• Journal of Life Science
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• v.19 no.10
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• pp.1396-1402
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• 2009

Hizikia fusiforme is a kind of edible brown seaweed that grows mainly in the northwest Pacific including Korea, Japan and China, and has been widely used as food in Korea. Induction of cyclooxygenase-2 (COX-2) expression and prostaglandin $E_2$ ($PGE_2$) production is thought to have beneficial immunomodulatory effects in acute and chronic inflammatory disorders. In this study, we investigated the effects of extracts of H. fusiforme on the expression of COX-2 and production of $PGE_2$ in U937 human pre-monocytic cell models. In U937 cells stimulated with phorbol 12-myristate 13-acetate (PMA) to mimic inflammation, methanol extract of H. fusiforme (MEHF) and ethanol extract of H. fusiforme (EEHF), but not water extract of H. fusiforme (WEHF), inhibited PMA-induced expression of both COX-2 protein and mRNA, which was associated with inhibition of $PGE_2$ production. To investigate the mechanism by which MEHF and EEHF inhibit COX-2 gene expression and $PGE_2$ production, we examined the activation of nuclear factor-kappaB (NF-$\kappa$B) in U937 cells. Pre-treatment with MEHF and EEHF significantly attenuated the PMA-induced IkappaB degradation and prevented nuclear translocation of NF-$\kappa$B. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-inflammatory activity of H. fusiforme.

• Bulletin of the Korean Chemical Society
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• v.31 no.2
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• pp.384-388
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• 2010

Human microsomal prostaglandin E synthase-1 (mPGES-1) catalyzes the conversion of prostaglandin $H_2$ ($PGH_2$) into prostaglandin $E_2$ ($PGE_2$). To establish a stable and efficient method to assess the activity of mPGES-1, a coupled enzyme assay system using mPGES-1, 15-hydroxyprostaglandin dehydrogenase (15-PGDH) and phosphomolybdic acid (PMA) was developed. In this assay system, $PGH_2$ was converted to $PGE_2$ by mPGES-1, and then $PGE_2$ was further transformed to the 15-keto-$PGE_2$ by 15-PGDH accompanying the production of NADH, which was easily detected by fluorescence spectrometry in a multi-well plate format. During the reaction, spontaneous oxidation of $PGH_2$ was prevented by PMA. Using this novel assay, the $K_m$ value of mPGES-1 for $PGH_2$ and the $IC_{50}$ value of the previously characterized inhibitor, MK-886, were determined to be 0.150 mM and $2.8\;{\mu}M$, respectively, which were consistent with the previously reported values. In addition, low backgrounds were observed in the multi-wall plate screening of chemical compounds.

• BMB Reports
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• v.38 no.6
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• pp.633-638
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• 2005

Biosynthesis of prostanoids is regulated by three sequential enzymatic steps, namely phospholipase $A_2$ enzymes, cyclooxygenase (COX) enzymes, and various lineage-specific terminal prostanoid synthases. Prostaglandin E synthase (PGES), which isomerizes COX-derived $PGH_2$ specifically to $PGE_2$, occurs in multiple forms with distinct enzymatic properties, expressions, localizations and functions. Two of them are membrane-bound enzymes and have been designated as mPGES-1 and mPGES-2. mPGES-1 is a perinuclear protein that is markedly induced by proinflammatory stimuli, is down-regulated by anti inflammatory glucocorticoids, and is functionally coupled with COX-2 in marked preference to COX-1. Recent gene targeting studies of mPGES-1 have revealed that this enzyme represents a novel target for anti-inflammatory and anti-cancer drugs. mPGES-2 is synthesized as a Golgi membrane-associated protein, and the proteolytic removal of the N-terminal hydrophobic domain leads to the formation of a mature cytosolic enzyme. This enzyme is rather constitutively expressed in various cells and tissues and is functionally coupled with both COX-1 and COX-2. Cytosolic PGES (cPGES) is constitutively expressed in a wide variety of cells and is functionally linked to COX-1 to promote immediate $PGE_2$ production. This review highlights the latest understanding of the expression, regulation and functions of these three PGES enzymes.

• ALGAE
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• v.27 no.1
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• pp.21-32
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• 2012

We report an oxidative burst triggered by prostaglandin $A_2(PGA_2)$ in the brown algal kelp Laminaria digitata, constituting the first such discovery in an alga and the second finding of an oxidative burst triggered by a prostaglandin in a living organism. The response is more powerful than the oxidative burst triggered by most other chemical elicitors in Laminaria. Also, it is dose-dependent and cannot be inhibited by diphenylene iodonium, suggesting that another source than NAD(P)H oxidase is operational in the production of reactive oxygen species. Despite the very strong oxidative response, rather few effects at other levels of signal transduction pathways could be identified. $PGA_2$ does not increase lipolysis (free fatty acids) in Laminaria, and only one oxylipin (15-hydroxyeicosatetraenoic acid; 15-HETE) was found to be upregulated in Laminaria. In a subsequent set of experiments in the genome model Ectocarpus siliculosus, none of 5 selected candidate genes, all established participants in various stress responses, showed any significant differences in their expression profiles.

