• Title, Summary, Keyword: GSH

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Effects of Ethanol Administration on Glutathione and Lipid Peroxide Levels in Rat Liver and Cerebellum (에탄을 공급이 흰쥐 조직중의 Glutathione 및 지질산화 수준에 미치는 영향)

  • 이정원
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.20 no.4
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    • pp.285-292
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    • 1991
  • The effects of acute and chronic ethanol administration on hepatic and cerebellar glutathione (GSH) statuses and lipid peroxide levels in rats were investigated. In the liver, chronic ethanol feeding (6.9 g/kg, per day) as 10% (v/v) drinking water for 4 weeks produced a slight decrease of total GSH and an increase in the ratio of GSSG/total GSH without change of GSSG (oxidized GSH). Lipid peroxide level however was not modified. Many other studies have shown the acute ethanol loading effect in the rat liver, that is moderate decrease of total GSH and elevation of lipid peroxide level. Relating to this, it was observed that total GSH in the plasma obtained from post. hepatic inferior vena cava was increased by acute ethanol injection (50 mmol/kg, i.p.). This increased hepatic efflux of GSH into blood, in addition to the promoted antioxidative utilization of GSH, could be suggested as one of the possible reasons for the decrease of hepatic GSH induced by ethanol load. In the cerebellum, acute ethanol load did not change the total GSH and GSSG, but increased the lipid peroxidation rate. In the chronic, neither GSH pattern nor lipid peroxidation rate was changed.

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Cloning of Genes for the Biosynthesis of Glutathione from E. coIi K-12 (E.coli K-12 균주로부터 글루타치온 합성 유전자의 클로닝)

  • 남용석;박영인;이세영
    • Microbiology and Biotechnology Letters
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    • v.19 no.6
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    • pp.575-582
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    • 1991
  • To increase the production of glutathione by the expression of recombinant gsh plasmids, two genes responsible for the biosynthesis of glutathione were isolated and cloned. To clone a gshI gene, the GS903 mutant strain, which is deficient in $\gamma$-glutamylcysteine synthetase activity, has been raised. A gshI gene was cloned using pBR322 plasmid as a 3.6 Kb PstI DNA fragment isolated from E. coli K-12 chromosomal DNA. Also a gshIl gene was cloned using pUC13 plasmid as a 2.2 Kb PstI-BamHI DNA fragment. To study the effects of plasmid copy number and passenger DNA size on the expression levels of the gsh genes, various recombinant plasmids containing different sets of genes were constructed. The expression levels of the gsh genes were increased approximately twice higher in pUC series plasmids than that in pBR322 plasmid. But the sizes of the passenger DNA containing the gsh genes in the vector plasmid did not affect on the expression levels of the gsh genes.

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Separation of Glutathione by Ion Exchange Chromatography (이온교환 크로마토그래픽을 이용한 Glutathione 분리)

  • 김정훈;손영종;구윤모
    • KSBB Journal
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    • v.13 no.1
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    • pp.77-82
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    • 1998
  • Glutathione(L-$\gamma$ -glutamyl-L-cysteinylglycine, GSH) produced by microbial enzymes was separated by a liquid chromatography. In order to select a resin which would bind GSH efficiently, a batch adsorption experiment was carried out with GSH solution and various resins at pH 8.0 GSH bound to Q-sepharose and QAE-sephadex among anion exchange resins, but the latter was found not to be suitable because of the reduction of resin volume at high salt concentration. Preliminary experiments using a standard solution were carried out to separate GSH. GSH and $\gamma$ -glutamylcysteine were separated from the other constituents by applying step gradient of salt(NaCl) concentration. GSH was successfully separated from $\gamma$ -glutamylcysteine by applying Tris buffer containing 35mM NaCl. Chromatographic separation behaviors for the enzymatic product was similar to that for the standard solution. Separation yields of GSH from the standard solution and enzymatic product solution were 72.6% and 84.4%, respectively.

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Selective Extraction and Quantification of Glutathione using Maleimide-Presenting Gold Nanoparticles

  • Oh, Hongseok;Lee, Jeongwook;Yeo, Woon-Seok
    • Bulletin of the Korean Chemical Society
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    • v.35 no.10
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    • pp.3047-3051
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    • 2014
  • In this paper, we describe a new method for the selective extraction and quantification of glutathione (GSH) using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and maleimide-presenting gold nanoparticles (Mal-AuNPs). Our strategy utilizes the Michael addition to selectively extract GSH, from chosen samples, onto the maleimide of Mal-AuNPs. After the extraction step, the GSH bound to the AuNPs was analyzed by MALDI-TOF MS in the presence of an internal standard which was prepared by reacting Mal-AuNPs with isotope-labeled GSH ($GSH^*$). The $GSH^*$ has the same structure as GSH but a higher molecular weight, and therefore, enables absolute quantification of GSH by comparing the mass signal intensities of the GSH- and $GSH^*$-conjugated alkanethiols. Our strategy was verified by analyzing GSH-spiked fetal bovine serum and NIH 3T3 cells.

