• Title/Summary/Keyword: GST-p

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Effects of Dietary Garlic Powder on GST-P Positive Foci and Glucose 6-Phosphatase Activity in Diethylnitrosamine-Initiated Rat Hepatocarcinogenesis

  • Seo, Jeong-Min;Park, Kyung-Ae;Yeo, Eui-Zu;Choi, Hay-Mie
    • BMB Reports
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    • v.32 no.3
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    • pp.259-265
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    • 1999
  • This study was designed to examine the anticarcinogenic effect of dietary supplementation with garlic powder on rat hepatocarcinogenesis. All rats were initiated by a single dose (200 mg/body weight) intraperitoneal injection of diethylnitrosamine (DEN), and three weeks later, subjected to two-thirds partial hepatectomy. Two weeks after initiation, four groups of rats were given experimental diets supplemented with 0 (control group), 0.5, 2.0, or 5.0% garlic powder for 6 weeks. Rats were sacrificed at eight weeks after initiation. The induction of placental glutathione S-transferase (GST-P) positive foci was significantly inhibited almost equally in all three groups fed garlic diets. Glucose 6-phosphatase (G6Pase) activity was increased in rats fed 0.5% and 2.0% garlic powder, and was negatively correlated with the number and area of GST-P positive foci. Thiobarbituric acid reactive substance (TBARS) contents were decreased in rats fed 2.0% and 5.0% garlic powder. Only 5.0% garlic powder supplementation significantly increased the glutathione content and the glutathione S-transferase activity, compared to the control group. Therefore, all levels of garlic powder, 0.5% to 5.0%, exerted an anti promotional effect during hepatocarcinogenesis. Dietary supplementation with garlic powder seemed to maintain microsomal membrane integrity by increasing G6Pase activities. Glutathione-dependent detoxifying enzymes did not seem to contribute to this protective effect directly. The present study suggests that garlic powder is effective in inhibiting the induction of GST-P positive foci, possibly by stabilizing the hepatic microsomal membrane.

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Effect of Timing of IPTG Addition on Expression of Turnip Mosaic Virus Coat Protein Gene in Escherichia Coli (IPTG의 첨가 시간이 대장균(Escherichia coli)에서 순무 모자이크 바이러스(TuMV)의 외피단백질 발현에 미치는 영향)

  • Kim, Su-Joong;Park, Won-Mok;Ryu, Ki-Hyun;Lee, Sang-Seon;Lee, Se-Yong
    • Korean Journal Plant Pathology
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    • v.13 no.4
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    • pp.248-254
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    • 1997
  • Expression vector (pGEX-Tu) for the coat protein (CP) gene of turnip mosaic virus Ca strain (TuMV-Ca) was constructed by incorporation of TuMV CP gene into pGEX-KG vector which had ${\beta}$-galactosidase gene and IPTG (isopropylthio-${\beta}$-D-galactoside) induction site. The results of ELISA and western hybridization indicated that optimal condition of the expression were when IPTG and western hybridization indicated that optimal condition of the expression were when IPTG induction was carried out on YTA medium with ampicillin in 2 hours after the E. coli seed inoculation ($A_{595}$=0.1/ml). TuMV CP gene was expressed with GST (Glutathion S-Transferase) gene fusion system, and the size of fusion protein was estimated to be 59kDa, for TuMV CP was 33 kDa and GST was 26 kDa.

