• Title/Summary/Keyword: Glutathione reductase

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Transformation of Orchardgrass (Dactylis glomerata L.) with Glutathione Reductase Gene (Glutathione Reductase 유전자의 도입에 의한 오차드그래스의 형질전환)

  • 이효신;배은경;김기용;원성혜;정민섭;조진기
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.21 no.1
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    • pp.21-26
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    • 2001
  • To develop transgenic orchardgrass resistant to reactive oxygen species produced from environmental stresses, a vector with the cytosolic glutathione reductase cDNA (BcGRl) from Chinese cabbage was constructed under the control of the cauliflower mosaic virus 35S promoter and was introduced into orchardgrass using Agrobacterium tumefaciens EHA101. Transgenic plants from hygromycin-selected calli of orchardgrass did not show any morphological difference from wild-type plants. The results of PCR amplification and genomic Southern blot analysis confirmed the integration of foreign gene into the chromosome of transgenic orchardgrass. Northern blot analysis with total RNA from leaves also confirmed the constitutive expression of BcGR1 in transgenic orchardgrass.

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Reductive Depolymerization of Bovine Thyroglobulin Multimers via Enzymatic Reduction of Protein Disulfide and Glutathiony­lated Mixed Disulfide Linkages

  • Liu Xi-Wen;Sok Dai-Eun
    • Archives of Pharmacal Research
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    • v.28 no.9
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    • pp.1065-1072
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    • 2005
  • The nascent thyroglobulin (Tg) multimer molecule, which is generated during the initial fate of Tg in ER, undergoes the rapid reductive depolymerization. In an attempt to determine the depolymerization process, various types of Tg multimers, which were generated from deoxy­cholate-treated/reduced Tg, partially unfolded Tg or partially unfolded/reduced Tg, were subjected to various GSH (reduced glutathione) reducing systems using protein disulfide isomerase (PDI), glutathione reductase (GR), glutaredoxin or thioredoxin reductase. The Tg multimers generated from deoxycholate-treated/reduced Tg were depolymerized readily by the PDI/GSH system, which is consistent with the reductase activity of PDI. The PDI/GSH-induced depolymerization of the Tg multimers, which were generated from either partially unfolded Tg or partially unfolded/reduced Tg, required the simultaneous inclusion of glutathione reductase, which is capable of reducing glutathionylated mixed disulfide (PSSG). This suggests that PSSG was generated during the Tg multimerization stage or its depolymerization stage. In particular, the thioredoxin/thioredoxin reductase system or glutaredoxin system was also effective in depolymerizing the Tg multimers generated from the unfolded Tg. Overall, under the net GSH condition, the depolymerization of Tg multimers might be mediated by PDI, which is assisted by other reductive enzymes, and the mechanism for depolymerizing the Tg multimers differs according to the type of Tg multimer containing different degrees and types of disulfide linkages.

Effect of Dantrolene Sodium on Tissue Sulfhydryl Groups and Glutathione in Rats (Dantrolene Sodium이 간 조직내 Sulfhydryl Group과 Glutathione에 미치는 영향)

  • Kim, Kwang-Kook;Paik, Kwang-Sea;Kang, Bok-Soon
    • The Korean Journal of Physiology
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    • v.19 no.2
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    • pp.155-160
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    • 1985
  • Dantrolene sodium(DS) is a long acting skeletal muscle relaxant which has been successfully used to control muscle spasticity in patients with various neurological disorders. However, its use is associated with hepatotoxicity. Tissue sulfhydryl group has many important roles for cellular integration and glutathione serves as a substrate for the detoxification metabolism. The purpose of this study were to investigate the effect of DS on tissue sulfhydrl group and glutathione content. Foully albino rats were divided into two groups ; saline treated (control) and DS treated groups. DS dissolved in saline was administered orally. All rats were sacrificed after 7. 14. 21 and 28 days of DS ana saline treatment by dacapitation ana liver was removed for the enzyme preparation. Total and nonprotein sulfhydryl were measured by the method of Sedlak and Lindsay (1968). Total glutathione content was assayed according to the method described by Tietze (1969) and glutathione reductase was assayed according to the method of Racker (1955), The results obtained are summarized as follows : DS administration significantly depressed the total, protein and nonprotein sulfhydryl content in liver. There were significant reduction of both total glutathione content and glutathione reductase activity in liver. On the basis of the above results it may be speculated that the toxicity of DS are well correlated with tissue sulfhydryl content and glutathione reductase activity.

