• Title/Summary/Keyword: NO/iNOS activity

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Anti-inflammatory Effect of Yongseollan on the LPS-activated RAW 264.7 Cells

  • Jo, Mi-Jeong;Lee, Byung-Wook;Eom, Dong-Myung;Lee, Jong-Rok;Hwangbo, Min;Jee, Seon-Young;Kim, Sang-Chan
    • Herbal Formula Science
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    • v.15 no.1
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    • pp.175-183
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    • 2007
  • This study was conducted to evaluate the inhibitory effects of Yongseollan(YSL) on the production of nitric oxide (NO) and the expression of inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-activated RAW264.7 cells. YSL is tropical plant originating from Mexico. The biological activity of this plant is not yet evaluated systematically. The aim of the present work is to investigate a potential anti-inflammatory activity of YSL. The RAW264.7 cells were cultured in D MEM/F12 medium for 24 hrs. After serum starvation, cells were treated with YSL for 1 hr, followed by stimulating NO production with a LPS. We found that YSL has an inhibitory effect on the production of NO, iNOS expression and $phospho-I{\kappa}B$ expression. YSL also inhibited tumor necrosis factor $(TNF)-{\alpha}$, interleukin (IL)-6, and $IL-1{\beta}$. Moreover, YSL inhibited cyclooxygenase (COX)-2 expression and prostanglandin E2 (PGE2). These findings showed that YSL could have some anti-inflammatory effects which might play a role in therapy in Gram-negative bacterial infections.

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Inhibitory Effect of Taraxci Herba Methanol Extract on Pro-inflammatory Mediator in Lipopolysaccharide;Activated Raw 264.7 cells

  • Jo, Mi-Jeong;Chu, Yan-Hui;Back, Young-Doo;Lee, Byung-Wook;Shin, Soon-Shik;Kwon, Young-Kyu;Kim, Sang-Chan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.4
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    • pp.907-913
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    • 2008
  • Taraxci Herba (TH; Pogongyoung in Korean) has been used in traditional oriental medicine for the treatment of various ailments. The biological activity of this plant is not yet evaluated systematically. This study was conducted to evaluate the inhibitory effects of TH on the production of nitric oxide (NO) and the expression of inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-activated Raw264.7 cells. The aim of the present work is to investigate a potential anti-inflammatory activity of TH. The Raw264.7 cells were cultured in DMEM medium for 24 h. After serum starvation for 12 h, the cells were treated with TH for 1 h, followed by stimulating NO production with LPS ($2{\mu}g/ml$). As result of this study, TH inhibited the levels of NO, PGE2, $TNF-{\alpha}$, IL-6 and $IL-1{\beta}$, and the expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) activated by LPS. These inhibitory effects were mediated though the inhibition of phosphorylation of inhibitory kappa B ($I{\kappa}B$). These findings showed that TH could have some anti-inflammatory effects.

Anti-Inflammatory Effect of Mangostenone F in Lipopolysaccharide-Stimulated RAW264.7 Macrophages by Suppressing NF-κB and MAPK Activation

  • Cho, Byoung Ok;Ryu, Hyung Won;So, Yangkang;Lee, Chang Wook;Jin, Chang Hyun;Yook, Hong Sun;Jeong, Yong Wook;Park, Jong Chun;Jeong, Il Yun
    • Biomolecules & Therapeutics
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    • v.22 no.4
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    • pp.288-294
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    • 2014
  • Mangostenone F (MF) is a natural xanthone isolated from Garcinia mangostana. However, little is known about the biological activities of MF. This study was designed to investigate the anti-inflammatory effect and underlying molecular mechanisms of MF in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. MF dose-dependently inhibited the production of NO, iNOS, and pro-inflammatory cytokines (TNF-${\alpha}$, IL-6, and IL-$1{\beta}$) in LPS-stimulated RAW264.7 macrophages. Moreover, MF decreased the NF-${\kappa}B$ luciferase activity and NF-${\kappa}B$ DNA binding capacity in LPS-stimulated RAW264.7 macrophages. Furthermore, MF suppressed the NF-${\kappa}B$ activation by inhibiting the degradation of $I{\kappa}B{\alpha}$ and nuclear translocation of p65 subunit of NF-${\kappa}B$. In addition, MF attenuated the AP-1 luciferase activity and phosphorylation of ERK, JNK, and p38 MAP kinases. Taken together, these results suggest that the anti-inflammatory effect of MF is associated with the suppression of NO production and iNOS expression through the down-regulation of NF-${\kappa}B$ activation and MAPK signaling pathway in LPS-stimulated RAW264.7 macrophages.

