• Title/Summary/Keyword: 3G cellular system

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The Butanol Fraction of Bitter Melon (Momordica charantia) Scavenges Free Radicals and Attenuates Oxidative Stress

  • Kim, Hyun Young;Sin, Seung Mi;Lee, Sanghyun;Cho, Kye Man;Cho, Eun Ju
    • Preventive Nutrition and Food Science
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    • v.18 no.1
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    • pp.18-22
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    • 2013
  • To investigate radical scavenging effects and protective activities of bitter melon (Momordica charantia) against oxidative stress, in vitro and a cellular system using LLC-$PK_1$ renal epithelial cells were used in this study. The butanol (BuOH) fraction of bitter melon scavenged 63.4% and 87.1% of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals at concentrations of 250 and $500{\mu}g/mL$, respectively. In addition, the BuOH fraction of bitter melon effectively scavenged hydroxyl radicals (${\cdot}OH$). At all concentrations tested, the scavenging activity of the BuOH fraction was more potent than that of the positive control, ascorbic acid. Furthermore, under the LLC-$PK_1$ cellular model, the cells showed a decline in viability and an increase in lipid peroxidation through oxidative stress induced by pyrogallol, a generator of superoxide anion ($O_2{^-}$). However, the BuOH fraction of bitter melon significantly and dose-dependently inhibited cytotoxicity. In addition, 3-morpholinosydnonimine (SIN-1), a generator of peroxynitrite ($ONOO^-$) formed by simultaneous releases of nitric oxide and $O_2{^-}$, caused cytotoxicity in the LLC-$PK_1$ cells while the BuOH fraction of bitter melon ameliorated oxidative damage induced by $ONOO^-$. These results indicate that BuOH fraction of bitter melon has protective activities against oxidative damage induced by free radicals.

Radical Scavenging Effect and Neuroprotective Activity from Oxidative Stress of Engelhardtia chrysolepis Leaf (Engelhardtia chrysolepis의 라디칼 소거능 및 신경세포의 산화 스트레스 보호효과)

  • Kim, Eun Jung;Lee, Ah Young;Choi, Soo Yeon;Seo, Hye Rin;Lee, Young A;Cho, Eun Ju
    • Korean Journal of Pharmacognosy
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    • v.47 no.3
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    • pp.251-257
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    • 2016
  • In this study, the radical scavenging activity and protective effect of ethanol extract from leaf of Engelhardtia chrysolepis HANCE (ECE) against oxidative stress were investigated under in vitro and cellular system. ECE showed strong radical scavenging activities in 1,1-diphenyl-2-picrylhydrazyl, hydroxyl(${\cdot}OH$) and nitric oxide(NO) radical as a concentration-dependent manner. Particularly, strong scavenging activity against the ${\cdot}OH$ and NO radical were observed with the $IC_{50}$ value of $1.30{\mu}g/ml$ and $12.61{\mu}g/ml$, respectively. Furthermore, the cellular oxidative stress was induced by amyloid beta($A{\beta}_{25-35}$) in C6 glial cells. The treatment of $A{\beta}_{25-35}$ to C6 glial cells generated high levels of reactive oxygen species(ROS) and declined cell viability. However, production of ROS was decreased by the treatment of ECE. In addition, the cell viability was significantly increased at each concentration(10, 25, $50{\mu}g/ml$) as dose-dependent manner. The Alzheimer's disease-related protein expressions in $A{\beta}_{25-35}$-treated C6 glial cells were analyzed. The ECE treatment inhibited expression of amyloid precursor protein(APP), C-terminal fragment-${\beta}(CTF-{\beta})$, ${\beta}$-site APP cleaving enzyme(BACE), phosphorylated tau(p-tau) proteins in C6 glial cells induced by $A{\beta}_{25-35}$. The present study indicated that ECE has strong radical scavenging activity and neuroprotective effect through attenuating oxidative stress.

