• Title/Summary/Keyword: NO production inhibitory effect

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Anti-inflammatory Effect of Castanopsis cuspidata Extracts in Murine Macrophage RAW 264.7 Cells (Murine Macrophage RAW 264.7 세포에서 구실잣밤나무 추출물의 항염증 효과)

  • Ko, Yeong-Jong;Song, Sang Mok;Hyun, Woo-Chol;Yang, Soo-Kyung;Song, Chang-Khil;Lee, Dong-Sun;Yoon, Weon-Jong
    • Korean Journal of Plant Resources
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    • v.27 no.5
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    • pp.439-446
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    • 2014
  • This study describes a preliminary evaluation of the anti-inflammatory activity of Castanopsis cuspidata extracts. C. cuspidata was extracted using 80% ethanol and then fractionated sequentially with n-hexane, dichloromethane, ethylacetate, and butanol. To screen for anti-inflammatory agents effectively, we first examined the inhibitory effect of the C. cuspidata extracts on the production of pro-inflammatory factors and cytokines stimulated with lipopolysaccharide. In addition, we examined the inhibitory effect of C. cuspidata extracts on pro-inflammatory mediators (NO, iNOS, COX-2) in murine macrophage RAW 264.7 cells. The amounts of protein levels were determined by immunoblotting. Of the sequential solvent fractions of C. cuspidata, the n-hexane, dichloromethane and ethylacetate fractions inhibited the mRNA expression of pro-inflammatory cytokines (IL-$1{\beta}$ and IL-6), production of NO, and the protein level of iNOS and COX-2. These results suggest that C. cuspidata may have significant effects on inflammatory factors and may be provided as a possible anti-inflammatory therapeutic plant.

Inhibition of NO Production by Ethanol Extract of Prunus mume Fruits in LPS-Stimulated RAW 264.7 Macrophages through Regulation of the Nrf2/HO-1 Signaling Pathway (LPS가 처리된 RAW 264.7 대식세포에서 Nrf2/HO-1 경로 조절을 통한 매실 추출물의 NO 생성 억제 효과)

  • Kang, Hye-Joo;Choi, Eun Ok;Jeong, Jin-Woo;Park, Shin-Hyung;Park, Cheol;Hong, Su Hyun;Shin, Soon Shik;Cheong, Jae-Hun;Choi, Yung Hyun
    • Herbal Formula Science
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    • v.25 no.1
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    • pp.1-10
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    • 2017
  • Objectives : The fruit of Prunus mume Siebold & Zucc. has been used as an alternative medicine and functional food in Korea and Japan for preventive and therapeutic purposes. However, its molecular actions and mechanism on anti-inflammatory activity have not been clearly investigated. The aim of this study was to clarify the anti-inflammatory activity of the ethanol extract of P. mume fruit (EEPM) in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells, and sought to understand the associated molecular mechanisms. Methods : Cytotoxicity was assessed by an MTT assay. The amount of nitric oxide (NO) production was determined by nitrite assay. The mRNA expression of inducible nitric oxide synthase (iNOS) was analyzed by RT-PCR. In addition, expression levels of iNOS, nuclear factor-erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) protein were detected by Western blotting. Results : Our data indicated that EEPM inhibited NO production in LPS-stimulated RAW264.7 cells in a concentration-dependent manner. At the mRNA and protein levels, EEPM suppressed LPS-induced iNOS expression. On the other hand, EEPM markedly enhanced HO-1 expression, which was associated with an induction and nuclear translocation of Nrf2. Moreover, the inhibitory effect of EEPM against LPS‑induced NO production was significantly enhanced by hemin, a HO-1 inducer; however, EEPM's effect on the production of NO was abolished by zinc protoporphyrin IX, a HO-1 inhibitor. Conclusion : The results suggest that EEPM can act as a suppressor agent on NO production through an activation of Nrf2/HO-1 signaling pathway, and may be a promising candidate for the treatment of inflammatory diseases.

