• Title/Summary/Keyword: nitric oxide synthase activity

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The Study of Anti-inflammtory Mechanism with Bee Venom on Human Synoviocytes (인체(人體) 활막세포(滑膜細胞)를 대상으로 한 봉양침액(蜂藥鍼液)의 염증(炎症) 치료(治療) 기전(機轉) 연구(硏究))

  • Bae, Chul-woo;Song, Ho-sueb
    • Journal of Acupuncture Research
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    • v.21 no.3
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    • pp.121-131
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    • 2004
  • Objective : The purpose of this study was investigation how the bee venom(BV) prevents inflammation in human cell. Methods : we induced inflammation on human synoviocyte cell by lipopolysaccharide(LPS) and sodium nitroprusside(SNP), treated the bee venom and melittin on this cell, surveyed the expression of Nisotric oxide(NO), inducible nitric oxide synthase(iNOS), Cyclooxygenease-2(COX-2), cytolic phospholipase $A_2(cPLA_2)$, Prostaglandin $E_2(PGE_2)$ and nuclear factor-${\kappa}B$(NF-${\kappa}B$), and got below conclusions. Results : Compared with control 1. Expressions of LPS-induced $PGE_2$(BV 1, $5{\mu}g/m{\ell}$) and SNP-induced PGE2(BV 0.5, 1, $5{\mu}g/m{\ell}$)were decreased significantly. 2. Expressions of LPS-induced NO(BV 0.5, 1, $5{\mu}g/m{\ell}$) and SNP-induced NO(BV 1, $5{\mu}g/m{\ell}$)were decreased significantly. 3. Expressions of LPS-induced COX-2(BV 1, $5{\mu}g/m{\ell}$) and SNP-induced COX-2(BV $5{\mu}g/m{\ell}$)were decreased significantly. 4. Expressions of LPS-induced iNOS(BV 0.5, 1, $5{\mu}g/m{\ell}$) and SNP-induced iNOS(BV $5{\mu}g/m{\ell}$) were meanless by all dose. 5. Expressions of LPS-induced $cPLA_2$(BV 1, $5{\mu}g/m{\ell}$) and SNP-induced cPLA2(BV 1, $5{\mu}g/m{\ell}$)were decreased significantly. 6. Expressions of LPS-induced NF-${\kappa}B$(BV $5{\mu}g/m{\ell}$, melittin $5{\mu}g/m{\ell}$) and SNP-induced NF-${\kappa}B$(BV 0.5, 1, $5{\mu}g/m{\ell}$, melittin 5, $10{\mu}g/m{\ell}$)were decreased significantly. 7. Expressions of LPS-induced NF-${\kappa}B$ binding activity (BV $1{\mu}g/m{\ell}$, melittin $5{\mu}g/m{\ell}$, melittin $5{\mu}g/m{\ell}$+ DTT 20mM) were decreased significantly. Conclusion : The bee venom treatments on synoviocyte showed significant changes in LPS and SNP induced NO, iNOS, COX-2, cPLA2, PGE2 and NF-${\kappa}B$, these results suggest that bee venom is effective to inflammations and establish the process of bee venom therapy, so we expect active use of bee venom to control the inflammation.

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Gastroprotective Activity of Curcumae Longae Rhizoma against Gastric Ulcer in Mice (위궤양 유발 마우스모델에서 강황(薑黃) 추출물의 위 보호 효과)