Gene Expression of Glutathione Peroxidase in $Saccharomyces$ $cerevisiae$ Treated with $N$-acetyl-L-cysteine and Gamma-rays ($Saccharomyces$ $cerevisiae$에서 $N$-acetyl-L-cysteine 처리와 감마선 조사에 따른 Glutathione Peroxidase 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.4
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    • pp.258-264
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    • 2011
  • Glutathione (GSH) has important roles in cellular defense against oxidative stress, 1) direct scavenging of reactive oxygen species (ROS), and 2) coenzyme of ROS scavenging enzyme like glutathione peroxidases (GPx). GSH peroxidase reduces free hydrogen peroxide to water using 2GSH. $N$-acetyl-L-cysteine (NAC), one of the antioxidants, is used as a precursor for intracellular GSH. In this study, relation of GSH, NAC, and GSH peroxidase was investigated through transcriptional expression of $GPX1$ and $GPX2$, which are GSH peroxidase encoding genes, in yeast cells treated with 0 mM to 20 mM of NAC or in combination with 100 Gy gamma-rays. The transcriptional expression of $GPX1$ and $GPX2$ was induced by NAC and 100 Gy gamma-rays. The gene expression of both GSH peroxidases was decreased with increasing concentrations of NAC in irradiated yeast cells. These results suggest that elevation of intracellular GSH by NAC and oxidative stress and ROS generated from gamma-rays induces expression of GSH peroxidase genes, and that NAC can protect the yeast cells against ROS generated from gamma-rays through direct scavenging of ROS and transcriptional activation of GSH peroxidase.

Construction Various Recombiant Plasmids for the Enhancement of Glutathione Production in E. coli. (E. coli에서 글루타치온 생산 증가를 위한 재조합 플라스미드의 구성)

  • 남용석;이세영
    • Journal of Life Science
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    • v.7 no.4
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    • pp.253-261
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    • 1997
  • In order to enhance glutathione production, various recombinant plasmids containing gshI and/or gshII genes isolated from E. coli K-12 were constructed and introduced into E. coli. Some plasmids contained one to three copies of gshI genes in pBR325 and others contained both gshI and genes for glutathione biosynthesis. $\gamma$-Glutamylcysteine synthetase activities of E, coli strains amplified tandem repeated gshI genes were dependent on the number of inserted gshI genes. The glutathione productivity of E. coli strains harboring various plasmids was investigated using an E. coli acetate kinase reaction as an ATP regenerating system. The glutathione productivity of E. coli strains harboring tandem repeated gshI genes was increased in proportion to the number of inserted gshI genes. By the introduction of gshII gene, the glutathione productivity of the E. coli was increased by two-fold compared with E. coli strain amplified gshI gene only. The enzymatic production of glytathione in E. coli was mainly affected by the increase of $\gamma$-glutamylcysteine synthetase activity. The highest glutathione productivity was obtained in E. coli strains harboring pGH-501 plasmid containing two copies of gshI and copy of gshII genes in pUC8 vector.

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Myrrha-induced Apoptosis in Human Cervical Carcinoma HeLa Cells (몰약(沒藥)이 자궁경부암세포(子宮經部癌細胞)(HeLa Cell)의 Apoptosis에 미치는 영향(影響))

