• The Journal of Korean Medicine
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• v.28 no.2 s.70
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• pp.155-165
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• 2007

Objective : The Angelica gigas Nakai ethanol extract (AGE) was investigated to compare nitric oxide (NO) production and $NF-{\kappa}B$ activity from RAW 264.7 cells, since NO and nuclear $factor-{\kappa}B$ $(NF-{\kappa}B)$ have been shown to be factors implicated in inflammatory disease. Method : AGE was prepared by extracting medicinal herb with 70% (v/v) ethanol solution. We investigated production of nitric oxide (NO) and inducible nitric oxide synthase (iNOS) gene expression by ARE in LPS-stimulated RAW 264.7 macrophage cells. We also investigated inhibition of LPS-induced activation of $NF-{\kappa}B$ on western blot. Result : LPS-induced RAW 264.7 cells increased NO production and iNOS expression. Upon treatment with AGE, nitrite production was significantly inhibited in a concentration-dependent manner compared to the untreated control. AGE inhibited this LPS-induced iNOS mRNA and protein in a dose-dependent manner. AGE markedly inhibited the expression of iNOS mRNA and protein at a concentration of 100 ${\mu}g/ml$. LPS-induced RAW 264.7 cells with AGE blocked inhibitory $factor-{\kappa}B{\alpha}$ degradation. Conclusion :This study shows that AGE seems to attenuate inflammation through inhibition of NO production and iNOS expression by blockade of $NF-{\kappa}B$ activation in LPS-stimulated RAW 264.7 cells.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.22 no.2
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• pp.19-38
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• 2009

Objective : The purpose of this study is to find out whether Herba Ephedrae, solely used, and Herbal medicine in which this was included, have inhibitory effects of Nitric Oxide(NO). Methods : We tested the inhibitory effects of Nitric Oxide(NO) with Herba Ephedrae and ten kinds of Herbal medicine combined with Herba Ephedrae(Worlbikachul-Tang, 越婢加朮湯; Mahaengkamsuk-Tang, 麻杏甘石湯; Shinbi-Tang, 神秘湯; Mahwangbujaseshin-Tang, 麻黃附子細辛湯; Euiin-Tang, 薏苡仁湯; Galgeun-Tang, 葛根湯; Mahaengeuigam-Tang, 麻杏薏甘湯; Mahwang-Tang, 麻黃湯; Socheongryong-Tang, 小靑龍湯; Gaemagakban-Tang, 桂麻各半湯) on RAW264.7 cells. Results and Conclusions : 1. We carried out MTT assay on Herba Ephedrae and those decoctions including this in order to determine whether they accommodate cotytoxicity. The results were that Worlbikachul-Tang, Mahaengkamsuk-Tang, Mahwangbujaseshin-Tang, Mahaengeuigam-Tang, Mahwang-Tang, Socheongryong-Tang and Gaemagakban-Tang showed no cytotoxicity on RAW264.7 with 0.1mg/ml and 0.5mg/ml dosages of decoctions but displayed cytotoxicity on the cell with 1mg/ml. Solely used Herba Ephedrae, Shinbi-Tang, Euiin-Tang and Galgeun-Tang exhibited cytotoxicity beyond the concentration of 0.5mg/ml. 2. Worlbikachul-Tang, Mahaengkamsuk-Tang, Shinbi-Tang, Mahwangbujaseshin-Tang, Euiin-Tang, Galgeun-Tang, Mahaengeuigam-Tang, Mahwang-Tang and Socheongryong-Tang showed inhibition of NO production but solely used Herba Ephedrae and Gaemagakban-Tang did not exhibit such reaction.