Studies on the Cellular Metabolism in Microorganisms as Influenced by Gamma-irradiation.(II) - On the Respiration Rate and Dehydrogenase Actibity in Yeast Cells Irradiated by $\gamma$-ray. (미생물의 세포생리에 미치는 전이방사선의 영향에 관한 연구 (제 2보) - 효모균의 산소호흡기및 탈수소효소능에 대한 $\gamma$-ray 의 영향)

  • 김종협
    • Korean Journal of Microbiology
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    • v.5 no.2
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    • pp.69-78
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    • 1967
  • Kim, Jong Hyup, (Div. of Biology, Atomic Energy Research Institute.) Studies on the Cellular Metabolism in Microorganisms as influenced by Gamma-irradiation(II). On respiration rate and dehydrogenase activity of yeast cells irradiated by gamma ray from cobalt-60. 1. Oxygen uptake rate of the gamma irraiated yeast cells had been measured with Warburg's manometer, and the $O_{2}$-uptake was compared with those of normal cells. The rate of endogetious respiration increases in its $O_2$-uptake at 150, 000 rentgen dose, and at higher rentoen doses it was decreased. Exogenous respiration begin to decrease in its O_2$-uptake at 5, 000r. doses of irradiation, further decrease with increasing of doses unproportionally. 2. It appears that plasma-membrane and nuclear membrane of yeast cells have changed and denatured by gamma-irradiation, as exogenous respiration of glucose had been decreased at a dose of 200, 000r's irradiation. 3. The activity of glucose, alcoholic, lactic, succinic and glutamic deliydrogenase (G.D.H., A.D.H., L.D.H., S.D.11., and GL.D.H.) in the gamma irradaited cells had been assayed by T.T.C.(Triphenyl tetrazolium chloride) method and spectrophotometry, the obtained results were compared with those of normal cells. 4. At a dose of and 10, 000 rentgens' irradiation of gamma ray, the activty of each debydrogenase (G.D.H., A.D.H., L.D.H., ) shows a sharp and highest peak in optical absorbalicy, but each abtivity of S.D.H and Gl.D.H shows its' maximum peak at a dose of 30, 000r. 5. The curve of each dehydrogenase activity was found to be rhythmical according to dose-rate of gamma irradiation. 6. Comparing with activity of debydrogenase each other, the maximum peak in optical absorbency can be arranged according to order as follows; glucose > alcoholoic > lactic > glutamic > succinic, this order is identical to the order of breakdown utility in respiration of normal yeast cells. 7. The activity of dehydrogenase experimented exhibit a resistance against gamma irradiation at lethal dose of cells, and the activity of dehydrogenase are found to be much resistant than those of respiratory system. We may assume that the membrane substrate of mitochondria or cytoplasm had been destructed by gamma-irradiation much more than that of dehydronase system.

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Antioxidative Activity and Component Analysis of Phellinus linteus Extracts (상황버섯 추출물의 항산화 및 성분분석)

  • Kim, A-Reum;Kim, Jung-Eun;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.37 no.4
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    • pp.309-318
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    • 2011
  • In this study, the antioxidative effect, antibacterial, inhibitory effects on tyrosinase, inhibitory effects on elastase and component analysis of Phellinus linteus (P. linteus) extracts were investigated. The ethyl acetate fraction of P. linteus extracts ($2.94\;{\mu}g/mL$) showed the highest free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$). Reactive oxygen species (ROS) scavenging activity ($OSC_{50}$) of P. linteus extracts on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system was investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate fraction ($0.0072\;{\mu}g/mL$) showed the most prominent ROS scavenging activity. The protective effects of extract/fractions of P. linteus extracts on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The P. linteus extracts showed cellular membrane protective effects in a concentration dependent manner ($5{\sim}50\;{\mu}g/mL$). The inhibitory effect ($IC_{50}$) on tyrosinase of P. linteus extract was the highest at 50 % ethanol extract ($6.34\;{\mu}g/mL$), and the inhibitory effect ($IC_{50}$) on elastase of P. linteus was the highest at ethyl acetate fraction ($14.08\;{\mu}g/mL$). TLC, HPLC chromatogram and LC/ESI-MS of the ethyl acetate fraction obtained from P. linteus extracts were identified interfungin A (PL RPT-1a). These results indicate that extract/fractions of P. linteus can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging ROS, and protect cellular membranes against ROS. Extract/fractions of P. linteus can be applicable to new cosmeceuticals for antioxidant, antiaging, antiwrinkle and whitening.

