• Title/Summary/Keyword: free radical concentrations

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Antioxidative Activity and Component Analysis of Prunella vulgaris L. Extract/Fractions (하고초 추출물의 항산화 활성 및 성분 분석)

  • Suh, Ji Young;Seong, Joon Seob;Yun, Mid Eum;Lee, Ye Seul;Ha, Ji Hoon;Park, Dong Soon;Park, Soo Nam
    • Journal of the Korean Applied Science and Technology
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    • v.33 no.4
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    • pp.647-657
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    • 2016
  • In this study, the antioxidative effects and active component analysis of 50% ethanol extract, ethyl acetate fraction and aglycone fraction obtained from Prunella vulgaris L. were investigated. The free radical scavenging activities ($FSC_{50}$) was investigated at 50% ethanol extract ($15.25{\mu}g/mL$), ethyl acetate fraction ($8.68{\mu}g/mL$), and aglycone fraction ($8.25{\mu}g/mL$) respectively. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) in $Fe^{3+}-EDTA/H_2O_2$ system using the luminol-dependent chemiluminescence assay was investigated at 50% ethanol extract ($4.68{\mu}g/mL$), ethyl acetate fraction ($1.00{\mu}g/mL$), and aglycone fraction($1.02{\mu}g/mL$) respectively. In the cellular protective effect against $^1O_2$ induced cellular damage of human erythrocytes, extract/fractions of P. vulgaris L. were increased in a concentration dependent manner($1{\sim}25{\mu}g/mL$). Especially, ${\tau}_{50}$ of aglycone fraction at concentrations of $25{\mu}g/mL$ showed the most protective effects at 337.9 min. It's showed nine times higher (+)-${\alpha}$-tocopherol (${\tau}_{50}=38.7min$) as typical antioxidant in the $^1O_2$-induced photohemolysis of human erythrocytes. TLC and HPLC were used to analyse active components in the ethyl acetate fraction and aglycone fraction of P. vulgaris L. In ethyl acetate fraction, caffeic acid, rosmarinic acid, quercetin 3-${\beta}$-D-glucoside, rutin, kaempferol-3-O-rutinoside, astragalin (kaempferol-3-O-glucoside) were identified. In aglycone fraction, caffeic acid, rosmarinic acid, quercetin, kaempferol were identified. These results indicated that extract/fraction of P. vulgaris L. is may be used in cosmetics industry as natural antioxidants by quenching and/or scavenging $^1O_2$ and other ROS, and protecting cellular membranes.

THE EFFECT OF CONCENTRATION AND APPLICATION TIME OF HYDROGEN PEROXIDE ON THE MICROTENSILE BOND STRENGTH OF RESIN RESTORATIONS TO THE DENTIN AT DIFFERENT DEPTHS (과산화수소의 농도와 적용시간이 상아질의 깊이에 따라 레진 수복물의 미세인장결합강도에 미치는 영향)

  • Son, Jeong-Lyong;Lee, Gye-Young;Kang, Yu-Mi;Oh, Young-Taek;Lee, Kwang-Won;Kim, Tae-Gun
    • Restorative Dentistry and Endodontics
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    • v.34 no.5
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    • pp.406-414
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    • 2009
  • The purpose of this study was to examine the effect of hydrogen peroxide at different application time and concentrations on the microtensile bond strength of resin restorations to the deep and the pulp chamber dentin. A conventional endodontic access cavity was prepared in each tooth, and then the teeth were randomly divided into 1 control group and 4 experimental groups as follows: Group 1, non treated; Group 2, with 20% Hydrogen peroxide ($H_2O_2$); Group 3, with 10% $H_2O_2$; Group 4, with 5% $H_2O_2$; Group 5, with 2.5% $H_2O_2$; the teeth of all groups except group 1 were treated for 20, 10, and 5min. The treated teeth were filled using a Superbond C&B (Sun medical Co., Shiga, Japan). Thereafter, the specimens were stored in distilled water at $37^{\circ}C$ for 24-hours and then sectioned into the deep and the chamber dentin. The microtensile bond strength values of each group were analyzed by 3-way ANOVA and Tukey post hoc test(p < 0.05). In this study, the microtensile bond strength of the deep dentin (D1) was significantly greater than that of the pulp chamber dentin (D2) in the all groups tested. The average of microtensile bond strength was decreased as the concentration and the application time of $H_2O_2$ were increased. Analysis showed significant correlation effect not only between the depth of the dentin and the concentration of $H_2O_2$ but also between the concentration of H202 and the application time(p < 0.05), while no significant difference existed among these three variables(p > 0.05). The higher $H_2O_2$ concentration, the more opened dentinal tubules under a scanning electron microscope(SEM) examination.