Anti-inflammatory Activity of Extracts from Ultra-Fine Ground Saururus chinensis Leaves in Lipopolysaccharide-Stimulated Raw 264.7 Cells

  • Kim, Dong-Hee;Cho, Jun-Hyo;Cho, Young-Je
    • Journal of Applied Biological Chemistry
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    • v.59 no.1
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    • pp.37-43
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    • 2016
  • Bioactive components of ultra-fine ground Saururus, the extraction yield increases when the leaves are ultra-fine ground. Comparison of normal-ground and ultra-fine ground Saururus chinensis leaves showed that the solid content and antiinflammatory activity of ultra-fine ground extracts was higher than that of normal-ground extracts. Lipopolysaccharide (LPS)-stimulated Raw 264.7 cells were treated with different concentrations of Saururus chinensis extract and the amount of nitric oxide (NO) was determined; LPS-treated cells produced 2 times more NO than cells that were not treated with LPS. Moreover, the NO production in cells treated with Saururus chinensis extract was inhibited in a concentration-dependent manner. Because the stimulant-induced NO production is regulated by the inducible nitric oxide synthase (iNOS), we measured the iNOS protein level to elucidate the mechanism by which the NO production was inhibited. We found that the amount of iNOS decreased dose-dependently. It was reduced by 53% at a Saururus chinensis extract concentration of $100{\mu}g/mL$. The protein expression of cyclooxygenase-2 (COX-2) in LPS-treated Raw 264.7 cells was inhibited by 31% at $100{\mu}g/mL$ of Saururus chinensis extract. Gel shift of the nuclear factor kappa B-DNA complex occurred in LPS-treated cells and the intensity of the band decreased gradually in a concentration-dependent manner. Ultra-fine ground Saururus chinensis extract had a concentration-dependent inhibitory effect on the production of prostaglandin $E_2$, tumor necrosis factor ${\alpha}$, interleukin $1{\beta}$ (IL-$1{\beta}$), IL-6, and IL-8 in LPS-treated Raw 264.7 cells, i.e., at $50{\mu}g/mL$ of Saururus chinensis extract, their levels were decreased by 53, 67, 52, 37, and 21% respectively.

The Effects of Cortex Mori on NO, $TNF-{\alpha}$ and $IL-1{\alpha}$ production by macrophage (상백피(桑白皮)가 대식세포의 NO, $TNF-{\alpha}$$IL-1{\alpha}$ 생산에 미치는 영향)

  • Ahn, Jae-Kyu;Ahn, Duk-Kyun;Cho, Jae-Chon
    • The Journal of Korean Medicine
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    • v.19 no.2
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    • pp.485-501
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    • 1998
  • Cortex Mori (Moros alba L.), the root bark of mulberry tree has been used as an autiphlogistic, diuretic and expectorant in herval medicine. Recently, a few papers reported that phenolic extract of Cortex Mori had the hypotensive, hypoglycemic, antiviral and anticancer effects, and hot water extract of Cortex Mori(CM) had inhibitory effect on the degranulation and histamine release from activated mast cells. These previous studies suggest a possibility that CM has an antidotal activity against inflammation which was mediated mainly by macrophage-secreting inflammatory factors. This study was performed to evaluate the influences of CM on carrageenan-induced edema in vivo and release of inflammatory mediators such as NO, TNF and IL-1 by macrophages stimulated with LPS or $IFN-{\gamma}$ in vitro. Subcutaneous injections of carrageenan into the mouse paw rapidly induced local edema by increasing vascular permeability, but single intraperitoneal injection of CM extract at 30 minutes before carrageenan suppressed the development of edema. NO and TNF production from macrophage stimulated by LPS or $IFN-{\gamma}$ were significantly suppressed, especially TNF secretion by up to 3-4 folds. LPS stimulated IL-1 production was also inhibited, but not significantly. Cell viability assay verified that the inhibition was not due to general cell toxicity. These results suggest that reduction of NO, TNF and IL-1 production may be one of the means by which CM prevent inflammation associated diseases.

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The Effects of bilobalide Extracted from Ginkgonis Folium on Inflammation (은행잎의 주성분인 bilobalide가 염증반응에 미치는 효과)

