• Toxicological Research
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• v.22 no.1
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• pp.39-45
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• 2006

As the antioxidant and free radical scavenger, glutathione (GSH) participates in the preservation of cellular redox status and defense against reactive oxygen species and xenobiotics. Glutamate-cysteine ligase (GCL; also known as ${\gamma}$-glutamylcysteine synthetase, EC 6.3.2.2) is the rate limiting enzyme in GSH synthesis. In the present study, the accurate method for determination of GCL activity in crude liver extracts was developed by measuring both ${\gamma}$-glutamylcysteine and GSH from cysteine in the presence of glutamate, glycine and an ATP-generating system. We added glycine to promote the conversion of ${\gamma}$-glutamylcysteine to GSH, and to minimize the possibility of ${\gamma}$-glutamylcysteine metabolism to cysteine and oxoproline by ${\gamma}$-glutamylcyclotransferase. We established optimal conditions and substrate concentrations for the enzyme assay, and verified that inhibition of GCL by GSH did not interfere with this assay. Therefore, this assay of hepatic GCL under optimal conditions could provide a more accurate measurement of this enzyme activity in the crude liver extracts.

• Journal of Ginseng Research
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• v.20 no.2
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• pp.173-178
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• 1996

Aerobic cells are normally protected from the damage of free radicals by antioxidative on , zymes such as superoxide dismutase (SOD), catalase, glutathione (GSH) peroxidase, GSH S- transferase and GSH reductase which scavenge free radicals as well as nonenzymatic antioxidants such as ceruloplasmin, albumin and nonprotein-SH including GSH. The effects of each component (ginsenoside $Rb_1$, $Rb_2$, Rc, Rd, Re, $Rb_1$, Rf, $Rh_1$ and $Rh_2$) of red ginseng on the antioxidative enzyme activities were investigated in the liver in order to screen antioxidative components of red ginseng. Ginsenoside $Rb_1$ and Rc showed a tendency to increase GSH peroxidase activity, while ginsenoside Rc significantly decreased Cu,Zn-SOD activity. Especially, ginsenoside $Rh_2$ significantly increased catalase activity. These results suggest that ginsenoside $Rh_2$ is an important active component among total saponins of red ginseng.

• 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.

• Journal of Applied Biological Chemistry
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• v.64 no.4
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• pp.363-368
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• 2021

The fruits of tomato (Lycopersicon esculentum) were extracted with 70% aqueous methanol (MeOH) and the concentrates were partitioned into ethyl acetate (EtOAc), n-butanol (n-BuOH), and water (H₂O) fractions. The repeated silica gel (SiO₂) and octadecyl silica gel column chromatographies for the EtOAc fraction, whose activity was confirmed, led to isolation of one flavone compound. Nuclear magnetic resornance, infrarad spectroscopy, and mass spectroscopy (MS) revealed the chemical structure of the isolated compound, 5,7,3'-trihydroxy-6,4',5'-trimethoxyflavone (1). LC-MS/MS analysis determined the content level of compounds 1 in the MeOH extract to be 4.02±0.12 ㎍/mg and in the TME-10 fraction to be 0.96±0.03 ㎍/mg. Through this study, the antioxidantive capacity was confirmed by demonstrating that the L. esculentum extract and their fractions showing an increase in glutathione mean and a decrease in glutathione heterogeneity uniformly raises the intracellular glutathione level.

• Environmental Analysis Health and Toxicology
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• v.24 no.4
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• pp.303-309
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• 2009

Hepatic antioxidant defense systems were examined in rats treated with tetrabromobisphenol-A (TBBPA), a brominated flame retardant, at the doses of 0, 250, 500 and 1,000 mg/kg for four weeks. Hepatic ratio of glutathione disulfide to glutathione (GSH) and levels of malondialdehyde, oxidative stress markers were not changed in rats treated with TBBPA. Hepatic expression of antioxidant enzymes including GSH peroxdiase-1 (GPX-1)/GSH reductase (GR), alpha-, mu- and pi-class glutathione-S-transferase (GST) and gamma-glutamylcysteine ligase catalytic subunit was determined using immunoblot analysis. Alpha-class GSTs, GPX-1 and GR levels were significantly decreased in rats treated with TBBPA at the dose of 500 or 1,000 mg/kg. These results show that TBBPA results in down-regulation of hepatic expression of antioxidant enzymes related with GSH, suggesting the liver in TBBPA-treated rats may be more sensitive to oxidants.

• Bulletin of the Korean Chemical Society
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• v.26 no.3
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• pp.433-439
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• 2005

To gain further insight on the relationship between structure and functions of glutathione S-transferase (GST), the three tyrosine 108 mutants, Y108A, Y108F, and Y108W, of human GST P1-1 were expressed in Escherichia coli and purified to electrophoretic homogeneity by affinity chromatography on immobilized GSH. The substitution of Tyr 108 with alanine resulted in significant decrease of the GSH-conjugation activity and the GSH peroxidase activity, but approximately 63% increase of steroid isomerase activity toward ${\Delta}^5$–[androstene 3,17-dione. On the other hand, the substitution of Tyr 108 with phenylalanine resulted in decreases of $k_{cat}\;and\;k_{cat}/K_m{^{EPNP}}$ by 2 orders of magnitude, suggesting that Tyr 108 residue of hGSTP1-1 are considered to be important for the catalysis and the binding of the epoxide substrates. The substitution of Tyr 108 with tryptophan resulted in significant decreases of the specific activities toward EPNP, cumene hydroperoxide and ${\Delta}^5$–ndrostene 3,17-dione, but approximately 2-fold increase on the enzyme-catalyzed addition of GSH to DCNB. We conclude from these results that Tyr 108 in hGST P1-1 plays very different roles depending upon the nature of the electrophilic substrates.

