• Title/Summary/Keyword: inducible NO synthase (iNOS)

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Convergence Studies of NO Homeostasis in Cellular Signalling (세포의 신호전달 과정에서 NO 항상성에 관한 융복합 연구)

  • Oh, Hee-Kyun;Do, Eun-Young;Park, Hae-Ryoung
    • Journal of Digital Convergence
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    • v.13 no.12
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    • pp.461-467
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    • 2015
  • Saussurea lappa is known for a variety of physiological activities as a component but has not known to show the effect of the cellular signaling pathway. We investigated the anti-inflammatory effects by Saussurea lappa ethanol extract on the LPS(lipopolysaccharide) induced nitric oxide (NO) production by RAW 264.7 cell line. It shows the expressions of iNOS and COX-2 at the transcriptional level (RT-PCR). The Saussurea lappa ethanol extract showed transcriptional expression levels of pro-inflammatory cytokine TNF- and IL-$1{\beta}$ induced by LPS(lipolysaccharide) in RAW264.7 cell line. Saussurea lappa ethanol extract reduced the LPS-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) at the RNA levels in a concentration-dependent manner. The finding that ethanol extract of Saussurea lappa has an influence on NO (nitric oxide) homeostasis in the study of the action mechanism on the macrophage-mediated inflammatory reaction was considered in terms of convergence. And it is to provide an important basis for the prevention and treatment of inflammatory diseases in the future.

Inhibitory Effect of Aqueous Extract from Lonicera japonica Flower on LPS-induced Inflammatory Mediators in RAW 264.7 Macrophages. (금은화 수용성 추출물의 LPS 유도 염증매개물 억제 효과)

  • Yun, Young-Gab;Kim, Gyu-Min;Lee, Sung-Jun;Ryu, Seong-Hun;Jang, Seon-Il
    • The Korea Journal of Herbology
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    • v.22 no.3
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    • pp.117-125
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    • 2007
  • Objective : Lonicera japonica (Caprifoliaceae) has long been used for treatment of infectious diseases in oriental countries. The aim of this study was to investigative the effect by which the aqueous extract from flower of L. japonica (LJFAE) inhibited the lipopolysaccharide (LPS)-induced inflammatory mediators in murine macrophages, RAW 264.7 cells Methods : The dried flowers of L. japonica were extracted with distilled water at $100^{\circ}C$ for 7 h. The extract was filtered through 0.45 ${\mu}m$ filter, freeze-dried. The dried extract was dissolved in Hank's balanced salt solution (HBSS) and filtered through 0.22 ${\mu}m$ filter before use. Accumulated nitrite, an oxidative product of nitric oxide (NO), was measured in the culture medium by the Griess reaction. The levels of prostaglandin E2 (PGE2), tumor necrosis factor-$\alpha$ (TNF-$\alpha$), interleukin-1$\beta$ (IL-1$\beta$), and IL-6 production, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression were measured by enzyme-linked immunosorbent assay and Western blot analysis. Results: LJFAE (10-400 ${\mu}g$/ml) per se had no cytotoxic effect in unstimulated macrophages, but LJFAE concentration-dependently reduced NO, PGE2, TNF-, IL-l, and IL-6 production and COX-2 activity caused by stimulation of LPS. The levels of iNOS and COX-2 protein expressions were markedly suppressed by the treatment with LJFAE in a concentration dependent manner. Conclusions : These results suggest that LJFAE suppress the NO and PGE2production in macrophages by inhibiting iNOS and COX-2 expression and these properties may contribute to the anti-inflammatory activity of Lonicera japonica.

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Anti-inflammatory effect of ethanol extract from Ecklomia cava on gram-positive bacteria(Peptidoglycan)-induced macrophages (그람-양성균(Peptidoglycan)으로 유도한 대식세포에서 감태 에탄올 추출물의 항염증 효과)

  • Kang, Ok-Hwa;Kim, Sung-Bae;Keum, Joon-Ho;Mun, Su-Hyun;Kim, Yong-Sik;An, Byung-Kwan;An, Hyeon-Jin;Kwon, Dong-Yeul
    • Herbal Formula Science
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    • v.19 no.1
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    • pp.195-205
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    • 2011
  • Objectives : Ecklonia cava is brown alga(Laminariaceae) which grows is sea, it has antioxidant, diarrhea and anticoagulant effect. In this study, the effect of ethanol extract of Ecklonia cava (EC) on peptidoglycan(PGN) -induced NO production in RAW 264.7 cells was investigated. Methods : In the present study, IL-6 production was measured using the enzyme-linked immunosorbent assay(ELISA), prostaglandin $\E_2$($\PGE_2$) production was measured using the EIA kit, and inducible NO synthase(iNOS), cyclooxygenase-2(COX-2), and mitogen-activated protein kinase(MAPK) activation, as determined by western blot analysis and reverse transcription -polymerase chain reaction(RT-PCR). Results : EC inhibited PGN-induced NO and IL-6 production. Consistent with these observations, the protein expression of iNOS and COX-2 were inhibited by EC. Moreover, EC suppressed the phosphorylation of extracellular signal-regulated kinase(ERK) 1/2 in PGN-induced RAW 264.7. Conclusions : These results suggest that EC has inhibitory effects on PGN-induced $\PGE_2$, NO, and IL-6 production, as well as the expressions of iNOS and COX-2 in the murine macrophage. These inhibitory effects occur through blockades on the MAPKs phosphorylation.

