• Title, Summary, Keyword: CuZn-SOD

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Effects of ischemic preconditioning, KATP channel on the SOD activation and apoptosis in ischemic reperfused skeletal muscle of rat (허혈양상화와 KATP 통로가 허혈후 재관류된 흰쥐의 골격근육에서 SOD 활성 및 apoptosis에 미치는 영향)

  • Abn, Dong-choon;Paik, Doo-jin;Yang, Hong-hyun
    • Korean Journal of Veterinary Research
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    • v.39 no.5
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    • pp.878-895
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    • 1999
  • Ischemic preconditioing (IPC), i.e., a preliminary brief episode of ischemia and reperfusion, has been shown to reduce the cell damage induced by long ischemia and reperfusion. Superoxide radical which is produced during reperfusion after ischemia was recognized as a factor of the ischemic injury and it is dismutated into $H_2O_2$ and $O_2$ by two types of intracellular superoxide dismutase (SOD), Cu,Zn-SOD in cytoplasm and Mn-SOD in mitochondria. Recently oxygen free radicals are suggested to induce the apoptosis, however mechanism of the reduced apoptosis by ischemic preconditioing was unknown, while many studies performed in mammalian heart indicated that ATP-sensitive $K^+$ ($K_{APT}$) channel activation related with the protective effects. The aim of present study is to investigate 1) whether IP upregulate the Cu,Zn-SOD and Mn-SOD activities, and 2) whether ischemic preconditioning decreases apoptosis via $K_{APT}$ channel activation in timely reperfused skeletal muscle after long ishemia. The experimental animals, Sprague-Dawley rats weighing 250~300g, were divided into 8 groups; 1) control group, 2) ischemic preconditioning only groups, 3) pinacidil, a $K_{APT}$ channel opener, treatment only groups, 4) glibenclamide, a $K_{APT}$ channel blocker, treatment only groups, 5) ischemia groups, 6) ischemia after IPC groups, 7) ischemia and pinacidil treatment groups, and 8) IP and ischemia after glibenclamide pretreatment groups. Animals of the control group were administered with the vehicle (DMSO) alone. Pinacidil (1mg/kg) was administered intravenously 5 minutes after initiation of ischemia, and glibenclamide (0.5mg/kg) was injected intravenously 20 minutes before IPC. In rats that were ischemic preconditioned, the left common iliac artery was occluded for 5 minutes followed by 5 minutes of reperfusion by three times using vascular clamp. Ischemia was done by occlusion of the same artery for 4 hours. The specimens of left rectus femoris muscle were obtained immediately (0 hour), 12 hours, 24 hours after drug administrations, IP or ischemia and reperfusion. The immunoreactivities of SOD and its alterations were observed by use of sheep antihuman Cu,Zn-SOD and Mn-SOD antibodies on the $10{\mu}m$ cryosections. The incidencies of apoptosis were observed by TUNEL methods with in situ apoptosis detection kit on $6{\mu}m$ paraffine section. The results obtained were as follows : 1. After IPC, immunoreactivities of Cu,Zn-SOD mainly in the small-sized fibers were increased by 24 hours, that of Mn-SOD at 0 hour and 24 hours. 2. No significant changes in immunoreactivities of SOD was observed in the pinacidil and in the glibenclamide treatment only groups, and in the ischemia only groups. 3. The immunoreactivities of the Cu,Zn-SOD were increased in the ischemia after IPC groups and the ischemia and pinacidil treatment groups. 4. The immunoreactivities of the Cu,Zn-SOD in the IPC and ischemia after glibenclamide pretreatment groups were not increased except for the 12 hours reperfusion group. But, Mn-SOD immunoreactivities were increased in the 0 hours, 12 hours and 24 hours after reperfusion. 5. In the control group, the IPC only groups, and the pinacidil treatment only groups, negative or trace apoptotic reactions were observed, but the positive apoptotic reaction occured in the glibenclamide treatment groups. 6. Moderate or many number of apoptosis were revealed in the ischemia groups, and also the IPC and ischemia after glibenclamide pretreatment group except for 12 hours and 24 hours after reperfusion. However, the incidence of apoptosis was decreased in the ischemia after IPC groups and in the ischemia and pinacidil treatment groups. 7. There is a coincidence between the increase of Cu,Zn-SOD immunoreactivities and the decrease of apoptosis in the presence of ischemia and reperfusion. These results suggest that the protective effects of ishemic preconditioing may related to the SOD activation, and the ischemic preconditioning decreases the apoptosis partially via $K_{APT}$ channel activation in timely reperfused rat skeletal muscle. It is also suggested that inhibition of apoptosis by IPC may related with the SOD activation.

