• Title/Summary/Keyword: Superoxide dismutases (SOD)

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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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Effects of Genistein on the Gene Expressions of Glutathione Peroxidases and Superoxide Dismutases in Ethanol-Treated Mouse Fetuses

  • Yon, Jung-Min;Lin, Chunmei;Jung, A-Young;Lee, Jong-Geol;Jung, Ki-Youn;Baek, In-Jeoung;Lee, Beom-Jun;Nam, Sang-Yoon;Yun, Young-Won
    • Journal of Embryo Transfer
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    • v.26 no.2
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    • pp.135-140
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    • 2011
  • Genistein is a product of naturally occurring isoflavones at relatively high levels in soybeans. The harmful effects of ethanol are attributed to the induction of biological processes which lead to an increase in the generation of reactive oxygen species in fetuses. In this study, we investigated the effects of genistein ($1{\times}10^{-8}$ and $1{\times}10^{-7}\;{\mu}g$/ml) on gene expressions of the representative cellular antioxidative enzymes in ethanol (1 ${\mu}l$/ml)-treated mouse fetuses during the critical period (embryonic days 8.5~10.5) of organogenesis using a semi-quantitative RT-PCR analysis. The mRNA levels of cytosolic glutathione peroxidase (GPx), phospholipid hydroperoxide GPx, cytosolic CU,Zn-superoxide dismutase (SOD), and mitochondrial SOD were significantly decreased in ethanol-treated fetuses. However, the mRNA levels of ethanol plus genistein-treated fetuses were significantly higher than those of ethanol alone fetuses. These results indicate that genistein can up-regulate the expressions of GPx and SOD mRNAs reduced by the ethanol treatment in fetuses.

A Study of the Generation of Transgenic Chickens That Express Human SOD-3 Protein (사람의 SOD-3 단백질을 발현하는 형질전환 닭 생산 연구)

  • Byun, S.J.;Park, C.;Kim, J.A.;Woo, J.S.;Lee, H.C.;Kim, T.Y.;Kim, S.H.;Seong, H.H.;Park, J.K.;Jeon, I.S.
    • Korean Journal of Poultry Science
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    • v.35 no.3
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    • pp.241-245
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    • 2008
  • Lentiviral vector system is efficient vehicles for the delivery of exogenous genes, and it is generally used in the generation of transgenic chickens. In this study, we used recombinant lentiviral vectors to generate transgenic chicks that express the human superoxide dismutase-3 gene driven by the chicken ovalbumin promoter. It is well known that superoxide dismutases(SODs) are believed to play a crucial role in protecting cells against oxygen toxicity. There are three forms of SOD proteins: cytosolic Cu-Zn SOD, mitochondrial Mn SOD, and extracellular SOD(SOD-3). The recombinant lentivirus containing the human SOD-3 gene was injected into the subgerminal cavity of freshly laid eggs. Subsequently, the embryos were incubated to hatch using phases II and III of the surrogate shell ex vivo culture system. From 341 injected embryos, the 78 chicks hatched after 21 days incubation. The hatched chicks were screened for the human SOD-3 gene by using PCR. Two of 47 male chickens that survived to sexual maturity contained the human SOD-3 gene in their semen. These results showed that our transgenic chicken generation system was completely established.

Molecular Cloning and Expression of a Cu/Zn-Containing Superoxide Dismutase from Thellungiella halophila