Effect of Frutus gardeniae herbal acupuncture on the rat model of ankle sprain pain (치자(梔子) 약침(藥鍼)이 백서(白鼠) 모델 족과 염좌(捻挫) 통증(痛症)에 미치는 영향(影響))

  • Koo, Sung-Tae;Cho, Myoung-Soo;Park, Sung-Sub;Kim, Young-Tae;Park, Kwi-Jong;Kim, Kyoung-Sik;Sohn, In-Cheul
    • Korean Journal of Acupuncture
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    • v.22 no.2
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    • pp.57-74
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    • 2005
  • Objective : Frutus gardeniae, seed of Gardenia jasminoides Ellis is one of the crude drugs used for the treatment of inflammatory condition in oriental medicine. Methodes : The present study aimed to examine the analgesic effect and anti-inflammatory effect of Frutus gardeniae extract (FGE) on a rat model of ankle sprain pain, and the relations between FGE-induced effect and endogenous nitric oxide (NO) and inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), and c-Fos protein expression in the spinal cord. As a chronic pain model, ankle sprain pain model was used to test the effect of FCE injection applied to acupuncture point. After the induction of ankle sprain, rats subsequently showed a reduced stepping force of the affected limb for at least the next 4 days. The reduced stepping force of the limb was presumably due to a painful knee. FGE dissolved in normal saline was injected several acupoints. Results : After the treatment, behavioral tests measuring stepping force were periodically conducted during the next 8 hours. FGE produced significant improvement of stepping force of the hindlimb affected by the ankle sprain lasting at least 4 hours. FGE produced the improvement of stepping force of the affected hindlimb in a dose-dependent manner. In addition, FGE injection showed inhibitory effect on the paw edema induced by ankle sprain. Both NO production and iNOS, COX-2 protein expression increased by ankle sprain were suppressed by FGE. FGE on combination with electroacupuncture (EA) produced more powerful and longer lasting improvement of stepping force of the hindlimb affected by the ankle sprain than either FGE or EA did. The present study suggest that FGE produces a potent analgesic effect on the ankle sprain pain model of the rat and that FGE-induced analgesia modulate endogenous NO through the suppression of iNOS/COX-2 protein expression.

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Effect of Daebangpungtang herbal acupuncture on the carrageenan-induced arthritis in rats (대방풍탕(大防風湯) 약침(藥鍼)이 Carrageenan으로 유도한 관절염에 미치는 영향(影響))

  • Koo, Sung-Tae;Hwang, Jae-Ho;Song, Mun-Yeong;Park, Yu-Ri;Sohn, In-Cheul;Kim, Kyoung-Sik
    • Korean Journal of Acupuncture
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    • v.22 no.3
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    • pp.83-104
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    • 2005
  • Objectives : DaeBangPungTang(DBPT) is one of the prescriptions used for the treatment of knee arthritis in oriental medicine. The present study aimed to examine the analgesic effect of DBPT on a rat model of carrageenan-induced arthritis, and the relations between DBPT-induced analgesia and endogenous nitric oxide(NO) and inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), and c-Fos protein expression in the spinal cord. Methods : Carrageenan-induced arthritis rat model was used to test the effect of DBPT as a chronic pain model. After the induction of arthritis, rats subsequently showed a reduced stepping force of the affected limb for at least tile next 4 days. The reduced stepping force of the limb was presumably due to a painful knee, since oral infection of indomethacin produced temporary improvement of weight bearing. DBPT dissolved in normal saline was minted several acupoints. After the treatment, behavioral tests measuring stepping force were periodically conducted during the next 12 hours. Results : DBPT produced significant improvement of stepping force of the hindlimb affected by the arthritis lasting at least 9 hours. The magnitude of this improvement was equivalent to that obtained after an oral injection of 3mg/kg of indomethacin and this improvement of stepping force was interpreted as an analgesic effect. DBPT produced the improvement of stepping force of the affected hindlimb in a dose-dependent manner. Both NO production and iNOS, COX-2 protein expression increased by arthritis were suppressed by DBPT. DBPT on combination with electroacupuncture (EA) produced more powerful and longer lasting improvement of stepping force of the hindlimb affected by the arthritis than either DBPT or EA did. Conclusion : The present study suggest that DBPT produces a potent analgesic effect on the chronic hee arthritis pain model in the rat and that DBPT-induced analgesia modulate endogenous NO through the suppression of iNOS/COX-2 protein expression.