$\beta$-Glucan Suppresses LPS-stimulated NO Production Through the Down-regulation of iNOS Expression and $NF{\kappa}B$ Transactivation in RAW 264.7 Macrophages

  • Yang, Jeong-Lye;Jang, Ji-Hyun;Radhakrishnan, Vinodhkumar;Kim, Yang-Ha;Song, Young-Sun
    • Food Science and Biotechnology
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    • v.17 no.1
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    • pp.106-113
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    • 2008
  • The antioxidant and anti-inflammatory protective effects of $\beta$-glucan from barley on RAW 264.7 murine macrophage cells induced by lipopolysaccharide (LPS) were examined. The RAW 264.7 murine macrophages were preincubated with various concentrations ($0-200\;{\mu}g/mL$) of $\beta$-glucan and stimulated with LPS to induce oxidative stress and inflammation. The $\beta$-glucan treatments were found to reduce thiobarbituric acid-reactive substance (TBARS) accumulation, and enhance glutathione levels and the activities of antioxidative enzymes, including superoxide dismutase (SOD), catalase, glutathione reductase, and glutathione peroxidase (GSH-px) in the LPS-stimulated macrophages as compared to the LPS-only treated cells. Nitric oxide (NO) production was significantly suppressed in a dose-dependent manner (p<0.05) with an $IC_{50}$ of $104\;{\mu}g/mL$. Further treatment with $\beta$-glucan at $200\;{\mu}g/mL$ suppressed NO production to 2% of the LPS-control, and suppressed the levels of inducible nitric oxide synthase (iNOS) protein and mRNA in a dose-dependent manner. The specific DNA binding activity of nuclear factor ${\kappa}B\;(NF{\kappa}B)$ was significantly suppressed by $\beta$-glucan treatment with an $IC_{50}$ of $220\;{\mu}g/mL$ in a dose-dependent manner. Finally, barley $\beta$-glucan ameliorates NO production and iNOS expression through the down-regulation of $NF{\kappa}B$ activity, which may be mediated by attenuated oxidative stress in RAW 264.7 macrophages.

Anti-oxidant, Anti-inflammation and Anti-microbial Effects of Hoangtonogak Plus Extracts (황토노각플러스 추출물의 항산화, 항염 및 항미생물 효능)

  • Cho, Jun-Hee;Lee, Ji-An
    • Journal of Convergence for Information Technology
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    • v.10 no.12
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    • pp.183-190
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    • 2020
  • This study evaluated the possibility of Hoangtonogak Plus extracts as a bioactive ingredients for cosmetic products. Methanol(MN) and hot-water(WN) extracts were analysed by DPPH/ABTS radical scavenging activity, FRAP value for anti-oxidant activity, MTT assay for cell viability, inhibition of NO production and iNOS protein expression for anti-inflammatory effect, paper disc diffusion method for anti-microbial activity against Staphylococcus aureus, Staphylococcus epidermidis and Escherichia coli.. The contents of total polyphenol of MN and WN extracts were 2.92±0.01 mgGAE/g and 1.67±0.02 mgGAE/g, respectively. DPPH, ABTS and FRAP values of MN extracts were higher than WN at each concentration. No significant cytotoxicity was observed in RAW264.7 cells. Furthermore, NO production of MN and WN at 1 mg/mL concentration was measured as 11.69 μM, 20.4 μM, respectively. In addition, MN extracts showed anti-microbial effect only on S. epidermidis. Also MN extracts suppressed iNOS protein level in a concentration-dependent manner. According to our results, the MN extracts demonstrated its potential as a natural source of antioxidant with anti-microbial and anti-inflammatory properties.

