• Title/Summary/Keyword: Glutathione synthetase

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Cysteine improves boar sperm quality via glutathione biosynthesis during the liquid storage

  • Zhu, Zhendong;Zeng, Yao;Zeng, Wenxian
    • Animal Bioscience
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    • v.35 no.2
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    • pp.166-176
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    • 2022
  • Objective: Sperm is particularly susceptible to reactive oxygen species (ROS) stress. Glutathione (GSH) is an endogenous antioxidant that regulates sperm redox homeostasis. However, it is not clear whether boar sperm could utilize cysteine for synthesis GSH to protect sperm quality from ROS damage. Therefore, the present study was undertaken to elucidate the mechanism of how cysteine is involved in protecting boar sperm quality during liquid storage. Methods: Sperm motility, membrane integrity, lipid peroxidation, 4-hydroxyIlonenal (4-HNE) modifications, mitochondrial membrane potential, as well as the levels of ROS, GSH, and, ATP were evaluated. Moreover, the enzymes (GCLC: glutamate cysteine ligase; GSS: glutathione synthetase) that are involved in glutathione synthesis from cysteine precursor were detected by western blotting. Results: Compared to the control, addition of 1.25 mM cysteine to the liquid storage significantly increased boar sperm progressive motility, straight-line velocity, curvilinear velocity, beat-cross frequency, membrane integrity, mitochondrial membrane potential, ATP level, acrosome integrity, activities of superoxide dismutase and catalase, and GSH level, while reducing the ROS level, lipid peroxidation and 4-HNE modifications. It was also observed that the GCLC and GSS were expressed in boar sperm. Interestingly, when we used menadione to induce sperm with ROS stress, the menadione associated damages were observed to be reduced by the cysteine supplementation. Moreover, compared to the cysteine treatment, the γ-glutamylcysteine synthetase (γ-GCS) activity, GSH level, mitochondrial membrane potential, ATP level, membrane integrity and progressive motility in boar sperm were decreased by supplementing with an inhibitor of GSH synthesis, buthionine sulfoximine. Conclusion: These data suggest that boar sperm could biosynthesize the GSH from cysteine in vitro. Therefore, during storage, addition of cysteine improves boar sperm quality via enhancing the GSH synthesis to resist ROS stress.

Gene Expression of Enzymes Related to Glutathione Metabolism in Anticancer Drug-resistant L1210 Sublines (항암제 내성 L1210세포의 Glutathione 대사 관련효소 유전자의 발현 양상)

  • Kim, Seong-Yong;Kim, Jae-Ryong;Kim, Jung-Hye
    • Journal of Yeungnam Medical Science
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    • v.12 no.1
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    • pp.32-47
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    • 1995
  • Glutathione(GSH) has a very important role in detoxification of cells and is closely related to antitumor drug-resistance of cancer cells. In order to evaluate the importance of glutathione metabolism in the drug-resistant cancer cells, the concentration of celluar GSH and activities of ${\gamma}$-glutamylcysteine synthetase(GCS), ${\gamma}$-glutamyl transpeptidase (GGT) and glutathione S-transferases(GST) in the adriamycin, vincristine, or cisplatin resistant L1210 (L1210AdR, L1210VcR, or L1210Cis) sublines were measured. Expression and amplification of GCS, GGT, and GST-${\pi}$ genes were also observed in the parent Ll210 and the drug-resistant Ll210 sublines. The concentration of GSH was increased 5.34 fold in L1210Cis, 2.83 fold in L1210VcR, and 1.78 fold in L1210AdR, compared to L1210. The activities of GCS and GGT were increased in drug-resistant L1210 sublines. The GST activity was increased in L1210VcR and L1210Cis but decreased in L1210AdR compared to Ll210. Expression of GCS, GGT, and GST-${\pi}$ genes were increased in the resistant L1210 sublines compare to the parent L1210 in northern blot analyses. Overexpression of GCS, GGT, and GST-${\pi}$ were observed in the resistant sublines, and the increases of the concentration of glutathione and the activities of GCS and GGT in the resistant sublines may be involved in a part of the drug-resistance in the resistant sublines.