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Isolation and Characterization of a Theta Glutathione S-transferase Gene from Panax ginseng Meyer

  • Kim, Yu-Jin;Lee, Ok-Ran;Lee, Sung-Young;Kim, Kyung-Tack;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.36 no.4
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    • pp.449-460
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    • 2012
  • Plants have versatile detoxification systems to encounter the phytotoxicity of the wide range of natural and synthetic compounds present in the environment. Glutathione S-transferase (GST) is an enzyme that detoxifies natural and exogenous toxic compounds by conjugation with glutathione (GSH). Recently, several roles of GST giving stress tolerance in plants have demonstrated, but little is known about the role of ginseng GSTs. Therefore, this work aimed to provide further information on the GST gene present in Panax ginseng genome as well as its expression and function. A GST cDNA (PgGST) was isolated from P. ginseng cDNA library, and it showed the amino acid sequence similarity with theta type of GSTs. PgGST in ginseng plant was induced by exposure to metals, plant hormone, heavy metals, and high light irradiance. To improve the resistance against environmental stresses, full-length cDNA of PgGST was introduced into Nicotiana tabacum. Overexpression of PgGST led to twofold increase in GST-specific activity compared to the non-transgenic plants, and the GST overexpressed plant showed resistance against herbicide phosphinothricin. The results suggested that the PgGST isolated from ginseng might have a role in the protection mechanism against toxic materials such as heavy metals and herbicides.

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.

Carpal Tunnel Syndrome in Stroke Patients According to the Degree of Spasticity in Median Nerve Cross-Sectional Area and Nerve Conduction Velocity and Comparison of Upper Extremity Function (수근관증후군 뇌졸중 환자에서 경직정도에 따른 정중신경 단면적과 신경전도속도 및 상지기능의 비교)

  • Kim, Tae-Gon;Jung, Dae-In;Kim, Kyung-Yoon
    • The Journal of the Korea Contents Association
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    • v.13 no.11
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    • pp.288-296
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    • 2013
  • The purpose of this study were carpal tunnel syndrome in stroke patients according to the degree of spasticity in the median nerve cross-sectional area, nerve conduction velocity, and to evaluate differences in upper extremity function. The subjects of this study was in adult patients with stroke 42 patients from 21 patients CTS group and 21 patients Non-CTS group were selected. Measurement of median nerve-cross sectional area, nerve conduction velocity, GST, FMAS, CTS-FSS was measured. The study results were each group between the unaffected side and the affected side CTS and Non-CTS group in each grade between groups unaffected side(p<.001), and affected side(p<.001) median nerve-cross sectional area, median motor and sensory nerve onset latency, there was a statistically significant difference. CTS and Non-CTS group between groups in each grade GST(p<.05), FMAS(p<.05), CTS-FSS(p<.001), there was a statistically significant difference. In this study, the carpal tunnel pathokinesiology ever presented by the contents of upper extremity functional training in stroke patients is one of the information that you need to consider when presented.

Potential Chemoprevention Activity of Pterostilbene by Enhancing the Detoxifying Enzymes in the HT-29 Cell Line

  • Harun, Zaliha;Ghazali, Ahmad Rohi
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6403-6407
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    • 2012
  • Detoxifying enzymes are present in most epithelial cells of the human gastrointestinal tract where they protect against xenobiotics which may cause cancer. Induction of examples such as glutathione S-transferase (GST) and its thiol conjugate, glutathione (GSH) as well as NAD(P)H: quinoneoxidoreductase (NQO1) facilitate the excretion of carcinogens and thus preventing colon carcinogenesis. Pterostilbene, an analogue of resveratrol, has demonstrated numerous pharmacological activities linked with chemoprevention. This study was conducted to investigate the potential of pterostilbene as a chemopreventive agent using the HT-29 colon cancer cell line to study the modulation of GST and NQO1 activities as well as the GSH level. Initially, our group, established the optimum dose of 24 hours pterostilbene treatment using MTT assays. Then, effects of pterostilbene ($0-50{\mu}M$) on GST and NQO1 activity and GSH levels were determined using GST, NQO1 and Ellman assays, respectively. MTT assay of pterostilbene ($0-100{\mu}M$) showed no cytotoxicity toward the HT-29 cell line. Treatment increased GST activity in the cell line significantly (p<0.05) at 12.5 and $25.0{\mu}M$. In addition, treatment at $50{\mu}M$ increased the GSH level significantly (p<0.05). Pterostilbene also enhanced NQO1 activity significantly (p<0.05) at $12.5{\mu}M$ and $50{\mu}M$. Hence, pterostilbene is a potential chemopreventive agent capable of modulation of detoxifiying enzyme levels in HT-29 cells.

