• Title/Summary/Keyword: antioxidative assay

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Effect of Interaction between Protocatechualdehyde Produced from Streptomyces lincolnensis M-20 and Copper Ions on Antioxidant and Pro-oxidant Activities (Streptomyces lincolnensis M-20 균주에서 생산된 Protocatechualdehyde와 구리 이온의 상호 작용이 항 산화 및 산화 촉진 활성에 미치는 영향)

  • Kim, Kyoung-Ja;Lee, Jae-Hun;Yang, Yong-Joon
    • Korean Journal of Microbiology
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    • v.50 no.1
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    • pp.22-26
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    • 2014
  • Protocatechualdehyde (PA) is phenolic compound having antioxidative and antitumor activities. PA was purified from supernatant of Streptomyces lincolnensis M-20. In the presence of copper ion, PA acted as pro-oxidant. The antioxidant activity was assessed with the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, and the pro-oxidant effect of PA on DNA damage as pBR322 plasmid DNA-cleaving agents in the presence of Cu(II) ions was investigated. The involvement of reactive oxygen species (ROS) in the DNA damage was confirmed by the inhibition of the DNA breakage by using glutathione (GSH), specific scavenger of ROS. When the increase in ROS reaches a certain level (the toxic threshold), it may trigger cell death. The formation of the PA/Cu(II) chelate complex was confirmed by reaction with ethylenediamine-tetraacetic acid (EDTA), a well-known chelating agent for metal ions, by using UV/Vis spectroscopic analysis.

Characterization of polysaccharide A-1 from Opuntia ficus-indica and it's protection effect on alcoholic induced hepatic oxidative stress (Opuntia ficus-indica 다당 A-1의 특성 및 알코올유도 간 산화스트레스의 보호 효과)

  • Ryu, Il-Hwan;Kwon, Ji-Wung;Lee, Eoh-Jin;Yun, Young-Gab;Kwon, Tae-Oh
    • Herbal Formula Science
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    • v.17 no.2
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    • pp.163-174
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    • 2009
  • Reactive oxygen species(ROS) can induce hepatotoxicity and trigger apoptosis in the liver. In this study, we investigated the sulfated polysaccharide A-1 from Opuntia ficus-indica against alcoholic oxidative stress in human liver Hep G2 cell. An antioxidant substance A-1 obtained from the enzymatic extract of Opuntia ficus-indica fruit was purified by DEAE-cellulose ion exchange and sephadex G-100 gel permeation chromatography. The purification yield and molecular weight were 14.3% and 1.8 KDa, respectively. The A-1 predominately contained arabinose, galactose, rhamnose and also sulfate group. The structure of A-1 was investigated by periodate oxidation, FT-IR spectroscopy, $^1H$-NMR spectroscopy. The A-1 mainly composed of alternating unit of ${\rightarrow}4$)-$\alpha$-L- Rapp-2-$SO_3^-$-$\alpha$-L-Galp-($1{\rightarrow}$ and branched linkage of $\beta$-D-Arbp- ($5{\rightarrow}$. The antioxidative activity was measured using the SOD, CAT activity and GSH assay, respectively. The expression of Nrf2 protein was analyzed by western blotting. The viable cell count analyzed by autofluorescence. Oxidative stress induced by ethanol(1 M) were dramatically reduced by A-1 treatment. A-1 also prevented cell death induced by oxidative stress. It also increased expression Nrf2 protein level. We concluded that sulfated polysaccharide A-1 from Opuntia ficus-indica effectively protect Hep G2 liver cell from alcoholic oxidative stress.

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Antioxidant and Genotoxic Inhibition Activity of Ethanol Extract from the Inonotus obliquus (차가버섯 분획물의 항산화활성 및 유전독성 억제효과)

  • 함승시;오상화;김영균;신광순;장현유;정국훈
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.7
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    • pp.1071-1075
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    • 2003
  • This study was performed to observe the antioxidative and genotoxic effect of the fractions from Inonotus obliquus using DPPH test and micronucleus assay. Stepwise fractionation of the ethanol extract from Inonotus obliquus was done by using hexane, chloroform, ethyl acetate, butanol and water to obtain effective fraction. Each fraction was tested in 1.5 ${\times}$ 10$^{-4}$ DPPH. Among six fractions, the ethyl acetate fraction showed the highest electron donating activities (46.5 $\mu\textrm{g}$/mL). The results on genotoxic effects on insoluble fractions and most of fractions showed cytotoxic effects more than 90% activity. These results suggest that some components contained in the Inonotus obliquus showed such activities and much more studies have to perform.

