• Title/Summary/Keyword: GSH

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A Study of in vitro Scavenging Reactions of Acrylamide with Glutathione Using Electrospray Ionization Tandem Mass Spectrometry

  • Cui, Sheng-Yun;Kim, Seung-Jin;Jo, Sung-Chan;Lee, Yong-Moon;Lee, Yong-Ill
    • Bulletin of the Korean Chemical Society
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    • v.26 no.8
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    • pp.1235-1240
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    • 2005
  • A combination of electrospray ionization and tandem mass spectrometry was used to characterize the scavenging reactions of acrylamide (AA) in the presence of glutathione (GSH) in vitro. In the presence of GSH, AA was deactivated effectively and scavenged by reactions consuming small amount of GSH. Reaction products and structural information were identified using collision-induced dissociation (CID) in an ion trap mass spectrometer. In the mixture of GSH and AA, significant increase in abundance of fragment ion peak was observed at m/z 233, which was identified as $[Cys-Glu]^+$, formed by the elimination of glycine moiety of GSH. GSH also contributes to the AA scavenging reaction by conjugating with AA through the sulfhydryl group in cysteine moiety. The probable scavenging reaction pathway of AA in the presence of GSH has been proposed based on the CID experimental data.

Effect of Reduced Glutathione on Non-Protein Sulfhydryl and Non-Protein Disulfide of the Mouse Liver in Vitro (Reduced Glutathione의 In Vitro 첨가(添加)가 마우스간조직(肝組織)의 내재(內在) NP-SH 및 NP-SS에 미치는 영향(影響))

  • Oh, Sang-You
    • The Korean Journal of Physiology
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    • v.7 no.2
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    • pp.9-16
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    • 1973
  • In view of the recent knowledge on the radioprotective action of reduced glutathione (GSH), the present study was designed the elucidate the effect of some concentrations of GSH on the levels of intrinsic non-protein sulfhydryl (NP-SH) and non-protein disulfide (NP-SS) of the mouse liver incubated at 4, 25 and 37C in vitro, respectively. The liver slice of the mouse was incubated at 4, 25 and 37C in the medium composed of 100 ml of Krebs-Ringer phosphate buffer (KRP) with the addition of 10, 20 and 30 mg of GSH, respectively. Measurement of NP-SH and NP-SS was made at 5, 30 and 60 min during the course of the incubation, and the results were compared with the controls which were incubated only in KRP medium, and the normal. The results thus obtained are summarized as follows: 1. When the mouse liver slice was incubated at 4C, the values of both NP-SH and NP-SS of the control and the group where 10 mg of GSH was added to the incubation medium were similar to those of the normal group, and the increase of NP-SH and NP-SS with the increased concentrations of GSH was not prominent. 2. When the liver slice was incubated in the concentrations of GSH 20 mg/100 ml KRP and GSH 30 mg/100 ml KRP at 25 C, the rate of increase of both NP-SH and NP-SS was proportional to the increase of GSH concentration. In the group where 10 mg of GSH was added to the incubation medium, the value of NP-SH and NP-SS reached the highest value at 30 min, but a tendency of decrease was observed at 60 min. 3. The rate of increase of NP-SH and NP-SS of the liver was most marked of all the group. studied when the incubation temperatuse was elevated to 37C, and the increase was proportional to the concentration of GSH and the incubation time.

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Effects of Guisinhwan on the Ovariectomized Rat Model of Postmenopausal Osteoporosis (귀신환이 난소적출 백서의 골다공증에 미치는 영향)

