• Title/Summary/Keyword: Inhibition of nitric oxide

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Inhibition of Nitric Oxide Synthesis by Methanol and Butanol Extracts of Euonymus Alatus (Thunb.) Sieb in Murine Macrophages

  • Lee Hyo-Hyun;Park Young-Soo;Kim Ra-Young;Kim Dong-Il;Lee Tae-Kyun
    • The Journal of Korean Medicine
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    • v.26 no.1 s.61
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    • pp.26-36
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    • 2005
  • Objective : Many traditional herbal remedies exhibit several beneficial effects including anti-inflammation. Euonymus alatus (Thunb.) Sieb (EA), known as Gui jun woo in Korea, has long been used in folk medicine to regulate Qi (bodily energy) and blood circulation, relieve pain, eliminate stagnant blood, and treat dysmenorrhea in oriental countries. The exact mechanism of the anti-inflammatory action of Euonymus alatus (Thunb.) Sieb (EA), however, has not been determined. Methods: Since there is increasing evidence that nitric oxide (NO) plays a crucial role in the pathogenesis of inflammatory diseases, this study was undertaken to address whether the methanol (MeOH) extract and its fractions of the bark of EA could modulate the expression of inducible NO synthase (iNOS) in thioglycollate-elicited murine peritoneal macrophages and murine macrophage cell line, RA W264.7 cells. Results: Stimulation of the peritoneal macrophages and RAW264.7 cells with $interferon-\gamma\;(IFN-\gamma)$ and lipopolysaccharide (LPS) resulted in increased production of NO in the medium. However, the butanol (BuOH) fraction of the MeOH extract of EA barks showed marked inhibition of NO synthesis in a dose-dependent manner. The inhibition of NO synthesis was reflected in the decreased amount of iNOS protein, as determined by Western blotting. The BuOH fraction did not affect the viability of RA W264.7 cells, as assessed by methylthiazol-2-yl-2, 5-diphenyl tetrazolium bromide (MTT) assay; rather, it reduced endogenous NO-induced apoptotic cell death via inhibition of NO synthesis in RAW264.7 cells. On the other hand, the MeOH and BuOH fraction showed no inhibitory effect on the synthesis of NO by RAW264.7 cells, when iNOS was already expressed by the stimulation with $IFN-\gamma$ and LPS. Conclusion: Collectively, these results demonstrate that the MeOH and BuOH fraction inhibits NO synthesis by inhibition of the induction of iNOS in murine macrophages.

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An analysis of the Gongjindan's ingredients and its efficacy on anti-oxidation (공진단(拱辰丹)의 성분 분석 및 갱산화(坑酸化) 작용에 미치는 영향)

  • Choi, Kum-Hee;Park, Chi-Sang
    • The Korea Journal of Herbology
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    • v.22 no.2
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    • pp.51-63
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    • 2007
  • Objectives : For the congenital feeble, the Gongjindan is useful medicine. The experiment is to estimate the value of the Gongjindan as therapeutic agent preventing against aging with an analysis of the ingredients and the bio-activating effects by enzymologic methods. Methods : General ingredients' of the Gongjindan's extract were analyzed first and the quantitative analysis of a reducing sugar, a soluble protein, an amino acid and minerals was made. The Gongjindan, which is extracted, concentrated, and freeze drying with water, ethanol and chloroform, have got applied for the experiment. The effects on electronic donating ability, SOD-like activity, nitric oxide inhibition, xanthine oxidase inhibition, whitening effect have been investigated in the physiological activity measurement of function experiment. Results : The contained amino acid, in order of high amount, is Arginine, Alanine, Glutamic acid, Proline and the contained free amino acid is Glutamic acid, Leucine, Lysine, Phenylalanine. The derivative of free amino acid is ${\gamma}-Aminoisobutyric$ acid, Phosphoserine, Taurine. And the Gongjindan is containing 13 species of minerals in order of Ca>K>Na>Mg>Fe>AI>Mn. Then, to assure of the Gongjindan's capability of anti-oxidation, these following subjects-polyphenol, electronic donating ability, SOD-like activity, nitric oxide inhibition, xanthine oxidase inhibition, tyrosinase inhibition- are analyzed and show high activity especially the most in ethanol extracts. Conclusion : With this analysis of ingredients, the Gongjindan is containing many materials functioning as anti-oxidation, anti-aging, anti-fatigue, neurotransmitter and immune agent. Moreover, In every water, ethanol, chloroform extracts, the Gongjindan's capability of anti-oxidation is confirmed so that we can apply to patients' treatment clinically.