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HepG2 세포의 산화적 손상에 대한 산삼 추출물의 보호효과 - DNA chip을 이용하여 -

  • Kim, Hyung-Seok;Park, Hee-Soo;Kwon, Ki-Rok
    • Journal of Pharmacopuncture
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    • v.10 no.1 s.22
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    • pp.121-135
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    • 2007
  • Objectives : This study was carried out to examine protective effect of wild ginseng extract on HepG2 human hepatoma cell line against tert-Butyl hydroperoxide (t-BHP)-induced oxidative damage. Methods : To evaluate protective effect of wild ginseng extract against t-BHP induced cytotoxicity, LDH level and activity of glutathione peroxidase and reductase were measured. Gene expression was also measured using DNA microarray. Results : Wild ginseng extract showed a significant protective effect against t-BHP-induced cytotoxicity in HepG2 cell line. It is not, however, related with the activities of glutathione peroxidase and glutathione reductase. Analysis of gene expression using DNA chip, demonstrated that 28 genes were up-regulated in t-BHP only group. Five genes - selenoprotein P, glutathione peroxidase 3, sirtuin 2, peroxiredoxin 2, serfiredoxin 1 homolog - may be related with the protective effect of wild ginseng extract. Conclusions : Based on the results, a protective effect of wild ginseng extract against t-BHP-induced oxidative damage in HepG2 cell line is not associated with the activities of glutathione peroxidase and glutathione reductase, but with the expression of selenoprotein P, glutathione peroxidase 3, sirtuin 2, peroxiredoxin 2, and serfiredoxin 1 homolog.

Health Risk Evaluation of the Vinylhouse Workers with Exposure to Pesticide (비닐하우스 재배농민의 농약에 의한 인체 위해성 연구)

  • Yang, Jae-Ho;Park, Jung-Han
    • Journal of agricultural medicine and community health
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    • v.19 no.2
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    • pp.107-118
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    • 1994
  • Health effects of pesticide among vinylhouse workers in Sangjoo County, Kyungpook Province were assessed by measuring cholinesterase, glutathion reductase, and methemoglobin. Activities of cholinesterases among vinylhouse workers and general farmers were 3.89U and 3.98U in serum and 5.29U and 5.50U in red blood cells, respectively. While levels of methemoglobin between vinylhouse workers(0.16%) and general farmers(0.17%) were very similar, glutathione reductase among vinylhouse workers were 8% lower than that of general farmers. Inhibition of RBC cholinesterase among vinylhouse workers was two times greater than general farmers, suggesting a greater exposure of vinylhouse workers to the organophosphate pesticides. Methemoglobin level among vinylhouse workers with more than or equal to 10 vinylhouse units was 13% higher than that of the workers with less than 10 units. Vinylhouse workers using protective gears during pesticide spray showed higher level of reduced glutathione and lower level of methemoglobin, as compared to the non-users. This indicates that protective gears play an important role against pesticide exposure. Vinylhouse workers practicing good personal hygiene showed a higher level of reduced glutathione, as compared to those with poor personal hygiene, indicating that personal hygiene is also an important factor in reducing pesticide exposure. The present study represents a first attempt to analyze cholinesterase, methemoglobin and glutathione reductase activities among the vinylhouse workers. While neither vinylhouse workers nor general farmers showed abnormal findings among biochemical parameters used in the present study, increased level of methemoglobin and decrease of glutathione reductase activity among vinylhouse workers suggest that these workers have been subject to a higher exposure of pesticide than general farmers.

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Melatonin Enhances Hepatic Glutathione-peroxidase Activity in Sprague-Dawley Rats

  • Kim, Choong-Yong;Yun, Choong-Soon;Park, Dae-Hun;Choi, Woo-Sung;Kim, Jin-Suk
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.2
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    • pp.221-224
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    • 1997
  • Effects of melatonin on hepatic glutathione-peroxidase (GSH-Px) and glutathione-reductase (GSH-reductase) activities were studied in Sprague-Dawley (SD) rats administered i.p. (10 mg/kg body weight) with melatonin during 15 days. The activity of cytosolic GSH-reductase in the liver was not changed by melatonin. However, melatonin injection increased significantly the activity of liver cytosolic GSH-Px activity compared with those in saline-treated rats. At the same time, plasma GSH-Px was also increased significantly in melatonin-treated rats. Since GSH-Px, a major antioxidative enzyme, removes $H_2O_2$ and lipid peroxides which are formed during lipid peroxidation from cellular membrane, such elevation of heptatic GSH-Px activity may contribute to the improvement of antioxidative effects under oxidative damage in the liver.

