• The Korean Journal of Physiology
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• 제5권1호
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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.

• 약학회지
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• 제53권6호
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• pp.314-320
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• 2009

We examined metabolic conversion of cysteine into glutathione (GSH) and taurine in rat liver under oxidative stress. Administration of tert-butylhydroperoxide (t-BHP) into the portal vein of male rats resulted in a rapid elevation of serum sorbitol dehydrogenase, alanine aminotransferase, and aspartate aminotransferase activities, which decreased gradually in 24 hr. Hepatic cysteine concentration was reduced in 3 hr, and recovered progressively, reaching a level greater than 200% of the normal value in 24 hr. GSH was increased both in liver and blood at 9 hr after t-BHP challenge, whereas hypotaurine or taurine was not altered. $\gamma$-Glutamylcysteine synthetase (GCS) activity was increased from 9 hr after t-BHP treatment, but protein expression of the GCS-heavy subunit was not changed in liver. Activity or expression of cysteine dioxygenase was not affected by t-BHP treatment. Taken together, these data show that an acute oxidant challenge to the rats may induce upregulation of cysteine availability and GCS activity, resulting in an enhancement of hepatic GSH synthesis, but the increased cysteine level does not stimulate taurine synthesis via cysteine sulfinate pathway. It is indicated that the regulation of GSH and taurine biosynthesis from cysteine is not solely dependent on the cysteine concentration in rat liver under oxidative stress.

• Bulletin of the Korean Chemical Society
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• 제28권5호
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• pp.772-776
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• 2007

In order to study the role of residue in the active site of glutathione S-transferase (GST), Arg13 residue in human GST P1-1 was replaced with alanine, lysine and leucine by site-directed mutagenesis to obtain mutants R13A, R13K and R13L. These three mutant enzymes were expressed in Escherichia coli and purified to electrophoretic homogeneity by affinity chromatography on immobilized GSH. Mutation of Arg13 into Ala caused a substantial reduction of the specific activity by 10-fold. Km GSH, Km DCNB and Km EPNP values of R13A were approximately 2-3 fold larger than those of the wild type. Mutation of Arg13 into Ala also significantly affected I50 values of S-methyl-GSH that compete with GSH and ethacrynic acid, an electrophilic substrate-like compound. These results appeared that the substitution of Arg13 with Ala resulted in significant structural change of the active site. Mutation of Arg13 into Leu reduced the catalytic activity by approximately 2-fold, whereas substitution by Lys scarcely affected the activity, indicating the significance of a positively charged residue at position 13. Therefore, arginine 13 participates in catalytic activity as mainly involved in the construction of the proper electrostatic field and conformation of the active site in human GST P1-1.

• Plant Biotechnology Reports
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• 제5권4호
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• pp.353-365
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• 2011

The present study investigates the possible regulatory role of exogenous nitric oxide (NO) in antioxidant defense and methylglyoxal (MG) detoxification systems of wheat seedlings exposed to salt stress (150 and 300 mM NaCl, 4 days). Seedlings were pre-treated for 24 h with 1 mM sodium nitroprusside, a NO donor, and then subjected to salt stress. The ascorbate (AsA) content decreased significantly with increased salt stress. The amount of reduced glutathione (GSH) and glutathione disulfide (GSSG) and the GSH/GSSG ratio increased with an increase in the level of salt stress. The glutathione S-transferase (GST) activity increased significantly with severe salt stress (300 mM). The ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), catalase (CAT) and glutathione peroxidase (GPX) activities did not show significant changes in response to salt stress. The glutathione reductase (GR), glyoxalase I (Gly I), and glyoxalase II (Gly II) activities decreased upon the imposition of salt stress, especially at 300 mM NaCl, with a concomitant increase in the $H_2O_2$ and lipid peroxidation levels. Exogenous NO pretreatment of the seedlings had little influence on the nonenzymatic and enzymatic components compared to the seedlings of the untreated control. Further investigation revealed that NO pre-treatment had a synergistic effect; that is, the pre-treatment increased the AsA and GSH content and the GSH/GSSG ratio, as well as the activities of MDHAR, DHAR, GR, GST, GPX, Gly I, and Gly II in most of the seedlings subjected to salt stress. These results suggest that the exogenous application of NO rendered the plants more tolerant to salinity-induced oxidative damage by enhancing their antioxidant defense and MG detoxification systems.

