• Title/Summary/Keyword: Reactive oxygen

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Antioxidative Effect and Active Component Analysis of Quercus salicina Blume Extracts (참가시나무의 항산화 효과 및 활성 성분 분석)

  • Lee, Hye-Jin;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.37 no.2
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    • pp.143-152
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    • 2011
  • In this study, the antioxidative activity, inhibitory effects on tyrosinase and elastase, and active components of Quercus salicina Blume extracts were investigated. Q. salicina Blume was extracted using 50 % ethanol, from which ethyl acetate and aglycone fractions were prepared. The DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging activity ($FSC_{50}$) of Q. salicina Blume aglycone fraction was the highest ($8.25\;{\mu}g$/mL). The luminol-dependent chemiluminescence assay showed that reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of Q. salicina Blume aglycone fraction on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system was the most prominent at $0.039\;{\mu}g$/mL. The protective effects of extract/fractions of Q. salicina Blume against the rose-bengal sensitized photohemolysis of human erythrocytes were increased in a concentration dependent manner ($1{\sim} 25\;{\mu}g$/mL). Especially, ${\tau}_{50}$ of aglycone fraction in $10 \;{\mu}g$/mL concentration showed the most protective effect at 259.9 min. The inhibitory effects ($IC_{50}$) on tyrosinase and elastase of Q. salicina Blume extracts were higher at aglycone fraction (respectively, $21.82 \;{\mu}g$/mL, $41.18\;{\mu}g$/mL). Active component analysis by TLC and HPLC showed quercetin, keampferol, catechin, quercitrin, isoquercitrin, hyperoside, and etc. These results indicate that Q. salicina Blume extract has strong antioxidative activity and can be used as antioxidant. Particularly, aglycone fraction of Q. salicina Blume showed superior antioxdative activity and high inhibitory effect on tyrosinase and elastase. Therefore, aglycone fraction of Q. salicina Blume could be applicable to new functional cosmetics.

Antioxidative Effects and Component Analysis of Graviola (Annona muricata) Leaf Extract/Fractions (그라비올라(Annona muricata) 잎 추출물 및 분획물의 항산화 활성과 성분 분석)

  • Park, So Hyun;Kim, Ji Min;Kim, Jin Hwan;Oh, Yoon Soo;Joo, Dong Hee;Lee, Eun Young;Shin, Hyuk Soo;Kim, A Rang;Lee, Sang Lae;Park, Soo Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.43 no.4
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    • pp.309-320
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    • 2017
  • In this study, the antioxidative effects and component analysis of 50% ethanol extract, ethyl acetate fraction and aglycone fraction obtained from Annona muricata leaf were investigated. Free radical scavenging activities were performed by 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, total antioxidant capacities were estimated using luminol-dependent chemiluminescence assay and $^1O_2$ quenching effects were estimated. Free radical scavenging activities ($FSC_{50}$) of 50% ethanol extract, ethyl acetate fraction and aglycone fraction were 45.6, 29.8 and $18.0{\mu}g/mL$, and total antioxidant capacities ($OSC_{50}$) were 4.4, 1.1 and $2.8{\mu}g/mL$, respectively. As a result of $^1O_2$ quenching experiment, ethyl acetate and aglycone fraction showed similar activities to L-ascorbic acid used as a comparative control. The cellular protective effects of 50% ethanol extract on the $^1O_2-induced$ cellular damage of human erythrocytes were exhibited at concentration-dependent ($5-50{\mu}g/mL$). TLC and HPLC were used to analyse components in the ethyl acetate fraction and aglycone fraction of Annona muricata leaf. In ethyl acetate fraction, rutin (quercetin-3-O-rutinoside), kaempferol-3-O-neohesperidoside, nicotiflorin (kaempferol-3-O-rutinoside), p-coumaric acid were identified. In aglycone fraction, kaempferol was identified. These results suggest that the extracts of Annona muricata leaf have the applicability as antioxidant cosmeceutical ingredients.