Glutathione S-transferase polymorphism of neonatal hyperbilirubinemia in Korean neonates (한국인 신생아 황달과 Glutathione S-transferase 다형성에 관한 연구)

  • Kang, Chang Seok;Hong, Seung Su;Kim, Ji Sook;Kim, Eun Ryoung
    • Clinical and Experimental Pediatrics
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    • v.51 no.3
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    • pp.262-266
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    • 2008
  • Purpose : Glutathione S-transferase (GST) is a polymorphic supergene family of detoxification enzymes that are involved in the metabolism of numerous diseases. Several allelic variants of GSTs show impaired enzyme activity and are suspected to increase the susceptibility to diseases. Bilirubin is bound efficiently by GST members. The most commonly expressed gene in the liver is GSTM1, and GSTT1 is expressed predominantly in the liver and kidneys. To ascertain the relationship between GST and neonatal hyperbilirubinemia, the distribution of the polymorphisms of GSTT1 and GSTM1 were investigated in this study. Methods : Genomic DNA was isolated from 88 patients and 186 healthy controls. The genotypes were analyzed by polymerase chain reaction (PCR). Results : The overall frequency of the GSTM1 null was lower in patients compared to controls (P=0.0187, Odds ratio (OR) =0.52, 95% confidence interval (CI), 0.31-0.88). Also, the GSTT1 null was lower in patients compared to controls (P=0.0014, OR=0.41, 95% CI=0.24-0.70). Moreover, the frequency of the null type of both, in the combination of GSTM1 and GSTT1, was significantly reduced in jaundiced patients (P=0.0008, OR=0.31, 95% CI=0.17-0.61). Conclusion : We hypothesized that GSTM1 and GSTT1 might be associated with neonatal hyperbilirubinemia. However, the GSTT1 and GSTM1 null type was reduced in patients. Therefore the null GSTT1, null GSTM1, and null type of both in the combination of GSTM1 and GSTT1 may be not a risk factor of neonatal jaundice.

In vivo Antioxidant Effects of Aralia elata Seemann Ethanol Extract Administered with Benzo($\alpha$) pyrene

  • Nam, Sang-Myung;Chung, Cha-Kwon
    • Preventive Nutrition and Food Science
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    • v.4 no.1
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    • pp.52-56
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    • 1999
  • This study has examined the effects of Aralia elata Seemann ethanol extract on antioxidant enzyme systems inrats along with benzo($\alpha$) pyrene(B(a)P) administration . The ethanol extract of Aralia elata Seemann (50mg/kg body wt.) was fed to rats for 4 weeks by stomach tubing. The extract administration increased antioxidant activities of glutathione sulfur transferase(GST) comparing to the control. also total superoxide dismutase(SOD) and Cu, Zn-SOD activities were stimulated. Catalase activities were increased by 50% with the extract feeding compared to the control . Combined administration of B($\alpha$)P and the extract increased GST activity in B($\alpha$) P group. Although total SOD acitivity was decreased , Cu, Zn-SOD was greately increased from 0.10unit to 0.18 unit and catalase activity also was increased compared to the group of B($\alpha$) P. GST activity in CLE group was 1.32 unit, increased by 33% comparing to the group CL of 0.99unit. Cu, Zn-SOD and catalase activities in thegroup fed high fat and ethanol extracts were increased by 25% and 39%, respectivley comparing to the group of high fat. In addition , total SOD was decreased but, Cu, Zn-SOD acitivity was increased from 0.09 unit to 0.18unit. Catalase activity was 76.05 unit in the group of B($\alpha$) P and extract comparing to 65.26 units in B($\alpha$)P group. Serum $\alpha$-tocopherol of rat was markedly increased by theextract. Administration of B9$\alpha$)P reduced $\alpha$-tocopherol levels in the serum, on the other hand, lard in the diet increased $\alpha$-tocopherol levels in the serum. The above results indicate that Aralia bud exerts antioxidant functions in vivo against B($\alpha$)P. Further research may be necessary for the identification fo the biologically active material.

