• Title/Summary/Keyword: NO의생성

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Comparative Study of the Biological Activity Effects of Manjakani (Quercus infectoria Olivier) Extract using Water and 80% Ethanol (열수 및 80% 에탄올로 추출한 만자카니(Quercus infectoria Olivier)의 생리활성 비교연구)

  • Lee, Hea-Jin;Kim, Dong-Han;Lee, Eun-Jin;Lim, Mi-Hye
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.1
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    • pp.124-132
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    • 2020
  • The purpose of this study was to investigate the biological activities such as cytotoxicity and anti-inflammation using Manjakani (Quercus infectoria Olivier) extract. Manjakani was extracted from hot DW and 80% ethanol. Cell viability was assessed using MTT assay on RAW 264.7 cells. Also, anti-inflammatory activities were measured through changes in the levels of nitric oxide (NO), prostaglandin E2 (PGE2), leukotrien B4 (LTB4), pro-inflammatory cytokines (IL-1β, IL-6 and tumor necrosis factor (TNF)-α) and transcription factor on LPS-induced RAW264.7 cells. The results confirmed that significant cytotoxicity does not appear in the concentration range of 1, 5, and 10 ㎍/㎖ of both extracts of this study. The production of NO was slowed by approximately MDE 37.2% and MEE 43.7% at 10 ㎍/㎖ concentration. Also, level of PGE2 and LTB4 was decreased MDE 30.9%/MEE 43.7% and MDE 37.1%/MEE 43.7%. In the case of inflammatory cytokine was reduced to MDE 38.8%/MEE 50.8% for IL-1β, MDE 35.0%/MEE 44.2% for IL-6 and MDE 31.9%/MEE 36.6% for TNF-α at 10 ㎍/㎖ concentration. The mRNA expression of NF-κB, iNOS and COX-2 significantly decreased by MDE 44.0%/MEE 16.0%, MDE 44.0%/MEE 55.0% and MDE 45.0%/MEE 40.0%, respectively, following the 10 ㎍/mL sample treatment when compared to the control. Both extracts were effective in anti-inflammatory activity. In addition, both extracts showed efficient changes of production of NO, PGE2, LTB4, pro-inflammatory cytokines and transcription factor. But MEE was found to have a higher inhibitory effect than MDE. In other words, Manjakani was showed significant biological activities showing anti-inflammation without cytotoxicity. These results will be provided as fundamental data for further development of the new health food and therapeutics related to the results above.

Antioxidant and anti-inflammatory activities of fruiting bodies of Dyctiophora indusiata (흰망태버섯 자실체의 메탄올 및 열수추출물의 항산화 및 항염증 활성)

  • Nguyen, Trung Kien;Shin, Do Bin;Lee, Kyung Rim;Shin, Pyung Gyun;Cheong, Jong Chun;Yoo, Young Bok;Lee, Min Woong;Jin, Ga-Heon;Kim, Hye Young;Im, Kyung Hoan;Lee, Tae Soo
    • Journal of Mushroom
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    • v.11 no.4
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    • pp.269-277
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    • 2013
  • Dictyophora indusiata is an edible mushroom belongs to Family Phallaceae of Phallales, Basidiomycota. The purpose of this study was to investigate the antioxidant and anti-inflammatory activities of methanol and hot water extracts prepared from fruiting bodies of Dictyophora indusiata. Besides measuring of 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, a reducing power and a chelating activity on ferrous ions were also measured to evaluate the antioxidant activity for those extracts. To measure the anti-inflammatory activities for the extracts, nitric oxide(NO) production from lipopolysaccharide(LPS) treated RAW 264.7 macrophage cells and carrageenan-induced acute hind paw edema of rats were investigated. The results showed that the extracts have excellent DPPH scavenging and chelating activity on the ferrous ions compared with positive control. The nitric oxide(NO) production in LPS-stimulated RAW 264.7 macrophage cells were decreased as we increased the concentration of the mushroom extracts. Significant reduction of paw edema of rats were observed at 2~6 h after treatment of methanol and hot-water extracts with 50 mg/kg concentration to the rats which are induced acute hind paw edema by carrageenan administration. Therefore, the experimental results suggested that methanol and hot-water extracts of Dictyophora indusiata fruiting bodies might be used for natural sources of antioxidant and anti-inflammatory agents.