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Inhibition of Nitric Oxide Production and Hyaluronidase Activities from the Combined Extracts of Platycodon grandiflorum, Astragalus membranaceus, and Schisandra chinensis (길경, 황기와 오미자 혼합추출물의 NO 억제활성과 Hyaluronidase 억제활성 효과)

  • Kang, Chang-Ho;Kwak, Dae Young;So, Jae-Seong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.6
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    • pp.844-850
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    • 2013
  • In this study, the optimal extraction conditions for three medicinal herbs as functional sources against inflammatory and arthritic diseases were developed. Traditional medicinal herbs were screened for their inhibition of hyaluronidase (HAse) activity and nitric oxide (NO) synthesis. For the screening of anti-inflammatory properties, ethanolic extracts of 53 species of traditional medicinal herb were examined. We confirmed that Astragalus membranaceus (A.R.), Schisandra chinensis (S.F.), and Platycodon grandiflorum (P.G.) inhibit NO production. For extraction from all three herbs simultaneously, an ethanol concentration of 95%, a 1:2:1 mixture ratio, and at 50 rpm mixing speed, for over 12 h and at $30^{\circ}C$ was the best condition for optimal extract yield and NO inhibition effects. HAse inhibition from the three herb extraction was three fold higher than single samples. The ethanol extracts were fractionated with various solvents (n-hexane, chloroform, ethyl acetate, n-butanol, and water). The ethyl acetate-soluble fraction of the herb mixture showed the highest extract yield (13%) and NO inhibition effects (73%). In conclusion, this study provides experimental evidence that a mixture of P.G., A.R., and S.F. could be used as a source of antioxidant ingredients in the food industry.

Anti-inflammatory Activity of Dichloromethane Extract of Auricularia auricula-judae in RAW264.7 Cells

  • Damte, Dereje;Reza, Md. Ahsanur;Lee, Seung-Jin;Jo, Woo-Sik;Park, Seung-Chun
    • Toxicological Research
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    • v.27 no.1
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    • pp.11-14
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    • 2011
  • The present study investigated the anti-inflammatory effects of dichloromethane extract of Auricularia auricula-judae. Dichloromethane extract of Auricularia auricula-judae inhibited Lipopolysaccharide (LPS)-induced nitric oxide (NO) production significantly in a dose-dependent manner in the concentration ${\geq}\;10\;{\mu}g/ml$ (p < 0.05). Furthermore, RT-PCR results of this study indicated that the extract markedly reduced the expressions of inflammatory cytokines (IL-6, TNF-$\alpha$ and IL-$1{\beta}$) mRNA in LPS-treated murine RAW 264.7 macrophages, which could possibly ameliorate the inflammation. Nevertheless, dichloromethane extract of Auricularia auricula-judae did not show complete inhibition of IL-6 mRNA expression. The inhibition of IL-$1{\beta}$ cytokine at protein level was also observed in a dose dependent manner. In conclusion, the current study revealed the previously unknown effect of dichloromethane ethyl extract of Auricularia auricula-judae inhibitions of the production of NO, IL-6, TNF-$\alpha$ and IL-$1{\beta}$ in LPS-stimulated macrophages.

