-2 mRNA expression but not iNOS mRNA expression. Assays of nitric oxide release correlated with the mRNA data, which showed selective inhibition of LPS-mediated nitric oxide production. Although the regulatory mechanisms need to be further investigated, these results suggest that astrocyte-derived ${\beta}ig-h3$ may contribute to protection of the CNS from immune-mediated damage via controlling microglial inflammatory responses.

• The Korea Journal of Herbology
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• v.27 no.4
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• pp.99-107
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• 2012

Objectives : The aim of this study was to authenticate whether fractionated extract of Rumex japonicus HOUTT. (RJ) has anti-inflammatory effects in mouse macrophage, RAW264.7 cells. Methods : Roots of RJ were extracted by methanol for 48hours. The methanol that gained was filtered and freeze dried. The methanol extract was dissolved in water and dichloromethane (DCM). After that, two layers were separated. Ethyl acetate (EA) added to the water layer and separated again. All the layers were filtered and freeze dried and the extracts were tested. Cytotoxic activity of extracts on RAW 264.7 cells was measured using MTS assay. The nitric oxide (NO) production was measured and proinflammatory cytokines and $PGE_2$ were measured by ELISA kit. The levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), I ${\kappa}$-B-${\alpha}$ and nuclear NF-${\kappa}B$ p65 expression were detected by western blot. Results : Our results indicated that DCM and EA extracts of RJ inhibited the LPS-induced NO, $PGE_2$ production and iNOS, COX-2 expression accompanied by an attenuation of TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 production in RAW 264.7 cells most effectively. DCM and EA extracts also had suppression effects of LPS-induced NF-${\kappa}B$ and MAPKs activation. Conclusions : This results demonstrate that fractionated extract of RJ has anti-inflammatory effects and among the fractioned extract, dichloromethane and ethyl acetate extract have best anti-inflammatory effects.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2003.11a
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• pp.77-77
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• 2003

Sepsis remains common surgical problems with high morbidity and mortality despite improvement in the management for septic patient. Although hepatocellular dysfunction occurs during sepsis, the mechanism responsible for this remains unclear. In sepsis, a state of severe oxidative stress is encountered, with host endogenous antioxidant defenses overcome. Therefore, the aim of this study was to determine the effect of a-tocopherol (AT) vasoregulatory gene expression during polymicrobial sepsis. Rats were subjected to polymicrobial sepsis by cecal ligation and puncture (CLP). AT (15 mg/kg) was intraperitonealy injected for 3 days prior to CLP. Blood samples were taken 24 h after CLP for measurement of the extent of hepatocellular damage. Liver samples were taken for RT-PCR analysis of mRNA for genes of interest: endothelin-l (ET-l), its receptors $ET_{A}$ and $ET_{B}$, nitric oxide synthases (iNOS and eNOS), cyclooxygenase-2 (COX -2), heme oxygenase-l (HO-l), and tumor necrosis factor-$\alpha$ (TNF-$\alpha$). The activities of serum alanine aminotransferase and lipid peroxidation level were significantly increased; an increase which was prevented by AT pretreatment. CLP significantly increased the mRNA levels of ET-1 and $ET_{B}$; an increase that was prevented by AT pretreatment. However, the level of $ET_{A}$ mRNA significantly decreased after CLP; a decrease that was not prevented by AT pretreatment. There were significant increases in the mRNA expression of iNOS, HO-l and COX -2 in CLP groups. This increase was prevented by AT pretreatment. The expression of eNOS and TNF-$\alpha$ mRNA significantly increased in CLP, which was not prevented by AT pretreatment. Our findings suggest that there was an imbalanced vasoregulatory gene expression in sepsis, and AT ameliorates this change through its free radical scavenging activity.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.3
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• pp.503-509
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• 2011

Jakyaktang(芍藥湯; JYT) exhibits potent anti-inflammatory activity in widely intestinal disease, but its mechanism was undisclosed. To elucidate the molecular mechanisms of JYT on pharmacological and biochemical actions in inflammation, we examined the effect of JYT on pro-inflammatory mediators in phorbol 12-myristate 13-acetate (PMA) plus A23187-induced mast cell and lipopolysaccharide (LPS)-stimulated macrophages. The investigation focused on whether JYT inhibited pro-inflammatory cytokines such as interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) in PMA plus A23187- induced HMC-1 cells and inflammatory madiators such as nitric oxide (NO), TNF-${\alpha}$, IL-6, iNOS, COX-2 in LPS-stimulated RAW 264.7 cells. We found that JYT inhibited LPS-induced NO, TNF-${\alpha}$ and IL-6 productions as well as the expressions of iNOS and COX-2. These results suggest that JYT has inhibitory effects on mast cell-mediated and macropage-mediated inflammation.