• The Korea Journal of Herbology
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• v.23 no.3
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• pp.127-134
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• 2008

Objectives : The present study was designed to investigate whether Ostericum koreanum (OK) could regulate lipopolysaccharide (LPS)-induced inflammatory response in vitro and in vivo. Methods : To evaluate of anti-inflammatory effect of OK, we examined Nitric oxide (NO), proinflammatory cytokines production in LPS-stimulated RAW264.7 cells. Furthermore, we checked molecular mechanism especially in the phosphorylation of mitogen-activated protein kinases (MAPKs) and the degradation of inhibitory kappa B a ($Ik-B{\alpha}$) using western blot and also investigated survival of mice in LPS-mediated endotoxin shock. Results : 1. Extract from OK itself have weak cytotoxic effect on RAW264.7 cells. Extract from OK inhibited LPS-induced NO, tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), interleukin $(IL)-1{\beta}$, IL-6 and IL-10 production in RAW264.7 cells. 2. OK inhibited the phosphorylation of MAPKs, such as p38, extracelluar signal-regulated kinase (ERK1/2) and c-Jun NH2-terminal kinase (JNK) and also the degradation of $I{\kappa}-B{\alpha}$ in the LPS-stimulated RAW264.7 cells 3. OK did not inhibit LPS-induced endotoxin shock. Conclusions : OK down-regulated LPS-induced NO and cytokines production through suppressing activation of MAPKs and degradation of $I{\kappa}-B{\alpha}$. Our results suggested that OK may be a beneficial drug against inflammatory diseases.

• YAKHAK HOEJI
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• v.51 no.1
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• pp.35-43
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• 2007

To investigate the immunomodulatory roles of cyclic AMP (CAMP) on macrophage- and T lymphocyte-mediated immune responses, CAMP elevating agents were employed and carefully re-examined under the activation conditions of the cells. Various inhibitors tested dose-dependently blocked tumor necrosis factor (TNF)-${\alpha}$ production with IC$_{50}$ values ranged from 0.04 to 300 ${\mu}$M. Of the inhibitors, cAMP-elevating agents showed lower cytotoxicity assessed by lactate dehydrogenase (LDH) release, suggesting less toxic and more selective. In particular co-treatment of dbcAMP with a protein kinase C inhibitor staurosporine displayed the synergistic inhibition of TNF-${\alpha}$ production. The modulatory effect of dbcAMP on TNF-${\alpha}$ and nitric oxide (NO) was significantly affected by treatment time of dbcAMP. Thus, post-treatment of dbcAMP (three hours before LPS) abrogated dbcAMP's inhibitory activity and rather enhanced TNF-${\alpha}$ level up to 60%. In contrast, additional NO production was shown at the co-treatment of dbcAMP with LPS. Unlike simultaneous treatment of phorbol 12-myristate 13-acetate (PMA) and interferon (IFN)-${\gamma}$co-treatment, the combination of dbcAMP with other NO-inducing stimuli did not show drastic overproduction of NO. cAMP elevating agents also diminished splenocyte proliferation stimulated by concanavalin (Con) A, phytohemaglutinin A (PHA) and lipopolysaccharide (LPS). In addition, dbcAMP but not rolipram strongly suppressed CD8$^+$ T cells (CTLL-2). Finally, cAMP elevating agents were differentially involved in regulating CD98-mediated cell-cell adhesion. Thus, dbcAMP and rolipram significantly enhanced the cell-cell adhesion, whereas forskolin blocked. Therefore, our results suggest that CAMP elevating agents participate in various immune responses mediated by macrophages and T cells with a different fashion depending on cellular environments and activation signals.

• Biomolecules & Therapeutics
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• v.20 no.1
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• pp.96-103
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• 2012

In the present study, the inhibitory effect of neem leaf extract (NLE) on lipopolysaccaride (LPS)-induced nitric oxide (NO) and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) production was examined both in vitro and in vivo. In vitro study revealed that NLE treatment ($100{\mu}g/ml$) inhibits LPS (100 ng/ml)-induced NO production by 96% and TNF-${\alpha}$ production by 32%. The reduction in NO production is probably conferred by the complete suppression of inducible nitric oxide synthase (iNOS) expression. Interestingly, in vivo NLE significantly improved the survival rate of mice in an experimental sepsis model. Administration of NLE (100 mg/kg) 24 h before LPS treatment (20 mg/kg) improved the survival rate of mice by 60%. The inhibition of plasma NO and TNF-${\alpha}$ production by NLE is likely to account for the improved survival of mice. Our results suggest that NLE may present a promising avenue in the development of therapeutic agents for the treatment of inflammatory diseases.