Effect of Ethanol Extracts from Defatted Perilla frutescens on LPS-induced Inflammation in Mouse BV2 Microglial Cells

  • Lee, Sung-Gyu;Kang, Hyun
    • Biomedical Science Letters
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    • v.24 no.4
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    • pp.398-404
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    • 2018
  • To evaluate the antioxidant and anti-neuroinflammatory effects of defatted Perilla frutescens extract (DPE) in lipopolysaccharide (LPS)-stimulated BV-2 microglial cells. Cell viabilities were estimated by MTT assay. LPS-stimulated BV-2 microglia were used to study the expression and production of inflammatory mediators such as nitric oxide (NO), inducible NO synthase (iNOS), Cyclooxygenase-2 (COX-2), and prostaglandin $E_2$ ($PGE_2$). Pretreatment with DPE prior to LPS treatment significantly inhibited excessive production of NO (10, 25, 50, 75, and $100{\mu}g/mL$) in a dose-dependent manner, and was associated with down regulation of expression of iNOS and COX-2. DPE also suppressed the LPS-induced increase in $PGE_2$ level (10, 25, 50, 75, and $100{\mu}g/mL$) in BV-2 cells. Therefore, DPE can be considered as a useful therapeutic and preventive approach for the treatment of several neurodegenerative diseases.

Antioxidant and NO-scavenging Activities of Acanthopanax senticosus var. subinermis Leaf Extracts Prepared Using Ethanol and Extrusion Processing

  • Lee, Sung-Hee;Oh, Hea-Young;Leem, Jae-Yoon;Yoon, Sun
    • Food Science and Biotechnology
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    • v.18 no.5
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    • pp.1124-1131
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    • 2009
  • Acanthopanax senticosus has been used in East Asia as a traditional medicine. The purpose of this study is the investigation of an extraction method for A. senticosus var subinermis (ASvS) leaves to improve their antioxidant and anti-inflammatory activities. Six extracts were prepared: extracted with water (W), ethanol (Eth), water or ethanol after ultra high pressure (WP, EthP), and water or ethanol after an extrusion process (WEx, EthEx). Diphenylpicrylhydrazyl (DPPH) radical scavenging activity and ferric reducing ability showed that all extracts had a significant antioxidant activity. In anti-inflammatory activities, Eth and EthEx significantly inhibited the release of lipopolysaccharide (LPS)-induced nitric oxide (NO), tumor necrosis factor (TNF)-${\alpha}$, and interleukin (IL)-$1{\beta}$ independent of cell viability in RAW 264.7 macrophage cells. Inducible NO synthase (iNOS) protein was significantly decreased by EthEx. These findings present that Eth and EthEx extracts of ASvS leaves have anti-inflammatory activities, and EthEx extract suppresses LPS-induced NO through the down-regulation of iNOS and pro-inflammatory cytokines.

Inhibitory Activity of Nitric Oxide Synthase and Peroxynitrite Scavenging Activity of Extracts of Perilla frutescens (들깨 잎 추출물의 Nitric Oxide Synthase 저해활성 및 Peroxynitrite 소거활성)

  • Kim, Jae-Yeon;Kim, Ji-Sun;Jung, Chan-Sik;Jin, Chang-Bae;Ryu, Jae-Ha
    • Korean Journal of Pharmacognosy
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    • v.38 no.2 s.149
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    • pp.170-175
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    • 2007
  • Activated microglia by neuronal injury or inflammatory stimulation overproduce nitric oxide (NO) by inducible nitric oxide synthase (iNOS) and reactive oxygen species (ROS) such as superoxide anion, resulting in neurodegenerative diseases. The toxic peroxynitrite (ONOO$^-$), the reaction product of NO and superoxide anion further contributes to oxidative neurotoxicity. We tried to evaluate the effects of two kinds of varieties of Perilla frutescens var japnica Hara on the NO production in lipopolysaccharide (LPS)-activated microglia. The perilla cultivars of Namcheondeulkkae (NC) and Boradeulkkae (BR) were developed by pure line from the local variety and by a cross between 'deulkkae' and 'chajogi', respectively. Spirit, hexane, chloroform and butanol fractions of the leaves of NC and BR inhibited the production of NO in LPS-activated microglia. The fractions of BR showed stronger activity than NC and the spirit extracts was the most potent in both cultivars. The solvent fractions of BR suppressed the expression of protein and mRNA of iNOS in LPS-activated microglial cells. Moreover, the extracts of NC and BR showed the activity of peroxynitrite scavenging in cell free bioassay system. These results imply that Namcheondeulkkae and Boradeulkkae might have neuroprotective activity through the inhibition of NO production by activated microglial cells and peroxynitrite scavenging activity.