  • Xu, Xiaojing;Zhou, Yijun;Wei, Shanjun;Ren, Dongtao;Yang, Min;Bu, Huahu;Kang, Mingming;Wang, Junli;Feng, Jinchao
    • Molecules and Cells
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    • v.27 no.4
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    • pp.423-428
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    • 2009
  • Superoxide dismutases (SODs) constitute the first line of cellular defense against oxidative stress in plants. SODs generally occur in three different forms with Cu/Zn, Fe, or Mn as prosthetic metals. We cloned the full-length cDNA of the Thellungiella halophila Cu/Zn-SOD gene ThCSD using degenerate RT-PCR and rapid amplification of cDNA ends (RACE). Sequence analysis indicated that the ThCSD gene (GenBank accession number EF405867) had an open reading frame of 456 bp. The deduced 152-amino acid polypeptide had a predicted molecular weight of 15.1 kDa, an estimated pI of 5.4, and a putative Cu/Zn-binding site. Recombinant ThCSD protein was expressed in Escherichia coli and assayed for SOD enzymatic activity in a native polyacrylamide gel. The SOD activity of ThCSD was inactivated by potassium cyanide and hydrogen peroxide but not by sodium azide, confirming that ThCSD is a Cu/Zn-SOD. Northern blotting demonstrated that ThCSD is expressed in roots, stems, and leaves. ThCSD mRNA levels increased by about 30-fold when plants were treated with sodium chloride (NaCl), abscisic acid (ABA), and indole-acetic acid (IAA) and by about 50-fold when treated with UVB light. These results indicate that ThCSD is involved in physiological pathways activated by a variety of environmental conditions.

Protective Effect of Ginsenoside R0 on Anoxic and Oxidative Damage In vitro

  • Jiang, Zhou;Wang, Yuhui;Zhang, Xiaoyun;Peng, Tao;Li, Yanqing;Zhang, Yi
    • Biomolecules & Therapeutics
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    • v.20 no.6
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    • pp.544-549
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    • 2012
  • To examine the neuroprotective effects of ginsenoside R0, we investigated the effects of ginsenoside R0 in PC12 cells under an anoxic or oxidative environment with Edaravone as a control. PC12 neuroendocrine cells were used as a model target. Anoxic damage or oxidative damage in PC12 cells were induced by adding sodium dithionite or hydrogen peroxide respectively in cultured medium. Survival ratios of different groups were detected by an AlamarBlue assay. At the same time, the apoptosis of PC12 cells were determined with flow cytometry. The putative neuroprotective effects of ginsenoside R0 is thought to be exerted through enhancing the activity of antioxidant enzymes Superoxide dismutases (SOD). The activity of SOD and the level of malondialdehyde (MDA) and intracellular reactive oxygen species (ROS), were measured to evaluate the protective and therapeutic effects of ginsenoside R0. Ginsenoside R0 treated cells had a higher SOD activity, lower MDA level and lower ROS, and their survival ratio was higher with a lower apoptosis rate. It is suggested that ginsenoside R0 has a protective effect in the cultured PC12 cells, and the protection efficiency is higher than Edaravone. The protective mechanisms of these two are different. The prevent ability of ginsenoside R0 is higher than its repair ability in neuroprotection in vitro.

Carbon Monoxide Ameliorates 6-Hydroxydopamine-Induced Cell Death in C6 Glioma Cells

  • Moon, Hyewon;Jang, Jung-Hee;Jang, Tae Chang;Park, Gyu Hwan
    • Biomolecules & Therapeutics
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    • v.26 no.2
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    • pp.175-181
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    • 2018
  • Carbon monoxide (CO) is well-known as toxic gas and intrinsic signaling molecule such as neurotransmitter and blood vessel relaxant. Recently, it has been reported that low concentration of CO exerts therapeutic actions under various pathological conditions including liver failure, heart failure, gastric cancer, and cardiac arrest. However, little has been known about the effect of CO in neurodegenerative diseases like Parkinson's disease (PD). To test whether CO could exert a beneficial action during oxidative cell death in PD, we examined the effects of CO on 6-hydroxydopamine (6-OHDA)-induced cell death in C6 glioma cells. Treatment of CO-releasing molecule-2 (CORM-2) significantly attenuated 6-OHDA-induced apoptotic cell death in a dose-dependent manner. CORM-2 treatment decreased Bax/Bcl2 ratio and caspase-3 activity, which had been increased by 6-OHDA. CORM-2 increased phosphorylation of NF-E2-related factor 2 (Nrf2) which is a transcription factor regulating antioxidant proteins. Subsequently, CORM-2 also increased the expression of heme oxygenase-1 and superoxide dismutases (CuZnSOD and MnSOD), which were antioxidant enzymes regulated by Nrf2. These results suggest that CO released by CORM-2 treatment may have protective effects against oxidative cell death in PD through the potentiation of cellular adaptive survival responses via activation of Nrf2 and upregulation of heme oxygenase-1, leading to increasing antioxidant defense capacity.