• Nutrition Research and Practice
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• v.17 no.1
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• pp.13-31
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• 2023

BACKGROUND/OBJECTIVES: Epigenetic regulation by nutrients can influence the development of specific diseases. This study sought to examine the effect of individual nutrients and nutrient families in the context of preventing chronic metabolic diseases via epigenetic regulation. The inhibition of lipid accumulation and inflammation by nutrients including proteins, lipids, vitamins, and minerals were observed, and histone acetylation by histone acetyltransferase (HAT) was measured. Correlative analyses were also performed. MATERIALS/METHODS: Nutrients were selected according to information from the Korean Ministry of Food and Drug Safety. Selected nutrient functionalities, including the attenuation of fatty acid-induced lipid accumulation and lipopolysaccharide-mediated acute inflammation were evaluated in mouse macrophage Raw264.7 and mouse hepatocyte AML-12 cells. Effects of the selected nutrients on in vitro HAT inhibition were also evaluated. RESULTS: Nitric oxide (NO) production correlated with HAT activity, which was regulated by the amino acids group, suggesting that amino acids potentially contribute to the attenuation of NO production via the inhibition of HAT activity. Unsaturated fatty acids tended to attenuate inflammation by inhibiting NO production, which may be attributable to the inhibition of in vitro HAT activity. In contrast to water-soluble vitamins, the lipid-soluble vitamins significantly decreased NO production. Water- and lipid-soluble vitamins both exhibited significant inhibitory activities against HAT. In addition, calcium and manganese significantly inhibited lipid accumulation, NO production, and HAT activity. CONCLUSIONS: Several candidate nutrients and their family members may have roles in the prevention of diseases, including hepatic steatosis and inflammation-related diseases (i.e., nonalcoholic steatohepatitis) via epigenetic regulation. Further studies are warranted to determine which specific amino acids, unsaturated fatty acids and lipid-soluble vitamins or specific minerals influence the development of steatosis and inflammatory-related diseases.

• The Journal of Korean Medicine
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• v.39 no.1
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• pp.22-34
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• 2018

Objectives: The purpose of this study is to investigate changes of the marker compounds and anti-inflammatory effect of Gamisoyo-san decoction (GMSYS) depending on storage temperature and periods. Methods: GMSYS was stored at room temperature or refrigeration for 12 months. According to storage temperature and periods, pH and sugar content of GMSYS were measured. To determine the marker compounds of GMSYS, high-performance liquid chromatography analysis was performed. To estimate the anti-inflammatory effect of GMSYS, LPS-induced pro-inflammatory mediators and cytokines were measured in RAW 264.7 cells. Results: There was no change in pH and sugar content depending on storage temperature and periods of GMSYS. The contents of gallic acid and mangiferin in both of room temperature and refrigerated decoctions reduced with increasing storage periods. Chlorogenic acid was time-dependently decreased in case of stored at room temperature. GMSYS significantly inhibited the LPS-induced production of nitric oxide, prostaglandin $E_2$ ($PGE_2$) and IL-6 in RAW 264.7 cells. These effects equally maintained up to 3 months at both of room temperature and refrigeration. Since 4 months, the inhibitory effect of GMSYS on LPS-induced $PGE_2$ production was time-dependently reduced, and the decrease in $PGE_2$ inhibitory effect of decoction stored at refrigeration was lower than that of stored at room temperature. Conclusions: Our results indicate that the anti-inflammatory effect of GMSYS are maintained up to 12 months, but it shows optimal efficacy up to 3 months. It is recommended to store in a refrigeration for short periods since some components decrease as storage periods becomes longer.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.11
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• pp.1507-1511
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• 2011

