• Title, Summary, Keyword: SOD

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Expression of a Cu-Zn Superoxide Dismutase Gene in Response to Stresses and Phytohormones in Rehmannia Glutinosa

  • Park, Myoung-Ryoul;Ryu, Sang-Soo;Yoo, Nam-Hee;Yu, Chang-Yeon;Yun, Song-Joong
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.5
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    • pp.270-275
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    • 2005
  • Superoxide dismutases (SOD) are metalloenzymes that convert $O_2^-\;to\;H_2O_2$. Rehmannia glutinosa is highly tolerant to paraquat-induced oxidative stress. The primary objective of this study was to characterize regulation of SOD gene expression in R. glutinosa in response to oxidative stresses and hormones. A full-length putative SOD clone (RgCu-ZnSOD1) was isolated from the leaf cDNA library of R. glutinosa using an expressed sequence tag clone as a probe. RgCu-ZnSOD1 cDNA is 777 bp in length and contains an open reading frame for a polypeptide consisted of 152 amino acid residues. The deduced amino acid sequence of the clone shows highest sequence similarity to the cytosolic Cu-ZnSODs. The two to three major bands with several minor ones on the Southern blots indicate that RgCu-ZnSOD1 is a member of a small multi-gene family. RgCuZnSOD1 mRNA was constitutively expressed in the leaf, flower and root. The expression of RgCu-ZnSOD1 mRNA was increased about 20% by wounding and paraquat, but decreased over 50% by ethylene and $GA_3$. This result indicates that the RgCu-ZnSOD1 expression is regulated differentially by different stresses and phytohormones at the transcription level. The RgCu-ZnSOD1 sequence and information on its regulation will be useful in investigating the role of SOD in the paraquat tolerance of R. glutinosa.

Fenofibrate decreases radiation sensitivity via peroxisome proliferator-activated receptor ${\alpha}$-mediated superoxide dismutase induction in HeLa cells

  • Liu, Xianguang;Jang, Seong-Soon;An, Zhengzhe;Song, Hye-Jin;Kim, Won-Dong;Yu, Jae-Ran;Park, Woo-Yoon
    • Radiation Oncology Journal
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    • v.30 no.2
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    • pp.88-95
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    • 2012
  • Purpose: The fibrates are ligands for peroxisome proliferator-activated receptor (PPAR) ${\alpha}$ and used clinically as hypolipidemic drugs. The fibrates are known to cause peroxisome proliferation, enhance superoxide dismutase (SOD) expression and catalase activity. The antioxidant actions of the fibrates may modify radiation sensitivity. Here, we investigated the change of the radiation sensitivity in two cervix cancer cell lines in combination with fenofibrate (FF). Materials and Methods: Activity and protein expression of SOD were measured according to the concentration of FF. The mRNA expressions were measured by using real time reverse-transcription polymerase chain reaction. Combined cytotoxic effect of FF and radiation was measured by using clonogenic assay. Results: In HeLa cells total SOD activity was increased with increasing FF doses up to 30 ${\mu}M$. In the other hand, the catalase activity was increased a little. As with activity the protein expression of SOD1 and SOD2 was increased with increasing doses of FF. The mRNAs of SOD1, SOD2, $PPAR{\alpha}$ and $PPAR{\gamma}$ were increased with increasing doses of FF. The reactive oxygen species (ROS) produced by radiation was decreased by preincubation with FF. The surviving fractions (SF) by combining FF and radiation was higher than those of radiation alone. In Me180 cells SOD and catalase activity were not increased with FF. Also, the mRNAs of SOD1, SOD2, and $PPAR{\alpha}$ were not increased with FF. However, the mRNA of $PPAR{\gamma}$ was increased with FF. Conclusion: FF can reduce radiation sensitivity by ROS scavenging via SOD induction in HeLa. SOD induction by FF is related with $PPAR{\alpha}$.