Antioxidative, Antimicrobial and Cytotoxic activities of Fagopyrum esculentum $M{\ddot{o}}ench$ Extract in Germinated Seeds (발아 메밀 추출물의 항산화.항균활성 및 세포독성)

  • Hwang, Eun-Ju;Lee, Sook-Young;Kwon, Su-Jung;Park, Min-Hee;Boo, Hee-Ock
    • Korean Journal of Medicinal Crop Science
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    • v.14 no.1
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    • pp.1-7
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    • 2006
  • This research was conducted to investigate the possibilities of usage of germinated-buckwheat (Fagopyrum esculentum $M{\ddot{o}}ench$) by examining antioxidative, antimicrobial and cytotoxic effects of extracts from different germinated root length of buckwheat. Antioxidant activity $(RC_{50})$ was shown higher in extracts of non-germinated seed $(50.41\;{\mu}g/mL)$ and root length 10 mm $(80.57\;{\mu}g/mL)$, 2 mm $(93.77\;{\mu}g/mL)$, 5 mm $(107.09\;{\mu}g/mL)$ than BHT $(163.96\;{\mu}g/mL)$ as a synthetic antioxidant. In antimicrobial activity, non-germinated and germinated seeds were formed inhibitory zone against S. aureus $(4{\sim}10\;mm)$, P. aeruginosa $(2{\sim}9\;mm)$ at the concentrations of $10{\sim}40\;mg/mL$ but B. subtilis, E. coli and S. typhimurium were not apparent antimicrobial activity. Extracts of germinated seed also decreased their antimicrobial activity compared to non-germinated seed extract. In addition, the growth of Calu-6 was inhibited of both 5 mm root length germinated and non-germinated seeds $(800\;{\mu}g/mL)$ as 95.12% and 87.15%, respectively, but these did not show any influence on cytotoxic effect against MCF-7 and Caco-2 cell lines. Extracts of 2 mm and 5 mm germinated seeds were also inhibited against Calu-6 and SNU-601 cell lines.

Anti-inflammatory Effect and Antioxidative Activities of Ingredients used in Bibimbab (비빔밥 재료의 항염증 및 항산화 효과)

  • Ko, Yu-Jin;Seol, Hui-Gyeong;Lee, Gyeong-Ran;Jeong, Gye-Im;Ryu, Chung-Ho
    • Journal of Life Science
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    • v.23 no.2
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    • pp.213-221
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    • 2013
  • Bibimbab (mixed rice) is a traditional Korean one-dish meal. This study was carried out to investigate the anti-inflammatory and antioxidative effects of raw and seasoned ingredients used in Bibimbab (Cucurbita moschata P., Platycodon grandiflorum A., Vigna radiata L., Porphyra yezonensis udea, Allium ampeloprasum L., Pterdium aguilinum, Raphanus sativus). Human mast cells (HMC-1) were pretreated with 70% ethanol extracts of Bibimbab and further cultured for an appropriate time after the addition of phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187. Cell viability was determined by an MTT assay. None of the ingredients showed cytotoxic effects at a concentration of 1.0 ${\mu}g/ml$. Anti-inflammatory effects were analyzed at 0.01, 0.1, and 1.0 mg/ml concentrations of the raw, seasoned ingredients of PMA. A23187 stimulated HMC-1. Among the various ingredients, seasoned A. ampeloprasum L. extract showed the highest inhibition of TNF-${\alpha}$ and IL-6 secretion (90% and 93%, respectively) at a concentration of 1.0 mg/ml. The R. sativus extract showed the highest inhibition (85%) of IL-8 secretion. DPPH analysis of the antioxidation properties of the ingredients showed that raw and seasoned A. ampeloprasum extracts exhibited the highest DPPH free radical scavenging activity (67.50 and 73.65%, respectively). These results suggest that seasoned ingredients used in Bibimbab have lower anti-inflammatory effects in relation to TNF-${\alpha}$ and IL-6 secretion than raw ingredients in PMA- and A23187-treated HMC-1. In addition, the seasoned ingredients showed a tendency to increase antioxidative activity. Therefore, the ingredients used in Bibimbab have potential as anti-inflammatory and antioxidation agents.