Cytoprotective Effects of Schisandrin A against Hydrogen Peroxide-induced Oxidative Stress in SW1353 Human Chondrocytes (SW1353 인간 연골세포에서 산화적 스트레스에 대한 schisandrin A의 세포 보호 효과)

  • Jeong, Jin-Woo;Choi, Eun Ok;Kwon, Da Hye;Kim, Bum Hoi;Park, Dong Il;Hwang, Hye Jin;Kim, Byung Woo;Choi, Yung Hyun
    • Journal of Life Science
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    • v.27 no.9
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    • pp.1070-1077
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    • 2017
  • Chondrocyte apoptosis induced by reactive oxygen species (ROS) plays an important role in the pathogenesis of osteoarthritis. Schisandrin A, a bioactive compound found in fruits of the Schisandra genus, has been reported to possess multiple pharmacological and therapeutic properties. Although several studies have described the antioxidant effects of analogues of schisandrin A, the underlying molecular mechanisms of this bioactive compound remain largely unresolved. The present study investigated the cytoprotective effect of schisandrin A against oxidative stress (hydrogen peroxide [$H_2O_2$]) in SW1353 human chondrocyte cells. The results showed that schisandrin A preconditioning significantly inhibited $H_2O_2-induced$ growth inhibition and apoptotic cell death by blocking the degradation of poly (ADP-ribose) polymerase proteins and down-regulating pro-caspase-3. These antiapoptotic effects of schisandrin A were associated with attenuation of mitochondrial dysfunction and normalization of expression changes of proapoptotic Bax and antiapoptotic Bcl-2 in $H_2O_2-stimulated$ SW1353 chondrocytes. Furthermore, schisandrin A effectively abrogated $H_2O_2-induced$ intracellular ROS accumulation and phosphorylation of histone H2AX at serine 139, a widely used marker of DNA damage. Thus, the present study demonstrates that schisandrin A provides protection against $H_2O_2-induced$ apoptosis and DNA damage in SW1353 chondrocytes, possibly by prevention of ROS generation. Collectively, our data indicate that schisandrin A has therapeutic potential in the treatment of oxidative disorders caused by overproduction of ROS.

Transfer of Isolated Mitochondria to Bovine Oocytes by Microinjection (미세주입을 이용한 난자로의 분리된 미토콘드리아 전달)

  • Baek, Sang-Ki;Byun, June-Ho;Kim, Bo Gyu;Lee, A ram;Cho, Young-Soo;Kim, Ik-Sung;Seo, Gang-Mi;Chung, Se-Kyo;Lee, Joon-Hee;Woo, Dong Kyun
    • Journal of Life Science
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    • v.27 no.12
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    • pp.1445-1451
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    • 2017
  • Mitochondria play a central role in energy generation by using electron transport coupled with oxidative phosphorylation. They also participate in other important cellular functions including metabolism, apoptosis, signaling, and reactive oxygen species production. Therefore, mitochondrial dysfunction is known to contribute to a variety of human diseases. Furthermore, there are various inherited diseases of energy metabolism due to mitochondrial DNA (mtDNA) mutations. Unfortunately, therapeutic options for these inherited mtDNA diseases are extremely limited. In this regard, mitochondrial replacement techniques are taking on increased importance in developing a clinical approach to inherited mtDNA diseases. In this study, green fluorescence protein (GFP)-tagged mitochondria were isolated by differential centrifugation from a mammalian cell line. Using microinjection technique, the isolated GFP-tagged mitochondria were then transferred to bovine oocytes that were triggered for early development. During the early developmental period from bovine oocytes to blastocysts, the transferred mitochondria were observed using fluorescent microscopy. The microinjected mitochondria were dispersed rapidly into the cytoplasm of oocytes and were passed down to subsequent cells of 2-cell, 4-cell, 8-cell, morula, and blastocyst stages. Together, these results demonstrate a successful in vitro transfer of isolated mitochondria to oocytes and provide a model for mitochondrial replacement implicated in inherited mtDNA diseases and animal cloning.

