• Title/Summary/Keyword: glutathione S

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Induction of Phase II Enzymes and Inhibition of Cytochrome P450 Isozymes by Chitosanoligosaccharides

  • SHON, YUN-HEE;NAM, KYUNG-SOO
    • Journal of Microbiology and Biotechnology
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    • v.15 no.1
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    • pp.183-187
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    • 2005
  • Abstract The cancer chemopreventive potential of chitosanoligosaccharides was investigated by measuring the induction of quinone reductase and glutathione S-transferase activities and inhibition of cytochrome P450 1A1, 2B1, and 2E1 activities. Chitosanoligosaccharide I (1-${\kappa}$Da${\kappa}$Da) significantly induced glutathione S-transferase activity with a maximal 1.5-fold increase at 500 ${\mu}$g/ml, while chitosanoligosaccharide II (3-${\kappa}$Da${\kappa}$Da) (500 ${\mu}$g/ml) strongly induced quinone reductase (p<0.01) and glutathione S-transferase (p<0.005) activities. The in vitro incubation of rat liver microsomes with chitosanoligosaccharides I and II (2.5, 5, 50, and 500 ${\mu}$g/ml) showed a dose-dependent inhibiton of cytochrome P450 1A1, 2B1, and 2E1 activities. Chitosanoligosaccharide II was a more potent inhibitor of cytochrome P450 2B1 activity than chitosanoligosaccharide I. Accordingly, these findings suggest that chitosanoligosaccharides are potential chemopreventive agents.

Lymphocyte DNA damage and plasma antioxidant status in Korean subclinical hypertensive patients by glutathione S-transferase polymorphism

  • Han, Jeong-Hwa;Lee, Hye-Jin;Choi, Hee Jeong;Yun, Kyung Eun;Kang, Myung-Hee
    • Nutrition Research and Practice
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    • v.11 no.3
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    • pp.214-222
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    • 2017
  • BACKGROUND/OBJECTIVES: Glutathione S-transferase (GST) forms a multigene family of phase II detoxification enzymes which are involved in the detoxification of xenobiotics by conjugating substances with glutathione. The aim of this study is to assess the antioxidative status and the degree of DNA damage in the subclinical hypertensive patients in Korea using glutathione S-transferase polymorphisms. SUBJECTS/METHODS: We examined whether DNA damage and antioxidative status show a difference between GSTM1 or GSTT1 genotype in 227 newly diagnosed, untreated (systolic blood pressure $(BP){\geq}130mmHg$ or diastolic $BP{\geq}85mmHg$) subclinical hypertensive patients and 130 normotensive subjects (systolic BP < 120 mmHg and diastolic BP < 80 mmHg). From the blood of the subjects, the degree of the DNA damage in lymphocyte, the activities of erythrocyte superoxide dismutase, the catalase, and the glutathione peroxidase, the level of glutathione, plasma total radical-trapping antioxidant potential (TRAP), anti-oxidative vitamins, as well as plasma lipid profiles and conjugated diene (CD) were analyzed. RESULTS: Of the 227 subjects studied, 68.3% were GSTM1 null genotype and 66.5% were GSTT1 null genotype. GSTM1 null genotype had an increased risk of hypertension (OR: 2.104, CI: 1.38-3.35), but no significant association in GSTT1 null genotype (OR 0.982, CI: 0.62-1.55). No difference in erythrocyte activities of superoxide dismutase, catalase, or glutathione peroxidase, and plasma TRAP, CD, lipid profiles, and GSH levels were observed between GSTM1 or GSTT1 genotype. Plasma levels of ${\alpha}-tocopherol$ increased significantly in GSTT1 wild genotype (P < 0.05); however, plasma level of ${\beta}-carotene$ increased significantly in GSTT1 null genotype (P < 0.01). DNA damage assessed by the Comet assay was significantly higher in GSTM1 null genotype than wild genotype (P < 0.05). CONCLUSIONS: These results confirm the association between GSTM1 null genotype and risk of hypertension as they suggest that GSTM1 null genotype leads to an increased oxidative stress compared with wild genotype.