Antimicrobial, Antioxidant and Cellular Protective Effects against Oxidative Stress of Anemarrhena asphodeloides Bunge Extract and Fraction (지모 뿌리 추출물과 분획물의 항균활성과 항산화 활성 및 세포보호 연구)

  • Lee, Yun Ju;Song, Ba Reum;Lee, Sang Lae;Shin, Hyuk Soo;Park, Soo Nam
    • Microbiology and Biotechnology Letters
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    • v.46 no.4
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    • pp.360-371
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    • 2018
  • Extracts and fractions of Anemarrhena asphodeloides Bunge were prepared and their physiological activities and components were analyzed. Antimicrobial activities of the ethyl acetate and aglycone fractions were $78{\mu}g/ml$ and $31{\mu}g/ml$, respectively, for Staphylococcus aureus and $156{\mu}g/ml$ and $125{\mu}g/ml$, respectively, for Pseudomonas aeruginosa. 1,1-Diphenyl-2-picrylhydrazyl free radical scavenging activities ($FSC_{50}$) of 50% ethanol extract, ethyl acetate fraction, and aglycone fraction of A. asphodeloides extracts were $146.2{\mu}g/ml$, $23.19{\mu}g/ml$, and $71.06{\mu}g/ml$, respectively. The total antioxidant capacity ($OSC_{50}$) in an $Fe^{3+}$-EDTA/hydrogen peroxide ($H_2O_2$) system were $17.5{\mu}g/ml$, $1.5{\mu}g/ml$, and $1.4{\mu}g/ml$, respectively. The cytoprotective effect (${\tau}_{50}$) in $^1O_2$-induced erythrocyte hemolysis was 181 min with $4{\mu}g/ml$ of the aglycone fraction. The ${\tau}_{50}$ of the aglycone fraction was approximately 4-times higher than that of (+)-${\alpha}$-tocopherol (${\tau}_{50}$, 41 min). Analysis of $H_2O_2$-induced damage of HaCaT cells revealed that the maximum cell viabilities for the 50% ethanol extract, ethyl acetate fraction, and aglycone fraction were 86.23%, 86.59%, and 89.70%, respectively. The aglycone fraction increased cell viability up to 11.53% at $1{\mu}g/ml$ compared to the positive control treated with $H_2O_2$. Analysis of ultraviolet B radiation-induced HaCaT cell damage revealed up to 41.77% decreased intracellular reactive oxygen species in the $2{\mu}g/ml$ aglycone fraction compared with the positive control treated with ultraviolet B radiation. The findings suggest that the extracts and fractions of A. asphodeloides Bunge have potential applications in the field of cosmetics as natural preservatives and antioxidants.

Effects of in vivo-stresses on the Activities of the Myocardial Antioxidant Enzymes and the Ischemia-Reperfusion Injury in Rat Hearts (스트레스성 자극에 의한 항산화효소 유도와 허혈/재관류 심장 보호효과)

  • 박종완;김영훈;김명석
    • Toxicological Research
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    • v.11 no.1
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    • pp.161-168
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    • 1995
  • It has been found that various stress challenges induce the myocardial antioxidant enzymes and produce an acquisition of the cellular resistance to the ischemic injury in animal hearts. Most of the stresses, however, seem to be guite dangerous to an animal's life. In the present study, therefore, we tried to search for safely applicable stress modalities which could lead to the induction of antioxidant enzymes and the production of myocardial tolerance to the ischemia-reperfusion injury. Male Sprague-Dawley rats (200-250 g) were exposed to various non-fatal stress conditions, i.e., hyperthermia (environmental temperature of $42^{\circ}C$ for 30 min, non-anesthetized animal), iramobilization (60 min), treadmill exercise (20 m/min, 30min), swimming (30 min), and hyperbaric oxyflenation (3 atm, 60 min), once a day for 5 days. The activities of myocardial antioxidant enzymes and the ischemia-reperfusion injury of isolated hearts were evaluated at 24 hr after the last application of the stresses. The activities of antioxidant enzymes, superoxide dismutase (SOD), catalase, glutathione peroxidase, glutathione reductase and glucose-6-phosphate dehydrogenase (G6PD), were assayed in the freshly excised ventricular tissues. The ischemia-reperfusion injury was produced by 20 min-global ischemia followed by 30 min-reperfusion using a Langendorff perfusion system. In swimming and hyperbaric oxygenation groups, the activities of SOD and G6PD increased significantly and in the hyperthermia group, the catalase activity was elevated by 63% compared to the control. The percentile recoveries of cardiac function at 30 min of the post-ischemic reperfusion were 55.4%, 73.4%, and 74.2% in swimming, the hyperbaric oxygenation and the hyperthermia groups, respectively. The values were significantly higher than that of the control (38.6%). In additions, left ventricular end-diastolic pressure and lactate dehydrogenase release were significantly reduced in the stress groups. The results suggest that the antioxidant enzymes in the heart could be induced by the apparently safe in vivo-stresses and this may be involved in the myocardial protection from the ischemia-reperfusion injury.