• Journal of Embryo Transfer
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• v.32 no.4
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• pp.337-342
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• 2017

The present study was aimed to determine the effects of green tea extract (GTE) and beta-mercaptoethanol (${\beta}-ME$) supplementation in boar sperm freezing extender on in vitro fertilization (IVF) and reactive oxygen species (ROS) and glutathione (GSH) levels of presumptive zygotes (PZs). Experimental groups were allocated into lactose egg yolk (LEY) without antioxidant (control), GTE (1,000 mg/l in LEY) and ${\beta}-ME$ ($50{\mu}M$ in LEY). In freezing, spermatozoa extended with LEY were cooled to $5^{\circ}C$ for 3 h and then kept at $5^{\circ}C$ for 30 min following dilution with LEY containing 9% glycerol and 1.5% Equex STM. The final sperm concentration was $1{\times}10^8/ml$. Spermatozoa were loaded into straws and frozen in nitrogen vapor for 20 min. For IVF, oocytes were matured in NCSU-23 medium and co-cultured with spermatozoa following thawing at $37^{\circ}C$ for 25 sec. At 12 h following IVF, IVF parameters (sperm penetration and monospermy) were evaluated. In addition, GSH and ROS levels of PZs were determined by Cell Tracker Blue CMF2HC and DCHFDA, respectively. IVF parameters did not show any significant difference among the experimental groups. GSH and ROS levels of PZs were not significantly different between groups. In conclusion, antioxidant supplementation in boar sperm freezing could not influence IVF parameters, ROS and GSH levels of PZs.

• The Korean Journal of Physiology
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• v.5 no.1
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• pp.43-49
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• 1971

In an attempt to better understand the effect of whole body X-irradiation on the spontaneous motility of the isolated mouse duodenum and to clarify the possible radioprotective notion of reduced glutathione (GSH), a whole body X-irradiation of 1,000r was given to albino mouse either singularly or immediately after injecting GSH intraperitoneally to mouse in the dose of 1mg per gm of body weight. The total length of contraction of the isolated duodenum was recorded on kymograph every five minutes for 60 minutes, and the comparison was made with the control (i.e., normal). The results thus obtained are summarized as follows: 1. The spontaneous motility of the isolated duodenum in the X·irradiated groups showed a significantly elevated pattern for the first 15 minutes comparing with the control. The motility, however, decreased after 15 minutes and remained so in the X-irradiated groups to the level of the non-irradiated control, but 12 hours post-irradiation group showed a significantly increased motility throughout the experiment comparing with the control. 2. When GSH was injected intraperitoneally Prior to the whole body X·irradiation with 1,000r, the spontaneous motility of the isolated duodenum of mouse showed a significantly decreased pattern for the first 10 and 15 minutes comparing with the X·irradiated group followed by the similar motility thereafter comparing with the control and X-irradiated groups. 3. The above results suggest that GSH is effective as a radioprotector in terms of the motility of the isolated mouse duodenum.

• Journal of The Korean Society of Grassland and Forage Science
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• v.43 no.2
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• pp.109-115
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• 2023

Ammonium (NH₄⁺) serves as a nitrogen source, but its elevated levels can hinder plant growth and production. Excess NH₄⁺ with α-ketoglutarate is assimilated into glutamate, a precursor of proline and glutathione (GSH). This study aimed to investigate the effects of excessive NH₄⁺ on the regulation of proline and GSH synthesis. Detached leaves from oilseed rape (Brassica napus L.) were fed with 0, 50, 100, 500, and 1000 mM NH₄Cl for 16 h. As the NH₄⁺ concentrations increased, the leaves exhibited progressive wilting and yellowing. Furthermore, total carotenoid and chlorophyll concentrations declined in response to all NH₄⁺ treatments, with the lowest levels observed in 1000 mM NH₄⁺ treatment. Hydrogen peroxide (H₂O₂) concentration showed a minor increase at low NH₄⁺ concentration (50 and 100 mM) treatments but a significant increase at high NH₄⁺ (500 and 1000 mM), which was consistent with the localization of H₂O₂. Amino acid concentrations increased with increasing in NH₄⁺ concentration, while the protein concentration displayed the opposite trend. Proline and cysteine concentrations exhibited a gradual increase in response to increasing NH₄⁺ concentrations. However, GSH concentrations rose only in the 50 mM NH₄⁺ treatment and decreased in the 500 and 1000 mM NH₄⁺ treatments. These results indicate that excessive NH₄⁺ is primarily assimilated into proline, while GSH synthesis is adversely affected.

• Mycobiology
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• v.30 no.4
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• pp.213-218
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• 2002

The induction of NAD(P)H: quinone oxidoreductase(QR), glutathione S-transferase(GST), and glutathione(GSH) levels in hepa1c1c7 cells(murine hepatoma) by waxy brown rice cultured with Phellinus igniarius to induce anticarcinogenic enzymes were measured. In addition, the inhibition of polyamines metabolism was tested with the growth of Acanthamoeba castellanii. The result shows that QR, GST activities, and GSH levels of experimental animals were increased much more by feeding the methanol extract of waxy brown rice cultured with Phellinus igniarius than those of the rats received the ethanol of uncultured brown rice. The growth of A. castellanii was inhibited mostly at 40 mg/3 ml concentration of methanol extract of waxy brown rice cultured with P. gniarius. The results suggested that waxy brown rice cultured with P. igniarius possess chemopreventive activity by inducing anticarcinogenic enzymes and inhibiting polyamine metabolism.