Antioxidant, anti-inflammatory and anti-acetylcholinesterase activities of fruiting bodies of Phellinus xeranticus (기와층버섯 자실체의 메탄올 및 열수추출물의 항산화, 항염증 및 아세틸콜린에스테라제 저해 활성)

  • Nguyen, Trung Kien;Shin, Do Bin;Lee, Kyung Rim;Shin, Pyung Gyun;Cheong, Jong Chun;Yoo, Young Bok;Lee, Min Woong;Jin, Ga-Heon;Kim, Hye Young;Im, Kyung Hoan;Lee, Tae Soo
    • Journal of Mushroom
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    • v.11 no.4
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    • pp.278-286
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    • 2013
  • Phellinus xeranticus is an medicinal mushroom belongs to Family Hymenochaetaceae of Polyporales, Basidiomycota. The purpose of this study was to investigate the antioxidant, anti-inflammatory and anti-acetylcholinesterase activities of methanol and hot water extracts prepared from fruiting bodies of Phellinus xeranticus. Besides measuring of 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, a reducing power and a chelating activity on ferrous ions were also measured to evaluate the antioxidant activity of the extracts. To measure the anti-inflammatory activities of the extracts, nitric oxide(NO) production from lipopolysaccharide (LPS) stimulated RAW 264.7 macrophage cells and carrageenan-induced acute hind paw edema of rats were investigated. The results showed that the extracts have excellent DPPH scavenging and chelating activity on the ferrous ions compared with positive controls. The nitric oxide (NO) production in LPS-induced RAW 264.7 macrophage cells were decreased as the concentration of the mushroom extracts increased. Significant reduction of paw edema of rats were observed at 2~6 h after treatment of methanol and hot-water extracts with 50 mg/kg concentration to the rats which are induced acute hind paw edema by carrageenan administration. The anti-acetylcholinesterase activity of the methanol extract of the mushroom showed 83.34% inhibition on AcHE which is lower than that of positive control galanthamine. The experimental results suggested that methanol and hot-water extracts of Phellinus xeranticus fruiting bodies might be used for good sources of antioxidant, anti-inflammatory and anti-acetylcholinesterase agents.

Anti-inflammatory Effects of Rebaudioside A in LPS-stimulated RAW264.7 Macrophage Cells (LPS에 의해 자극된 RAW264.7 대식세포에서 Rebaudioside A의 항염 효과)

  • Cho, Uk Min;Hwang, Hyung Seo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.43 no.2
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    • pp.157-164
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    • 2017
  • Stevia (Stevia rebaudiana) is a perennial plant of the genus Stevia, originated in South America. It stores many forms of glycosides, mainly stevioside and rebaudioside A, in which steviol is the basic structure. Steviol glycosides, widely used as sweeteners, are superior to sugar in sweetness. Recently, it has been reported that steviol glycosides are involved not only in the skin whitening and anti-inflammatory effect but also in enhancing skin barrier function through tight junction regulation. Thus, we examined anti-inflammatory effect of rebaudioside A and tried to identify its potential for improving atopic dermatitis as cosmetic ingredients. To investigate the anti-inflammatory effect, cell viability and mRNA expression level of inflammation-related cytokines were measured using mouse macrophage RAW264.7 cells. Cell counting kit 8 (CCK-8) assay was carried out to measure cell viability and the maximum concentration without cytotoxicity was set to $250{\mu}M$. A quantitative real-time RT-PCR method was used for the study of the inflammatory suppression of rebaudioside A. Rebaudioside A inhibited expression of inducible nitric oxide synthase (iNOS) up to 47% and COX-2 up to 41% compared to LPS treated condition. NO synthesis was decreased by rebaudioside A. Also, mRNA expression of interleukin (IL)-$1{\alpha}$, IL-$1{\beta}$ and IL-6 in LPS-stimulated RAW264.7 cells was decreased to 40%, 45% and 59%, respectively, as a concentration-dependent manner. In conclusion, rebaudioside A inhibited the inflammatory response by regulation of cytokine gene expression. From these results, we expect that steviol glycoside, such as rebaudioside A, can be used as a material for improving atopic dermatitis in the future.