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Inhibitory Effect of Water Extract from Dojuksan on LPS-induced Proinflammatory Cytokines Production in RAW 264.7 Cells (LPS로 자극한 RAW 264.7 세포에서 염증성세포활성물질 생산에 미치는 도적산(導赤散) 물 추출의 억제 효과)

  • Kim, Ji-Eun;Kim, Sung-Bae;Kang, Ok-Hwa;Shin, In-Sik;Kang, Suk-Hoon;Lee, Seung-Ho;Kwon, Dong-Yeul
    • The Korea Journal of Herbology
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    • v.28 no.3
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    • pp.53-60
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    • 2013
  • Objectives : DojukSan is known to be effective for treating a urinary diseases and stomatitis. However, there has been a lack of studies regarding the effects of Dojuksan on the inflammatory activities and effector inflammatory disease mechanism about macrophage before is not known. To elucidate the molecular mechanisms of Dojuksan water extract (DJS) on pharmacological and biochemical actions in inflammation, we examined the effect of DJS on pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated macrophages. Methods : In the present study, pro-inflammatory cytokine production was determined by performing enzyme-linked immunosorbent assay, reverse transcription polymerase chain reaction, and western blot analysis to measure the activation of MAPKs. Cells were treated with 200 ng/mL of LPS 1 h prior to the addition of DJS. Cell viability was measured by MTS assay. The investigation focused on whether DJS inhibited nitric oxide (NO) and prostaglandin E2 ($PGE_2$) productions, as well as the expressions of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-6 (IL-6) and mitogen-activated protein kinases (MAPKs) in LPS-stimulated RAW 264.7 cells. Results : We found that DJS inhibited LPS-induced NO, $PGE_2$ and IL-6 productions as well as the expressions of iNOS and COX-2. Furthermore, DJS suppressed the LPS-induced phosphorylation of p38 MAPK and c-Jun NH2-protein kinase (JNK). Conclusions : These results suggest that DJS has inhibitory effects on LPS-induced $PGE_2$, NO, and IL-6 production, as well as the expressions of iNOS and COX-2 in the murine macrophage. These inhibitory effects occur through blockades on the MAPKs phosphorylation.

Inhibitory Effects of Flavonoids Isolated from the Leaves of Stewartia koreana on Nitric-oxide Production in LPS-stimulated RAW 264.7 Cells (노각나무 잎에서 분리된 플라보노이드에 의한 대식세포에서 산화질소 생성 억제효과)

  • Lee, Seung-Su;Bang, Myun-Ho;Park, Se-Ho;Chung, Dae-kyun;Yang, Seun-Ah
    • Journal of Life Science
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    • v.28 no.5
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    • pp.509-516
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    • 2018
  • Five phenolic compounds were isolated from the ethyl acetate fraction of leaves from Stewartia koreana, and their nitric-oxide (NO) inhibitory activities were measured to identify the major active constituents responsible for the efficacy of the extract against inflammatory reactions. These five compounds were quercetin (1), quercitrin (2), hyperin (3), quercetin-3-O-(6"-O-galloyl)-${\beta}$-D-galactopyranoside (4), and kaempferol 3-O-[2",6"-di-O-(trans-p-coumaroyl)]-${\beta}$-D-glucopyranoside (5). Among the separated compounds in the EtOAc fraction, compounds 4 and 5 were isolated for the first time, and no study has yet reported their anti-inflammatory effects. The compounds were identified by spectroscopic analysis, and the isolated compounds showed significant NO inhibitory effects in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Compound 5 showed the most potent inhibitory effect (63.35% inhibition) against LPS-induced NO production compared to that of compound 1 (17.17%), compound 2 (5.0%), compound 3 (3.92%), and compound 4 (6.32%) at $10{\mu}g/ml$ concentration. NO production was inhibited by suppressing the protein expression of inducible nitric-oxide synthase in LPS-stimulated RAW 264.7 macrophages. These results indicate that kaempferol 3-O-[2",6"-di-O-(trans-p-coumaroyl)]-${\beta}$-D-glucopyranoside might be the major active compound responsible for the anti-inflammatory effects of S. koreana.