  • Park, Jong-Kyu;Jo, Ok-Hyon;Kim, Song-Baeg;Cho, Han-Baek
    • The Journal of Korean Obstetrics and Gynecology
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    • v.19 no.1
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    • pp.97-110
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    • 2006
  • Purpose : To address the ability of Myrrha (MY) to induce cell death, we investigated the effect of MY on apoptosis. In human cervical carcinoma HeLa cells, apoptosis occurred following MY exposure in a dose-dependent manner. Methods : We have tested several kinds of anti-oxidants to investigate the MY-induced apoptotic mechanism. Among the anti-oxidants, N-acetyl cysteine(NAC) or reduced glutathione (GSH) protects MY-induced apoptosis. NAC is an aminothiol and synthetic precursor of intracellular cysteine and GSH. To confirm the role of GSH in MY-induced apoptosis, methionine and cystathionine-glutathione extrusion inhibitors were treated in the presence of MY. Results : NAC, GSH, methionine or cystathionine led to protective effect against MY-induced apoptosis in HeLa cells. The GSH and GSH-associated reagents regulate MY-induced cytochrome c release and the resultant caspase-3 activation. Furthermore, the two specific inhibitors of carrier-mediated GSH extrusion, methionine and cystathionine demonstrate GSH extrusion occurs via a specific mechanism. While decreasing GSH extrusion and protecting against MY-induced apoptosis, methionine and cystathionine failed to exert anti-apoptotic activity in cells previously deprived of GSH. Conclusion : the target of the protection is indeed GSH extrusion. This shows that the protective effect is achieved by forcing GSH to stay within the cells during apoptogenic treatment. All this evidence indicates the extrusion of GSH precedes andis responsible for the apoptosis, probably by altering the intracellular redox state, thus giving a rationale for the development of redox-dependent apoptosis in MY-treated human cervical carcinoma HeLa cells.

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AMPLIFICATION OF MERCURY TOXICITY BY GLUTATHIONE DEPLETION IN V79 CELLS

  • Yisook Nam;Chung, An-Sik
    • Toxicological Research
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    • v.9 no.2
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    • pp.159-166
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    • 1993
  • The treatmene of V79 cells with diethyl maleate (DEM) led to decrease in glutathione (GSH) level as increasing DEM concentration. Mercuric chloride, treated for 6 hrs with 2ng/ml, affected the GSH metabolizing enzymes glutathione S-transferase (GST) and glutathione peroxidase (GSP), dropping their activities to 60% and 75%, respectively, though not so much in GSH level(80%). However, the toxic effects of mercuric chloride on those enzymes and GSH level were both amplified when the Hg2+ treatment was combined with the preceding DEM treatment.

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PROTECTIVE EFFECT OF SELENIUM ON GLUTATHIONE METABOLISM BY MERCURY TOXICITY IN THE CHO CELLS

  • Byun, Boo-Hyung;Cho, Su-Jung;Chung, An-Sik
    • Toxicological Research
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    • v.7 no.2
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    • pp.141-149
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    • 1991
  • The treatment with 5ng/ml of mercuric chloride caused time-dependent decreases, and in the activities of GSH S-transferase and GSH-peroxidase, and in the concentration of GSH in CHO cells. Three hours after treatment of $Hg^{2+}$, the activity of GSH S-transferase was decreased to almost half value of control group and the activity of GSH-peroxidase was reduced significantly at 6 hr after treatment. The concentration of GSH was decreased 2 hr after treatment of $Hg^{2+}$ and was decreased to nearly half value of control group 3 hr after treatment.

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Effects of Glucagon and Insulin on Glutathione Homeostasis: Role of Cellular Signaling Pathways and Glutathione Transport System (Glucagon과 insulin이 glutathione 항상성에 미치는 영향: 세포신호전달체계 및 glutathione transport system의 역할)

  • Kim, Bong-Hee;Oh, Jung-Min;Yun, Kang-Uk;Kim, Chung-Hyeon;Kim, Sang-Kyum
    • Environmental health and toxicology
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    • v.22 no.3
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    • pp.227-233
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    • 2007
  • It has been reported that hepatic glutathione (GSH) levels are decreased in diabetic patients, and glucagon increases hepatic efflux of GSH into blood. The signaling pathways responsible for mediating the glucagon effects on GSH efflux, however, are unknown. The signaling pathways involved in the regulation of GSH efflux in response to glucagon and insulin were examined in primary cultured rat hepatocytes. The GSH concentrations in the culture medium were markedly increased by the addition of glucagon, although cellular GSH levels are significantly decreased by glucagon. Insulin was also increased the GSH concentrations in the culture medium, but which is reflected in elevations of both cellular GSH and protein. Treatment of cells with 8-bromo-cAMP or dibutyryl-cAMP also resulted in elevation of the GSH concentrations in the culture medium. Pretreatment with H89, a selective inhibitor of protein kinase A, before glucagon addition markedly attenuated the glucagon effect. These results suggest that glucagon changes GSH homeostasis via elevation of GSH efflux, which may be responsible for decrease in hepatic GSH levels observed in diabetic condition. Furthermore, the present study implicates cAMP and protein kinase A in mediating the effect of glucagon on GSH efflux in primary cultured rat hepatocytes.