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Changes of Growth and Antioxidative Enzyme(SOD, APX, GR) Activities of Spinach Beet(Beta vulgaris var. cicla) Under Saline Condition (염 환경하에서 근대(Beta vulgaris var. cicla)의 생장과 항산화효소(SOD, APX, GR)의 활성변화)

  • 배정진;추연식;송승달
    • Journal of Life Science
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    • v.13 no.5
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    • pp.658-667
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    • 2003
  • Antioxidative enzymes (superoxide dismutase; SOD, ascorbate peroxidase; APX, glutathione reductase; GR) play major roles in scavenging mechanism of reactive oxygen species which were involved in various stress conditions including salt. In order to investigate the relation between their growth responses (dry weight) and the changes of antioxidative enzymes activity, salt-tolerant spinach beet having 15cm of shoot length were treated with various salt levels (0, 50, 200, 1000 mM NaCl) for 24 hours. Spinach beet exhibited an increase in the activity of antioxidative enzymes by salt, the maximal activity at 200 mM NaCl and the lowest activity at 50 mM NaCl in 2 hrs. after treatments. As a result of PAGE, it has been confirmed that spinach beet contained 3 isoforms (Fe-SOD, CuZn-SOD and Mn-SOD) of SOD and main isoform was CuZn- SOD form. In case of APX, isoforms of the low molecular weight(No. 7, 8) were showed strong expression especially at 200 and 400 mM NaCl treatment. Meanwhile, GR did not show specific pattern of isoforms among the salt treatments. Especially, in case of 50 mM treatment, plant showed the lowest activity of SOD with the best growth, a low enzyme activity was induced by inactivation of the Mn-SOD. Therefore, we suggested that the decrease of SOD activity at a low salt level (50 mM NaCl) or the increase of enzyme activity at a high salt level (200 mM NaCl) may be related to expression of the Mn-SOD isoform. These antioxidative enzymes showed the increase of activity in a short time by salt addition. So, it is considered that spinach beet copes effectively with a stressful condition such as salt by operating effective antioxidative defense mechanism rapidly under high salt level.

Antioxidant Effect of Rubi Fructus on TM4 Sertoli Cells (남성생식세포 Sertoli cell에 미치는 복분자(覆盆子)의 항산화 효과)

  • Kim, Young Joo;Chang, Mun Seog;Park, Seong Kyu
    • Herbal Formula Science
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    • v.26 no.2
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    • pp.103-111
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    • 2018
  • Objectives : The purpose of this study was to examine the antioxidant effects of the extract of Rubi Fructus on TM4 Sertoli cells. Methods : The extract was studied for diphenyl-picryl-hydrazyl (DPPH) radical scavenging activity and cell viability assays on Sertoli cells. In addition, hydrogen peroxide-induced oxidative stress on Sertoli cells were examined by MTT assay. The antioxidant enzyme of Cu/Zn SOD, Mn SOD, catalase protein expression on Sertoli cells were also measured. Results : The results showed that the extract scavenged DPPH radical dose-dependent manner. The extract showed no cytotoxicity at concentration of 1, 5, 10, 50, $100{\mu}g/ml$. The hydrogen peroxide-induced cytotoxicity of Sertoli cells was protected to 88.3% by the extract at concentration of $100{\mu}g/ml$. Cu/Zn SOD and Mn SOD protein expression were significantly increased on Sertoli cells, but catalase protein expression was not significantly changed. Conclusions : In conclusion, the extract of Rubi Fructus has antioxidant effects on Sertoli cells and protect male reproductive system against oxidative stress.

Cloning and Characterization of the Cu,Zn Superoxide Dismutase (SOD1) cDNA from the Spider, Araneus ventricosus

  • Choi Young Soo;Choo Young Moo;Li Jianhong;Sohn Hung Dae;Jin Byung Rae
    • International Journal of Industrial Entomology
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    • v.10 no.1
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    • pp.73-77
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    • 2005
  • A Cu,Zn superoxide dismutase (SOD1) cDNA was cloned from the spider, Araneus ventricosus. The A. ventricosus SOD1 (AvSOD1) cDNA contains an open reading frame of 495 bp encoding 165 amino acid polypeptide with a predicted molecular mass of 17,114 Da and pI of 6.55, and possesses the typical metal binding ligands of six histidines and one aspartic acid common to SOD1s. The deduced amino acid sequence of the AvSOD1 cDNA showed $51\%$ identity to Ceratitis capitata SOD1, and $50\%$ to SOD1 sequences of both Drosophila melanogaster and Chymomyza amoena. Northern blot analysis revealed the presence of AvSOD1 transcripts in all tissues examined.