Anti-inflammatory and Antioxidant Effects of Spiraea prunifolia Sieb. et Zucc. var. simpliciflora Nakai in RAW 264.7 Cells (조팝나무 뿌리 열수 추출물이 RAW264.7 세포에서 미치는 항산화 및 항염증 활성)

  • Sim, Mi-Ok;Lee, Hyun Joo;Jang, Ji Hun;Lee, Hyo Eun;Jung, Ho-Kyung;Kim, Tae-Muk;No, Jong hyun;Jung, Jakyun;Jung, Da Eun;Cho, Hyun-Woo
    • Korean Journal of Plant Resources
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    • v.30 no.4
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    • pp.335-342
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    • 2017
  • Spiraea prunifolia Sieb. et Zucc. var. simpliciflora Nakai (SSN) has been used for the anti-inflammation in traditional folk medicine. To compare water and methanol extracts of SSN, we analyzed major components using LC IT TOF MS. The major components of hot water extract were identified as caffeic acid and p-coumaric acid, but methanol extract was not well established. However, methanol extract was detected with less polarity compounds compared to hot water extract. Next, we investigated the inhibitory effects of SSN water extract on the lipopolysaccharide (LPS)-induced inflammatory response or $H_2O_2-induced$ oxidative stress in Raw 264.7 macrophage cells. SSN strongly suppressed the production of nitric oxide in LPS-induced inflammatory response without cytotoxcity. The SSN possessed free radical scavenging activities such as DPPH ($IC_{50}=320.2{\mu}g/m{\ell}$), ABTS ($IC_{50}=124.0{\mu}g/m{\ell}$), and superoxide anion radical ($IC_{50}=122.6{\mu}g/m{\ell}$). The total phenol and flavonoid content of SSN was 56.7 mg/g, and 15.1 mg/g, respectively. Furthermore, SSN decreased the $H_2O_2-induced$ cytotoxicity by enhancing the cell viability, and SSN significantly reduced the intracellular reactive oxygen species (ROS) level. Therefore, SSN may be recommended as an effective strategy to prevent and/or treat various inflammation and ROS-induced diseases.

Pharmacological Activity of Chaga Mushroom on Extraction Conditions and Immunostimulating Polysaccharide (추출조건에 따른 차가버섯 생리활성 및 면역활성 다당)

