• Bulletin of the Korean Chemical Society
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• v.31 no.1
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• pp.27-30
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• 2010

A series of thioureas were synthesized as inhibitors of NO production in lipopolysaccharide-activated macrophages. We investigated the effect of lipophilic moiety and N-substituents of the thioureas on the activity. Phenoxazine and carbazole-containing derivatives revealed higher activity than indole-containing thioureas. The appropriate spacer between lipophilic tail and thiourea head and methyl substituent at N3 position of thiourea brought beneficial effect on the inhibition of NO production. Among prepared compounds, phenoxazine-containing derivative 2a was the most potent with $2.32 {\mu}M$ of $IC_{50}$ value. RT-PCR analysis suggested that the prepared thioureas inhibited NO production through the suppression of iNOS mRNA expression.

• YAKHAK HOEJI
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• v.58 no.1
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• pp.47-52
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• 2014

Clematis mandshurica (Ranunculaceae) has traditionally been used as a remedy for antidiuretic, antifungal, rheumatic conditions and alleviate pain. We carried out to evaluate the anti-oxidative effect, anti-inflammatory effect and anti-melanogenic effect of ethanol extract and solvent fractions of Clematis mandshurica. The ethanol extract and the dichloromethane fraction of Clematis mandshurica showed an anti-oxidative effect in DPPH assay, the inhibitory activity of nitric oxide (NO) production in lipopolysaccharide (LPS) activated RAW 264.7 cell, and melanin synthesis and tyrosinase activity of B16F10 melanoma cells. They reduced NO production and melanin content in a dose-dependent manner at concentrations of $2.5{\sim}10{\mu}g/ml$. They also suppressed iNOS and tyrosinase protein and m-RNA expressions dose dependently, assayed by western blot analysis and RT-PCR experiment.

• Bulletin of the Korean Chemical Society
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• v.34 no.5
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• pp.1339-1344
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• 2013

We have synthesized several glycosyl ergosterols and 5,6-dihydroergosterols (DHE) and examined their effects on production of nitric oxide (NO) and iNOS protein expression in LPS-treated RAW264.7 macrophage cells. Our results showed that DHE derivatives inhibited production of NO and iNOS protein expression more strongly than ergosterol derivative. Especially, DHE-Glc exhibited most potent inhibitory activity without cytotoxicity up to the concentration of $100{\mu}M$.

• Biomolecules & Therapeutics
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• v.7 no.3
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• pp.210-215
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• 1999

Nitric oxide (NO) can mediate numerous physiological processes, including vasodilation, neurotransmission, cytotoxicity, secretion and inflammatory response. The regulation of NO production by inducible NO synthase (iNOS) is considered to be the possible target of the development of anti-inflammatory agent, based on the observation that NO can activate cyclooxygenase, which results in the synthesis of prostaglandins. In an effort to screen new inhibitor of NO production from about 352 species of herbal extracts, we found 9 species with 50% or more inhibitory effect on NO production. Especially, the dose-dependent inhibition of NO production in lipopolysaccharide-treated macrophages by two of the herbal extracts (Artemisiae asiaticae Herba and Saussureae Radix) was due to the decrease in the expression of iNOS.

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

• The Journal of Korean Obstetrics and Gynecology
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• v.27 no.3
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• pp.12-27
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• 2014

Objectives: This study was performed to evaluate the effect of Guibi-tang water extract (GB) on osteoporosis. Methods: We examined the effect of GB on osteoclast differentiation using murine pre-osteoclastic RAW 264.7 cells treated with receptor activator of nuclear factor kappa-B ligand (RANKL). The effect of GB on osteoclast was measured by counting TRAP (+) multinucleated cells and measuring TRAP activity. The mRNA expressions of osteoclastogenesis-related genes (Cathepsin K, MMP-9, TRAP, NFATc1, MITF, TNF-${\alpha}$, IL-6, COX-2) were measured by real-time PCR. We examined the effect of GB on osteoblast proliferation, ALP activity, bone matrix protein synthesis and collagen synthesis using murine calvarial cell. Results: GB decreased the number of TRAP (+) multinucleated cells and inhibited TRAP activity in RANKL-stimulated RAW 264.7 cell. GB decreased the expression of genes related osteoclastogenesis such as Cathepsin K, MMP-9, TRAP, NFATc1, MITF, COX-2 in RANKL-stimulated RAW 264.7 cell. But GB did not decrease the expression of iNOS and increased the expression of TNF-${\alpha}$, IL-6 in RANKL-stimulated RAW 264.7 cell. These genes (iNOS, TNF-${\alpha}$, IL-6) are thought to be related with the inflammatory bone destruction. GB increased cell proliferation of rat calvarial cell and also increased ALP activity in rat calvarial cell. GB did not increase bone matrix protein synthesis but increased collagen synthesis in rat calvarial cell. Conclusions: This study suggests that GB may be effective in treating osteoporosis by inhibiting osteoclast differentiation and its related gene expression and by increasing osteoblast proliferation.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.2
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• pp.385-389
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• 2008

