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

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Effect of Sagunja-tang on Immune Function of Mouse Immune Cells (四君子湯이 免疫機能에 미치는 影響)

  • Lee, Sang-Hyun;Jung, Myung;Lim, Kyu-Sang;Yun, Yong-Gab
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.28 no.3
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    • pp.14-29
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    • 2015
  • Objectives : The extract of Sagunja-tang has been traditionally used for restorative treatment of constitutional weakness, vascular and immune disorder, and nervous disease in Oriental country. This study investigated the regulatory effects of Sagunja-tang on the expression, production, and activity of immune mediators.Methods : In this study, the extract of Sagunja-tang was prepared by extracting with distilled water at 100$^{\circ}C$ for 2.5h. The extract was freeze-dried following filtration through 0.45${{\mu}m}$ filter. The extract was dissolved in Hank's balanced salt solution (HBSS) and filtered again through 0.45${{\mu}m}$ filter before use. The level of nitrite, an oxidative product of nitric oxide(NO) was measured in the culture medium by the Griess reaction. The levels of prostaglandin E2(PGE2), Th1 cytokines (IFN-${\gamma}$, IL-2) and Th2 cytokines(IL-4, IL-5, IL-13) were measured by enzyme-linked immunosorbent assay and the protein levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression were determined by Western blot analysis. Also examined the effects of the extract on T-cell proliferation and cytotoxic activity of natural killer cells.Results : In this investigation, Production levels of Th2 cytokines (IL-4, IL-5, IL-13) was inhibited in a dose dependent manner by treatment with the extract. I also found that the extract increased T-cell proliferation and cytotoxic activity of natural killer cells in a dose-dependent manner.Conculsions : These results suggest that the water extract of Sagunja-tang may be useful for a therapeutic drug against a sickly constitution and immune diseases, probably by regulating the production of immune mediators.

Antioxidant and anti-inflammatory activity of parts of Rhus javanica L. (붉나무의 부위 별 항산화 및 항염증 활성)

  • Choi, Ji-Soo;Han, Sang-Don;Jang, Tae-Won;Lee, Seung-Hyun;Park, Jae-Ho
    • Journal of Applied Biological Chemistry
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    • v.62 no.2
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    • pp.195-202
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    • 2019
  • Rhus javanica L. is Anacardiaceae plant distributed in East Asia. We evaluated the antioxidant activity and antiinflammatory effect of leaf, branch, root of ethyl acetate fraction from R. javanica. To confirm effective each extraction, The antioxidant activity was evaluated using 1,1-Diphenyl-2-picryl-hydrazyl and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) scavenging activity assays, and the anti-inflammatory activity was evaluated based on inhibitory activities on the protein and mRNA expression of iNOS and COX-2 in LPS-induced RAW264.7 cells. The phenolic compounds content of each extract was analyzed with Folin reagents and HPLC/PDA method. The gallic acids were identified and quantified. The roots of R. javanica showed strong antioxidant activity. Its total phenolic compounds content were higher than the orders. In addition, anti-inflammatory activity inhibited the protein and mRNA expression of nitric oxide production factor, following the same pattern as contents of phenolic compounds included gallic acid and its antioxidant activity. In conclusion, R. javanica showed effective antioxidant and anti-inflammatory activity. Especially, the roots were evaluated to be highly valuable as a natural resource for reducing inflammation.

Extracts of Allium fistulosum Attenuates Pro-Inflammatory Action in the Lipopolysaccharide-Stimulated BV2 Microglia Cells (Lipopolysaccharide에 의한 BV2 세포의 염증 반응에 대한 파 추출물의 저해 활성)