Anti-Helicobacter and Anti-inflammatory Effects of Sohamhyungtang in Helicobacter pylori-Infected Human Gastric Epithelial AGS cells

  • Won, SangBum;Yim, Dongsool;Choi, SungSook
    • Natural Product Sciences
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    • v.23 no.3
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    • pp.175-182
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    • 2017
  • This study evaluated the anti-Helicobacter and anti-inflammatory effects of Sohamhyungtang (SHHT). The minimum inhibitory concentration (MIC) of SHHT against Helicobacter pylori (H. pylori) was determined by the agar dilution method. Expression of the H. pylori cagA gene in the presence of SHHT was determined by quantitative real-time polymerase chain reaction (qRT-PCR). Inhibition of H. pylori urease by SHHT was determined by the phenol-hypochlorite assay. Antiadhesion activity of SHHT was measured by urea-phenol red reagent. Inhibition of nitric oxide (NO) production in AGS cells was measured with Griess reagent. Inducible nitric oxide synthase (iNOS) and IL-8 mRNA expression in AGS cells which were infected with H. pylori was determined by qRT-PCR. IL-8 level was measured by enzyme-linked immunosorbent assay (ELISA). The MIC of SHHT was $100{\mu}g/mL$ and the expression of cagA gene was decreased about 25 folds in the presence of SHHT. H. pylori urease was inhibited 90% by SHHT. SHHT inhibited H. pylori adhesion on AGS cell in a concentration dependent manner. mRNA expression of iNOS and IL-8 and the production of NO and IL-8 were significantly decreased in the presence of SHHT. In conclusion, SHHT showed anti-Helicobacter activity and has potent anti-inflammatory effect on H. pylori-induced inflammation in human gastric epithelial AGS cells.

Neuroprotective effects of paeoniflorin against neuronal oxidative stress and neuroinflammation induced by lipopolysaccharide in mice

  • Meng, Hwi Wen;Lee, Ah Young;Kim, Hyun Young;Cho, Eun Ju;Kim, Ji Hyun
    • Journal of Applied Biological Chemistry
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    • v.65 no.1
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    • pp.23-31
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    • 2022
  • Oxidative stress and neuroinflammation play important roles in the pathogenesis of Alzheimer's disease (AD). This study investigated the protective effects of paeoniflorin (PF) against neuronal oxidative stress and neuroinflammation in lipopolysaccharide (LPS)-induced mice. The brains of LPS-injected control group showed significantly increased neuroinflammation by activating the nuclear factor kappa B (NF-κB) pathway and increasing inflammatory mediators. However, administration of PF significantly attenuated oxidative stress by inhibiting lipid peroxidation, nitric oxide levels, and reactive oxygen species production in the brain; PF at doses of 5 and 10 mg/kg/day downregulated the expression of NF-κB pathway-related proteins and significantly decreased inflammatory mediators including inducible nitric oxide synthase and cyclooxygenase-2. Moreover, the levels of brain-derived neurotrophic factor and its receptor, tropomycin receptor kinase B, were significantly increased in PF-treated mice. Furthermore, acetylcholinesterase activity and the ration of B-cell lymphoma 2 (Bcl-2)/Bcl-2 associated X were significantly reduced by PF in the brains of LPS-induced mice, resulting in the inhibition of cholinergic dysfunction and neuronal apoptosis. Thus, we can conclude that administration of PF to mice prevents the development of LPS-induced AD pathology through the inhibition of neuronal oxidative stress and neuroinflammation, suggesting that PF has a therapeutic potential for AD.

Pharmacological potential of Cordyceps militaris with enhanced Cordycepin production for anti-inflammatory and tumor cell anti-proliferative applications

  • Ha, Si-Young;Jung, Ji-Young;Yang, Jae-Kyung
    • Journal of Mushroom
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    • v.20 no.3
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    • pp.93-101
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    • 2022
  • Cordyceps militaris mycelium extracts containing high amounts of cordycepin were evaluated in vitro for their anti-inflammatory and tumor cell growth-inhibitory activities. All extracts dose dependently inhibited the increased production of inflammatory mediators including reactive oxygen species (ROS), nitric oxide (NO), and 𝛽-hexosaminidase in lipopolysaccharide (LPS)-stimulated inflammatory cells. All extracts were evaluated for anti-proliferative activity against normal RBL-2H3 cells and diverse types of cancer cell lines, including HCT, MC5-7, U-87MG, AGS, and A549 cells. The extract showed the strongest growth inhibition (IC50 = 28.13 ㎍/mL) relative to vehicle-treated control cells against fibrosarcoma (MC5-7). We have demonstrated anti-inflammatory activity of C. militaris via inhibition of NO, ROS production, and 𝛽-hexosaminidase release in activated cells. C. militaris mycelium extract was also evaluated mechanistically and found to exert six types of anti-cancer activity, confirming its pharmacological potential. Our study suggests C. militaris use as a potential source of anti-inflammatory and anti-cancer agents. C. militaris may also be considered a functional food.