• Biomolecules & Therapeutics
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• v.22 no.3
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• pp.200-206
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• 2014

N-(p-Coumaryol) tryptamine (CT), a phenolic amide, has been reported to exhibit anti-oxidant and anti-inflammatory activities. However, the underlying mechanism by which CT exerts its pharmacological properties has not been clearly demonstrated. The objective of this study is to elucidate the anti-inflammatory mechanism of CT in lipopolysaccharide (LPS)-challenged RAW264.7 macrophage cells. CT significantly inhibited LPS-induced extracellular secretion of pro-inflammatory mediators such as nitric oxide (NO) and $PGE_2$, and protein expressions of iNOS and COX-2. In addition, CT significantly suppressed LPS-induced secretion of pro-inflammatory cytokines such as TNF-${\alpha}$ and IL-$1{\beta}$. To elucidate the underlying anti-inflammatory mechanism of CT, involvement of MAPK and Akt signaling pathways was examined. CT significantly attenuated LPS-induced activation of JNK/c-Jun, but not ERK and p38, in a concentration-dependent manner. Interestingly, CT appeared to suppress LPS-induced Akt phosphorylation. However, JNK inhibition, but not Akt inhibition, resulted in the suppression of LPS-induced responses, suggesting that JNK/c-Jun signaling pathway significantly contributes to LPS-induced inflammatory responses and that LPS-induced Akt phosphorylation might be a compensatory response to a stress condition. Taken together, the present study clearly demonstrates CT exerts anti-inflammatory activity through the suppression of JNK/c-Jun signaling pathway in LPS-challenged RAW264.7 macrophage cells.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.6
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• pp.1320-1331
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• 2009

Oxidative stress and inflammation are important events in the development of chronic inflammatory diseases including arthritis, atherosclerosis, diabetes, hypertension. Cynomorium songaricum (CS) has been used as a traditional Korean herbal medicine, and it is currently used in traditional clinics to treat frequent urination, spermatorrhea, weakness of the sinews and constipation in the folk medicine. The aim of this study was to determine whether fractionated extracts of CS inhibit free radical generation such as DPPH radical, superoxide radical, nitric oxide and peroxynitrite, production of nitrite an index of NO, $PGE_2$, iNOS, COX-2 and pro-inflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. Cytotoxic activity of extracts on RAW 264.7 cells was measured using 5-(3-caroboxymeth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Our results indicated that the most superior extract which scavenged DPPH radical, reactive oxygen species (ROS) and RNS was CS ethyl acetate extract (CSEA). Moreover, CSEA significantly inhibited the LPS-induced NO, $PGE_2$ production and iNOS, COX-2 expression accompanied by an attenuation of TNF-$\alpha$, IL-$1{\beta}$ and IL-6 formation in macrophages. Furthermore, CSEA treatment also blocked LPS-induced intracellular ROS production and the activation of NF-${\kappa}B$. These findings indicate that CSEA inhibits the production of pro-inflammatory mediators and cytokines via the suppression of ROS production and NF-${\kappa}B$ activation. Take together, these results indicate that CSEA has the potential for use as an natural anti-oxidant and an agent of anti-chronic inflammatory diseases.

• The Korea Journal of Herbology
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• v.24 no.2
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• pp.49-56
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• 2009

Objectives : Inflammation is important event in the development of vascular diseases including hypertension, atherosclerosis, and restenosis. Bodusan (BDS) was a traditional Korean herbal medicine and widely used in treatment of gastrointestinal complaint and stomach ulcer. The aim of this study was to determine whether BDS and its components inhibit production of nitrite, PGE2 and proinflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. Methods : Cytotoxic activity of BDS and its components on RAW 264.7 cells was using 5-(3caroboxymcrophages. eth-oxyphenyj)-2H-tetra-zolium inner salt (MTS) assay. The nitric oxide (NO) production was measured by Griess reagent system. And proinflammatory cytokines were measured by ELISA kit. The levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression were detected by western blot. Results : Our results indicated that BDS and its components significantly inhibited the LPS-induced NO and PGE2 production. Moreover. BDS and its components inhibited iNOS and COX-2 expression accompanied by an attenuation of TNF-${\alpha}$, IL-11${\beta}$, IL-6 and MCP-1 formation in macrophages. Conclusions: These results indicate that BDS and its components have potential as an anti-inflammatory agent.