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Genomewide Expression Profile of Forsythia Suspensa on Lipopolysaccaride-induced Activation in Microglial Cells

  • Sohn, Sung-Hwa;Ko, Eun-Jung;Kim, Yang-Seok;Shin, Min-Kyu;Hong, Moo-Chang;Bae, Hyun-Su
    • Molecular & Cellular Toxicology
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    • v.4 no.2
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    • pp.113-123
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    • 2008
  • Microglia, which is the primary immune effector cells in the central nervous system, constitutes the first line of defense against infection and injury in the brain. The goal of this study was to determine the protective (anti-inflammation) mechanisms of forsythia suspense (FS) on LPS-induced activation of BV-2 microglial cells. The effects of FS on gene expression profiles in activated BV-2 microglial cells were evaluated using microarray analysis. BV-2 microglial cells were cultured in a 100mm dish $(1{\times}10^7/dish)$ for 24hr and then pretreated with $1{\mu}g/mL$ FS or left untreated for 30 min. Next, $1{\mu}g/mL$ LPS was added to the samples and the cells were reincubated at $37^{\circ}C$ for 30 min, 1hr, and 3hr. The gene expression profiles of the BV-2 microglial cells varied depending on the FS. The oligonucleotide microarray analysis revealed that MAPK pathway-related genes such as Mitogen activated protein kinase 1 (Mapk1), RAS protein activator like 2 (Rasal2), and G-protein coupled receptor 12 (Gpr12) and nitric oxide biosynthesis-related genes such as nitric oxide synthase 1 (neuronal) adaptor protein (Nos1ap), and dimethylarginine dimethylaminohydrolase 1 (Ddah1) were down regulated in FS-treated BV-2 microglial cells. FS can affect the MAPK pathway and nitric oxide biosynthesis in BV-2 microglial cells.

Physiological and Phylogenetic Analysis of Burkholderia sp. HY1 Capable of Aniline Degradation

  • Kahng, Hyung-Yeel;Jerome J. Kukor;Oh, Kye-Heon
    • Journal of Microbiology and Biotechnology
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    • v.10 no.5
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    • pp.643-650
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    • 2000
  • A new aniline-utilizing microorganism, strain HY1 obtained from an orchard soil, was characterized by using the BIOLOG system, an analysis of the total cellular fatty acids, and a 16S rDNA sequence. Strain HY1 was identified as a Burkholderia species, and was designated Burkholderia sp. HY1. GC and HPLC analyses revealed that Burkholderia sp. HY1 was able to degrade aniline to produce catechol, which was subsequently converted to cis,cis-muconic acid through an ortho-ring fission pathway under aerobic conditions. Strain HY1 exhibited a drastic reduction in the rate of aniline degradation when glucose was added to the aniline media. However, the addition of peptone or nitrate to the aniline media dramatically accelerated the rate of aniline degradation. A fatty acid analysis showed that strain HY1 was able to produce lipids 16:0 2OH, and 11 methyl 18:1 ${\omega}7c$ approximately 3.7-, 2.2-, and 6-fold more, respectively, when grown on aniline media than when grown on TSA. An analysison the alignment of a 1,435 bp fragment. A phylogenetic analysis of the 16S rDNA sequence based on a 1,420 bp multi-alignment sowed of the 16s rDNA sequence revealed that strain HY1 was very closely related to Burkholderia graminis with 95% similarity based that strain HY1 was placed among three major clonal types of $\beta$-Proteobacteria, including Burkholderia graminis, Burkholderia phenazinium, and Burkholderia glathei. The sequence GAT(C or G)${\b{G}}$, which is highly conserved in several locations in the 16S rDNA gene among the major clonal type strains of $\beta$-Proteobacteria, was frequently replaced with GAT(C or G)${\b{A}}$ in the 16S rDNA sequence from strain HY1.