  • Oh, Min Hyuck;Kim, Min Ju;Shin, Mi-Rae;Park, Hae-Jin;Seo, Bu-Il;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.35 no.3
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    • pp.17-24
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    • 2020
  • Objectives : The objective of this study was to evaluate the gastric protective effect of Curcuma Longae Rhizoma (CLR) in 150 mM HCl/60% ethanol induced gastric ulcer (GU) in mice. Methods : Forty ICR mice were divided into five groups (n=8/Group): Nor group; Normal, Veh group; GU control, SC group; GU + sucralfate 10 mg/kg, CL; GU + CLR 30% ethanol extract 100 mg/kg, CH group; GU + CLR 30% ethanol extract 200 mg/kg. Then, mice were orally administered with 150 mM HCl/60% ethanol and caused GU. After 1 hr, mice were sacrificed, and blood and stomach tissue were collected. Results : CLR showed significance scavenging effects in 1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) radical scavenging activities (DPPH IC50; 78.18 ± 0.60 ㎍/㎖, ABTS IC50; 55.91 ± 1.86 ㎍/㎖). CLR significance reduce inflammatory-related factors such as cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1β), and interleukin-6 (IL-6) via nuclear factor kappa B (NF-κB) inactivation. In addition, the activation of nuclear factor erythroid2-related factor 2 (Nrf2) significantly led to up-regulation of anti-oxidant enzymes including factors heme oxygenase-1 (HO-1), super oxide dismutase (SOD), and glutathione peroxidase-1/2 (GPx-1/2). Conclusions : Our discovery provides that CLR possesses anti-oxidant and anti-inflammatory effects. Hence, CLR may ameliorate the development of gastric ulcer though the inhibition of NF-κB inflammatory pathway and the elevation of Nrf2 anti-oxidant pathway.

Anti-inflammatory Activity of Antimicrobial Peptide Zophobacin 1 Derived from the Zophobas atratus (아메리카왕거저리 유래 항균 펩타이드 조포바신 1의 항염증활성)

  • Shin, Yong Pyo;Lee, Joon Ha;Kim, In-Woo;Seo, Minchul;Kim, Mi-Ae;Lee, Hwa Jeong;Baek, Minhee;Kim, Seong Hyun;Hwang, Jae Sam
    • Journal of Life Science
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    • v.30 no.9
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    • pp.804-812
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    • 2020
  • The giant mealworm beetle, Zophobas atratus (Coleoptera: Tenebrionidae) has been used as a protein source for small pets and mammals. Recently, it was temporarily registered in the list of the Food Code. We previously performed an in silico analysis of the Zophobas atratus transcriptome to identify putative antimicrobial peptides and identified several antimicrobial peptide candidates. Among them, we assessed the antimicrobial and anti-inflammatory activities of zophobacin 1 that was selected bio-informatically based on its physicochemical properties against microorganisms and mouse macrophage Raw264.7 cells. Zophobacin 1 showed antimicrobial activities against microorganisms without inducing hemolysis and decreased the nitric oxide production of the lipopolysaccharide-induced Raw264.7 cells. Moreover, ELISA and Western blot analysis revealed that zophobacin 1 reduced expression levels of pro-inflammatory enzymes such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). We also investigated expression of pro-inflammatory cytokines (interleukin-6 and interleukin-1β) production through quantitative real time-PCR and ELISA. Zophobacin 1 markedly reduced the expression level of cytokines through the regulation of mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B (NF-κB) signaling. We confirmed that zophobacin 1 bound to bacterial cell membranes via a specific interaction with lipopolysaccharides. These data suggest that zophobacin 1 could be promising molecules for development as antimicrobial and anti-inflammatory therapeutic agents.

Anti-inflammatory Activity of Antimicrobial Peptide Protaetiamycine 2 Derived from the Protaetia brevitarsis seulensis (흰점박이꽃무지 유래 항균 펩타이드 프로테티아마이신 2의 항염증활성)