  • Park, Shin-Hyoung;Kim, Jung-In;Jeong, Yong-Kee;Choi, Yung-Hyun
    • Journal of Life Science
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    • v.21 no.6
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    • pp.796-804
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    • 2011
  • Microglia are central nervous system (CNS)-resident professional macrophages that function as the principal immune cells responding to pathological stimulations in the CNS. Activation of microglia, induced by various pathogens, protects neurons and maintains homeostasis in the CNS, but severe activation causes inflammatory responses secreting various neurotoxic molecules such as nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) and pro-inflammatory cytokines. Allium fistulosum, a member of the onion family, is mainly cultivated for consumption, as well as medicinal use in Oriental medicine. It has been reported that A. fistulosum has various biological effects such as anti-oxidant, anti-platelet aggregation, anti-fungus and anti-cholesterol synthesis, however there has been no research about the anti-inflammatory effects of A. fistulosum extracts. In this study, it was undertaken to explore the functions of A. fistulosum as a suppressor of neuronal inflammation by using BV2 microglia cells. As a result, it was found that four kinds of extracts of A. fistulosum effectively reduced the expressions of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) at both mRNA and protein levels, and also attenuated pro-inflammatory cytokines such as tumor necrosis alpha (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$) and interleukin-6 (IL-6) at the mRNA level in BV2 stimulated by lipopolysaccharide (LPS). In addition, the extracts of A. fistulosum attenuated the release of NO markedly, as well as resulting in slight decreases of TNF-${\alpha}$ and IL-6 production, the effects of which were most significant when treated with ethyl alcohol extract from the whole A. fistulosum. In conclusion, the data indicated that the anti-inflammatory actions of A. fistulosum against BV2 microglia cells is through the down-regulation of iNOS, COX2 and pro-inflammatory cytokines such as TNF-${\alpha}$ and IL-6, and these effects are expected to help in the protection of nerve tissues by suppressions of neuronal inflammation in various neurodegenerative diseases.

Extracelluar Signal-Regulated Kinase-Dependent Nitric Oxide Production from Macrophage-Like Cells by Lactic Acid Bacteria

  • Byeon, Se-Eun;Yoo, Dae-Sung;Lee, Jae-Hwi;Kim, Suk;Rhee, Man-Hee;Park, Hwa-Jin;Cho, Jae-Youl
    • Biomolecules & Therapeutics
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    • v.17 no.3
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    • pp.276-281
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    • 2009
  • Lactic acid bacteria (LAB) are considered as probiotics with immunostimulatory property. In this study, we investigated the molecular mechanism of its immunostimulating potency on macrophages using combined preparation of LAB (cpLAB). cpLAB is able to strongly stimulate nitric oxide (NO) production as well as inducible NO synthase (iNOS) expression from macrophage-like RAW264.7 cells. The cpLAB-induced NO release seemed to be mediated by extracellular signal-regulated kinase (ERK) but not p38 and C-Jun N-terminal kinase (JNK), since U0126, an ERK inhibitor, clearly suppressed NO production. cpLAB significantly diminished the binding of toll like receptor (TLR)-2 antibody up to 25%, implying that cpLAB-mediated activation of macrophages may be required for the functional activation of TLR-2, but not TLR-4. Therefore, our data suggest that cpLAB may directly allow macrophages to immunostimulating potency via activation of TLR-2 and ERK.

Anti-inflammatory effect of enzymatic hydrolysates from Styela clava flesh tissue in lipopolysaccharide-stimulated RAW 264.7 macrophages and in vivo zebrafish model