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Hepatic Detoxification and Antioxidant Activity in Sea-urchin Roe and Ethanol Extract of Roe (성게 부위별 및 그 추출물의 간 해독과 항산화 활성 효과)

  • Lee, Seung-Joo;Ha, Wang-Hyun;Choi, Hye-Jin;Cho, Soon-Yeong;Choi, Jong-Won
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.43 no.5
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    • pp.428-436
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    • 2010
  • Sea-urchins (Anthocidaris crassispina) are widely distributed in the East Sea of Korea. The aim of this study was to evaluate the hepatoprotective effects of sea-urchin roe on bromobenzene (BB)-induced liver damage in rats. The antioxidative and detoxifying properties of sea-urchin roe in BB-poisoned rat liver was examined by chemical analysis of serum aminotransferase (AST, ALT), glutathione S-transferase (GST), $\gamma$-glutamylcystein synthetase, glutathione reductase, epoxide hydrolase, amino-N-demethylase (AD), aniline hydrolase (AH) enzyme activity, as well as lipid peroxide and glutathione contents. Sea-urchin roe inhibited the increase of serum AST, ALT enzyme activity. Increasing lipid peroxide contents and AD and AH activities were significantly decreased in ethanol extract of sea-urchin roe. GST, $\gamma$-glutamylcystein synthetase, glutathione reductase and epoxide hydrolase enzyme activities increased in sea-urchin roe-fed group, compared with the BB-treated group. These results suggest that sea-urchin roe facilitates recovery from liver damage by enhancing antioxidative defense mechanisms and hepatic detoxication metabolism.

The Study of Pretreated GE-132 on the Hepatic Glutathione S-Transferase Activity in Rat (유기게르마늄(GE-132) 이 Glutathone S-Transferase활성에 미치는 영향)

  • Kim, Seok-Hwan;Park, Eun-Sook;Jo, Tae-Hyun;Choi, Jong-Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.4
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    • pp.581-586
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    • 1994
  • The study was initiated elucidate the mechanism by examining the effect of GE-132 on hepatic glutathione S-transferase (GST) activity. Activity of GST increased with dose-dependent manner in hepatic cytosolic fraction of GE-132 treatment rats. Double reciprocal plotting gave Vmax value 1.4 fold increase by the treatment of GE-132(100mg/kg, p.o.for 6 weeks) compared with control group, but did not change Km value. Ethacryinc acid (85mg/kg, once a day, i.p) was injected to control rat, the GST activity decreased remarkably . However, GE-132 pretreated group, the effect caused by ethacrynic acid was markedly reduced. And activity of ${\gamma}$-glutamylcys- teine synthetase was not changed either by GE-132 treatment , but the activity of glutathione reudctase increased significantly. Decreasing properties of ethacrynic acid decreased level of hepatic glutathione , which was restored to same degree by GE -132 pretreatment . GE-132 protective effect on ethacrynic acid-induced mortality. It is concluded that the efect of GE-132 is partly mediated by increase in hepatic GST activity.

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Reduction of Hepatic Glutathione by Acute Taurine Treatment in Male Mice (숫컷 생쥐에서 타우린 투여에 의한 간내 글루타치온의 감소)

  • 이선영;곽혜은;김영철
    • YAKHAK HOEJI
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    • v.47 no.4
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    • pp.218-223
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    • 2003
  • Effect of taurine treatment on metabolism of glutathione (GSH) was studied in adult male ICR mice. An acute injection of taurine (250 mg/kg, ip) resulted in a significant decline of hepatic GSH level at t = 6 hr, but plasma GSH level was not altered. The activity of GSH-related enzyme in liver, such as GSH peroxidase, GSSG reductase, GSH S-transferases, ${\gamma}$-glutamylcysteine synthetase or ${\gamma}$-glutamyltranspeptidase, was not affected by taurine at t = 2.5 or 6 hr. Plasma cysteine and cystine levels were elevated rapidly following taurine treatment. Hepatic cysteine level was decreased by taurine, reaching a level approximately 70% of control at t = 4 and 6 hr. In conclusion, the results indicate that an acute dose of taurine decreases hepatic GSH level by reducing the availability of cysteine, an essential substrate for synthesis of this tripeptide in liver. It is also suggested that taurine may decrease the cysteine uptake by competing with this S-amino acid for a non-specific amino acid transporter.