Bioaccumulation, alterations of metallothionein, and antioxidant enzymes in the mullet Mugil cephalus exposed to hexavalent chromium

  • Min, Eun Young;Ahn, Tae Young;Kang, Ju-Chan
    • Fisheries and Aquatic Sciences
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    • v.19 no.4
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    • pp.19.1-19.7
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    • 2016
  • A laboratory experiment was conducted to determine hexavalent chromium ($Cr^{6+}$) accumulation in the mullet and investigate $Cr^{6+}$ toxicity using a panel of biomarkers including metallothioneins (MTs), glutathione (GSH), glutathione S-transferase (GST), and superoxide dismutases (SODs) for 4 weeks. $Cr^{6+}$ bioaccumulation in all tissues, except muscle, was consistently time- and dose-dependent. The accumulation of $Cr^{6+}$ for 4-week exposures was in the following order: $kidney{\approx}liver$ > $intestine{\approx}gill$ > spleen > muscle. Compared with the control, $Cr^{6+}$ bioaccumulation was increased in ${\geq}200{\mu}g\;L^{-1}$ groups (P < 0.05). An independent relation was observed between accumulation factors (AFs) and exposure concentration. But AFs increased with exposure time. In the liver and gill, GST and SOD differed from the control at a high $Cr^{6+}$ concentration at 2 and 4 weeks (P < 0.05). This study indicated that the gills were as sensitive as the liver to $Cr^{6+}$ toxicity. However, the latter appeared to influence largely on the organism's adaptive response to $Cr^{6+}$, since $Cr^{6+}$ may elevate GSH and MT levels by enhancing the hepatic uptake of metal in the mullet.

Extract from Edible Red Seaweed (Gelidium amansii) Inhibits Lipid Accumulation and ROS Production during Differentiation in 3T3-L1 Cells

  • Seo, Min-Jung;Lee, Ok-Hwan;Choi, Hyeon-Son;Lee, Boo-Yong
    • Preventive Nutrition and Food Science
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    • v.17 no.2
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    • pp.129-135
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    • 2012
  • GPAR{elidium (G.) amansii is a red alga widely distributed in the shallow waters around East Asian countries. We investigated the effect of G. amansii on lipid accumulation and ROS (Reactive Oxygen Species) production in 3T3-L1 cells. G. amansii extracts dose-dependently inhibited lipid formation and ROS generation in cultured cells. Our results showed that anti-adipogenic effect of G. amansii was due to the reduction in mRNA expressions of PPAR${\gamma}$(peroxisome proliferator-activated receptor-${\gamma}$) and aP2 (adipocyte protein 2). G. amansii extracts significantly decreased mRNA levels of a ROS-generator, NOX4 (nicotinamide adenine dinucleotide phosphate hydrogen oxidase 4), and increased the protein levels of antioxidant enzymes including SOD1/2 (superoxide dismutases), Gpx (glutathione peroxidase), and GR (glutathione reductase), which can lead to the reduction of ROS in the cell. In addition, the G. amansii extract enhanced mRNA levels of adiponectin, one of the adipokines secreted from adipocytes, and GLUT4, glucose uptake protein. Taken together, our study shows that G. amansii extract inhibited lipid accumulation and ROS production by controlling adipogenic signals and ROS regulating genes.

Comparison of Physiological Activities of Radish Bud (Raphanus sativus L.) according to Extraction Solvent and Sprouting Period (추출용매 및 발아시기에 따른 무순 추출물의 생리활성 비교)