Quality Characteristics and Antioxidant Activity of Mulberry Leaf Tea Fermented by Monascus pilosus (Monascus pilosus로 발효시킨 뽕잎차의 품질특성과 항산화능)

  • Lee, Sang-Il;Lee, Ye-Kyung;Choi, Jong-Keun;Yang, Seung-Hwan;Lee, In-Ae;Suh, Joo-Won;Kim, Soon-Dong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.5
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    • pp.706-713
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    • 2012
  • This study was carried out to investigate the contents of monacolin K and citrinin, along with the sensory quality and antioxidant activity of mulberry leaf tea fermented by $Monascus$ $pilsous$ (FMM). Total monacolin K content of FMM was 0.058%, but citrinin was not detected. Redness of brewed FMM was remarkably higher than that of unfermented mulberry leaf tea (UFM). In sensory evaluation of brewed FMM, while astringent taste and savory taste were lower, flavor, color, and overall acceptability were significantly higher than those of UFM. Total polyphenol contents of UFM and FMM were 83.1 and 23.61 mg/g (dry basis), total flavonoid contents of UFM and FMM were 17.96 and 3.99 mg/g (dry basis), respectively. Xanthine oxidase (XO) inhibitory activity and superoxide dismutase (SOD)-like activity of FMM were lower than those of UFM. Electron-donating ability and ferric-reducing antioxidant power of FMM were slightly lower than those of UFM. However, the antioxidant activities of FMM per polyphenol content were markedly higher than those of UFM. These results suggest that FMM may scavenge excessive reactive oxygen spices (ROS) via inhibition of XO and SOD-like activity. Furthermore, FMM demonstrated relatively higher acceptability and antioxidant ability along with functionality of $Hongguk$ (red yeast rice), and therefore could be utilized to prevent various ROS-induced diseases.

Spectral Response of $TiO_{2}$/Se : Te Heterojunction for Color Sensor (컬러센서를 위한 $TiO_{2}$/Se : Te 이종접합의 스펙트럼 응답)

  • Woo, Jung-Ok;Park, Wug-Dong;Kim, Ki-Wan;Lee, Wu-Il
    • Journal of Sensor Science and Technology
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    • v.2 no.1
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    • pp.101-108
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    • 1993
  • $TiO_{2}$/Se : Te heterojunction for color sensor has been fabricated by RF reactive sputtering and thermal evaporation methods onto glass substrate. The optimum deposition condition of $TiO_{2}$ films was such that RF power was 120 W, substrate temperature was $100^{\circ}C$, oxygen concentration was 50%, working pressure was 50 mTorr for the $TiO_{2}$ film thickness of $1000{\AA}$. In this case, the optical transmittance of $TiO_{2}$ film at 550 nm-wavelength was 85%, resistivity was $2{\times}10^9{\Omega}{\cdot}cm$, refractive index was 2.3, and optical bandgap was 3.58 eV. The composition ratio of 0 to Ti by AES analysis was 1.7. When $TiO_{2}$ films were annealed at $400^{\circ}C$ for 30 min. in $O_{2}$ ambient, the optical transmittance of $TiO_{2}$ films at the wavelength range of $300{\sim}580$ nm was improved from 0 to 25%. When Se : Te films were annealed at $190^{\circ}C$ for 1 min., photosensitivity under illumination of 1000 lux was 0.75. The optical bandgap of Se : Te films was 1.7 eV. The structures of Se : Te films were the hexagonal with (100) and (110) orientation. The spectral response of a-Se was improved by the addition of Te, especially in the long wavelength region. The $TiO_{2}$/Se : Te heterojunction showed wide spectral response, and more improved one than that of a-Si film in the blue light region.

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Effect of Nicotinic Acid on Sperm Characteristic and Oocyte Development after In Vitro Fertilization using Cryopreserved Boar Semen