Zanthoxylum rhetsa Stem Bark Extract Inhibits LPS-induced COX-2 and iNOS expression in RAW 264.7 Cells via the NF-${\kappa}B$ Inactivation

  • Thu, Nguyen Bich;Trung, Trinh Nam;Ha, Do Thi;Khoi, Nguyen Minh;Than, Nguyen Viet;Soulinho, Thipthaviphone;Nam, Nguyen Hai;Phuong, Tran Thi;Bae, Ki-Hwan
    • Natural Product Sciences
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    • v.16 no.4
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    • pp.265-270
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    • 2010
  • The methanol extract of Zanthoxylum rhetsa (MZRR) were evaluated for its ability to suppress the formation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. MZRR presented an inhibition of LPS-induced production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) in RAW 264.7 macrophages. Western blotting and RT-PCR analyses demonstrated that MZRR significantly inhibited the protein and mRNA expressions of iNOS and COX-2 in LPS-activated macrophages in a dose-dependent manner. LPS-induced COX-2, iNOS, and nuclear factor kappa beta (NF-${\kappa}B$) activity were also decreased in the presence of MZRR. The production of tumor necrosis factor-$\alpha$ (TNF-$\alpha$), the mRNA expression levels of pro-inflammatory cytokines, including TNF-$\alpha$ and IL-$1{\beta}$, were reduced after MZRR administration in a dose dependent-manner. These results suggest that the MZRR extract involved in the inhibition of iNOS and COX-2 via the NF-${\kappa}B$ pathway, revealing a partial molecular basis for anti-inflammatory properties of the MZRR extract.

Anti-Inflammatory Activities of (+)-Afzelechin against Lipopolysaccharide-Induced Inflammation

  • In-Chul Lee;Jong-Sup Bae
    • Biomolecules & Therapeutics
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    • v.32 no.4
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    • pp.467-473
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    • 2024
  • In this study, we investigated the potential protective effects of (+)-afzelechin (AZC), a natural compound that is derived from Bergenia ligulata, on lipopolysaccharide (LPS)-induced inflammatory responses. AZC is known to have antioxidant, anticancer, antimicrobial, and cardiovascular protective properties. However, knowledge regarding the therapeutic potential of AZC against LPS-induced inflammatory responses is limited. Thus, we investigated the protective attributes of AZC against inflammatory damage caused by LPS exposure. We examined the effects of AZC on heme oxygenase (HO)-1, cyclooxygenase (COX)-2, and inducible nitric oxide synthase (iNOS) in LPS-activated human umbilical vein endothelial cells (HUVECs). In addition, the effects of AZC on the expression of iNOS, tumor necrosis factor (TNF)-α, and interleukin (IL)-1β were analyzed in the lung tissues of LPS-injected mice. Data revealed that AZC promoted the production of HO-1, inhibited the interaction between luciferase and nuclear factor (NF)-κB, and reduced the levels of COX-2/PGE2 and iNOS/NO, thereby leading to a decrease in the signal transducer and activator of transcription (STAT)-1 phosphorylation. Moreover, AZC facilitated the nuclear translocation of Nrf2, increased the binding activity between Nrf2 and the antioxidant response elements (AREs), and lowered the expression of IL-1β in the LPS-treated HUVECs. In the animal model, AZC significantly reduced the expression of iNOS in the lung tissue structure and the TNF-α level in the bronchoalveolar lavage fluid. These findings demonstrate that AZC possesses anti-inflammatory properties that regulate iNOS through the inhibition of both NF-κB expression and p-STAT-1. Consequently, AZC has potential as a future candidate for the development of new clinical substances for the treatment of pathological inflammation.