  • Han, Jin-Hee;Moon, Hye-Kyung;Chung, Shin-Kyo;Kang, Woo-Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.4
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    • pp.549-556
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    • 2015
  • This study extracted radish bud (Raphanus sativus L.) and investigated its nitrite scavenging activity, superoxide dismutases (SOD)-like activity, tyrosinase inhibition activity, xanthine oxidase inhibition activity, and angiotensin-converting-enzyme (ACE) inhibition activity according to extraction solvent and sprouting period. For nitrite scavenging activity, each extract recorded its highest level of 81.44~89.71% at pH 1.2. Radish bud extracts on sprouting days 4 and 8 showed greater scavenging activities than those on sprouting day 12 at pH 1.2 and pH 4.0. There were differences in scavenging activity according to extraction solvent based on water extract exhibiting improved scavenging activity. Ethanol extract recorded scavenging activity of 16.12% at pH 6.0, which was similar to those of ethanol and methanol radish bud extracts on sprouting day 12. SOD-like activity of radish bud extracts was in the range of 4.57~27.05%. For comparison purposes, SOD-like activity of L-ascorbic acid was 52.15%, which was higher than that of radish bud extracts. Acetone and methanol extracts showed high SOD-like activities on sprouting day 8. SOD-like activity of radish bud extracts on sprouting day 12 significantly decreased to 4.57~15.59%. Radish bud extracts recorded good tyrosinase inhibitory activities on sprouting 8 and 12, whereas methanol extracts recorded the greatest tyrosinase inhibitory activity at 62.65~84.89%. Radish bud extracts recorded xanthine oxidase inhibition activity of 21.26~29.52% on sprouting day 4, and acetone extracts showed the highest level of xanthine oxidase inhibition activity. Xanthine oxidase inhibitory activity tended to decrease with sprouting period compared early on. ACE inhibitory activity was in the range of 12.48~51.78% according to sprouting period and extraction solvent. Ethanol extracts on sprouting day 8 showed the highest ACE inhibitory activity of 51.78%. These results will hopefully contribute to research into the identification of materials and development of products for natural functional foods.

The Effects of LR3 and SP6 Acupuncture on Renal Damage in Streptozotocin-induced Diabetic Mice (태충·삼음교의 침 자극이 Streptozotocin으로 유발된 당뇨쥐의 신장 손상에 미치는 영향)

  • Lee, Cho In;Lee, Hyun Jong;Lee, Yun Kyu;Lim, Seong Chul;Kim, Jae Soo
    • Journal of Acupuncture Research
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    • v.32 no.3
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    • pp.41-51
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    • 2015
  • Objectives : This study was performed to investigate the effects of $LR_3$ and $SP_6$ acupuncture on renal damage in streptozotocin(STZ)-induced diabetic mice. Methods : ICR male mice were stabilized for a week and divided into four groups: a normal mice group(N), no-acupuncture diabetic mice group(Control), $LR_3$ acupuncture diabetic mice group($LR_3$), and $SP_6$ acupuncture diabetic mice group($SP_6$). Diabetes was experimentally induced by intraperitoneal injection of STZ(150 mg/kg) in citrate buffer(pH 4.5). For two weeks, $LR_3$ and $SP_6$ acupunctures were administered bilaterally at each point once a day. After two weeks, the animals' weight was measured and they underwent a laparotomy. Serum glucose and blood urea nitrogen(BUN) were measured from the blood taken from the heart. We measured glucose, reactive oxygen species(ROS), peroxynitrite($ONOO^-$) and thiobarbituric acid reactive substances(TBARS) in the kidney and compared expression levels of superoxide dismutases(SOD), glutathione peroxidase(GPx), nuclear factor-kappa B(NF-${\kappa}B$), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase(iNOS) and Interleukin-1 beta(IL-$1{\beta}$). Results : BUN significantly decreased in $LR_3$, $SP_6$ compared to the control group. $LR_3$ showed significantly decreased glucose compared to the control group. $LR_3$, $SP_6$ significantly decreased in ROS and $ONOO^-$ compared to the control group. $LR_3$ significantly decreased in TBARS compared to the control group. $SP_6$ significantly increased in expressions of SOD-1, catalase, and GPx compared to the control group. $LR_3$, $SP_6$ significantly decreased in COX-2 compared to the control group. $SP_6$ significantly decreased in IL-$1{\beta}$ compared to the control group. Conclusions : This study suggests that $LR_3$ acupuncture may be effective in controlling glucose and lipid peroxidation and that $SP_6$ acupuncture may have anti-oxidative and anti-inflammatory effects on renal damage in STZ-induced diabetic mice.