Effects of zinc bearing palygorskite supplementation on the growth performance, hepatic mineral content, and antioxidant status of broilers at early age

  • Yang, Weili;Chen, Yueping;Cheng, Yefei;Wen, Chao;Zhou, Yanmin
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.7
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    • pp.1006-1012
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    • 2017
  • Objective: This study was conducted to investigate effects of zinc (Zn) bearing palygorskite (ZnPal) supplementation on growth performance, hepatic mineral content, and antioxidant status of broilers at early age. Methods: A total of 240 1-day-old Arbor Acres broiler chicks were allocated into 5 treatments with 6 replicates of 8 chicks each. Birds in 5 treatments were fed a basal diet supplemented with 0 (Control group; Analyzed Zn content: 81 mg/kg), 20, 40, 60, and 80 mg/kg Zn as ZnPal for 21 days, respectively. Blood, liver and intestinal mucosa were collected at 21 days of age. Results: Treatments did not affect growth performance of broilers during the 21-day study (p>0.05). The contents of hepatic Zn and magnesium (Mg) were linearly increased (p<0.001) by ZnPal supplementation. ZnPal inclusion linearly (p = 0.007) reduced malondialdehyde (MDA) concentration in serum. The activity of total superoxide dismutase (T-SOD) in liver increased linearly (p = 0.001) with concentration of ZnPal in diet. ZnPal inclusion linearly (p = 0.036) and quadratically (p = 0.005) increased T-SOD activity, and linearly (p = 0.012) increased copper/zinc superoxide dismutase (Cu/Zn SOD) activity in jejunal mucosa. The maximum responses of hepatic and jejunal antioxidant enzymes activities (T-SOD and Cu/Zn SOD) were found when supplementing the basal diet with 60 mg/kg Zn as ZnPal. Furthermore, ZnPal supplementation quadratically (p = 0.001) increased Cu/Zn SOD activity in ileal mucosa, and its maximum activity was observed in the diet supplemented with 20 mg/kg Zn as ZnPal. Conclusion: ZnPal supplementation did not alter growth performance of broilers. Dietary ZnPal inclusion could increase concentrations of hepatic trace minerals (Zn and Mg) and inhibit lipid peroxidation by reducing serum MDA accumulation, with the optimal dosage of Zn from ZnPal being 80 mg/kg diet (analyzed Zn content in the diet: 165 mg/kg), and 60 mg/kg Zn as ZnPal (analyzed Zn content in the diet: 148 mg/kg) was the optimum dosage for broilers to achieve maximum antioxidant enzyme activities.

Induction of antioxygenic enzymes as defense systems in plant cells against low temperature stress : (II) $Mn^{+2}-induced$ SOD activation and enhancement of cold tolerance in rice seedlings (식물의 냉해에 대한 생체방어기구로서 항산소성 효소의 유도 : (II) $Mn^{+2}$이온에 의한 세포내 SOD의 활성화와 벼 유묘의 내냉성 향상)

  • Hahn, Chang-Kyun;Kim, Jong-Pyung;Jung, Jin
    • Applied Biological Chemistry
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    • v.34 no.2
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    • pp.168-173
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    • 1991
  • The uptake of $Mn^{+2}$, a metal cofactor Mn-SOD, by rice seedings resulted in not only a substantial increase in SOD activity in leaf tissues of the plants, but also a significant enhancement of their cold tolerance : the relative extent of the cold tolerance appeared to accord with relative level of the SOD activity. In contrast, $Fe^{+3},\;Cu^{+2}$ and $Zn^{+2}$, which are the cofactors of Fe-SOD and Cu/Zn-SOD, were found to be ineffective for increasing the SOD activity as well as for improving the chilling-resistant capacity of the plants. The results suggest that Mn-SOD, which is most likely induced by its substrate(superoxide) and activated by the presence of $Mn^{+2}$a at high level, is the enzyme acting as an active component of the defense system against low temperature stress in rice plants. In addition, the application of abscisic acid which has been know to protect to some extent certain plants from chilling injury brought about an increase in SOD activity in rice tissues, providing another affirmative information for the crucial role of SOD under the circumstance of cold stress in plants.

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Evaluation of Zinc and Copper Status in Korean College Women (일부 여대생의 구리와 아연 영양상태 평가)

  • 김정혜
    • Journal of Nutrition and Health
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    • v.32 no.3
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    • pp.277-286
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    • 1999
  • This study was conducted to assess dietary intake and nutritional status of zinc and copper in Korean college women. Dietary survey was conducted by 24-hour recall method and fasting serum samples were collected from 111 apparently healthy subjects. Intake levels of zinc and copper were calculated using newly developed database for Zn & Cu of Korea food. Serum levels of Zn, Cu and activities of ALP, EC-SOD were measured from fasting serum sample. Mean daily zinc and copper intakes were 6.72mg/day(56.0% RDA) and 1.11mg/day respectively. Mean values of serum ALP activity, zinc and copper concentration were 43.9U/L, 14.8umol/1, 15.5umol/1and these values were mostly within normal range. EC-SOD activitis of the subjects were low and had no correlation with intake or serum levels of Zn, Cu. In conclusion, these results show that zinc and copper intake of Koran college women are lower than those from other counties but higher than those of adults in rural area of Korea. Their serum levels of Zn, Cu, ALP are relatively normal. These results indicate that marginal deficiency of Zn and Cu may be quite prevalent in these subjects but serum indicators measured may not be sensitive enough to detect such marginal deficiency. Further study in needed to develop a biochemical index sensitive enough to evaluate Zn and Cu status.