Peroxynitrite Scavenging Activity and its Mechanism of Cheonga-hwan (청아환의 Peroxynitrite 제거 활성 및 기전)

  • 김성호;정지천
    • The Journal of Korean Medicine
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    • v.23 no.4
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    • pp.55-63
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    • 2002
  • Objectives: Peroxynitrite ($ONOO^{-}$), formed from the reaction of superoxide <${\cdot}O_2^{-}$) and nitric oxide (NO), is a cytotoxic species that can oxidize several cellular components such as proteins, lipids and DNA. It has been implicated in diseases such as aging process, Alzheimer's disease, rheumatoid arthritis, cancer and arteriosclerosis. Due to the lack of endogenous enzymes responsible for $ONOO^{-}$ inactivation, developing a specific $ONOO^{-}$ scavenger is of considerable importance. The aim of this study was to evaluate $ONOO^{-}$ scavenging activity and its mechanism in Cheonga-hwan (CAH). Methods: The $ONOO^{-}$ scavenging activity in CAH was assayed by measuring oxidized dihydrorhodamine 123 (DHR 123) by fluorescence. The scavenging efficacy was expressed as $IC_{50}$, showing the concentration of each sample required to cause 50% inhibition of DHR 123 oxidation. In a separate study, the protective effect of CAR on $ONOO^{-}$-induced nitration of bovine serum albumin (BSA) was investigated using immunoassay with a monoclonal anti-nitrotyrosine antibody, and a horseradish peroxidase-conjugated anti-mouse secondary antibody from sheep. Results: CAH showed potent scavenging activities of $ONOO^{-}$, NO and ${\cdot}O_2^{-}$. The data demonstrated that CAH led to decreased $ONOO^{-}$-mediated nitration of tyrosine through electron donation. CAH showed significant inhibition on nitration of bovine serum albumin by $ONOO^{-}$ in a dose-dependent manner. Conclusions: CAH can be developed as an effective peroxynitrite scavenger for the prevention of the $ONOO^{-}$ involved diseases.

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Protocatechuic Acid Content and Physiological Activities of Chaenomeles sinensis Extracts Prepared with Different Methods (모과의 추출조건에 따른 Protocatechuic Acid 함량 변화 및 활성 평가)

  • Lee, Jeong Ho;Im, So Yeon;Lee, Wang Ro
    • Korean Journal of Pharmacognosy
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    • v.51 no.3
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    • pp.199-206
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    • 2020
  • The aim of this study was to investigate the potential of Chaenomeles sinensis as a content of protocatechuic acid and physiological activities. The analyzed the content of protocatechuic acid of Chaenomeles sinensis water extract (CSW) according to different extract temperature and time for extraction. It was analyzed as 0.14±0.00 mg/g in the extract at 100℃ for 3 h. Protocatechuic acid content increased as extraction time and temperature increased. The activity of the CSW at 100℃ for 3 h was measured. As a result of measuring DPPH, ABTS radical scavenging activity and SOD activity, antioxidant activity was found and α-glucosidase inhibition activities. Cell viability was shown for RAW 264.7 cells, and NO (nitric oxide) production was also inhibited. Finally, content of protocatechuic acid increased with increasing extraction time and extraction temperature, and exhibited antioxidant activity, inhibition of NO production, and anti-diabetic activity. It is believed that the Chaenomeles sinensis is likely to be developed as a functional material.