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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Comparison of enzyme activities of the native and N-terminal 6xHis-tagged Fe supreoxide dismutase from Streptomyces subrutilus P5 (Streptomyces subrutilus P5의 천연 Fe superoxide dismutase와 N-말단 6xHis-태그가 결합된 Fe superoxide dismutase의 활성비교)

  • Park, Joong-ho;Kim, Jae-heon
    • Korean Journal of Microbiology
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    • v.52 no.2
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    • pp.230-235
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    • 2016
  • This study was carried out to analyze the differences in enzyme activity and stability between the native Fe superoxide dismutase (FeSOD) and the 6xHis-tagged superoxide dismutase (6xHis-FeSOD) of Streptomyces subrutilus P5. The optimum pHs for both native FeSOD and 6xHis-FeSOD were 7, while the pH range of the activity was narrower for the 6xHis-FeSOD. The native FeSOD was stable at pH 4-9, but the 6xHis-FeSOD lost its stability at pH > 9. The temperatures of the optimum activities were same for both types of enzymes. However, the heat stability of the 6xHis-FeSOD was clearly decreased; even at $20^{\circ}C$ the enzyme lost the activity after 360 min. In contrast, the native FeSOD was stable after 720 min at below $40^{\circ}C$. $H_2O_2$ inhibition was occurred already at 0.5 mM for the 6xHis-tagged enzyme. Therefore, from the results that the 6xHis-FeSOD retained the enzyme activity at pH 6-7 and $20-40^{\circ}C$, it can be assumed that the protein structure became destabilized under different storage conditions and sensitive to the enzyme inhibitor.

Bidirectional Regulation of Manganese Superoxide Dismutase (MnSOD) on the Radiosensitivity of Esophageal Cancer Cells

  • Sun, Guo-Gui;Hu, Wan-Ning;Wang, Ya-Di;Yang, Cong-Rong;Lu, Yi-Fang
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3015-3023
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    • 2012
  • The mitochondrial antioxidant protein manganese superoxide dismutase (MnSOD) may represent a new type of tumor suppressor protein. Overexpression of the cDNA of this gene by plasmid or recombinant lentiviral transfection in various types of cancer leads to growth suppression both in vitro and in vivo. We previously determined that changes in MnSOD expression had bidirectional effects on adriamycin (ADR) when combined with nitric oxide (NO). Radiation induces free radicals in a manner similar to ADR, so we speculated that MnSOD combined with NO would also have a bidirectional effect on cellular radiosensitivity. To examine this hypothesis, TE-1 human esophageal squamous carcinoma cells were stably transfected using lipofectamine with a pLenti6-DEST plasmid containing human MnSOD cDNA at moderate to high overexpression levels or with no MnSOD insert. Blastidicin-resistant colonies were isolated, grown, and maintained in culture. We found that moderate overexpression of MnSOD decreased growth rates, plating efficiency, and increased apoptosis. However, high overexpression increased growth rates, plating efficiency, and decreased apoptosis. When combined with NO, moderate overexpression of MnSOD increased the radiosensitivity of esophageal cancer cells, whereas high MnSOD overexpression had the opposite effect. This finding suggests a potential new method to kill certain radioresistant tumors and to provide radioresistance to normal cells.

Nucleus-DNA Damage and Different Response of Plant Cells to Paraquat in Relation to Enzyme Activity of Superoxide Dismutase. (Superoxide dismutase의 활성차이에 따른 식물세포의 paraquat에 대한 반응과 핵 DNA 손상 검정)

  • 권순태;이명현;오세명;정도철;김길웅
    • Journal of Life Science
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    • v.14 no.4
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    • pp.614-619
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    • 2004
  • This study was undertaken to investigate the different responses of cultured plant cells to paraquat treatment and nucleus-DNA damage in relation to enzyme activity of superoxide dismutase (SOD). Furthermore, this study was also carried out to understand the antioxidative mechanism of plant cells to environmental stress. We selected two different species of plant cultured cells, Ipomoea batatas as high-SOD species and Lonicera japonica as low-SOD species. The total activity and specific activity of SOD in a chlorophyllous cell of I. batatas were 3,736 unit/gㆍfresh weight and 547 unit/mgㆍprotein, respectively, and those in L. japonica were 23 unit/gㆍfresh weight and 13 unit/mgㆍprotein, respectively SOD activity in chlorophyllous I. batatas cells reached its maximum level at 10 to 15 days after subculture, whereas that in L. japonica remained at a very low SOD level during the whole period of subculture. In comparison to L. japonica, I. batatas, a high-SOD species, showed high tolerance to paraquat 10 and 50 mg/l treatment in terms of cell viability and electrolyte leakage. Based on the result of comet assay, the nucleus-DNA damage of two species by paraquat 50 mg/l treatment was not significantly different. However, I. batatas cells repaired their damaged DNA more effectively than the cells of the low-SOD species, L. japonica.