This study investigated the anti-inflammatory effect of Erigeron annuus L. flower (EAF) methanol extract. We examined the involvement of heme oxygenase-1 (HO-1) in the inhibitory activities of EAF methanol extract on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. Cell viability and NO assays were performed. In addition, inducible nitric oxide synthase (iNOS) and HO-1 expressions were detected by Western blotting and blocking HO-1 activity on NO production. EAF methanol extract (25, 50, 100, 200 ${\mu}g$/mL) significantly inhibited LPS-stimulated NO production (p<0.05; 12.82, 9.61, 6.83, 2.52 ${\mu}m$) in a concentration-dependent manner. EAF methanol extract also reduced the expression of iNOS protein. The EAF methanol extract induced the expression of HO-1 in a dose-dependent manner. Blockage of HO-1 activity by zinc protoporphyrin suppressed EAF methanol extract-induced reductions in the production of NO. The present results suggest that EAF methanol extract has a potent anti-inflammatory effect in RAW264.7 macrophages through the induction of HO-1.

• Food Science of Animal Resources
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• v.32 no.5
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• pp.635-643
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• 2012

As an appraisal for the application of a new starter culture, more than 200 lactic acid bacteria strains were isolated from raw milk and healthy human feces. The strains showing excellent growth and acid production ability in 10% skim milk media were selected and identified as Lactobacillus casei based on the results of their API carbohydrate fermentation patterns, as well as 16S rDNA sequence analysis. To assess the effect of L. casei strains on irritable bowel disease (IBD), the inhibitory effect of the selected strains against the nitric oxide (NO) production of lipopolysaccharide (LPS)-stimulated RAW 264.7 cells was measured. Among the tested L. casei strains, L. casei MCL was observed to have the greatest NO inhibitory activity. Additionally, L. casei MCL was found to inhibit mRNA expression of pro-inflammatory cytokines (interleukin-$1{\beta}$, IL-6, TNF-${\alpha}$), as well as cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) involved in pathophysiologic processes such as inflammation. The mRNA expression of anti-inflammatory cytokines, including IL-10 and transforming growth factor-$1{\beta}$ (TGF-${\beta}$) of L. casei MCL, was confirmed using quantitative real-time PCR. In conclusion, L. casei MCL showed decreases in the expression of pro-inflammatory cytokines and up-regulated expression of the anti-inflammatory cytokine.

• Ocean and Polar Research
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• v.43 no.3
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• pp.113-126
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• 2021

In the present study, the halophyte C. falcatum extract and its solvent fractions (n-hexane, 85% aqueous methanol, n-butanol, and water) were evaluated for antioxidant and anti-inflammatory activities. Antioxidative ability was measured by DPPH radical, intracellular reactive oxygen species (ROS) and peroxynitrite scavenging, DNA oxidation inhibition, and ferric reducing antioxidant power (FRAP). For DPPH radical and peroxynitrite scavenging, DNA oxidation inhibition, and FRAP, 85% aq.MeOH and n-BuOH fractions showed significant scavenging activity. For production of intracellular ROS in HT-1080 cells, 85% aq.MeOH fraction showed the highest scavenging activity. In addition, anti-inflammatory activity was also assessed by measuring the inhibitory effect against mRNA expression of pro-inflammatory factors (NO, IL-1β, IL-6 and COX-2) in LPS-stimulated Raw 264.7 macrophages. For NO production, crude extract exhibited a strong inhibitory effect at a concentration of 100 ㎍/ml. For mRNA expression of pro-inflammatory cytokines (IL-1β, IL-6, and COX-2), n-BuOH greatly suppressed expression levels of IL-1β and IL-6 at 100 ㎍/ml concentration while 85% aq. MeOH fraction significantly inhibited that of COX-2 even at 100 ㎍/ml. These results suggest that C. falcatum may be used as a potential source for the development of a natural antioxidant or anti-inflammatory agent.