  • Ko, Seok-Chun;Jeon, You-Jin
    • Nutrition Research and Practice
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    • v.9 no.3
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    • pp.219-226
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    • 2015
  • BACKGROUND/OBJECTIVES: In this study, potential anti-inflammatory effect of enzymatic hydrolysates from Styela clava flesh tissue was assessed via nitric oxide (NO) production in lipopolysaccahride (LPS) induced RAW 264.7 macrophages and in vivo zebrafish model. MATERIALS/METHODS: We investigated the ability of enzymatic hydrolysates from Styela clava flesh tissue to inhibit LPS-induced expression of pro-inflammatory mediators in RAW 264.7 macrophages, and the molecular mechanism through which this inhibition occurred. In addition, we evaluated anti-inflammatory effect of enzymatic hydrolysates against a LPS-exposed in in vivo zebrafish model. RESULTS: Among the enzymatic hydrolysates, Protamex-proteolytic hydrolysate exhibited the highest NO inhibitory effect and was fractionated into three ranges of molecular weight by using ultrafiltration (UF) membranes (MWCO 5 kDa and 10 kDa). The above 10 kDa fraction down-regulated LPS-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), thereby reducing production of NO and prostaglandin $E_2$ ($PGE_2$) in LPS-activated RAW 264.7 macrophages. The above 10 kDa fraction suppressed LPS-induced production of pro-inflammatory cytokines, including interleukin $(IL)-1{\beta}$, IL-6, and tumor necrosis factor $(TNF)-{\alpha}$. In addition, the above 10 kDa fraction inhibited LPS-induced phosphorylation of extracellular signal-regulated kinases (ERKs), c-Jun N-terminal kinase (JNK), and p38. Furthermore, NO production in live zebrafish induced by LPS was reduced by addition of the above 10 kDa fraction from S. clava enzymatic hydrolysate. CONCLUSION: The results of this study suggested that hydrolysates derived from S. clava flesh tissue would be new anti-inflammation materials in functional resources.

The Efficiency of Deer Antler Herbal Acupuncture on Modulation and Prevention of IL-1 Mediated Activation in Rat Chondrocytes at a Receptor Level

  • Kim, Woo-Young;Lee, Seung-Deok;Kim, Kyung-Ho;Baek, Seung-Tae;Kim, Kap-Sung
    • Journal of Acupuncture Research
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    • v.23 no.2
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    • pp.113-123
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    • 2006
  • Objectives : Deer antler Herbal-Acupuncture (DHA) solution represents one of the most commonly used medicine to treat rheumatoid arthritis. But, mechanisms of its antiarthritic activities are still poorly understood. Identification of common DHA aqua-acupuncture capable of affording protection or modulating the onset and severity of arthritis may have important human health implications. Results : We determined if DHA could prevent the binding of $IL-1{\beta}$ to its cellular receptors. DHA addition to rat chondrocytes treated with $IL-1{\beta}$ or with reactive oxygen species(ROS) prevents the activation of proteoglycan synthesis. After treatment with $IL-1{\beta}$, DHA increased the expression of mRNA encoding the type II $IL-1{\beta}$ receptor. These results emphasize the potential role of two regulating proteins of the $IL-1{\beta}$ signaling pathway that could account for the beneficial effect of DHA in osteRArthritis. The present study also identifies a novel mechanism of DHA-mediated anti-inflammatory activity. Conclusion : It is shown that DHA inhibits both $IL-1{\beta}-$ and $TNF-{\alpha}-induced$ NO production in normal human articular chondrocytes. The observed suppression of IL-1-induced NO production is associated with inhibition of inducible NO synthase(iNOS) mRNA and protein expression. In addition, DHA also suppresses the production of IL-1-induced cyclooxygenase-2 and IL-6. The constitutively expressed cyclooxygenase-1, however, was not affected by the sugar. These results demonstrate that DHA expresses a unique range of activities and identifies a novel mechanism for the inhibition of inflammatory processes.

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Anti-Inflammatory Effect of Chondrus nipponicus Yendo Ethanol Extract on Lipopolysaccharide-Induced Inflammatory Responses in RAW 264.7 Cells (LPS로 유도된 RAW 264.7 세포에 대한 가락진두발 에탄올 추출물의 항염증 효과)

  • Kim, Min-Ji;Bae, Nan-Yong;Kim, Koth-Bong-Woo-Ri;Park, Ji-Hye;Park, Sun-Hee;Jang, Mi-Ran;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.2
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    • pp.194-201
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    • 2016
  • The anti-inflammatory activity of ethanol extract from Chondrus nipponicus Yendo (CNYEE) was investigated by measuring production of a lipopolysaccharide-induced inflammatory response mediator. CNYEE had no cytotoxic effects on proliferation of macrophages compared to the control. CNYEE significantly inhibited (over 50%) NO production at $50{\mu}g/mL$, with inhibitory effects on expression levels of cytokines such as interleukin (IL)-6, tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and IL-$1{\beta}$. In particular, IL-6 inhibitory activity of CNYEE was higher than 70% at $100{\mu}g/mL$. CNYEE also reduced protein expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and nuclear factor (NF)-${\kappa}B$ in a dose-dependent manner. CNYEE also significantly reduced phosphorylation of p38, extracellular signal-regulated kinase, and c-Jun N-terminal kinase. Therefore, these results suggest that CNYEE may have anti-inflammatory effects by modulating the NF-${\kappa}B$ and mitogen-activated protein kinases signaling pathways and may be used as an anti-inflammatory therapeutic material.