HO-1 Induced by Cilostazol Protects Against TNF-${\alpha}$-associated Cytotoxicity via a PPAR-${\gamma}$-dependent Pathway in Human Endothelial Cells

  • Park, So-Youn;Bae, Jin-Ung;Hong, Ki-Whan;Kim, Chi-Dae
    • The Korean Journal of Physiology and Pharmacology
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    • v.15 no.2
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    • pp.83-88
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    • 2011
  • A large body of evidence has indicated that induction of endogenous antioxidative proteins seems to be a reasonable strategy for delaying the progression of cell injury. In our previous study, cilostazol was found to increase the expression of the antioxidant enzyme heme oxygenase-1 (HO-1) in synovial cells. Thus, the present study was undertaken to examine whether cilostazol is able to counteract tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-induced cell death in endothelial cells via the induction of HO-1 expression. We exposed human umbilical vein endothelial cells (HUVECs) to TNF-${\alpha}$ (50 ng/ml), with or without cilostazol ($10{\mu}M$). Pretreatment with cilostazol markedly reduced TNF-${\alpha}$-induced viability loss in the HUVECs, which was reversed by zinc protoporphyrine IX (ZnPP), an inhibitor of HO-1. Moreover, cilostazol increased HO-1 protein and mRNA expression. Cilostazol-induced HO-1 induction was markedly attenuated not only by ZnPP but also by copper-protoporphyrin IX (CuPP). In an assay measuring peroxisome proliferator-activated receptor-${\gamma}$ (PPAR-${\gamma}$) transcription activity, cilostazol directly increased PPAR-${\gamma}$ transcriptional activity which was completely abolished by HO-1 inhibitor. Furthermore, increased PPAR-${\gamma}$ activity by cilostazol and rosiglitazone was completely abolished in cells transfected with HO-1 siRNA. Taken together, these results indicate that cilostazol up-regulates HO-1 and protects cells against TNF-${\alpha}$-induced endothelial cytotoxicity via a PPAR-${\gamma}$-dependent pathway.

Isolation and Structure Determination of Antioxidants from the Root of Paeonia lactiflora (작약(芍藥)(Paeonia lactiflora) 뿌리로부터 항산화활성 물질의 분리)

  • Bang, Myun-Ho;Song, Jung-Choon;Lee, Sang-Yang;Park, Nam-Kyu;Baek, Nam-In
    • Applied Biological Chemistry
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    • v.42 no.2
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    • pp.170-175
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    • 1999
  • On the purpose of development of novel antioxidative compounds from natural sources, 38 plants expected to show antioxidant activity have been examined concerning DPPH radical scavenging activity. Among them, thirteen plants, including Paeoniae radix, the root of Paeonia lactiflora, exhibited the activity. In order to isolate active component, the root was extracted in 80% aqueous MeOH and solvent fractionated with EtOAc, n-BuOH and water, successively. Silica gel column chromatographies of the EtOAc and n-BuOH fraction exhibiting antioxidant activity. were repeatedly carried out with monitoring by DPPH assay to afford three active compounds. On the basis of spectral data and the chemical characteristics, the structures of the compounds were determined as (+)-catechin, $1,2,3,4-tetragalloyl-6-digalloyl-{\beta}-D-glucose$ and $1,2,3,4,6-penta-galloyl-{\beta}-D-glucose$.