Effects of Dietary Supplementation of Mulberry Leaves and Silkworm Excreta Extracts on Weight Performance, Blood Characteristics and Cecal Microflorae of Chicks (뽕잎과 잠분 추출물 급여가 육계의 생산성, 혈액 성상 및 맹장 내 미생물군에 미치는 영향)

  • Han, Sang-Hoon;Kim, Dong Wook;Ji, Sang Yoon;Hong, Seong Koo;Kim, Sang-Ho;Lee, Heui-Sam
    • Journal of Sericultural and Entomological Science
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    • v.50 no.2
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    • pp.150-160
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    • 2012
  • The object of this study was to evaluate the effects of dietary supplementation of mulberry leaves and silkworm excreta ethanol extracts on weight performances, blood characteristics, cecal microflorae of chickens. Two hundred forty male broiler chicks(Ross) were fed diets for five weeks containing 0.1%(MLA) and 1%(MLB) of mulberry leaves ethanol extracts, and 0.1%(SEA) and 1%(SEB) of silkworm excreta ethanol extracts. Weight performance did show no significant difference in all test groups which were fed with supplementation of mulberry leaves and silkworm excreta ethanol extracts. They showed better weight gain and feed conversion than the negative control group which was fed only with forage without antibiotics. ABTS(2'-azine-bis[3-ethylbenzothiazoline-6-sulfonic acid]) test was conducted to investigate free radical scavenging activity of blood in tested groups. ABTS scavenging activities of tested groups were higher than control groups in significant level, though there was no significant difference(P = 0.396). Specifically, MLB group showed the highest scavenging activity. Blood-level concentration of MDA, which is an indicator of lipid peroxidation, was also decreased in tested groups and the lowest level was observed in SEA(P = 0.001). As storage time increased at $4^{\circ}C$, muscle-level MDA concentrations of all tested groups were generally increased and significant difference was obsereved between tested groups and controls in total increase of MDA concentration($P=4.417{\times}10^{-3}$). In cecal microflorae, SEA and SEB showed decreased total microbe population compared to NC($P=6.462{\times}10^{-5}$) and even to PC. Supplementation of mulberry leave and silkworm excreta ethanol extract did show a similar inhibition effect against Salmonella sp., furthermore, MLB did enhanced the growth of Lactobacillus sp.($P=3.636{\times}10^{-7}$). In summary, ethanol extract of silkworm excreta may be a potential alternative of antibiotics for chicks. In addition, both of ethanol extracts supplementation to broiler chicks would be very useful not only to improve antioxidant effect of blood but also to suppress lipid peroxidation without any loss of weight performance in poultry farming.

Antioxidant, Antimicrobial and Cytoprotective Effects of the Extract and Its Fraction Obtained from Rhizomes of Belamcanda chinensis (L.) DC (범부채 뿌리 추출물 및 분획물의 항산화, 항균 및 세포 보호 효과)

  • Song, Ba Reum;Lee, Sang Lae;Lee, Yun Ju;Shin, Hyuk Soo;Park, Soo Nam
    • Applied Chemistry for Engineering
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    • v.29 no.6
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    • pp.772-781
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    • 2018
  • In this study, we investigated antioxidant, antimicrobial and cytoprotective effects of 50% ethanol extract and ethyl acetate fraction from rhizomes of Belamcanda chinensis (L.) DC. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activities ($FSC_{50}$) of the 50% ethanol extract and ethyl acetate fraction were 621.5 and $253.0{\mu}g/mL$, respectively. Total antioxidant capacities ($OSC_{50}$) of the extract and fraction were 13.6 and $3.0{\mu}g/mL$, respectively. Minimum inhibitory concentrations (MIC) of the ethyl acetate fraction for Staphylococcus aureus and Candida albicans were 156, $1,250{\mu}g/mL$, respectively, indicating similar or higher levels of those of using methyl paraben. Cytoprotective effects of the 50% ethanol extract against $^1O_2$-induced cellular damage (${\tau}_{50}$) showed in a dose dependent manner at 4 to $64{\mu}g/mL$. ${\tau}_{50}$ of the 50% ethanol extract, ethyl acetate fraction and (+)-${\alpha}$-tocopherol at $16{\mu}g/mL$ were 36.4, 45.0 and 45.8 min respectively, and the ethyl acetate fraction showed cytoprotective effects similar to (+)-${\alpha}$-tocopherol. In ultraviolet B radiation-induced HaCaT cell damage, the ethyl acetate fraction decreased intracellular reactive oxygen species (ROS) up to 45.9% at $8{\mu}g/mL$. Also in $H_2O_2$-induced HaCaT cell damage, the ethyl acetate fraction significantly increased the cell viability at $0.5{\sim}8.0{\mu}g/mL$. As a result of chemical analyses of the ethyl acetate fraction, the presence of flavonoids and polyphenol such as irisflorentin, irigenin, tectorigenin, resveratrol, iridin and tectoridin were identified. In conclusion, the extract/fraction from rhizomes of B. chinensis can be applied as a natural antioxidant and antimicrobial material to cosmetics.

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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