  • Baek, Gil-Hun;Jeong, Heon-Sang;Kim, Hoon;Yoon, Taek-Joon;Suh, Hyung-Joo;Yu, Kwang-Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.10
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    • pp.1378-1387
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    • 2012
  • To investigate the pharmacological activity of chaga mushroom (Inonotus obliquus) on extraction conditions, chaga was extracted using water (reflux at $50^{\circ}C$, decoction over $90^{\circ}C$, pressure at $121^{\circ}C$) or ethanol (reflux at 50, 70, or $90^{\circ}C$). When water extract was further fractionated into crude polysaccharide (IO-CP), yields of IO-CP (4.8~16.8%) were higher than those of ethanolic extracts (IO-E, 1.9~2.7%) at increased temperature. For antioxidant activity, crude polysaccharide (IO-CP-121) obtained by pressurized extraction showed the highest polyphenolic and flavonoid contents (35.10 mg TAE/g and 18.48 mg QE/g, respectively) as well as DPPH and ABTS free radical scavenging activities (26.08 and 27.99 mg AEAC/100 mg, respectively). Meanwhile, IO-CP-D (decoction) and IO-CP-50 (reflux) had more potent mitogenic effects (2.10- and 1.95-fold of saline control at 100 ${\mu}g/mL$) as well as intestinal immune system modulating activities (6.30- and 5.74-fold) compared to IO-CP-121, whereas ethanolic extracts showed no activity. Although no IO-CP showed cytotoxicity against RAW 264.7 cells at 0.1 mg/mL, IO-CP-121 significantly inhibited TNF-${\alpha}$ and NO production as pro-inflammatory factors in LPS-stimulated RAW 264.7 cells (29.2 and 63.5%, respectively). Ethanolic extracts also showed no cytotoxicity at 0.1 mg/mL, whereas inhibition of TNF-${\alpha}$ and NO production was significantly low compared to that of IO-CP-121. In addition, active IO-CP-D was further fractionated into an unadsorbed (IO-CP-I) and seven adsorbed fractions (IO-CP-II~VIII) by DEAE-Sepharose CL-6B column chromatography in order to isolate immunostimulating polysaccharide. IO-CP-II showed the most potent mitogenic effect and macrophage stimulating activity (4.51- and 1.64-fold, respectively). IO-CP-II mainly contained neutral sugars (61.86%) in addition to a small amount of uronic acid (2.96%), and component sugar analysis showed that IO-CP-II consisted mainly of Glc, Gal, and Man (molar ratio of 1.00:0.55:0.31). Therefore, extraction conditions affect the physiological activity of chaga, and immunostimulating polysaccharide fractionated from chaga by decoction is composed mainly of neutral sugars.

A Study of the Anti-inflammatory Effect of Protein Derived from Tenebrio molitor Larvae (알칼리 법으로 추출한 갈색거저리 유충 단백질의 항염증 효능)

  • Seo, Minchul;Lee, Hwa Jeong;Lee, Joon Ha;Baek, Minhee;Kim, In-Woo;Kim, Sun Young;Hwang, Jae-Sam;Kim, Mi-Ae
    • Journal of Life Science
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    • v.29 no.8
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    • pp.854-860
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    • 2019
  • This study investigated the optimum pH conditions for efficient extraction of protein from defatted Tenebrio molitor (TM) larvae. We examined the anti-inflammatory effect of protein derived from defatted TM larvae obtained by an alkaline extraction method. Six extraction pH values (7, 8, 9, 10, 11, and 12) and three precipitation pH values (2, 4, and 6) were used. The protein content, browning degree, and recovery yield of the protein obtained under each pH condition were determined. For efficient extraction of protein from defatted TM larvae, a combination of an extraction pH of 9 and precipitation pH of 4 resulted in a 32.4% recovery yield based on the extraction value and degree of browning. To determine whether the protein ameliorated inflammation by inhibition of macrophage activation by lipopolysaccharides (LPS), we measured nitric oxide (NO), cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS) expression in LPS-stimulated raw 264.7 macrophage cells. The protein markedly inhibited the production of NO without cytotoxicity and reduced the expression level of COX-2 and iNOS protein through the regulation of mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B ($NF-{\kappa}B$) signaling. These results suggested that protein derived from TM larvae could have potential applications in anti-inflammatory therapeutic agents and protein supplements.