• Journal of Ginseng Research
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• 제24권3호
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• pp.113-117
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• 2000

적변인삼의 원인 물질을 구명코저 인삼뿌리의 적변현상과 항산화 활성과의 상관관계를 조사하기 위하여 적변뿌리와 건전뿌리의 각 부위별 항산화 활성을 조사하였다. 적변삼의 표피와 피층조직의 superoxide dismutase(SOD) 활성(units/mg protein)은 각각 126.9와 79.6으로 정상뿌리에 비해 각각 3.5배, 2.0배 높았다. 적변뿌리의 catalase활성은 약 1.6배 높았으나, peroxidase 활성은 오히려 약간 낮았다. 적변뿌리 각 조직의 DPPH 자유라디칼 포착활성은 정상뿌리에 비해 약 2.0배 높았다. 정상뿌리의 ascorbate 함량은 조직에 따라 약간의 차이를 나타내었지만(166~240 $\mu$g/g fr. wt.), 산화형dehydroascorbate(DHA) 함량이 전체 ascorbate 함량의 약 97~79%를 차지하였다. 적변뿌리의 전체 ascorbate함량은 정상뿌리와 비슷하였지만 환원형 ascorbate의 함량은 조직에 따라 다르지만 정상뿌리에 비해 3.5~7.5배 높았다. 적변뿌리의 표피조직, 피층조직, 중심주 조직의 전체 glutathione 함량은 정상조직에 비해 각각 7.3배, 4.8배, 1.2배 높았다. 적변화에 따른 환원형 glutathione(GSH)과 산화형 glutathione(GSSG)의 함량비는 전체 glutathione 함량변화와 비슷하였다. 이상의 결과는 인삼의 적변조직은 어떤 외부스트레스에 의해 과다하게 발생되는 활성산소종을 제거하기 위하여 SOD 활성, 환원형 ascorbate 함량, 환원형 glutathione 함량이 크게 증가하는 것으로 사료된다.

• 생약학회지
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• 제32권4호통권127호
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• pp.269-273
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• 2001

Ethanol extracts from Xanthii Fructus (XFE) and Prunellae Spica (PSE) were investigated for the effects on the induction of cancer chemoprevention-associated enzymes. The following effects were measured: (a) induction of quinone reductase (QR) (b) induction of glutathione S-transferase (GST) (c) reduced glutathione (GSH) level. XFE and PSE were potent inducers of quinone reductase activity in Hepa1c1c7 murine hepatoma cells. Glutathione levels were increased with XFE and PSE. In addition, glutathione S-transferase activity was increased with XFE. However, GST activity was not increased with PSE. These results suggest that XFE and PSE have chemopreventive potentials by inducing quinone reductase and increasing GSH levels.

• Archives of Pharmacal Research
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• 제29권2호
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• pp.172-177
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• 2006

Hepatotoxic potential of 2, 3-dibromopropene (2, 3-DBPE) and its conjugation with glutathione (GSH) were investigated in male ICR mice. Treatment of mice with 20, 50, and 100 mg/kg of 2, 3-DBPE for 24 h caused elevation of serum alanine aminotransferase and aspartate aminotransferase activities. The hepatic content of GSH was not changed by 2, 3-DBPE. Meanwhile, the GSH content was slightly reduced when mice were treated with 2, 3-DBPE for 6 h and significantly increased 12 h after the treatment. Subsequently, a possible formation of GSH conjugate of 2, 3-DBPE was investigated in vivo. After the animals were treated orally with 20, 50, and 100 mg/kg of 2, 3-DBPE, the animals were subjected to necropsy 6, 12, and 24 h later. A conjugate of S-2-bromopropenyl GSH was identified in liver and serum treated with 100 mg/kg of 2, 3-DBPE by using liquid chromatography-electrospray ionization tandem mass spectrometry. The protonated molecular ions $[M+H]^+$ of S-2-bromopropenyl GSH were observed at m/z 425.9 and 428.1 in the positive ESI spectrum with a retention time of 6.35 and 6.39 min, respectively. In a time-course study in livers following an oral treatment of mice with 100 mg/kg of 2, 3-DBPE for 6, 12, and 24 h, the 2, 3-DBPE GSH conjugate was detected maximally 6 h after the treatment. The present results suggested that 2, 3-DBPE-induced hepatotoxicity might be related with the production of its GSH conjugate.