SO42- Uptake and Assimilation in Forage Rape (Brassica napus) (유채의 SO42- 흡수 및 동화에 관한 연구)

  • Li, Ming;Jin, Yu-Lan;Lee, Bok-Ryu;Li, Lu-Shen;Kim, Dae-Hyun;Kim, Tae-Hwan
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.28 no.2
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    • pp.75-80
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    • 2008
  • To investigate the sulfate utilization efficiency that has been examined in rape (Brassica napus L.) cultivars (cv. Akela and Colosse). During 72h of treatment, in two cultivars, ${SO_4}^{2-}$ uptake was significantly higher in complete S condition (2.0 mM ${SO_4}^{2-}$) than that of the S-deficient condition (2.0 mM ${SO_4}^{2-}$). In young leaves, ATP sulfurylase activity increased after S-deficient treatment. However, in old leaves, ATP sulfurylase activity was not significantly changed in Akela. Glutathione concentration in young leaves significantly increased in all cultivars examined under S-deficient condition. The rate of increase in glutathigne concentration caused by S-deficiency treatment was higher in Akela. However, in old leaves, the glutathione concentration in Akela significantly decreased. The results suggest that ${SO_4}^{2-}$ uptake and ATP sulfurylase in rape plants were species specific, and that ${SO_4}^{2-}$ uptake was highly related to its assimilation into S containing compound such as glutathione.

Effect of Cysteine and Glutathione on Motility Index and Morphology in Human Spermatozoa (Cysteine 및 Glutathione이 사람정자의 운동성지수와 정자형태에 미치는 영향)

  • 윤정임;한만희;전은숙;허영문;이종인;이규승
    • Journal of Embryo Transfer
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    • v.15 no.3
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    • pp.211-218
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    • 2000
  • This experiment was conducted to investigate the effect of the cysteine and glutathione on the motility index and morphology of human spermatozoa at the sperm processing in vitro. After treating the sperm with medium containing cysteine and glutathione, we measured the motility index and morphology at 0.5 h and 24 h. 1. Following the sperm culture for 0.5 h after treating the sperm with the medium containing 0, 1, 5, 10 mM cysteine, curvilinear velocity (VCL) was significantly (p<0.05) higher in control than that in all treatments. And straight-line velocity (VSL) was high at 1 mM and average path velocity (VAP) was low at 5 mM and 10 mM. But the motility (MOT) and morphology (NOM) were not different between control and all treatments. Following the sperm culture for 24 h, the MOT was significantly high in treatment groups (58.9, 74.4 and 62.3%), compared with that in control(28.7%) and the VCL was also high in treatment groups (31.4, 37.9, and 34.0 ${\mu}{\textrm}{m}$/s), compared with that in control (21.3 ${\mu}{\textrm}{m}$/s). The VSL (18.4, 21.7, and 18.9 ${\mu}{\textrm}{m}$/s) was significantly higher than control (10.7 ${\mu}{\textrm}{m}$/s) and the VAP (20.3, 24.7, and 21.4 ${\mu}{\textrm}{m}$/s) in treatments was also compared with that in control (12.6 ${\mu}{\textrm}{m}$/s). The NOM was not difference between control and treatments. 2 Following the sperm culture for 0.5 after treating the sperm with the medium containing 0, 1, 5, 10 mM glutathione, the MOT, VCL, VSL, VAP, and NOM were not different between control and treatments. Following the sperm culture for 24 h, the MOT was higher in treatment groups (82.9, 83.6, 83.4%) than in control (51.1%) and the VCL was higher in treatment groups (50.9, 51.3, and 49.4 ${\mu}{\textrm}{m}$/s) than control (34.1 ${\mu}{\textrm}{m}$/s). The VSL was also higher in treatment (17.1 ${\mu}{\textrm}{m}$/s) and the VAP was also higher in treatment groups (30.1, 32.5, and 29.7 ${\mu}{\textrm}{m}$/s) than in control (19.8 ${\mu}{\textrm}{m}$/s). The NOM was not different between control and treatments.