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Studies on Synthetic 1,2-Benzothiazine Anti-inflammatory Agents: Pharmacological Effect and the Expression of Xenobiotic-metabolizing Enzymes (1,2-Benzothiazine계열 새로운 항염진통제에 대한 약리작용 및 대사효소발현 유형의 연구)

  • 김상건;조주연;권순경;이은방
    • YAKHAK HOEJI
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    • v.44 no.4
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    • pp.300-307
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    • 2000
  • Expression of xenobiotic-metabolizing enzymes can be altered by xenobiotics, which represents changes in the production of reactive metabolic intermediates as well as toxicities in tissues. Metabolic intermediates derived from xenobiotics are considered to produce the reactive oxygen species including drug free radicals and hydroxyl free radicals, which would be ultimately responsible for drug-induced toxicities. The effects of 1,2-benzothiazine anti-inflammatory agents on the expression of xenobiotic-metabolizing enzymes including major cytochrome P450s, microsomal epoxide hydrolase (mEH) and glutathione S-transferase (GST) were studied in the liver with the aim of providing the part of information on potential production of reactive metabolites and hepatotoxicity by the agents. The synthetic compounds 24, 36 and 39 exhibited anti-inflammatory effects in rats as assessed by the Randall-Selitto method. The anti-inflammatory effect was detected as early as at 30 min after gavaging the agents with the ED5O being noted at 80 mg/kg, which was comparable to that of ibuprofen. Treatment of rats with each compound (100 mg/kg, 3d) resulted in no significant induction in the immunochemically-detectable cytochromes P45O 1A1/2, P450 2B1/2, P45O 2 Cl1 and P45O 2El. Changes in the mEN expression were also minimal, as evidenced by both Western blot and Northern blot analyses. Hepatic GST expression was slightly increased by the agents: GST Ya protein and mRNA expression was ~1.5-fold increased after treatment with compounds 24 and 39, whereas GST Yb1/2 and Yc1/2 mRNA levels were elevated 2- to 3-fold. In summary the effects of the synthetic 1,2-benzothiazines on the expression of major P45O, mEH and G57 were not significant, providing evidence that metabolic activation of the agents, potential drug interaction and hepatotoxicity would be minimal.

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Molecular Cloning of a Delta-class Glutathione S-transferase Gene from Bombus ignitus

  • Park, Jong-Hwa;Yoon, Hyung-Joo;Gui, Zhong Zheng;Jin, Byung-Rae;Sohn, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.18 no.1
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    • pp.28-32
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    • 2009
  • We describe here the cloning and characterization of a cDNA encoding the glutathione S-transferase (GST) from the bumblebee Bombus ignitus. The Delta-class B. ignitus GST (BiGSTD) gene spans 1668 bp and consists of four introns and five exons that encode 216 amino acid residues with a calculated molecular weight of approximately 24561 Da and a pI of 8.03. The N-terminal domain of BiGSTD has a conserved Ser residue, as well as conserved Lys, Pro, Glu, Ser and Tyr residues that are involved in the GSH-binding site of GST. The BiGSTD showed 60% protein sequence identity to the Bombyx mori GSTT1, 58% to Musca domestica GST, 57% to Drosophila melanogaster GST, and 55% to Anopheles gambiae GST1. BiGSTD was close to the insect-specific Delta class of GSTs in a phylogenetic tree. Northern blot analysis showed that BiGSTD is highly expressed in the fat body and midgut, and less so in the muscles of B. ignitus worker bees.