Changes in physiologically active ingredients and anti-inflammatory properties of underutilized wild vegetables by complex fermentation using beneficial microorganisms (유용미생물에 의한 저이용 산채류의 복합발효 중 생리활성 성분 및 항염증 변화)

  • Sang-Hyeob Sim;Ha KyoungChoi;Da Eun Lee;Soo Chang Na;Dae Il Hwang;Hyo Bin Oh;Yi Teak Lim;Tae-Young Kim;Dae-Woon Kim
    • Food Science and Preservation
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    • v.31 no.2
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    • pp.287-297
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    • 2024
  • It was confirmed that complex fermentation (CF) was more efficient than single-strain fermentations in inducing changes in the contents of phenolic compounds of Maclura tricuspidate and Pyrus Montana Nakai. A mixture of Maclura tricuspidata, Pyrus montana Nakai, Platycodon grandiflorum and Codonopsis lanceolata were fermented in CF using Aspergillus shirousamii (koji), yeast, and lactic acid bacteria (LAB) for 24 days, and the pH, °Brix, total acidity, anti-oxidant activity, polyphenol content, nitric oxide (NO), and Western blotting of inducible nitric oxide synthase (iNOS), cyclo-oxygenase-2 (COX-2), and tumor necrosis factor-𝛼 (TNF-𝛼) of the sample were determined. There was no significant change in pH and total acidity. °Brix significantly decreased from day 6 onwards. HPLC confirmed that the concentrations of chlorogenic acid, 4-hydrobenzoic acid, vanillic acid, and caffeic acid significantly increased from day 18 during the fermentation. Additionally, DPPH, ABTS radical scavenging activity, total phenol, and total flavonoid were confirmed to be increased until 18 days. NO was significantly inhibited from day 6, along with significant inhibition of iNOS, COX-2, and TNF-a. In conclusion, this study confirmed that CF of low-use (or underutilized) wild vegetables enhances phenolic compounds. It effectively suppresses NO, iNOS, COX-2, and TNF-𝛼, markers of inflammation-related pathogenesis. Altogether, our results suggest that CF of the above plants has a potential anti-inflammatory effect.

The Effect of Nitric Oxide Donor or Nitric Oxide Synthase Inhibitor on Oxidant Injury to Cultured Rat Lung Microvascular Endothelial Cells (산화질소 공여물과 산화질소 합성효소 길항제가 백서 폐미세혈관 내피세포 산화제 손상에 미치는 영향)

  • Chang, Joon;Michael, John R.;Kim, Se-Kyu;Kim, Sung-Kyu;Lee, Won-Young;Kang, Kyung-Ho;Yoo, Se-Hwa;Chae, Yang-Seok
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.6
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    • pp.1265-1276
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    • 1998
  • Background : Nitric oxide(NO) is an endogenously produced free radical that plays an important role in regulating vascular tone, inhibition of platelet aggregation and white blood cell adhesion to endothelial cells, and host defense against infection. The highly reactive nature of NO with oxygen radicals suggests that it may either promote or reduce oxidant-induced cell injury in several biological pathways. Oxidant injury and interactions between pulmonary vascular endothelium and leukocytes are important in the pathogenesis of acute lung injury, including acute respiratory distress syndrome(ARDS). In ARDS, therapeutic administration of NO is a clinical condition providing exogenous NO in oxidant-induced endothelial injury. The role of exogenous NO from NO donor or the suppression of endogenous NO production was evaluated in oxidant-induced endothelial injury. Method : The oxidant injury in cultured rat lung microvascular endothelial cells(RLMVC) was induced by hydrogen peroxide generated from glucose oxidase(GO). Cell injury was evaluated by $^{51}$chromium($^{51}Cr$) release technique. NO donor, such as S-nitroso-N-acetylpenicillamine(SNAP) or sodium nitroprusside(SNP), was added to the endothelial cells as a source of exogenous NO. Endogenous production of NO was suppressed with N-monomethyl-L-arginine(L-NMMA) which is an NO synthase inhibitor. L-NMMA was also used in increased endogenous NO production induced by combined stimulation with interferon-$\gamma$(INF-$\gamma$), tumor necrosis factor-$\alpha$(TNF-$\alpha$), and lipopolysaccharide(LPS). NO generation from NO donor or from the endothelial cells was evaluated by measuring nitrite concentration. Result : $^{51}Cr$ release was $8.7{\pm}0.5%$ in GO 5 mU/ml, $14.4{\pm}2.9%$ in GO 10 mU/ml, $32.3{\pm}2.9%$ in GO 15 mU/ml, $55.5{\pm}0.3%$ in GO 20 mU/ml and $67.8{\pm}0.9%$ in GO 30 mU/ml ; it was significantly increased in GO 15 mU/ml or higher concentrations when compared with $9.6{\pm}0.7%$ in control(p < 0.05; n=6). L-NMMA(0.5 mM) did not affect the $^{51}Cr$ release by GO. Nitrite concentration was increased to $3.9{\pm}0.3\;{\mu}M$ in culture media of RLMVC treated with INF-$\gamma$ (500 U/ml), TNF-$\alpha$(150 U/ml) and LPS($1\;{\mu}g/ml$) for 24 hours ; it was significantly suppressed by the addition of L-NMMA. The presence of L-NMMA did not affect $^{51}Cr$ release induced by GO in RLMVC pretreated with INF-$\gamma$, TNF-$\alpha$ and LPS. The increase of $^{51}Cr$ release with GO(20 mU/ml) was prevented completely by adding 100 ${\mu}M$ SNAP. But the add of SNP, potassium ferrocyanate or potassium ferricyanate did not protect the oxidant injury. Nitrite accumulation was $23{\pm}1.0\;{\mu}M$ from 100 ${\mu}M$ SNAP at 4 hours in phenol red free Hanks' balanced salt solution. But nitrite was not detectable from SNP upto 1 mM The presence of SNAP did not affect the time dependent generation of hydrogen peroxide by GO in phenol red free Hanks' balanced salt solution. Conclusion : Hydrogen peroxide generated by GO causes oxidant injury in RLMVC. Exogenous NO from NO donor prevents oxidant injury, and the protective effect may be related to the ability to release NO. These results suggest that the exogenous NO may be protective on oxidant injury to the endothelium.