Acacia ferruginea Inhibits Tumor Progression by Regulating Inflammatory Mediators-(TNF-α, iNOS, COX-2, IL-1β, IL-6, IFN-γ, IL-2, GM-CSF) and Pro-Angiogenic Growth Factor-VEGF

  • Sakthivel, Kunnathur Murugesan;Guruvayoorappan, Chandrasekaran
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3909-3919
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    • 2013
  • The aim of the present investigation was to evaluate the effect of A ferruginea extract on Dalton's lymphoma ascites (DLA) induced tumours in BALB/c mice. Experimental animals received A ferruginea extract (10 mg/kg.b.wt) intraperitoneally for 14 consecutive days after DLA tumor challenge. Treatment with extract significantly increased the life span, total white blood cell (WBC) count and haemoglobin (Hb) content and decreased the level of serum aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), gamma glutamyl transferase (${\gamma}$-GT) and nitric oxide (NO) in DLA bearing ascites tumor models. In addition, administration of extract significantly decreased the tumour volume and body weight in a DLA bearing solid tumor model. The levels of pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-${\alpha}$), interleukin-1 beta (IL-$1{\beta}$), interleukin-6 (IL-6) and granulocyte monocyte-colony stimulating factor (GM-CSF), as well as pro-angiogenic growth factors such as vascular endothelial growth factor (VEGF) and inducible nitric oxide synthase (iNOS) were elevated in solid tumour controls, but significantly reduced by A ferruginea administration. On the other hand, the extract stimulated the production of interleukin-2 (IL-2) and interferon-gamma (IFN-${\gamma}$) in animals with DLA induced solid tumours. Increase in $CD4^+$ T-cell population suggested strong immunostimulant activity for this extract. GC/MS and LC/MS analysis showed quinone, quinoline, imidazolidine, pyrrolidine, cyclopentenone, thiazole, pyrazole, catechin and coumarin derivatives as major compounds present in the A ferruginea methanolic extract. Thus, the outcome of the present study suggests that A ferruginea extract has immunomodulatory and tumor inhibitory activities and has the potential to be developed as a natural anticancer agent.

5β-Hydroxypalisadin B isolated from red alga Laurencia snackeyi attenuates inflammatory response in lipopolysaccharide-stimulated RAW 264.7 macrophages

  • Wijesinghe, W.A.J.P.;Kang, Min-Cheol;Lee, Won-Woo;Lee, Hyi-Seung;Kamada, Takashi;Vairappan, Charles S.;Jeon, You-Jin
    • ALGAE
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    • v.29 no.4
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    • pp.333-341
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    • 2014
  • In this study, four compounds isolated from the red alga Laurencia snackeyi were evaluated for their potential anti-inflammatory effect in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. These compounds were tested for their inhibitory effects on nitric oxide (NO) production in LPS-stimulated RAW 264.7 cells. Since $5{\beta}$-hydroxypalisadin B showed the best activity it was further tested for the production of prostaglandin-$E_2$ ($PGE_2$), expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), the release of pro-inflammatory cytokines tumor necrotic factor-alpha (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), and interleukin-6 (IL-6). $5{\beta}$-Hydroxypalisadin B significantly reduced the $PGE_2$ release and suppressed the iNOS and COX-2 expression in LPS-stimulated RAW 264.7 cells. It also significantly reduced the release of pro-inflammatory cytokines TNF-${\alpha}$, IL-$1{\beta}$, and IL-6. These findings provide the first evidence of anti-inflammatory potential of $5{\beta}$-hydroxypalisadin B isolated from the red alga L. snackeyi and hence, it could be exploited as an active ingredient in pharmaceutical, nutraceutical and functional food applications.

The Anti-Inflammatory Effect of IH-901 in HT-29 Cells

  • Lee, Seung-Min;Kim, Ki-Nam;Kim, Yu-Ri;Kim, Hye-Won;Shim, Boo-Im;Lee, Seung-Ho;Bae, Hak-Soon;Kim, In-Kyoung;Kim, Meyoung-Kon
    • Molecular & Cellular Toxicology
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    • v.3 no.4
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    • pp.254-261
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    • 2007
  • 20-O-($\beta$-D-Glucopyranosyl)-20 (S)-protopanaxadiol (IH-901) is one of the major metabolites of ginsenosides from Panax ginseng, and is suggested that IH-901 has been associated with various pharmacological and physiological activities. In this study, we demonstrate that IH-901 induced anti-inflammation in HT-29 human colon adenocarcinoma cells. Our results showed that IH-901 inhibited cell proliferation of HT-29 in a time- and dose-dependent manner. We also found that IH-901 was significantly decreased expression of iNOS compared with non-treated. We observed effect of IH-901 related with inflammatory genes using by cDNA microarray. We were known that the 34 inflammatory genes such as E2F, CDK6, TNF-$\alpha$, and PKC were down-regulated. Thus, these results suggest that IH-901 may have a potential preventive factor to improving cancer induced by chronic inflammation.