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Protective influence of selenium on nitrite-induced methemoglobinemia in rabbits (가토(家兎)에서 nitrite에 의한 methemoglobinemia에 미치는 selenium의 영향(影響))

  • Kim, Jin-sang;Han, Jeong-hee;Kim, Kye-soo
    • Korean Journal of Veterinary Research
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    • v.31 no.1
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    • pp.41-47
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    • 1991
  • The protective of influences of sodium selenitc ($Na_2SeO_3$) against the methemoglobinemia with sodium nitrite were investigated on hemoglobin, methemoglobin, glutathione peroxidase and NADH-methemoglobin reductase activity in rabbits which were given 0,1,3 and 9ppm sodium selenite of drinking water for a week. Dietary selenium did not alter total hemoglobin in the blood of rabbits. Selenium was found to decrease nitrite-induced methemoglobin in a dose-dependent manner. The glutathione peroxidase activity was also increased by selenium in all the experimental groups. However, the NADH-methemoglobin reductase activity by selenite did not show significant differences as concerns the methemoglobinemia. These results showed that selenium could inhibit nitrite-induced methemoglobinemia. Its influence of inhibition is suggested that the effect of the reduction of methemoglobin was greatly stimulated by glutathione peroxidase activity.

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Effect of GE-132 on the Hepatic Bromobenzene Metabolizing Enzyme System in Rats (유기게르마늄(GE-132)이 Bromobenzene의 대사계에 미치는 영향)

  • 김석환;조태현;최종원
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.6
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    • pp.702-708
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    • 1993
  • The study was attempted to elucidate the mechanism of GE-132(100mg/kg, p.o. for 6 weeks) on the metabolism of bromobenzene (460mg/kg, i.p. bid, for 2 days), which has potent carcinogenicity, mutagenicity and hepatotoxicity. It showed that activities of cytochrome p-450, aminopyrine demethylase and aniline hydroxylase, which have epoxide generating property, were not changed by GE-132 treatment. On the other hand, epoxide hydrolase was not changed but that glutathione S-transferase was significantly increased by GE-132 treatment. And also ${\gamma}-glutamylcysteine$ synthetase was not changed following the GE-132 treatment, but the activity of glutathione reductase was significantly increased. The level of hepatic glutathione which was decreased by bromobenzene recovered markedly by GE-132 pretreatment. It is concluded that the mechanism for the observed effect of GE-132 on bromobenzene metabolism is due to the induction of glutathione S-transferase.

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Transformation of Chinese Cabbage Glutathione Reductase (GR) gene into Lettuce (Lactuca sativa L.) with Particle Bombardment (유전자총을 이용한 상추 내로의 배추 Glutathione Reductase (GR)유전자의 도입)

  • 정재동;이부자;이효신;김창길
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.6
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    • pp.475-478
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    • 2000
  • The cytosolic glutathione reductase(GR) gene of chinese cabbage was introduced into Lettuce (Lactuca sativa L.) with particle bombardment method. When cotyledon explants were treated with osmoticum-conditioning medium (0.6 M sorbitol/mannitol) 4 hours prior to and 16 hours after bombardment, it was identified by GUS assay that this condition was the most efficient one for introduction of foreign genes into cotyledon tissue of lettuce with particle bombardment. The stable integration of a GR gene was confirmed by the PCR analysis. 0.3, 0.6, 1.5 kbp PCR fragments of transgenes were obtained by three types of primers designed on the basis of GR sequence. To know whether the expression of the GR gene of pBKs-GR 1 can be stably maintained in the next generation, T$_2$selfing seeds obtained from the transformed mother plants were sowed on MS medium supplemented with 200 mg/L kanamycin sulfate. 70% of seedlings showed resistance to kanamycin.

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