• Title/Summary/Keyword: sod gene

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Expression of the Antioxidant Enzyme and Apoptosis Genes in in vitro Maturation lin vitro Fertilization of Porcine Embryos

  • H. Y. Jang;H. S. Kong;Park, K. D.;G. J. Jeon;Lee, H. K.;B. K. Yang
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.47-47
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    • 2003
  • The present study was conducted to determine the expression of the antioxidant enzyme(CuZn-SOD, Mn-SOD and GPX and apoptosis gene(caspase-3) for in vitro culture in in vitro maturation and in vitro fertilization(IVM/IVF) embryos in porcine. Porcine embryos derived from IVM/IVF were cultured in NCSU23 medium under 5% $CO_2$ in air at 38.5$^{\circ}C$. The patterns of gene expression for several antioxidant enzyme and apoptosis genes during preimplantion porcine embryo development were examined by the modified semi-quantitative single cell reverse transcriptase- polymerase chain reaction (RT-PCR). Preimplantation porcine embryos produced by IVM/IVF have expressed mRNAs for CuZn-SOD and GPX, whereas transcripts for Mn-SOD have not detected at any developmental stages. Expression of caspase-3 mRNA was detected at 2 cell, 8 cell, 16 cell and morula stages. The fas ligand transcripts were detected in porcine blastocyst. These results suggest that various antioxidant enzymes and apoptosis genes play crucial roles in in vitro culture of porcine IVM/IVF embryos.

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The Role of Heat Shock Protein 25 in Radiation Resistance

  • Lee Yoon-Jin;Lee Su-Jae;Bae Sangwoo;Lee Yun-Sil
    • Environmental Mutagens and Carcinogens
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    • v.25 no.2
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    • pp.51-59
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    • 2005
  • Overexpression of HSP25 delayed cell growth, increased the level of $p21^{waf}$, reduced the levels of cyclin D1, cylcin A and cdc2, and induced radioresistance in L929 cells. We demonstrated that extracellular regulated kinase (ERK) and MAP kinase/ERK kinase (MEK) expressions as well as their activation (phospho-forms) were inhibited by hsp25 overexpression. To confirm the relationship between ERK1/2 and hsp25-mediated radioresistance, ERK1 or ERK2 cDNA was transiently transfected into the hsp25 overexpressed cells and their radioresistance was examined. HSP25-mediated radioresistance was abolished by overexpression of ERK2, but not by overexpression of ERK1. Alteration of cell cycle distribution and cell cycle related protein expressions (cyclin D, cyclin A and cdc2) by hsp25 overexpression were also recovered by ERK2 cDNA transfection. Increase in Bc1-2 protein by hsp25 gene transfection was also reduced by subsequent ERK2 cDNA-transfection. In addition, HSP25 overexpression reduced reactive oxygen species (ROS) and increased expression of manganese superoxide dismutase (MnSOD) gene. Increased activation of NF-kB (IkB degradation) was also found in hsp25-overexpressed cells. Moreover, transfection of hsp25 antisense gene abrogated all the HSP25-mediated phenomena. To further elucidate the exact relationship between MnSOD induction and NF-kB activation, dominant negative $I-kB\alpha(I-kB\alpha-DN)$ construction was transfected to HSP25 overexpressed cells. $I-kB\alpha-DN$ inhibited HSP25 mediated MnSOD gene expression. In addition, HSP25 mediated radioresistance was blocked by $I-kB\alpha-DN$ transfection. Blockage of MnSOD with antisense oligonucleotides in HSP25 overexpressed cells, prevented apoptosis and returned the ERK1/2 activation to the control level. From the above results, we suggest for the first time that reduced oxidative damage by HSP25 was due to MnSOD-mediated down regulation of ERK1/2.

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Structural Characteristics of NiSOD from Streptomyces seoulensis

  • Lee, Jin-Won;Wuerges, Jochen;Carugo, Kristina Djinovic;Kang, Sa-Ouk
    • Proceedings of the Korean Biophysical Society Conference
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    • 2002.06b
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    • pp.50-50
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    • 2002
  • The heterologous expression of sodN gene from Streptomyces seoulensis in Streptomyces lividans together with the gel filtration and sedimentation equilibrim data indicated that the quaternary structure of NiSOD is homohexamer, which is novel among SODs, not the previously reported homotetramer. The EPR spectrum of $^{61}$ Ni (I = 3/2) substituted NiSOD showed a clear resolved hyperfine structure at g=2.016, unambiguously identifying that the EPR signal from NiSOD is due to Ni.(omitted)

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Antioxidant Activity of Ethyl acetate Fraction of Berberis koreana Palibin in Caenorhabditis elegans (매자나무 Ethyl acetate 분획물의 예쁜 꼬마선충 내의 항산화 효과)