  • Jung, Je-Ryong;Kil, Ki-Jung
    • The Korea Journal of Herbology
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    • v.30 no.1
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    • pp.85-93
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    • 2015
  • Objectives : Bilobalide (BIL) is a predominant sesquiterpene trilactone constituent that accounts for a partial portion of the standardized Ginkgonis Folium extract, which has been widely used to treat a variety of neurological disorders involving cerebral ischemia and neurodegeneration. In this study, it was tested whether BIL exhibits anti-inflammatory activities on inflammation response, or not. Methods : To elucidate the molecular mechanisms of BIL on pharmacological and biochemical actions in inflammation, we examined the effect of BIL on pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated macrophages. The investigation was focused on how BIL affect on inflammation-related mediators including various signals such as nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), inducible NO synthase(iNOS), cyclooxygenase-2(COX-2), interleukin-6(IL-6), tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), mitogen-activated protein kinases(MAPKs) and nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$) in LPS-stimulated RAW 264.7 cells. Results : We found that BIL inhibited LPS-induced NO, $PGE_2$, IL-6 and $TNF-{\alpha}$ productions as well as the expressions of iNOS and COX-2. Furthermore, BIL suppressed the LPS-induced phosphorylation for MAPK activation. Conclusions : These results suggest that BIL has inhibitory effects on LPS-induced $PGE_2$, NO, IL-6 and $TNF-{\alpha}$ production, as well as the expressions of iNOS and COX-2 in the murine macrophage. It seems that these inhibitory effects occur by blocking the phosphorylation of MAPKs for activation. Then, BIL suppressed the activation of nuclear factor $NF-{\kappa}B$ in nucleus. These observations suggest that BIL has anti-inflammatory effect by inhibiting.

Efficacy Evaluation of Extracts from Different Parts of Buxus Koreana (회양목 부위별 추출물의 효능평가)

  • Kim, Yong-Min;Kim, Myong-Ki
    • Journal of Convergence for Information Technology
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    • v.10 no.11
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    • pp.272-278
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    • 2020
  • This study was conducted to confirm the component and efficacy evaluation of Buxus Koreana by different part. Leaf, stem, root, and fruit were extracted with 70% ethanol, and the contents of polyphenols and flavonoids were analyzed, and the efficacy against anti-inflammatory, moisturizing and wrinkles were confirmed. The contents of polyphenols and flavonoids of B. Koreana were the highest in fruit extracts at 77.32 and 51.91 mg/g, respectively. As a result of confirming the NO production inhibitory effect, the root extracts showed an inhibitory effect of 29.6%, and iNOS significantly decreased as the concentration of the extracts increased compared to the LPS treatment group. As a result of checking the mRNA expression level of MMP1, it was found to be effective in the root and stem extracts. Through the efficacy evaluation of B. Koreana the possibility of use as a cosmetic material was confirmed.

Reconsideration of Classical Antibiotic Lincomycin: Anti-inflammatory Effect in LPS-stimulated RAW 264.7 Cells

  • Yang, Eun-Jin;Lee, Nari;Hyun, Chang-Gu
    • Microbiology and Biotechnology Letters
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    • v.48 no.3
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    • pp.366-372
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    • 2020
  • Since, side effects of antibiotics are frequently emphasized these days, their use is gradually diminishing, and alternative drugs are being developed. We have sought to reintroduce them as raw materials for human health as conventional 'weapons' that have been retired after their historical duties. In this study, we investigated the anti-inflammatory effects of lincomycin (LIN), on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Our findings show that LIN potently inhibited production of LPS-induced proinflammatory mediators, such as nitric oxide (NO) and prostaglandin E2 (PGE2), without cytotoxicity. Consistent with these findings, LIN strongly decreased protein expression levels of inducible NO synthase (iNOS) and cyclooxygenase (COX-2). Furthermore, LIN reduced pro-inflammatory cytokines such as tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β. To further elucidate the mechanisms of these inhibitory effects of LIN, we studied LPS-induced IκB-α degradation, and mitogen-activated protein kinase (MAPK) phosphorylation. LIN suppressed downregulation of inhibitory κB (IκB-α) degradation, and the phosphorylation of the c-Jun N-terminal kinase (JNK) pathway. Based on these results, we suggest that LIN may be considered a potential candidate as an anti-inflammatory cosmetic or a medicine for human health.