Nitric oxide (NO) has been suggested to play an important role in endotoxin-mediated shock and inflammation. In this study, we investigated the effect of Rosa laevigata Michx. (Rosaceae) on the production of NO and the molecular mechanism of its action. Rosa laevigata inhibited NO generation and iNOS expression in LPS-stimulated murine macrophages. Activity of nuclear $factor{-\kappa}B\;(NF{-\kappa}B)$ and the degradation of $I{\kappa}B-{\alpha}$ were suppressed by Rosa laevigata. Furthermore, extracellular signal-stimulated kinase (ERK), which is known to be involved in $NF{-\kappa}B$ activation, is inhibited by Rosa laevigata. These results suggest that Rosa laevigata could exert its anti-inflammatory actions by suppressing the synthesis of NO through inhibition of $NF{-\kappa}B$ activity.

• Advances in Traditional Medicine
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• v.7 no.5
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• pp.509-517
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• 2008

Gyejijakyakjimo-tang is a multi-herbal formula that is composed of nine medicinal herbs. Gyejijakyakjimo-tang has been reported to have antipyretic and analgesic effects. Gyejijakyakjimo-tang has traditionally been used for goat and rheumatoid arthritis. However, analgesic and antiinflammatory effects of Gyejijakyakjimo-tang has not been clarified yet. In this study, we investigated the analgesic and anti-inflammatory effect of the aqueous extract of Gyejijakyakjimo-tang. We evaluated the aqueous extract of Gyejijakyakjimo-tang on Lipopolysaccharide (LPS)-induced inflammation in murine raw 264.7 macrophage cells. For this study, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, reverse transcription-polymerase chain reaction (RT-PCR), prostaglandin $E_2$ ($PGE_2$) immunoassay, and nitric oxide (NO) detection were performed. Gyejijakyakjimo-tang suppressed $PGE_2$ synthesis and NO production by inhibiting the LPS-induced expressions of COX-2 and iNOS mRNA in murine raw 264.7 macrophage cells. These results show that Gyejijakyakjimo-tang has the analgesic and anti-inflammatory effect by mostly suppressing COX-2 and iNOS expressions, and resulting in the inhibition of $PGE_2$ synthesis and NO production.

• Journal of Korean Medicine Rehabilitation
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• v.24 no.2
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• pp.31-40
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• 2014

Objectives Trachelospermi caulis, known as Nak-Suk-Deung in Korea, is the dried leafy stem of Trachelospermum asiaticum var. intermedium Nakai, and climbing stems and branches of Trachelospermum sdisyivum var, intermedium nakai or Apocyanaceae. Trachelospermi caulis has antipyretic and analgesic activity. It has traditionally been used as a folk remedy in Korea for the treatment of various infla mMatory diseases, including rheumatoid arthritis. The purpose of this study was to evaluate the Effects of Trachelospermum caulis extract on SNP-induced infla mMatory responses in rabbit HIG-82 synovial membrane cells. Methods Anti-infla mMatory effects of the extract of Trachelospermum caulis were investigated using rabbit HIG-82 synovial membrane cells. For this study, 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, western blot analysis, PGE2 i mMunoassay, and NO detection were conducted. Results The aqueous extract of Trachelospermum caulis exerted cytotoxicity and suppressed PGE2 synthesis and NO production in rabbit HIG-82 synovial membrane cells. The aqueous extract of Trachelospermum caulis also inhibited the SNP-induced expressions of COX-2, iNOS, and TNF-$\alpha$ in rabbit HIG-82 synovial membrane cells. Conclusions These results showed that the extract of Trachelospermum caulis exerts the anti-infla mMatory effect by suppressing COX-2, iNOS, and TNF-$\alpha$ expressions in the synovial membrane cells.

• Proceedings of the PSK Conference
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• 2003.04a
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• pp.213.1-213.1
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• 2003

Alpha-viniferin was isolated from Carex humilis (Cyperaceae), and showed anti-inflammatory effects on carrageenin or histamine-induced paw edema in mice. To understand mode of the anti-inflammatory action. effects of alpha-viniferin on cyclooxygenase (COX)-2, iNOS, oxygen radicals and proinflammatory cytokines have been analyzed. Alpha-viniferin showed selective inhibitory effect with an IC50 value of 5 $\mu\textrm{m}$ on COX-2 activity but showed weak inhibitory effect on the synthesis of COX-2 transcript which was identified by RT-PCR. (omitted)