  • Ji Woo Choi;Jun Hyeong Kim;Jae Hyeok Lee;Dae Keun Kim
    • Korean Journal of Pharmacognosy
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    • v.54 no.2
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    • pp.66-71
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    • 2023
  • Ethyl acetate (EA) soluble fraction of the Berberis amurensis (Berberidaceae) methanol extract showed the potent DPPH radical scavenging activity through Caenorhabditis elegans model system. The EA fraction was measured for the activity of superoxide dismutase (SOD), catalase, and oxidative stress tolerance by using C. elegans along with reactive oxygen species (ROS) level. In addition, SOD-3 expression was conducted using a transgenic strain (CF1553) to confirm that the regulation of the stress response gene is responsible for the increased stress tolerance of C. elegans treated by the EA fraction. As a result, the EA soluble fraction of B. amurensis increased SOD and catalase activity, and decreased ROS accumulation in a dose-dependent manner. Furthermore, the EA fraction-treated CF1553 worm showed higher SOD-3::GFP intensity than the control worm.

Effect of Sopyung-tang Extract on Insulin Secretion and Gene Expression in RIN-m5F Cells (소평탕(消平湯)이 RIN-m5F 세포에서 인슐린 분비 및 유전자 발현에 미치는 영향)

  • Youn, Sung-Sik;Cho, Chung-Sik
    • The Journal of Internal Korean Medicine
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    • v.31 no.1
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    • pp.25-39
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    • 2010
  • Background : At high glucose levels in $\beta$-cells, cell viability and insulin secretion are decreased by glucotoxicity. Sopyung-tang(SPT) had an effect on blood glucose level decrease and antioxidant enzyme activities in streptozotocin-induced diabetic rats. Objectives : This study performed a series of experiment to verify the effects of SPT extract on the cell viability, antioxidant enzyme activities, insulin secretion and insulin mRNA expression at hyperglycemic states of RIN-m5F. Methods : After treatment at various concentrations of SPT added to the RIN-m5F cells, cell viability by MTT assay, free radical-scavenging activity, SOD activity and insulin secretion were measured. Additionally, insulin-related gene expression was measured using real-time RT-PCR. Results : Compared to the control group, SPT extract showed considerable effects on RIN-m5F cell viability, DPPH radical-scavenging activity, superoxide dismutase (SOD) activity, insulin secretion and insulin-related gene expression. Conclusions : This study showed that SPT extract has an effect on $\beta$-cell cell viability, insulin secretion and insulin-related gene expression. Thus, SPT extract may be used for treatment of diabetes and its complications. Further mechanism studies of SPT seem to be necessary on the glucotoxicity and oxidative stress.

Oxidative damage biomarker levels according to Mn-SOD and GST gene polymorphisms in preschool children (Mn-SOD와 GST 유전자 다형성에 따른 유아의 산화손상지표의 분포)

  • Shin, You-kyung;Choi, Ji-Won;Oh, Se-Young;Chung, Jayong
    • Journal of Nutrition and Health
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    • v.48 no.6
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    • pp.468-475
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    • 2015
  • Purpose: Genetic polymorphisms in antioxidant defense and detoxification genes may modulate the levels of oxidative stress biomarkers. Methods: A total of 301 healthy preschool-aged children in the Seoul and Kyung-gi areas were recruited. DNA was extracted from blood for genotyping of manganese superoxide dismutase (Mn-SOD) Val16Ala, glutathione S-transferase (GST) P1 Ile105Val, GSTT1 present/null, and GSTM1 present/null polymorphisms by PCR-restriction fragment length polymorphism or multiplex PCR analyses. In addition to a questionnaire survey, the levels of urinary 8-hydroxyl-2-deoxiguanosine (8-OHdG) and plasma malondialdehyde (MDA) were measured by ELISA. Results: Significantly higher urinary 8-OHdG concentrations were observed in GSTP1 Ile/Val + Val/Val genotype (p = 0.030), and tended to be higher in Mn-SOD Val/Val genotype (p = 0.065). On the other hand, exposure to environmental tobacco smoking (ETS) and interaction between ETS and gene polymorphisms did not significantly influence either urinary 8-OHdG concentrations or serum MDA. Conclusion: Based on our findings, GSTP1 Ile/Val gene polymorphisms might modulate the levels of oxidative stress biomarkers in healthy preschool children.