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Antioxidative Effects and Component Analysis of Extracts of the Rumex acetosa L. (수영 전초 추출물의 항산화 활성 평가 및 성분 분석)

  • Jeong, Yoo Min;Kim, Ho Jae;Lee, Su Hyun;Jang, Do Yun;Choi, Yae Chan;Min, Na Young;Gong, Bong Ju;Park, Soo Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.40 no.4
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    • pp.391-402
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    • 2014
  • In this study, the antioxidative effects and component analysis for the extracts of Rumex acetosa L. were investigated. All experiments were performed with 50% ethanol extract, ethyl acetate fraction and aglycone fraction obtained from dried R. acetosa L.. Free radical scavenging activities (1,1-diphenyl-2- picrylhydrazyl) size of, in the order of aglycone fraction > ethyl acetate fraction > 50% ethanol extract, aglycone fraction ($45.10{\mu}g/mL$) showed the highest radical scavenging activity. Reactive oxygen species (ROS) scavenging activity (total antioxidant capacity, $OSC_{50}$) on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system was also, in the order of ethyl acetate fraction> aglycone fraction> 50% ethyl acetate fraction, ethyl acetate fraction ($2.68{\mu}g/mL$) was shown a great antioxidant capacity. The total antioxidant capacity of the ethyl acetate fraction was found to be greater than L-ascorbic acid, known as a typical hydrophilic antioxidant ($6.88 {\mu}g/mL$). The cellular protective effects of R. acetosa L. extracts on the $^1O_2$-induced cellular damage of human erythrocytes were exhibited at all concentration-dependent ($1{\sim}25{\mu}g/mL$). Especially, aglycone fraction (${\tau}_{50}$, 104.80 min) in $25{\mu}g/mL$ showed the most protective effect among extracts. Components of the ethyl acetate fraction obtained from R. acetosa L. extracts were analyzed by TLC, HPLC chromatogram, LC/ESI-MS/MS. As a result, the ethyl acetate fraction contained several flavonoids, such as orientin, isoorientin, vitexin, isovitexin. These results indicate that the R. acetosa L. extracts can be used as antioxidants in biological systems, particularly skins exposed to UV radiation by quenching and/or scavenging $^1O_2$ and other ROS. Thus, the extracts of R. acetosa L. could be applicable to new anti-aging cosmeceutical ingredients.

Antioxidative Activity and Component Analysis of Cayratia japonica Extract (오렴매 추출물의 항산화 활성, 성분 분석)

  • Yang, Hee-Jung;Kim, Eun-Hee;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.34 no.2
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    • pp.117-127
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    • 2008
  • In this study, the antioxidative effects, inhibitory effects on elastase, and components of Cayratia japonica extracts were investigated. The free radical(1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities($FSC_{50}$) of extract/fractions of Cayratia japonica were in the order: 50% ethanol extract(114.3 ${\mu}g/mL$)${\mu}g/mL$)${\mu}g/mL$). Reactive oxygen species(ROS) scavenging activities($OSC_{50}$) of some Cayratia japonica extracts in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activities were deglycosylated flavonoid aglycone fraction($OSC_{50},\;3.30{\mu}g/mL$)<50% ethanol extract(1.21 ${\mu}g/mL$)${\mu}g/mL$). Ethyl acetate fraction showed the most prominent scavenging activity. The protective effects of extract/fractions of Cayratia japonica on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Cayratia japonica extracts suppressed photohemolysis in a concentration dependent manner($1{\sim}25{\mu}g/mL$), particularly deglycosylated flavonoid aglycone fraction exhibited the most prominent celluar protective effect(${\tau}_{50}$, 175.05min at 25 ${\mu}g/mL$). Aglycone fractions obtained from the deglycosylation reaction of ethyl acetate fraction among the Cayratia japonica extracts, showed 2 bands in TLC and 2 peaks in HPLC experiments(360 nm). Two components were identified as luteolin(composition ratio, 47.50%), apigenin(52.50). TLC chromatogram of ethyl acetate fraction of Cayratia japonica extract revealed 3 bands and HPLC chromatogram showed 4 peaks, which were identified as luteolin-7-O-${\beta}$-D-glucopyranoside(composition ratio, 11.14%), apigenin-7-O-${\beta}$-D-glucuronopyranoside(15.38%), luteolin(23.55%) and apigenin(49.92%) in the order of elution time. The inhibitory effect of aglycone fraction on elastase($IC_{50},\;70.5{\mu}g/mL$) was very high. These results indicate that extract/fractions of Cayratia japonica can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. And component analysis of Cayratia japonica extract and antioxidative effects could be applicable to new cosmetics.