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Manipulation of Antioxidative Mechanism in Chloroplasts

  • Kwon, Suk-Yoon;Lee, Haeng-Soon;Kwak, Sang-Soo
    • Proceedings of the Botanical Society of Korea Conference
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    • pp.79-84
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    • 1999
  • Oxidative stress is one of the major environmental stresses to plants. Reactive oxygen species (ROS) generated during metabolic processes damage cellular functions and consequently lead to cell death. Fortunately plants have in vivo defense system by which the ROS is scavenged by enzymes such as superoxide dismutase (SOD) and ascorbate peroxidase (APX). In attempts to understand the protection mechanism of plant against oxidative stress, we developed transgenic tobacco (Nicotiana tabacum cv. Xanthi) plansts thet expressed both SOD and APX in chloroplast using Agrobacterum-mediated transformation and evaluated their protection capabilities against methyl viologen (MV, paraquat) -mediated oxidative damage. Three double transformants (CAI, CA2, and CA3) expressed the chimeric CuZnSOD and chimeric APX in chloroplast, and one transformant (AM) expressed the chimeric APX and chimeric MnSOD in chloroplast. In addition, we obtained three lines of transformants (C/Al, C/A2, and A/C) that expressed the APX and SOD than control plants, and more resistant to oxidative stress caused by MV. TRansformants (C/A and A/C) overexpressing MnSOD, CuZnSOD and APX at the same time showed the highest resistance to MV-mediated oxidative stress among the transformants.

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Expression of Lily Chloroplastic Cu,Zn Superoxide Dismutase Enhances Resistance to Erwinia carotovora in Potatoes

  • Kim, Mi-Sun;Kim, Hyun-Soon;Kim, Yoon-Sik;Baek, Kwang-Hyun;Moon, Jae-Sun;Choi, Do-Il;Joung, Hyouk;Jeon, Jae-Heung
    • The Plant Pathology Journal
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    • v.23 no.4
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    • pp.300-307
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    • 2007
  • Previously, a chloroplast-localized Cu,Zn superoxide dismutase (chCu,ZnSOD) was isolated from lily and the sense- and antisense- sequences of the lily chCu,ZnSOD were used to transform potato plants. Two selected lines, the sense- and anti-sense strand of transgenic plants, were further characterized for resistance to Erwinia carotovora, which is a severe pathogen affecting potato plants. Only the sense-strand transgenic potato, which contained less $O_2^{.-}$ and more $H_2O_2$ than wild-type and antisense-strand transgenic plants, showed increased resistance to E. carotovora. Additional studies using $O_2^{.-}$ or $H_2O_2$ scavengers in wild-type, sense-strand, and antisense-strand transgenic plants suggest that resistance to E. carotovora is induced by reduced $O_2^{.-}$ and is not influenced by $H_2O_2$. To the best of our knowledge, this report is the first study suggesting that resistance to E. carotovora is enhanced by reduced $O_2^{.-}$, and not by increased amounts of $H_2O_2$.

Protective Effects of Antoxidant Enzymes of Candida albicans against Oxidative Killing by Macrophages

  • Kim, Hye-Jin;Na, Byoung-Kuk;Kim, Moon-Bo;Park, Duk-Young;Song, Chul-Yong
    • Journal of Microbiology
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    • v.37 no.2
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    • pp.117-122
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    • 1999
  • Protective roles of antioxidant enzymes, copper-zinc superoxide dismutase (CuZnSOD), manganese superoxide dismutase (MnSOD), and catalase of Candida albicans against exogenous reactive oxygens and oxidative killing by macrophages were investigated. The initial growth of C. albicans was inhibited by reactive, oxygen-producing chemicals such as hydrogen peroxide, pyrogallol, and paraquat, but it was restored as the production of antioxidant enzymes were increased. The growth inhibition of C. albicans by reactive, oxygen-producing chemicals was reduced by treating the purified candidal SOD and catalase. Also, in the presence of SOD and catalase, the oxidative killing of C. albicans by macrophages was significantly inhibited. These results suggest that antioxidant enzymes, CuZnSOD, MnSOD, and catalase of C. albicans may play important roles in the protection of C. albicans not only from exogenous oxidative stress but also from oxidative killing by macrophages.

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