• Archives of Pharmacal Research
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• 제22권4호
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• pp.384-390
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• 1999

Thirty-four glutathione conjugates of 5,8-dimethoxy-1,4-naphthoquinones (DMNQ) were synthesized and their structure was determined. The yield of GSH conjugate was dependent on size of alkyl group; the longer the size of alkyl group was, the lower was the yield. It was also found that the length of alkyl side chain influenced the chemical shift of quinonoid protons; the quinonoid protons of 2-glutathionyl DMNQ derivatives with R=H to propyl, 6.51-6.59 ppm vs. other ones with R=butyl to heptyl, 6.64-6.68 ppm. this was explained to be due to a folding effect of longer alkyl group. Glutathione (GSH) reacted with DMNQ derivative first to form a 1,4-adduct (2- or 3-glutathionyl-1,4-dihydroxy-5,8-dimethoxynaphthalenes) and then the adduct was autooxidized to 2- or 3-glutathionyl-DMNQ derivatives. Moreover, GSH reduced DMNQ derivatives to their hydrogenated products. It was suggested that such an organic reaction might play an important role for a study of metabolism or toxicity of DMNQ derivative sin the living cells.

• Journal of Microbiology
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• 제43권4호
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• pp.375-380
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• 2005

Autonomous ultradian metabolic oscillation (T$\simeq$50 min) was detected in an aerobic chemostat culture of Saccharomyces cerevisiae. A pulse injection of GSH (a reduced form of glutathione) into the culture induced a perturbation in metabolic oscillation, with respiratory inhibition caused by $H_2S$ burst pro-duction. As the production of $H_2S$ in the culture was controlled by different amino acids, we attempted to characterize the effects of GSH on amino acid metabolism, particularly with regard to branched chain and sulfur-containing amino acids. During stable metabolic oscillation, concentrations of intra-cellular glutamate, aspartate, threonine, valine, leucine, isoleucine, and cysteine were observed to oscil-late with the same periods of dissolved $O_2$ oscillation, although the oscillation amplitudes and maximal phases were shown to differ. The methionine concentration was stably maintained at 0.05 mM. When GSH (100 $\mu$M) was injected into the culture, cellular levels of branched chain amino acids increased dramatically with continuous $H_2S$production, whereas the cysteine and methionine concentrations were noticeably reduced. These results indicate that GSH-dependent perturbation occurs as the result of the promotion of branched chain amino acid synthesis and an attenuation of cysteine and methionine synthesis, both of which activate the generation of $H_2S$. In a low sulfate medium containing 2.5 mM sulfate, the GSH injections did not result in perturbations of dissolved $O_2$ NAD(P)H redox oscillations without burst $H_2S$ production. This suggests that GSH-dependent perturbation is intimately linked with the metabolism of branched-chain amino acids and $H_2S$ generation, rather than with direct GSH-GSSG redox control.

• The Korean Journal of Physiology and Pharmacology
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• 제23권6호
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• pp.519-528
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• 2019

Mitochondrial dysfunction is closely associated with reactive oxygen species (ROS) generation and oxidative stress in cells. On the other hand, modulation of the cellular antioxidant defense system by changes in the mitochondrial DNA (mtDNA) content is largely unknown. To determine the relationship between the cellular mtDNA content and defense system against oxidative stress, this study examined a set of myoblasts containing a depleted or reverted mtDNA content. A change in the cellular mtDNA content modulated the expression of antioxidant enzymes in myoblasts. In particular, the expression and activity of glutathione peroxidase (GPx) and catalase were inversely correlated with the mtDNA content in myoblasts. The depletion of mtDNA decreased both the reduced glutathione (GSH) and oxidized glutathione (GSSG) slightly, whereas the cellular redox status, as assessed by the GSH/GSSG ratio, was similar to that of the control. Interestingly, the steady-state level of the intracellular ROS, which depends on the reciprocal actions between ROS generation and detoxification, was reduced significantly and the lethality induced by $H_2O_2$ was alleviated by mtDNA depletion in myoblasts. Therefore, these results suggest that the ROS homeostasis and antioxidant enzymes are modulated by the cellular mtDNA content and that the increased expression and activity of GPx and catalase through the depletion of mtDNA are closely associated with an alleviation of the oxidative stress in myoblasts.