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Bioassay-Guided Isolation and Identification of Compounds from Arecae Pericarpium with Anti-inflammatory, Anti-oxidative, and Melanogenesis Inhibition Activities

  • Indriana, Amelia;Lee, Kyoung Jin;Kim, Yeong Shik
    • Natural Product Sciences
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    • v.22 no.3
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    • pp.193-200
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    • 2016
  • This study describes the anti-inflammatory, anti-oxidant, and melanogenesis inhibition activities of methanol extract and various organic solvent fractions of Arecae Pericarpium. We examined the inhibition of lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 cells, 1,1-diphenyl-2-picrylhydrazine (DPPH) scavenging activity, mushroom tyrosinase inhibition activity and melanin contents. The study showed that, among all tested fractions, methylene chloride fraction showed the strongest inhibition of LPS-induced NO production in RAW 264.7 cells ($IC_{50}$ value $8.89{\mu}g/mL$) and DPPH radical scavenging activity ($EC_{50}$ value $21.39{\mu}g/mL$). Methylene chloride and ethyl acetate fractions similarly inhibited mushroom tyrosinase activity. Methanol extract exhibited strongest reduction of melanin content in B16F10 melanoma cells. Based on the bioactivity assay results, methylene chloride and ethyl acetate fractions were further separated. Eight phenolic compounds were isolated, which are dimeric syringol (1), catechol (2), 4-hydroxybenzaldehyde (3), vanillin (4), 4-hydroxyacetophenone (5), apocynin (6), protocatechuic acid (7) and 4-hydroxybenzoic acid (8). Among the isolated compounds tested, catechol showed the strongest inhibition of LPS-induced NO production in RAW 264.7 cells. Catechol also showed the concentration-dependent NF-${\kappa}B$ inhibition activity. Arecae Pericarpium might have potentials to be developed as anti-inflammatory agent or dermatological product for skin-whitening agent.

Anti-inflammatory effect of lycopene in SW480 human colorectal cancer cells

  • Cha, Jae Hoon;Kim, Woo Kyoung;Ha, Ae Wha;Kim, Myung Hwan;Chang, Moon Jeong
    • Nutrition Research and Practice
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    • v.11 no.2
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    • pp.90-96
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    • 2017
  • BACKGROUND/OBJECTIVES: Although the antioxidative effects of lycopene are generally known, the molecular mechanisms underlying the anti-inflammatory properties of lycopene are not fully elucidated. This study aimed to examine the role and mechanism of lycopene as an inhibitor of inflammation. METHODS/MATERIALS: Lipopolysaccharide (LPS)-stimulated SW 480 human colorectal cancer cells were treated with 0, 10, 20, and $30{\mu}M$ lycopene. The MTT assay was performed to determine the effects of lycopene on cell proliferation. Western blotting was performed to observe the expression of inflammation-related proteins, including nuclear factor-kappa B ($NF-{\kappa}B$), inhibitor kappa B ($I{\kappa}B$), mitogen-activated protein kinase (MAPK), extracellular signal-related kinase (ERK), c-jun NH2-terminal kinase (JNK), and p38 (p38 MAP kinase). Real-time polymerase chain reaction was performed to investigate the mRNA expression of tumor necrosis factor ${\alpha}$ ($TNF-{\alpha}$), interleukin-1 beta ($IL-1{\beta}$), interleukin-6 (IL-6), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). Concentrations of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) were determined via enzyme-linked immunosorbent assays. RESULTS: In cells treated with lycopene and LPS, the mRNA expression of $TNF-{\alpha}$, $IL-1{\beta}$, IL-6, iNOS, and COX-2 were decreased significantly in a dose-dependent manner (P < 0.05). The concentrations of $PGE_2$ and NO decreased according to the lycopene concentration (P < 0.05). The protein expressions of $NF-{\kappa}B$ and JNK were decreased significantly according to lycopene concertation (P < 0.05). CONCLUSIONS: Lycopene restrains $NF-{\kappa}B$ and JNK activation, which causes inflammation, and suppresses the expression of $TNF-{\alpha}$, $IL-1{\beta}$, IL-6, COX-2, and iNOS in SW480 human colorectal cancer cells.