  • Lee, Joon Ha;Baek, Minhee;Lee, Hwa Jeong;Kim, In-Woo;Kim, Sun Young;Seo, Minchul;Kim, Mi-Ae;Kim, Seong Hyun;Hwang, Jae Sam
    • Journal of Life Science
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    • v.29 no.11
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    • pp.1218-1226
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    • 2019
  • The white-spotted flower chafer Protaetia brevitarsis seulensis is a medicinally beneficial and important edible insect species. We previously performed an in silico analysis of the Protaetia brevitarsis seulensis transcriptome to identify putative antimicrobial peptides and then tested their antimicrobial and hemolytic activities. These peptides had potent antimicrobial activities against bacteria and yeast without inducing hemolysis. In the present study, the cationic antimicrobial peptide, protaetiamycine 2, was selected for further assessment of its anti-inflammatory properties in mouse macrophage Raw264.7 cells. Protaetiamycine 2 treatment of Raw264.7 cells suppressed LPS-induced nitric oxide production and reduced the expression of inducible nitric oxide synthase and cyclooxygenase-2, as determined by real-time PCR and western blotting. The expression of proinflammatory cytokines ($TNF-{\alpha}$, IL-6, and $IL-1{\beta}$) was also attenuated through the MAPKs and $NF-{\kappa}B$ signaling. We also confirmed that protaetiamycine 2 bound to bacterial cell membranes by a specific interaction with LPS. Collectively, these data obtained from LPS-induced Raw264.7 cells indicated that protaetiamycine 2 could have both antimicrobial and anti-inflammatory properties.

Two new triterpenoid saponins derived from the leaves of Panax ginseng and their antiinflammatory activity

  • Li, Fu;Cao, Yufeng;Luo, Yanyan;Liu, Tingwu;Yan, Guilong;Chen, Liang;Ji, Lilian;Wang, Lun;Chen, Bin;Yaseen, Aftab;Khan, Ashfaq A.;Zhang, Guolin;Jiang, Yunyao;Liu, Jianxun;Wang, Gongcheng;Wang, Ming-Kui;Hu, Weicheng
    • Journal of Ginseng Research
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    • v.43 no.4
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    • pp.600-605
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    • 2019
  • Background: The leaves and roots of Panax ginseng are rich in ginsenosides. However, the chemical compositions of the leaves and roots of P. ginseng differ, resulting in different medicinal functions. In recent years, the aerial parts of members of the Panax genus have received great attention from natural product chemists as producers of bioactive ginsenosides. The aim of this study was the isolation and structural elucidation of novel, minor ginsenosides in the leaves of P. ginseng and evaluation of their antiinflammatory activity in vitro. Methods: Various chromatographic techniques were applied to obtain pure individual compounds, and their structures were determined by nuclear magnetic resonance and high-resolution mass spectrometry, as well as chemical methods. The antiinflammatory effect of the new compounds was evaluated on lipopolysaccharide-stimulated RAW 264.7 cells. Results and conclusions: Two novel, minor triterpenoid saponins, ginsenoside $LS_1$ (1) and 5,6-didehydroginsenoside $Rg_3$ (2), were isolated from the leaves of P. ginseng. The isolated compounds 1 and 2 were assayed for their inhibitory effect on nitric oxide production in LPS-stimulated RAW 264.7 cells, and Compound 2 showed a significant inhibitory effect with $IC_{50}$ of $37.38{\mu}M$ compared with that of NG-monomethyl-L-arginine ($IC_{50}=90.76{\mu}M$). Moreover, Compound 2 significantly decreased secretion of cytokines such as prostaglandin $E_2$ and tumor necrosis factor-${\alpha}$. In addition, Compound 2 significantly suppressed protein expression of inducible nitric oxide synthase and cyclooxygenase-2. These results suggested that Compound 2 could be used as a valuable candidate for medicinal use or functional food, and the mechanism is warranted for further exploration.

A Methanol Extract of Adansonia digitata L. Leaves Inhibits Pro-Inflammatory iNOS Possibly via the Inhibition of NF-κB Activation