  • Kim, Yu-Jin;Lee, Sang-Hee;Lee, Yeon-Ju;Oh, Hae-In;Cheong, Hee-Tae;Yang, Boo-Keun;Lee, Seunghyung;Park, Choon-Keun
    • Journal of Embryo Transfer
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    • v.30 no.1
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    • pp.7-15
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    • 2015
  • The objective of this study was to investigate the efficiency of nicotinic acid on sperm cryosurvival and fertilization ability in frozen-thawed boar semen. Boar semen was collected by glove-hand method and was frozen using freezing solution treated to 0, 5, 10 and 20 mM of nicotinic acid. The frozen sperm for sperm characteristic analysis was thawed such as viability, acrosome reaction, and mitochondrial integrity. The frozen-thawed sperm was estimated by SYBR14/PI double staining for viability, FITC-PNA/PI double staining for acrosome reaction and Rhodamine123/PI double staining for mitochondrial integrity using a flow cytometry. The embryo was estimated in vitro development and DCFDA staining for reactive oxygen species assessment. As results, frozen-thawed sperm viability was significantly higher in 5 and 10 mM ($61.1{\pm}1.5%$,$64.7{\pm}2.0%$) of nicotinic acid than other groups (0 mM, $52.1{\pm}2.3%$; 20 mM, $47.8{\pm}5.1%$, P<0.05). The live sperm with acrosome reaction was significantly higher in 5 and 10 mM of nicotinic acid ($26.1{\pm}1.8%$, $24.9{\pm}1.5%$) than other groups (0 mM, $35.3{\pm}0.8%$; 20 mM, $36.5{\pm}1.9%$, P<0.05). The live sperm with mitochondrial integrity was significantly higher in 5 and 10 mM ($84.2{\pm}3.6%$, $88.4{\pm}2.3%$) of nicotinic acid than other groups (0 mM, $77.3{\pm}4.4%$; 20 mM, $73.3{\pm}3.6%$, P<0.05). Blastocyst rate of in vitro development was significantly higher in 10 mM ($17.0{\pm}1.3%$) of nicotinic acid than other groups (0 mM, $9.4{\pm}0.5%$; 5mM, $12.6{\pm}0.8%$; 20 mM, $5.0{\pm}1.0%$, P<0.05). Moreover, total cell number was higher in 5 and 10 mM ($53.6{\pm}2.9%$, $57.9{\pm}2.8%$) of nicotinic acid than other groups (0 mM, $41.0{\pm}1.4%$; 20 mM, $23.2{\pm}2.8%$, P<0.05). Hydrogen peroxide in embryos was lower in 5 mM nicotinic acid ($0.7{\pm}0.1%$) than other groups (0 mM, $1.0{\pm}0.1%$; 10mM, $0.9{\pm}0.0%$; 20 mM, $1.4{\pm}1.0%$, P<0.05). In conclusion, nicotinic acid-treated semen improves cryosurvival and quality of spermatozoa. Also, the fertilized oocytes with nicotinic acid improve quality of embryo and blastocyst formation.

Anti-Oxidative and Anti-Inflammatory Activities of Euptelea Pleiosperma Ethanol Extract (Euptelea pleiosperma 에탄올 추출물의 항산화 및 항염증 활성)

  • Jin, Kyong-Suk;Park, Jung Ae;Lee, Ji Young;Kang, Ji Sook;Kwon, Hyun Ju;Kim, Byung Woo
    • Microbiology and Biotechnology Letters
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    • v.42 no.2
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    • pp.170-176
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    • 2014
  • In this study, the anti-oxidative and anti-inflammatory activities of Euptelea pleiosperma ethanol extract (EPEE) were evaluated using in vitro assays and cell culture model systems. EPEE possessed a more potent scavenging activity against 1,1-diphenyl-2-picryl hydrazyl than the ascorbic acid used as a positive control. EPEE effectively suppressed lipopolysaccharide (LPS), in addition to hydrogen peroxide induced reactive oxygen species on RAW 264.7 cells. Furthermore, EPEE induced the expression of the anti-oxidative enzyme heme oxygenase 1 (HO-1) and its upstream transcription factor, nuclear factor-E2-related factor 2 (Nrf2), dose and time dependently. The modulation of HO-1 and Nrf2 expression might be regulated by mitogen-activated protein kinases and phosphatidyl inositol 3 kinase/Akt as their upstream signaling pathways. On the other hand, EPEE inhibited LPS induced nitric oxide (NO) formation without cytotoxicity. Suppression of NO formation was the result of the down regulation of inducible NO synthase (iNOS) by EPEE. Suppression of NO and iNOS by EPEE may be modulated by their upstream transcription factor, nuclear factor ${\kappa}B$, and AP-1 pathways. Taken together, these results provide important new insights into E. pleiosperma, namely that it possesses anti-oxidative and anti-inflammatory activities, indicating that it could be utilized as a promising material in the field of nutraceuticals.