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Glycoprotein Isolated from Morus indica Linne Has an Antioxidative Activity and Inhibits Signal Factors Induced by Bisphenol A in Raw 264.7 Cells (뽕잎 당단백질의 항산화능과 Raw 264.7 세포에 있어서 bisphenol A에 유도된 신호전달인자의 억제)

  • Shim, Jae-Uoong;Lee, Sei-Jung;Oh, Phil-Sun;Lim, Kye-Taek
    • Korean Journal of Food Science and Technology
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    • v.39 no.2
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    • pp.209-216
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    • 2007
  • The present study investigated anti-oxidative and anti-inflammatory activity of glycoprotein isolated from Morus Indica Linne (MIL glycoprotein). We found that MIL glycoprotein has a molecular weight of 32 kD and consists of carbohydrate (40.03%) and protein (59.97%), and that it has a strong scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical $({\cdot}OH)$, and superoxide anion $(O_2{\cdot}\;^-)$ radicals. In addition, MIL glycoprotein had a stable character and an optimal DPPH radical scavenging activity in the alkaline and neutral pH solution, and up to at 105. However, the results indicated that it has a minimal scavenging activity in the metal ionic solution ($Ca^{2+}$, $Mn^{2+}$, and $Mg^{2+}$) in the presence of EDTA. In addition, we further investigated whether MIL glycoprotein scavenges oxygen radicals and blocks inflammation-related signals in the bisphenol A (BPA)-stimulated Raw 264.7 cells. The results in this study showed that it has a character to scavenge the productions of reactive oxygen species (ROS) and nitric oxide (NO) dose-dependently. Also it blocked the activities of inflammation-related signals such as nuclear factor-kappa B ($NF-{\kappa}B$) and inducible nitric oxide synthase (iNOS). For example, it had an inhibitory effect on the activation of $NF-{\kappa}B$ (p50) and iNOS proteins at 200 ${\mu}g/mL$ MIL glycoprotein. Here, we speculate that MIL glycoprotein is one of natural antioxidants and of modulators of the BPA-induced inflammation.