  • Ayele, Yihunie;Kim, Jung-Ah;Park, Eunhee;Kim, Ye-Jung;Retta, Negussie;Dessie, Gulelat;Rhee, Sang-Ki;Koh, Kwangoh;Nam, Kung-Woo;Kim, Hee Seon
    • Biomolecules & Therapeutics
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    • v.21 no.2
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    • pp.146-152
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    • 2013
  • This study examined the total polyphenol content of eight wild edible plants from Ethiopia and their effect on NO production in Raw264.7 cells. Owing to its relatively high polyphenol concentration and inhibition of NO production, the methanol extract of Adansonia digitata L. leaf (MEAD) was subjected to detailed evaluation of its antioxidant and anti-inflammatory effects. Antioxidant effects were assessed by measuring free-radical-scavenging activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and oxygen-radical-absorbance capacity (ORAC) assays, while anti-inflammatory effects were assessed by measuring inducible nitric oxide synthase (iNOS) expression in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. In the ORAC assay, MEAD was 10.2 times more potent than vitamin C at eliminating peroxyl radicals. In DPPH assay, MEAD also showed a strong ROS scavenging effect. MEAD significantly inhibited iNOS activity ($IC_{50}=28.6{\mu}g/ml$) of LPS-stimulated Raw264.7 cells. We also investigated the relationship between iNOS expression and nuclear factor kappa B (NF-${\kappa}B$) activation. MEAD inhibited $I{\kappa}B{\alpha}$ degradation and NF-${\kappa}B$ translocation from the cytosol to the nucleus in LPS-induced RAW264.7 cells without significant cytotoxic effects, as confirmed by MTT assay. These results suggest that MEAD inhibits anti-inflammatory iNOS expression, which might be related to the elimination of peroxyl radicals and thus the inhibition of $I{\kappa}B{\alpha}$-mediated NF-${\kappa}B$ signal transduction.

Anti-Inflammatory Effect of Ethanol Extract from Grateloupia elliptica Holmes on Lipopolysaccharide-Induced Inflammatory Responses in RAW 264.7 Cells and Mice Ears (LPS로 유도된 RAW 264.7 세포와 마우스 귀 조직에 대한 참도박(Grateloupia elliptica Holmes) 에탄올 추출물의 항염증 효과)

  • Bae, Nan-Young;Kim, Min-Ji;Kim, Koth-Bong-Woo-Ri;Ahn, Na-Kyung;Choi, Yeon-Uk;Park, Ji-Hye;Park, Sun-Hee;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.8
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    • pp.1128-1136
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    • 2015
  • This study investigated the anti-inflammatory effects of ethanol extract from Grateloupia elliptica Holmes (GEHEE) on the lipopolysaccharide-induced inflammatory response. Anti-inflammatory effects were detected by enzyme-linked immunosorbent assay, Western blotting, and immunohistochemistry. There were no cytotoxic effects on proliferation of macrophages treated with GEHEE compared to the control. GEHEE remarkably suppressed NO and pro-inflammatory cytokines (interleukin-6, tumor necrosis $factor-{\alpha}$, and $interleukin-1{\beta}$) production and reduced expression of inducible nitric oxide synthase, cyclooxygenase-2, and nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) proteins in a dose-dependent manner. GEHEE also significantly reduced activation of mitogen-activated protein kinases (MAPKs). The formation of edema in mouse ears was reduced at the highest dose compared to the control. GEHEE also reduced dermal thickness and mast cell numbers based on histological analysis. These results suggest that GEHEE exerts significant anti-inflammatory activity via inhibition of $NF-{\kappa}B$ and MAPKs activation and may be a potential anti-inflammatory therapeutic material.

In vitro and in vivo anti-inflammatory activities of Korean Red Ginseng-derived components