High Glucose Induces Apoptosis through Caspase-3 Dependent Pathway in Human Retinal Endothelial Cell Line (인간망막 내피세포주에서 고농도 포도당이 caspase-3 경로를 통해 세포자연사 유도)

  • Seo, Eun-Sun;Chae, Soo-Chul;Kho, Eun-Gyeong;Lee, Jong-Bin
    • Korean Journal of Environmental Biology
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    • v.27 no.1
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    • pp.66-72
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    • 2009
  • Diabetic Retinopathy (DR) is a leading cause of blindness among adults in the western countries. Hyperglycemia is a condition, that induces apoptotic cell death in a variety of cell types in diabetes, but the mechanism remains unclear. The aim of the study is to understand the effects of high Glucose on Human Retinal Endothelial Cells. Retinal endothelial cells were cultured in Iscove's Modified Dulbecco's Medium (IMDM) containing 5, 25 and 50 mM Glucose, incubated for 24, 36 and 48 hours in humidified 5 % CO$_2$ incubator at 37$^{\circ}C$. Human Retinal Endothelial Cell Line (HREC) were characterized for morphology with different treatment by phase contrast microscopic analysis. Number of dead and viable cells was counted by trypan blue exclusion and supported by MTT assay. The intracellular Hydrogen peroxide (H$_2$O$_2$), a Reactive Oxygen Species (ROS) generation in high glucose conditions was assessed by FOX II assay and apoptosis by caspase-3 assay. The high glucose treated cells undergoing DNA fragmentation was witnessed by Agarose gel electrophoresis. We found that the cells incubated with 25 and 50 mM glucose containing medium for 48 hours altered the morphology of the cell, induced apoptosis and DNA fragmentation. The dead cell number were high in 25 and 50 mM when compared to the cells incubated with 5 mM glucose for 24, 36, and 48 hours. Also, the H$_2$O$_2$ levels and the activity of caspase-3 were increased in high glucose treated cells. Conclusions/interpretation: Our results demonstrated that elevated glucose induces apoptosis in cultured HREC. The hyperglycemia-induced increase in apoptosis may be dependent on caspase activation. The association between ROS generation and caspase-3 activation on high glucose treated cells is yet to be investigated.

Neuroprotective effects of Momordica charantia extract against hydrogen peroxide-induced cytotoxicity in human neuroblastoma SK-N-MC cells (산화적 스트레스에 대한 여주 (Momordica charantia) 추출물의 항산화 효과 및 세포사멸 억제 기전을 통한 신경세포보호효과)

  • Kim, Kkot Byeol;Lee, Seonah;Heo, Jae Hyeok;Kim, Jung hee
    • Journal of Nutrition and Health
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    • v.50 no.5
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    • pp.415-425
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    • 2017
  • Purpose: Many studies have suggested that neuronal cells protect against oxidative stress-induced apoptotic cell death by polyphenolic compounds. We investigated the neuroprotective effects and the mechanism of action of Momordica charantia ethanol extract (MCE) against $H_2O_2-induced$ cell death of human neuroblastoma SK-N-MC cells. Methods: The antioxidant activity of MCE was measured by the quantity of total phenolic acid compounds (TPC), quantity of total flavonoid compounds (TFC), and 2,2-Diphenyl-1-pycrylhydrazyl (DPPH) radical scavenging activity. Cytotoxicity and cell viability were determined by CCK-8 assay. The formation of reactive oxygen species (ROS) was measured using 2,7-dichlorofluorescein diacetate (DCF-DA) assay. Antioxidant enzyme (SOD-1,2 and GPx-1) expression was determined by real-time PCR. Mitogen-activated protein kinases (MAPK) pathway and apoptosis signal expression was measured by Western blotting. Results: The TPC and TFC quantities of MCE were 28.51 mg gallic acid equivalents/extract g and 3.95 mg catechin equivalents/extract g, respectively. The $IC_{50}$ value for DPPH radical scavenging activity was $506.95{\mu}g/ml$ for MCE. Pre-treatment with MCE showed protective effects against $H_2O_2-induced$ cell death and inhibited ROS generation by oxidative stress. SOD-1,2 and GPx-1 mRNA expression was recovered by pre-treatment with MCE compared with the presence of $H_2O_2$. Pre-treatment with MCE inhibited phosphorylation of p38 and the JNK pathway and down-regulated cleaved caspase-3 and cleaved PARP by $H_2O_2$. Conclusion: The neuroprotective effects of MCE in terms of recovery of antioxidant enzyme gene expression, down-regulation of MAPK pathways, and inhibition apoptosis is associated with reduced oxidative stress in SK-N-MC cells.