Enhancement of potency and stability of human extracellular superoxide dismutase

  • Kim, Sunghwan;Kim, Hae-Young;Kim, Jung-Ho;Choi, Jung-Hye;Ham, Won-Kook;Jeon, Yoon-Jae;Kang, Hara;Kim, Tae-Yoon
    • BMB Reports
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    • v.48 no.2
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    • pp.91-96
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    • 2015
  • Cells express several antioxidant enzymes to scavenge reactive oxygen species (ROS) responsible for oxidative damages and various human diseases. Therefore, antioxidant enzymes are considered biomedicine candidates. Among them, extracellular superoxide dismutase (SOD3) had showed prominent efficacy against asthma and inflammation. Despite its advantages as a biomedicine, the difficulty in obtaining large quantity of active recombinant human SOD3 (rhSOD3) has limited its clinical applications. We found that a significant fraction of over-expressed rhSOD3 was composed of the inactive apo-enzyme and its potency against inflammation depended on the rate of metal incorporation. Also, purified rhSOD3 was unstable and lost its activity very quickly. Here, we suggest an ideal preparative method to express, purify, and store highly active rhSOD3. The enzymatic activity of rhSOD3 was maximized by incorporating metal ions into rhSOD3 after purification. Also, albumin or polyethylene glycol prevented rapid inactivation or degradation of rhSOD3 during preparative procedures and long-term storage.

Oxygen Toxicity of Superoxide Dismutase-Deficient Saccharomyces cerevisiae by Paraquat (Paraquat에 의해 유도된 Superoxide Dismutase 결핍 효모의 산소 독성)

  • 김지면;남두현용철순허근
    • KSBB Journal
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    • v.10 no.5
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    • pp.561-567
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    • 1995
  • Using superoxide dismutase (SOD)-deficient mutants of Saccharomyces cerevisiae, the oxygen toxicity induced by paraquat was studied. In aerobic culture condition, yeasts lacking MnSOD (milochondrial SOD) showed more significant growth retardation than CuZnSOD (cytoplasmic SOD)-deficient yeasts. However, not so big differences in growth pattern of those mutants compared with wild type were observed under anaerobic condition. When exposed to paraquat, the growth of yeasts lacking CuZnSOD was severely affected by higher than 0.01mM of paraquat in culture medium. By the analysis of several cellular components ivolved in free radical generating and scavenging system, it was found that, under aerobic condition, the content of lipid peroxides in cell membrane as well as cellular activity of glutathion peroxidase of CuZnSOD-deficient mutants was increased in the presence of paraquat, although significant decrease of catalase activity was observed in those stratns. In MnSOD-deficient yeast, however, increment in cellular activity of glutathion peroxldase and catalase by paraquat was observed without any deterioration of membrane lipid. It implies that the lack of mitochondrial SOD could be compensated by both of glutathion peroxldase and catalase, but that only glutathion peroxidase might act for CuZnSOD in cytoplasm. In contrast, all of SOD-deficient mutants showed a significant decrease in catalase activity, but slight increase in the activities of glutathion peroxidase, when cultivated anaerobically in the medium containing paraquat. Nevertheless, any significant changes of lipid peroxides in cell membranes were not observed during anaerobic cultivation of SOD-deficient mutants. It suggests that a little amount of free radicals generated by paraquat under anaerobic condition could be sufficiently overcome by glutathion peroxidase but not by catalase.