Superoxide Dismutase Gene Expression in the Endotoxin-Treated Rat Lung (내독소에 의한 백서 폐장의 Superoxide Dismutase 유전자 발현에 관한 연구)

  • Yoo, Chul-Gyu;Suh, Gee-Young;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.3
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    • pp.215-221
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    • 1994
  • Background: It is well known that oxygen free radicals(OFR) play a vital role in the various type of acute lung injury. Among various antioxidant defense mechanisms, the superoxide dismutases(SOD) are thought to be the first line of antioxidant defense by catalyzing the dismutation of two superoxide radicals to yield hydrogen peroxide and oxygen. Eukaryotic cells contain two types of intracellular SOD : cytosolic, dimeric copper/zinc- containing enzyme(CuZnSOD) and mitochondrial, tetrameric manganese-containing enzyme(MnSOD). The purpose of this study is to evaluate the time-dependent gene expression of MnSOD and CuZnSOD in the endotoxin-treated rats, and to compare with the manifestations of LPS-induced acute lung injury in rats. Methods: Total RNA from rat lung was isolated using single step phenol extraction 0, 1, 2, 4, 6, 12, 18, 24 hours after E. coli endotoxin injection(n=3, respectively). RNA was separated by formaldehyde-containing 1.2% agarose gels elctrophoresis, transblotted, baked, prehybridized, and hybridized with $^{32}P$-labeled cDNA probes for rat MnSOD and CuZnSOD, which were kindly donated by Dr. Ho(Duke University, Durham, NC, USA). The probes were labeled by nick translation. Blots were washed and autoradiography were quantitated using laser densitometry. Equivalent amounts of total RNA/gel were assessed by monitoring 28S and 18S rRNA. Results: Endotoxin caused a rise in steady-state MnSOD mRNA levels by 4h with peak mRNA accumulation by 6h. Continued MnSOD mRNA expression was observed at 12h. CuZnSOD mRNA expression was observed from 1h to 24h with peak levels by 18h. Conclusion: These results suggest that SOD palys an important defensive role in the endotoxin-induced acute lung injury in rats.

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Gene Expression Patterns of the Endogenous Antioxidant Enzymes in Linuron-Treated Rat Ventral Prostates after Castration

  • Yon, Jung-Min;Lin, Chunmei;Lee, Yoon-Bok;Lee, Beom-Jun;Yun, Young-Won;Nam, Sang-Yoon
    • Journal of Embryo Transfer
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    • v.27 no.2
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    • pp.101-105
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    • 2012
  • Linuron is a pesticide with a weak anti-androgenic property, which impacts male reproductive organs. In this study, to clarify whether linuron affects the cellular antioxidant system of ventral prostate, gene expression patterns of the representative antioxidant enzymes such as glutathione peroxidase (GPx), selenoprotein P (SePP), and superoxide dismutase (SOD) were investigated in the rat ventral prostates exposed to linuron using real-time RT-PCR analyses. Sprague-Dawley rats castrated at 6 weeks old were treated with linuron (25, 50, or 100 mg/kg per oral) daily for 10 days after testosterone propionate administration (0.4 mg/kg) subcutaneously. As compared to normal control animals, mRNA levels of phospholipid hydroperoxide GPx (PHGPx), SePP, and Mn SOD significantly increased in the prostates exposed to linuron (25, 50, and 100 mg/kg). However, cytosolic GPx (100 mg/kg) and Cu/Zn SOD (25, 50, and 100 mg/kg) mRNA levels significantly decreased in the ventral prostates. These results indicate that linuron upregulates the expressions of PHGPx, SePP, and Mn SOD mRNAs, but down-regulates the expressions of cytosolic GPx and Cu/Zn SOD in rat prostates, suggesting that linuron may have dual effects in the cellular antioxidant system of prostate.

Cell Survival and Expression of Superoxide Dismutase and Catalase Genes in Saccharomyces cerevisiae Treated with N-acetyl-L-cysteine and Ionizing Radiation (Saccharomyces cerevisiae에서 이온화 방사선과 N-acetyl-L-cysteine 처리에 따른 세포 생존과 Superoxide Dismutase와 Catalase 유전자 발현)

  • Park, Ji-Young;Baek, Dong-Won;Nili, Mohammad;Kim, Jin-Kyu
    • Korean Journal of Environmental Biology
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    • v.29 no.1
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    • pp.61-67
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    • 2011
  • N-acetyl-L-cysteine (NAC) having a thiol, a precursor for glutathione (GSH), is known as one of the antioxidants. NAC used as a radioprotector against ionizing radiation (IR)-induced injury and damage. The aim of this study was to evaluate the radioprotective effects of NAC against IR-induced cell damage in Saccharomyces cerevisiae and the antioxidative effect of NAC on transcriptional level of yeast antioxidant enzyme genes such as superoxide dismutase (SOD) and catalase. In the present study, yeast cells were pretreated with various concentrations of NAC and/or irradiated with various doses of gamma rays. The cell viability was measured by counting the cell forming unit (CFU). The quantitative real-time PCR was performed for analysis of gene expression of SOD and catalase. The viability of irradiated cells was not improved by pretreatment with NAC. Ionizing radiation with 100 Gy highly induced the gene expression of antioxidant enzymes. In the irradiated group with NAC pretreatment, the gene expression of SOD and catalase was gradually reduced with the increased concentrations of NAC. These results indicate that NAC can act as a useful antioxidant to scavenge reactive oxygen species in vivo, but does not protect cells against IR-induced cell death in S. cerevisiae.