Anti-Inflammatory Effects of Annona muricata Leaf Ethanol Extracts (그라비올라(Annona muricata) 잎 에탄올 추출물의 항염증 효과)

  • Cho, Eun-Ji;Lee, Joeng Hee;Sung, Nak-Yun;Byun, Eui-Hong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.6
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    • pp.681-687
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    • 2017
  • This study was focused on the anti-inflammatory activities of Annona muricata leaf ethanol extracts (AME). Inflammation of macrophages was induced by lipopolysaccharide (LPS) treatment, and various inflammation-mediated factors [cytokines and nitric oxide (NO)] were measured. AME treatment significantly reduced LPS-induced NO, cytokine levels [interleukin (IL)-6, tumor necrosis $factor-{\alpha}$ and $IL-1{\beta}$], and expression of inducible NO synthase and cyclooxygenase-2 in a dose-dependent manner. Mechanical studies showed that AME treatment inhibited activation of mitogen-activated protein kinase and nuclear factor $(NF)-{\kappa}B$ in macrophages treated with LPS. From these results, AME treatment strongly inhibits LPS-induced inflammation through inhibition of $NF-{\kappa}B$ activation, suggesting AME could be a potential candidate for treatment of inflammatory disease as a nutraceutical drug.

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.

Effects of Citrus limon Extract on Oxidative Stress-Induced Nitric Oxide Generation and Bovine Teeth Bleaching

  • Jeong, Soon-Jeong
    • Journal of dental hygiene science
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    • v.21 no.2
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    • pp.96-103
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    • 2021
  • Background: We aimed to investigate the effect of Citrous limon extract (CLE) on oxidative stress-induced cytotoxicity and nitric oxide (NO) generation and the tooth bleaching effect of CLE as a substitute for hydrogen peroxide (H2O2) and determine the feasibility and application of CLE as a safe and effective natural tooth bleaching agent. Methods: The protective effect of CLE on H2O2-induced cytotoxicity in Raw264.7 macrophages was investigated by the MTT assay. The inhibitory effect of CLE on the generation of H2O2-induced NO was confirmed by the NO assay, and the changes in inducible nitric oxide synthase (iNOS) protein expression were confirmed by western blotting. Stained bovine teeth were treated with/without 15% and/or 35% CLE and H2O2, 15% sodium bicarbonate (NaHCO3) for 3 hours, and were irradiated with/without bleaching light (BL) for 15 minutes. The color change of the treated bovine tooth surface was measured using a colorimeter. Results: The viability of Raw264.7 cells treated with each concentration of CLE and 500 μM H2O2 significantly increased as CLE increased, and NO generation and iNOS protein expression were significantly reduced in cells treated with 300 ㎍ CLE+/500 μM H2O2+ and 300 ㎍ CLE+/500 μM H2O2+/150 ㎍ NaHCO3+. The bleaching effect of 35% CLE+ was higher than that of 15% CLE+ and 15% NaHCO3+, and the effect was similar to that of 15% H2O2+. The 35% CLE+/15% NaHCO3+ showed the greatest bleaching effect and was higher than that of the groups irradiated with the BL. The greatest bleaching effect was observed with 35% CLE+/15% NaHCO3+, followed by 35% H2O2+/BL+. Conclusion: CLE inhibited oxidative stress-induced cytotoxicity and NO generation in Raw264.7 cells and, could replace H2O2, which causes side effects and risks in teeth breaching treatment. It showed greatest teeth bleaching effect when combined with NaHCO3. CLE is an effective and safe natural tooth bleaching substitute.