PULMONARY XENOBIOTIC CONJUGATION IN THE ISOLATED PURFUSED RABBIT LUNG AND IN VITRO: EFFECT OF ETHANOL

  • Yang, C.Mierha;Carlson, Gary P.
    • Toxicological Research
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    • v.7 no.2
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    • pp.191-208
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    • 1991
  • Pulmonary conjugation pathways may be important for the metabolism of xenobiotics introduced via airways of systemically. The objective of this study was to determine the pulmonary conjugating capacity in both the isolated perfused rabbit lung (IPRL) and in vitro, and the ability of ethanol to alter the above. The IPRL was capable of conjugating glutathione (GSH) with either 1-chloro-2,4-dinitrobenzene (CDNB) of 1,2-epoxy-(p-nitrophenoxy) propane(ENP). The pulmonary GSH conjugation with ENP was inhibited by cibacron blue, indicating the presence of glutathione-S-transferase (GST) u and/or classes, but it was not altered by buthionine sulfoximine, a selective inhibitor of Gamma-glutamylcysteine synthetase.

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Potential for Efficient Synthesis of GSH Utilizing GCS1 and GLR1 Mutant Strains of Candida albicans

  • Jaeyoung SON;Min-Kyu KWAK
    • The Korean Journal of Food & Health Convergence
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    • v.10 no.2
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    • pp.7-11
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    • 2024
  • Glutathione (GSH) is a vital compound composed of glutamic acid, cysteine, and glycine, crucial for cellular functions including oxidative stress defense and detoxification. It has widespread applications in pharmaceuticals, cosmetics, and food industries due to its antioxidant properties and immune system support. Two primary methods for GSH synthesis are enzymatic and microbial fermentation. Enzymatic synthesis is efficient but costly, while microbial fermentation, particularly using yeast strains like Candida albicans, offers a cost-effective alternative. This study focuses on genetically modifying C. albicans mutants, specifically targeting glutathione reductase (GLR1) and gamma-glutamylcysteine synthetase (GCS1) genes, integral to GSH synthesis. By optimizing these mutants, the research aims to develop a model for efficient GSH production, potentially expanding its applications in the food industry.

Effect of Terminalia chebula on Physiological Activity in Mice (가자(Terminalia chebula) 추출물이 마우스의 생리활성에 미치는 영향)

  • 박종옥;이승은
    • Journal of Life Science
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    • v.14 no.1
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    • pp.148-153
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    • 2004
  • In this study, we investigated the effect of water, extract of Terminalia Chebula (TC) on physiological activity in mice. TC water extract showed hemagglutination against several different types of red blood cells. $LD_{50}$ of TC extract was 390 mg/kg (po). Treatment of TC water extract orally administered 200, 300 mg/kg daily for one week. Hepatic cytosolic enzymes, xanthine oxidase and aldehyde oxidase activities were significantly increased comparison with normal group. Treatment of TC water extract increased hepatic malondialdehyde (MDA) formation, and reduced glutathione content. We also found that the decreased activities of glutathione S-transferase and glutathione reductase but was not affected activities of $\gamma$-glutamylcysteine synthetase after treatment of TC water extract. These results suggested that increase of the hepatic lipid peroxide is caused by glutathione reduction.

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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Protective Effect of Marine Natural Products on the Hepatic Lipid Peroxidation in Acetaminophen-treated Rats (아세트아미노펜 유도 흰쥐에서 수산생물자원 추출물의 in vivo 간보호작용)

  • Choe, Jong-Won;Park, Jong-Cheol
    • YAKHAK HOEJI
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    • v.40 no.5
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    • pp.574-581
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    • 1996
  • The study was initiated to elucidate the protective mechanism by examining in vivo effect of some marine natural products, Styela plicata, Ecklonia stolonifera and Pachymeniopsis elliptica on acetaminophen-induced lipid peroxidation. The methanol extract of S. plicata prevented acetaminophen (800mg/kg, i.p.)-induced hepatotoxicity in rats as evidenced by the decreased formation of lipid peroxide. But the methanol extracts of E. stolonifera and P. elliptica were not affected on the formation of lipid peroxidation. The activities of cytochrome P-450, animopyrine N-demethylase and aniline hydroxylase were not changed by the treatment with S. plicata in comparison with acetaminophen-teated group. In acetaminophen-treated control rats, the glutathione S-transferase activity was decreased markably. However. in S. plicata pretreated group, the effect caused by acetaminophen was markably reduced. A-cetaminophen decreased the level of hepatic, glutathione, which was restored to same degree by S. plicata pretreatment. And activity of ${\gamma}$-glutamylcystein synthetase was not changed by S. plicata pretreatment, but the activity of glutathione reductase was increased significantly.

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