Antioxidative Activities of Methanol Extracts from Different Parts of Chrysanthemum zawadskii (구절초의 부위별 메탄올 추출물의 항산화활성)

  • Chung, Hai-Jung;Jeon, In-Sook
    • Food Science and Preservation
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    • v.18 no.5
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    • pp.739-745
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    • 2011
  • The major objective of this study was to investigate the antioxidant activities of methanolic extracts from different parts (flower, leaf stem, and root) of Chrysanthemum zawadskii by employing various in-vitro assay systems. The extraction yields from the flower, leaf stem, and root were 18.347, 12.93, and 11.33-----, respectively. The total polyphenol content was highest in the flower (17.16 mg/100 g) and lowest in the root (11.33 mg/100 g). The antioxidant activities were raised within creasing amounts of extracts, and the extracts from the flower showed the highest effect on the superoxide anion radical scavenging, metal chelating on ferrous ions and reducing power. In addition, the leaf stem also showed good antioxidant activity in various systems. These results suggest that the methanolic extracts from the flower and leaf stem possess excellent antioxidant activities and may thus serve as potential sources of natural antioxidants.

Screening of Traditional Medicines for Antioxidative and Anti-proliferative Effects on Rat Mesangial Cells (한약재 추출물의 항산화 및 사구체혈관간세포 증식 억제활성 탐색)

  • Sohn, Eun-Hwa;Jang, Seon-A;Woo, Han Goo;Koo, Hyun Jung;Han, Hyo-Sang;Kang, Se Chan
    • Korean Journal of Plant Resources
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    • v.26 no.5
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    • pp.652-657
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    • 2013
  • In the present study, anti-oxidative and the RMC proliferation inhibitory propeties of 80% ethanol extracts from 63 kinds of traditional medicines were investigated. Inhibitory effects of RMC proliferation were showed that Dalbergia odorifera T. Chen., Melia azedarach Linn$\acute{e}$ and Hydnocarpus anthelmintica Pierre. Among them Hydnocarpus anthelmintica Pierre had the highest anti-oxidative activity ($ORAC_{PE\;value}=1.6$, DPPH = 81.1), but Dalbergia odorifera T. Chen. and Melia azedarach Linn$\acute{e}$ had no effects. These results suggest that the Hydnocarpus anthelmintica Pierre could prevent or protect from kidney disease as antioxidant and anti-proliferative agent for RMC.

Isolation of a Natural Antioxidant, Dehydrozingerone from Zingiber officinale and Synthesis of lts Analogues for Recognition of Effective Antioxidant and Antityrosinase Agents

  • Kuo, Ping-Chung;Damu, Amooru G.;Cherng, Ching-Yuh;Jeng, Jye-Fu;Teng, Che-Ming;Lee, E-Jian;Wu, Tian-Shung
    • Archives of Pharmacal Research
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    • v.28 no.5
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    • pp.518-528
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    • 2005
  • In the present study, the antioxidative and inhibitory activity of Zingiber officinale Rosc. Rhizomes-derived materials (on mushroom tyrosinase) were evaluated. The bioactive co mponents of Z. officinale rhizomes were characterized by spectroscopic analysis as zingerone and dehydrozingerone, which exhibited potent antioxidant and tyrosinase inhibition activities. A series of substituted dehydrozingerones [(E)-4-phenyl-3-buten-2-ones] were prepared in admirable yields by the reaction of appropriate benzaldehydes with acetone and the products were evaluated in terms of variation in the dehydrozingerone structure. The synthetic analogues were examined for their antioxidant and antityrosinase activities to probe the most potent analogue. Compound 26 inhibited Fe$^{2+}$-induced lipid peroxidation in rat brain homogenate with an IC$_{50}$ = 6.3${\pm}$0.4 ${\mu}$M. In the 1,1-diphenyl- 2-picrylhydrazyl (DPPH) radical quencher assay, compounds 2, 7, 17, 26, 28, and 29 showed radical scavenging activity equal to or higher than those of the standard antioxidants, like ${\alpha}$-tocopherol and ascorbic acid. Compound 27 displayed superior inhibition of tyrosinase activity relative to other examined analogues. Compounds 2, 17, and 26 exhibited non-competitive inhibition against oxidation of 3,4- dihydroxyphenylalanine (L-DOPA). From the present study, it was observed that both number and position of hydroxyl groups on aromatic ring and a double bond between C-3 and C-4 played a critical role in exerting the antioxidant and antityrosinase activity.