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Resistance Functions of Woody Landscape Plants to Air Pollutants (I) - SOD Activity - (조경수목(造景樹木)의 대기오염물질(大氣汚染物質)에 대한 방어기능(防禦機能) (I) - SOD 활성(活性)을 중심으로 -)

  • Kim, Myung Hee;Lee, Soo Wook
    • Journal of Korean Society of Forest Science
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    • v.81 no.2
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    • pp.164-176
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    • 1992
  • This study was conducted to determine the toxic effects of air pollutants on landscaping trees, Pinus densiflora, Pinus koraierasis, Ginkgo biloba, Liriodeytdron tulipifera, Platanus occidentalis and their resistance to the pollutant toxicity in urban and industrial regions of Seoul and Taejon, Korea. Total sulfur content and superoxide dismutase activity were analysed in tree foliage of Pinus densiflora, Pinzes koraiensis, Ginkgo biloba, Liriodendron tulipifera, Platanus occidentalis. In addition, SOD activity was analyzed in the foliage of tree seedlings, i.e. Pinus densijlora, Pinus koraiensis, Ginkgo biloba, Liriodendron tulipifera, with the lurnigation of $SO_2$ in gas chamber 4 hours a day for six days. In all species total sulfur content and SOD activity had a positive correlation. Air pollutants accumulated in tree tissues were supposed to enhance the enzyme activity like SOD providing with the resistance mechanisms. Trees under the air pollution stress increased enzyme activity to develop internal self-resistance against pollutants, but after a critical point enzyme-activity decreased gradually and resulting in injury after all, Deciduous trees had greater filtration capacity than conifers and coniferous trees showed greater resistance against air pollutants than deciduous species. Foliage SOD activity was higher in polluted area than in unpolluted area for most species. Coniferous species and mature trees had higher SOD activity than deciduous seedlings. Especially Pinus koraiensis, Ginkgo biloba and Plcatanus occidentalis had higher SOD activity than other species. The tree species with the high SOD activity showed strong resistance against air pollutants. In 2nd-year needles of Pinus densiflora seedlings and current and 2nd-year needles of Pinus koraiensis seedlings containing high native SOD activity, SOD activity increased with the increase of $SO_2$ level. But in seedlings containing low native SOD activity, SOD activity increased at 0.5ppm $SO_2$ level while it decreased at 1.5 and 2.5ppm $SO_2$. Changes of SOD activity was different between species and in most species SOD seemed to participate in resistance mechanism.

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Role of Inducibility of Superoxide Dismutases and Metallothionein of Mouse Lungs by Paraquat in Aging (Paraquat에 의한 생쥐 폐의 Superoxide Dismutases와 Metallothionein의 유도능과 노화와의 관계)

  • Lee, Tae-Bum;Park, Yoo-Hwan;Choi, Cheol-Hee
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.5
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    • pp.579-590
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    • 2001
  • Background : The aging process may be induced, at least in part, by reactive oxygen species(ROS). It has been thought that the lung could be a good source of ROS because it has a high oxygen tension. In the present study, we invetigated the inducibility of the first and last lines against oxidative stress, superoxide dismutases(CujZn-SOD and Mn-SOD) as a scavenger of ${O_2}^-\;{\cdot}$ and metallothionein(MT) as a scavenger of $OH{\cdot}$, respectively, in mouse lungs with age. Methods : Oxidative stress was induced by paraquat, an intracellular superoxide generator, at 1, 4, 8, and 12 months of age and then SODs and MT mRNAs were determined by RT-PCR method. Results : The steady-state level of Mn-SOD mRNA increased from 1 to 8 months but decreased thereafter. However, Mn-SOD mRNA was not induced by paraquat after 1 month. On the other hand, there was no change in the steady-state level of Cu/Zn-SOD mRNA, which decreased abruptly at 12 months of age. Additionally, Cu/Zn-SOD mRNA was not induced by paraquat at any age. There was no change in the steady-state level of MT mRNA with age whereas its inducibility by paraquat was intact at all ages. Conclusion : These results indicate that lack of induction of SODs with age may be one of the causative factors in the aging process while induction of MT may play an important role in the defense against oxidative stress. It is therefore implicated that the tissue antioxidant/prooxidant balance could be one of determinants of mean life span.

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