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Effects of the Methanol Extract of the Leaves of Brassica juncea and Its Major Component, Isorhamnetin $3-O-{\beta}-D-Glucoside$, on Hepatic Drug Metabolizing Enzymes in Bromobenzene-treated Rats

  • Hur, Jong-Moon;Choi, Jong-Won;Park, Jong-Cheol
    • Food Science and Biotechnology
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    • v.16 no.3
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    • pp.439-443
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    • 2007
  • The effects of the methanol extract of the leaves of Brassica juncea and isorhamnetin $3-O-{\beta}-D-glucopyranoside$, major compound isolated from the ethyl acetate fraction of this plant on hepatic lipid peroxidation and drug-metabolizing enzymes, were evaluated in rats treated with bromobenzene. The extract and isorhamnetin $3-O-{\beta}-D-glucopyranoside$ of oral administration did not show any significant effects on activities of aminopyrine N-demethylase and aniline hydroxylase, enzymes forming toxic epoxide by bromobenzene as well as on glutathione content. However, both methanol extract and isorhamnetin $3-O-{\beta}-D-glucopyranoside$ significantly recovered the decreased activities of glutathione s-transferase and epoxide hydrolase, and also reduced the lipid peroxide level in rats treated with bromobenzene. From the results, the protections of this plant against bromobenzene-induced hepatotoxicity are thought to be via enhancing the activities of epoxide hydrolase and glutathione s-transferase, enzymes removing toxic epoxide, and reducing the lipid peroxide level.

Cancer Chemopreventive Potential of Scenedesmus spp. Cultured in Medium Containing Bioreacted Swine Urine

  • Shon, Yun-Hee;Nam, Kyung-Soo;Kim, Mi-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.158-161
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    • 2004
  • Scenedesmus spp. were cultured for 51 days in newly developed medium, KEP I (Kim and Ecopeace: initials of corresponding author and environmental company) made with Bacterio-Mineral-Water (3%, v/v) that had been bio-reacted with swine urine medium to 10% (v/v) Bold's Basal medium, and investigated for cancer chemopreventive potential by measuring the induction of quinone reductase (QR), glutathione S-transferase (GST), and reduced glutathione (GSH), and inhibition of cytochrome P450 (CYP) 1A1 activity. The activitives of QR and GST of Scenedesmus spp. cultured in KEP I medium were increased by 3.0-fold and 1.5-fold, respectively. However, Scenedesmus spp. cultured in control medium (CT) increased the activitives of QR and GST by 1.8-fold and 1.3-fold, respectively. Scenedesmus spp. in KEP I medium strongly inhibited CYP 1Al activity. These results show that Scenedesmus spp. in KEP I medium has cancer chemopreventive potential and may be a candidate for further development as a chemopreventive agent.

Induction of Glutathione S-transferase and NAD(P)H:Quinone Reductase by Astragali Radix Aqua-acupuncture Solution (황기(黃耆) 약침액(藥鍼液)의 Glutathione S-transferase 와 NAD(P)H: Quinone Reductase 유도)

  • Ryu Jun-Seon;Lim Jong-Kook
    • Korean Journal of Acupuncture
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    • v.18 no.1
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    • pp.21-26
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    • 2001
  • Induction of phase II enzymes such as quinone reductase (QR) or glutathione S-transferase (GST) is considered a major mechanism of protection against initiation of carcingenesis. This study was desinged to investigate the potential of Astragali Radix Aqua-acupuncture Solution (ARAS) to induce phase II enzymes and glutathione (GSH) in murine hepatoma cells grown in microtiter plate wells. ARAS was potent inducers of QR activity. ARAS was induced about 2.6-fold at concentration of $5{\times}$. In addition, GST activity was increased with ARAS. GSH levels were increased about 1.2-fold with ARAS at concentration of $0.1{\times}$. These results suggested that ARAS may act as blocking agents against carcinogenesis by induction of phase II marker enzymes.

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Expression of Human p53 Gene as Glutathione S-transferase Fusion Proteins in Escherichia coli (사람의 p53 유전자와 Glutathione S-Transferase와의 융합 단백질의 대장균에서의 발현)