  • Baek, Kwang-Soo;Yi, Young-Su;Son, Young-Jin;Yoo, Sulgi;Sung, Nak Yoon;Kim, Yong;Hong, Sungyoul;Aravinthan, Adithan;Kim, Jong-Hoon;Cho, Jae Youl
    • Journal of Ginseng Research
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    • v.40 no.4
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    • pp.437-444
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    • 2016
  • Background: Although Korean Red Ginseng (KRG) has been traditionally used for a long time, its anti-inflammatory role and underlying molecular and cellular mechanisms have been poorly understood. In this study, the anti-inflammatory roles of KRG-derived components, namely, water extract (KRG-WE), saponin fraction (KRG-SF), and nonsaponin fraction (KRG-NSF), were investigated. Methods: To check saponin levels in the test fractions, KRG-WE, KRG-NSF, and KRG-SF were analyzed using high-performance liquid chromatography. The anti-inflammatory roles and underlying cellular and molecular mechanisms of these components were investigated using a macrophage-like cell line (RAW264.7 cells) and an acute gastritis model in mice. Results: Of the tested fractions, KGR-SF (but not KRG-NSF and KRG-WE) markedly inhibited the viability of RAW264.7 cells, and splenocytes at more than 500 mg/mL significantly suppressed NO production at $100{\mu}g/mL$, diminished mRNA expression of inflammatory genes such as inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor-${\alpha}$, and interferon-${\beta}$ at $200{\mu}g/mL$, and completely blocked phagocytic uptake by RAW264.7 cells. All three fractions suppressed luciferase activity triggered by interferon regulatory factor 3 (IRF3), but not that triggered by activator protein-1 and nuclear factor-kappa B. Phospho-IRF3 and phospho-TBK1 were simultaneously decreased in KRG-SF. Interestingly, all these fractions, when orally administered, clearly ameliorated the symptoms of gastric ulcer in HCl/ethanol-induced gastritis mice. Conclusion: These results suggest that KRG-WE, KRG-NSF, and KRG-SF might have anti-inflammatory properties, mostly because of the suppression of the IRF3 pathway.

Comparison Study of Immunomodulatory Activity of Polysaccharide and Ethanol Extracted from Sargassum fulvellum (참모자반 조다당 추출물과 에탄올 추출물의 대식세포 및 비장세포 활성 비교)

  • Byun, Eui-Hong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.11
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    • pp.1621-1628
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    • 2015
  • The immune system plays an important role in maintaining and protecting human health. In the present study, comparison of immuno-modulatory activities between polysaccharides (SFP) and ethanol (SFE) extracts separated from Sargassum fulvellum in macrophages and murine splenocytes were investigated. Immuno-modulatory activities of macrophages were estimated based on cell proliferation, nitric oxide (NO), inducible NO synthase (iNOS), and cytokine production in RAW 264.7 macrophage cells, and lipopolysaccharide was used as a positive control. SFP and SFE treatment did not affect cytotoxicity in RAW 264.7 macrophage cells, and SFP treatment significantly increased NO and cytokine production ($TNF-{\alpha}$, IL-6, and $IL-1{\beta}$), whereas SFE did not contribute to the increase in NO and cytokine production. In the case of splenocytes, SFP treatment increased splenocyte proliferation and also highly increased production of Th-1 type cytokines (IL-2 and $IFN-{\gamma}$) than those of SFE. Through this study, we confirmed that immuno-modulatory activities of Sargassum fulvellum may be due to polysaccharide extracts and this can be a potential nutraceutical.

NO and Cytokine Production due to Crysochroa fulgidissima (비단벌레(Crysochroa fulgidissima) 에탄올추출물의 NO 증강 및 염증인자억제활성)

  • Ahn, Mi-Young;Kim, Soon-Ja;Jeong, Hye-Kyoung;Seo, Yun-Jung;Park, Hae-Cheol;Lee, Young-Bo;Kim, Mi-Aae
    • Korean journal of applied entomology
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    • v.50 no.3
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    • pp.227-233
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    • 2011
  • Crysochroa fulgidissima (Bidan-beole, Spanish fly) is traditionally used as a crude drug and insecticide in the East Asia and Korea, respectively. This study investigated the effect of ethanol extract of C. fulgidissima on the NO production activity. The C. fulgidissima extract was a potent inducer of NO production in CPAE cells and a stimulator of endothelial nitric oxide synthase in a dose-dependent manner. This study also evaluated the anti-inflammatory activity of this extract by determining the level of ICAM-1, VCAM-1, and prostaglandin $E_2$ from HUVEC cells. Although C. fulgidissima extract was a potent inducer of NO production in the CPAE cells, it showed weak inhibitory effects on vascular endothelial growth factor (VEGF) production in HUVEC cells. HPLC and GC-MS analysis of the ethanol extract of C. fulgidissima revealed the presence of cantharidin.