• The Journal of Internal Korean Medicine
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• v.30 no.2
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• pp.288-297
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• 2009

Objective : Inflammatory cytokines have a close relationship to insulin dependent diabetes mellitus (IDDM). The inhibitory effect of Smilacis Glabrae Rhizoma (SGR) were examined on production of nitric oxide (NO), prostaglandin $E_2$ $(PGE_2)$, synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and NF-${\kappa}$B activation in Raw 264.7 cells. Methods: Raw 264.7 cells were pretreated with SGR(20, 50, 100 ${\mu}g$/ml), and then cultured with lipopolysaccharides (LPS). Cell viability was measured by MTT assay; inhibition of NO, $PGE_2$, and TNF-${\alpha}$ production were measured by Griess reagent and enzyme-linked immunosorbent assay(ELISA). Induction of COX-2 and iNOS were determined by western blotting analysis. Inhibition of NF-${\kappa}$B was measured by immunofluorescence assay (IFA). Results: SGR inactivated NF-${\kappa}$B, and inhibited the production of NO, iNOS, and $PGE_2$. Inhibition of COX-2 and TNF-${\alpha}$ could not be confirmed. Conclusions: From the above result. SGR was found to have an anti-inflammatory effect of inhibition of NO, iNOS, and $PGE_2$ production via inhibition of NF-${\kappa}$B.

• Natural Product Sciences
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• v.15 no.4
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• pp.229-233
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• 2009

The essential oil fraction was obtained from the leaves and flowers of Artemisia iwayomogi (Compositae) by steam distillation, and its main component, vulgarone B, was isolated by column chromatography. RAW 264.7 cells were used to investigate the anti-inflammatory properties of A. iwayomogi and vulgarone B. Cell viability was determined by MTT assay after treatment with various dilutions of the compounds. In addition, several assays were used to determine the effects of A. iwayomogi essential oil components on immune stimulation. Nitric oxide production in cells activated with lipopolysaccharide (LPS) was evaluated by reaction with Griess reagent. Both vulgarone B and the essential oil fraction of A. iwayomogi inhibited the production of nitric oxide. The effects on various cytokines released from the cells were also measured using ELISA. The production of prostaglandin $E_2$ was significantly decreased by treatment with A. iwayomogi oils. LPS-induced IL-$1{\beta}$ and IL-6 production were also decreased in a dose-dependent manner, but no significant effect on TNF-${\alpha}$ was observed at the concentrations tested. Finally, Western blot analysis revealed that A. iwayomogi oils reduced the levels of COX-2 and iNOS.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2003.11a
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• pp.102-102
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• 2003

The inhibitors of prostaglandin biosynthesis and nitric oxide production by corresponding inducible isozyme have been considered as potential anti-inflammatory and cancer chemopreventive agents. In our continuous search for cancer chemopreventive agents from natural products, we have evaluated the inhibitory potential of PGE$_2$and NO production in lipopolysaccharide (LPS)-induced mouse macrophage RAW264.7 cells. As a result, pinosylvin (3,5-dihydroxy-trans-stilbene), a stilbenoid, mainly found from the heartwood and leaves of the Pinus sylvestris, showed potential inhibitory activity of LPS-induced PGE$_2$and NO production in a dose-dependent manner. Pinosylvin also suppressed the LPS-induced iNOS protein expression. Further study revealed that pinosylvin exhibited antioxidant activity by the DPPH free radical scavenging potential and inhibitory effect of xanthine oxidase activity. In addition, pinosylvin inhibited COX -2 overexpressed human colon cancer cell (HT-29) growth in a time- and dose-dependent manner. These findings suggest that pinosylvin might be a promising candidate for developing cancer chemopreventive agent.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.29 no.4
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• pp.24-33
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• 2016

Objectives : This study is to investigate the anti-inflammatory and anti-oxidant effects of Tongbi-san extract (TS) on RAW264.7 macrophages using by cell cytotoxicity, Nitric Oxide (NO) and Prostaglandin $E_2$ ($PGE_2$) production and 1,1-diphenyl-2-picryl ghdrazyl (DPPH) free radical scavenging capability. Methods : Cell cytotoxicity was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The production of NO was measured by Griess assay. The production of $PGE_2$ was measured by immunoassay. And the anti-oxidant activity was measured by the DPPH method. Results : TS did not increased significantly compared to the TS untreated group in the cell cytotoxicity. TS inhibited NO and $PGE_2$ production in lipopolysaccharide-stimulated RAW 264.7 cells. TS had the DPPH free radical scavenging capability. Conclusion : The anti-inflammtory and anti-oxidant effects of TS may be use for a treatment of anti-inflammatory diseases.

• Journal of Nutrition and Health
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• v.36 no.2
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• pp.109-116
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• 2003

This study investigated the effects of high amylose starch (HAS) consumption on gut functions in male Sprague-Dawley rats. Experimental animals were fed an diet containing HAS for 4 weeks (0, 125, 250, 500 g/kg diet). Stool weights, transit time, the pH of cecum, Bifidobacterium growth, short chain fatty acid production, and prostaglandin E$_2$production in colon mucus were measured. HAS intake did not affect body weight gain or food efficiency ratio during experimental period. There were no significant differences in kidney weight, epididymal fat pad weights or spleen weights, but the weights of the liver and thymus were significantly lower in the HAS100 group. The length of the large intestine, the weights of the cecum wall and cecum contents, and stool weights significantly increased through HAS intake. But transit time was not affected by the experimental diet. Although Bifidobacterium growth in the cecum increased through the HAS intake dose dependently, there were significant differences in the HAS50 and HAS100 groups. HAS intake increased the production of short chain fatty acid in the cecum contents. In particular, acetate and butyrate concentrations grew significantly. And the production of prostaglandin E$_2$in the colon mucus significantly decreased through HAS intake. These results demonstrate that high amylose starch intake significantly improves gut function.

• Proceedings of the Korean Society of Toxicology Conference
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• 2001.10a
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• pp.132-132
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• 2001

Proinflamamtory mediators such as prostaglandins (PGs), cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS) are known to be key mediators in pathogenesis of inflammatory diseases. Capsaicin, the major ingredient of hot pepper, is considered to elicit anti-inflammatory property. In this study, the effect of capsaicin on the prostaglandin E$_2$(PGE$_2$) production was investigated in murine peritoneal macrophages.(omitted)