  • 오상진
    • Korean Journal of Microbiology
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    • v.31 no.4
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    • pp.279-285
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    • 1993
  • Alterations of the p53 gene arc among the most frequent genetic changes in human cancer and often result in increased levels of p53 protein within the malignant cells. Detection of accumulated p53 protein can be a useful prognostic tool in human cancer. In order to make polyclonal antibodies for immunohistochemical screening. human p53 gene was expressed in E. coli in the form of GST (glutathione S-transfi.:rase) fusion proteins. Two p53 gene fragments. which were N('()I small fragment encoding amino acid residues of 1-151-: and Ncol large fragment of 159-393. were subeloned into the unique BamHI site present within the pGEX-2T vector using BamHI linker and recombinant plasmids pGTNS and pGTNL were constructed. respectively. The p53 cDNA fragment (from pC53-$SN_3$,) encoding amino acid 38-145 (proline at residue 72) was amplified by polymerase chain reaction(PCR). The amplified DNA was digested with BamHI and Prull and inserted into the BamHI-Smal sites of pG EX-2T and recombinant plasmid pGTBP was constructed. After IPTG induction of these plasmids for 4 hours. fusion proteins were purified from E. coli extracts with glutathione Sepharose beads. The bound proteins were resolved by 10% SDS-polyacrylamide gel electrophoresis and the molecular weights were 54 kDa. 53 kDa and 40 kDa. respectively. Approximately one milligram of fusion proteins were purified from 1 -liter cultures.

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A Phi Class Glutathione S-transferase from Oryza sativa (OsGSTF5): Molecular Cloning, Expression and Biochemical Characteristics

  • Cho, Hyun-Young;Lee, Hae-Joo;Kong, Kwang-Hoon
    • BMB Reports
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    • v.40 no.4
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    • pp.511-516
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    • 2007
  • A glutathione S-transferase (GST) related to the phi (F) class of enzymes only found in plants has been cloned from the Oryza sativa. The GST cDNA was cloned by PCR using oligonucleotide primers based on the OsGSTF5 (GenBank Accession No. $\underline{AF309382}$) sequences. The cDNA was composed of a 669-bp open reading frame encoding for 223 amino acids. The deduced peptide of this gene shared on overall identity of 75% with other known phi class GST sequences. On the other hands, the OsGSTF5 sequence showed only 34% identity with the sequence of the OsGSTF3 cloned by our previous study (Cho et al., 2005). This gene was expressed in Escherichia coli with the pET vector system and the gene product was purified to homogeneity by GSH-Sepharose affinity column chromatography. The expressed OsGSTF5 formed a homo-dimer composed of 28 kDa subunit and its pI value was approximately 7.8. The expressed OsGSTF5 displayed glutathione conjugation activity toward 1-chloro-2,4-dinitrobenzene and 1,2-epoxy-3-(p-nitrophenoxy)propane and glutathione peroxidase activity toward cumene hydroperoxide. The OsGSTF5 also had high activities towards the herbicides alachlor, atrazine and metolachlor. The OsGSTF5 was highly sensitive to inhibition by S-hexylGSH, benastatin A and hematin. We propose from these results that the expressed OsGSTF5 is a phi class GST and appears to play a role in the conjugation of herbicide and GPOX activity.

Effects of Arsenic (AsIII) on Lipid Peroxidation, Glutathione Content and Antioxidant Enzymes in Growing Pigs

  • Wang, L.;Xu, Z.R.;Jia, X.Y.;Jiang, J.F.;Han, X.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.5
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    • pp.727-733
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    • 2006
  • This experiment was conducted to investigate the effect of arsenic ($As^{III}$) on lipid peroxidation, glutathione content and antioxidant enzymes in growing pigs. Ninety-six Duroc-Landrace-Yorkshire crossbred growing pigs (48 barrows and 48 gilts, respectively) were randomly assigned to four groups and each group was randomly assigned to three pens (four barrows and four gilts). The four groups received the same corn-soybean basal diet which was supplemented with 0, 10, 20, 30 mg/kg As respectively. Arsenic was added to the diet in the form of $As_2O_3$. The experiment lasted for seventy-eight days after a seven-day adaptation period. Malondialdehyde (MDA) levels, glutathione (GSH) contents and superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione-S-transferase (GST) activities were analyzed in serum, livers and kidneys of pigs. The results showed that pigs treated with 30 mg As/kg diet had a decreased average daily gain (ADG) (p<0.05) and an increased feed/gain ratio (F/G) (p<0.05) compared to the controls. The levels of MDA significantly increased (p<0.05), and the contents of GSH and the activities of SOD, CAT, GPx, GR and GST significantly decreased (p<0.05) in the pigs fed 30 mg As/kg diet. The results indicated that the mechanism of arsenic-induced oxidative stress in growing pigs involved lipid peroxidation, depletion of glutathione and decreased activities of some enzymes, such as SOD, CAT, GPx, GR and GST, which are associated with free radical metabolism.