Modulation of Cellular Immune Response by Inosiplex (Inosiplex에 의한 세포성 면역반응의 변화)

  • Lee, Hern-Ku;Lee, Jeong-Ho;Kim, Hak-Kun;Ha, Tai-You
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.2
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    • pp.251-259
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    • 1986
  • This study was performed to assess the effect of inosiplex(ISP) on the resistance of mice Candida albicans infection, the migration of chicken leukocytes, the production of leukocyte migration inhibitory factor(LIF), and the cell-mediated immunity(CMI) to lepomin in multibacillary lepromatous leprosy patients. The treatment with ISP before or on the time of infection with C. albicans had no or deliterious effect, and treatment with ISP after infection had no effect on the recovery of C. albicans from the kidneys of mice. The migratory ability of chicken leukocytes and the production of LIF from splenocytes of mice were not affected by ISP treatment. However, ISP decreased the migration of chicken leukocytes in vitro, and this decrease was dose-dependent. The therapy of lepromatous leprosy patients with ISP for 10 or 30 days clearly showed the increase of the significant positive rate of Mitsuda skin test to lepromin. The immune recovery as a result of the therapy was found to be the best in the group of patients treated for 30 days. This results suggest that (1) the effect of ISP in renal candidiasis can vary depending on the time of treatment relative to infection, (2) ISP can primarily change the migratory ability of chicken leukocytes but does not affect the production of LIF in mice, and (3) the classical therapy combined with ISP can reinforce or restore the defences of lepromatous leprosy patients against Mycobacterium leprae.

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Inhibitory Effect of Extract of Teucrium veronicoides on the Production of Inflammatory Cytokines (곽향 추출물의 염증성세포활성물질 억제효과)

  • Bae, Gi-Sang;Park, Hee-Je;Kim, Do-Yun;Seo, Sang-Wan;Park, Kyung-Bae;Kim, Byung-Jin;Song, Je-Moon;Lee, Kyung-Yong;Na, Chul;Shin, Byung-Chul;Park, Sung-Joo;Song, Ho-Joon;Hwang, Sung-Yeon
    • The Korea Journal of Herbology
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    • v.23 no.3
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    • pp.119-125
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    • 2008
  • Objectives : The purpose of this paper was to investigate the anti-inflammatory effects of extract from Teucrium veronicoides (TV) on the RAW 264.7 cells. Methods : To evaluate of anti-inflammatory of TV, we examined the cytokine productions on lipopolysacchride (LPS)-induced RAW 264.7 cells and also inhibitory mechanisms using Western blot. Furthermore, We examined LPS-induced endotoxin shock. Results : 1. Extract from TV itself does not have any cytotoxic effect. 2. Extract from TV reduced LPS-induced Nitric oxide (NO),interleukin (IL)-1b, IL-6 and IL-10, tumor necrosis factor-a (TNF-a) production in RAW 264.7 cells. 3. TV inhibited the activation of mitogen-activated protein kinases (MAPKs) such as p38, extracelluar signal-regulated kinase (ERK 1/2) and c-Jun NH2-terminal kinase (JNK) and also the degradation of inhibitory kappa B a (Ik-Ba) in the LPS-stimulated RAW 264.7 cells. 3. TV slightly increased the duration of survival after LPS-induced endotoxin shock. Conclusions : TV down-regulated LPS-induced NO and cytokines production, which could provide a clinical basis for anti-inflammatory properties of TV.

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Inhibitory Effect of Brassica rapa in Ovalbumin-Stimulated Experimental Asthmic Mice

  • Bae, Eun-Ah;Min, Seung-Won;Kim, Nam-Jae;Bang, Myung-Ho;Baek, Nam-In;Han, Eun-Joo;Chung, Hae-Gon;Kim, Dong-Hyun
    • Natural Product Sciences
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    • v.13 no.2
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    • pp.114-117
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    • 2007
  • To evaluate antiallergic effect of Brassica rapa Metzg (BR, Family Cruciferae), which is widely used as a food source, its anti-asthmic effect was investigated. BR reduced IgE level in the blood of ovalbumin (OVA)-induced asthmic mice. BR also inhibited IgE and ploinflammatory cytokine IL-6 productions in trachea of OVA-induced mice, but did not inhibit IL-4 production. BR did not inhibit mRNA expression of proinflammatory cytokines IL-6 and IL-4 in IgE-induced RBL-2H3 cells. BR and its isolated components 10-undecenoic acid 2-methoxy methyl ester and galactosyl diglyceride potently inhibited the NO production of RAW264.7cells induced by lipopolysaccharide. These findings suggest that the anti-asthmic effect of BR may be due to the inhibition of the biosynthesis of proinflarnmatory cytokine IL-6 in macrophage and BR can improve IgE-induced allergic disease asthma.