  • Kim Tae Hee;Kang Young Keum;Ahn Kyu Hwan;Kim Song Baeg;Cho Han Baek;Yoo Sim Keun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.5
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    • pp.1476-1484
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    • 2004
  • Aging is characterized by an age-dependent reduction in bone density. Pronounced bone loss by postmenopausal estrogen deficiency induces a bad effect on dynamic balance of bone metabolism. When the bone density is reduced to the level below the fracture threshold, the risk for fracture is greatly increased. The purpose of this study is to examine what are the effects of the Guisinhwan(GSH : 歸腎丸) on the ovariectomized rat model of postmenopausal osteoporosis. The results of the experiment are as follows: Body weight in control group showed significant increase in comparison with sham, but that in GSH-treated showed no change in comparison with control. The level of serum albumin in control group showed significant decrease in comparison with sham. That in GSH-treated was slightly decreased in comparison with control. The level of serum ALP activity in control group showed significant increase in comparison with sham, but that in GSH-treated was significantly decreased in comparison with control. The level of serum phosphorous, calcium, GOT, GPT, T3 did not significant change among the three groups. The level of serum estrogen in control group showed slightly decreased in comparison with sham, but that in GSH-treated showed no change in comparison with control. The level of serum T4 in control group showed significant increase in comparison with sham, but that in GSH-treated was significantly decreased in comparison with control. Trabecular bone area as well as trabecular thickness in control group showed significant decrease In comparison with sham. Those in GSH-treated showed significant increase in comparison with control. Trabecular number did not significant change among the three groups. Trabecular separation only in GSH-treated showed significant decrease in comparison with control. Osteoclast number in control group showed significant increase in comparison with sham. That in GSH-treated showed significant decrease in comparison with control. Oseoblast surface in control group showed significant increase in comparison with sham, but that in GSH-treated showed no change in comparison with control. According to the above these results, GSH has shown to be capable of preventing and curing osteoporosis caused by old-aged and postmenopause.

PROTECTIVE ACTION OF N-ACETYLCYSTEINE AGAINST HEPATOTOXIC AGENTS IN ISOLATED RAT LIVER CELLS

  • Park, Soo-Hee;Dong, Mi-Sook;Kang, Dong-Chul;Lee, Ki-Wan;Cha, Young-Nam
    • Toxicological Research
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    • v.3 no.2
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    • pp.129-141
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    • 1987
  • Hepatocytes isolated from rats which have been pretreated with phenobarbital (80 mg/kg for 3 days), were able to take up N-acetylcysteine from surrounding medium and were able to synthesize the reduced glutathione ($GSH^{\ast}-3$) intracellularly. The N-acetylcysteine is quickly deacetylated after the uptake and increases the pool size of cysteine, which was very low initially (5 nmol/$10^6$ cells). From this increased intracellular cysteine pool, GSH was synthesized. Freshly isolated rat hepatocytes contained a high level of GSH (30 nmol/$10^6$ cells), but upon incubation with the diethylmaleate, it was markedly decreased (10 nmol/$10^6$ cells). The hepatocytes with depleted GSH have lost viability upon incubations with acetaminophen (5mM) and paraquat (2 mM). However, when the N-acetylcysteine (1 mM) was added to this incubation condition, these chemical induced hepatocellular necrosis were prevented for longer durations. This N-acetylcysteine dependent protective effect against the hepatotoxic chemicals was lost by adding methionine sulfoximine (10 mM), an inhibitor of GSH biosynthesis. Both the carbontetrachloride (5 mM) and chioroform (5 mM) added to the incubation medium caused rapid losses of GSH and cell viability, even without the prior depletion of cellular GSH. However, again, if the 1mM N-acetylcysteine was supplemented, the rates of losses of GSH and cell viability were retarded in both cases. Even though large amounts of the added N-acetylcysteine was present in the cell, N-acetylcysteine conjugate of acetaminophen was not formed. Instead, only large amounts of GSH conjugate of the drug was produced. Thus, it is concluded that the added N-acetylcysteine is taken up and utilized for resynthesis of GSH. In turn, this resynthesized GSH contributes to the protection against cytotoxicity inducible with hepatotoxic drugs.

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BIOACTIVATION OF DIBROMOETHANE BY CONJUGATION WITH GLUTAHIONE

  • Kim, Dong-Hyun
    • Toxicological Research
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    • v.7 no.2
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    • pp.231-238
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    • 1991
  • The pesticide and carcinogen ethylene dibromide(EDB) is metabolized both by cytosolic GSH S-transferase and by microsomal mixed function oxygenase. Cytochrome P-450 IIE1 appears to be major enzyme to metabolize EDB.EDB is activated to a mutagen by enzymatic conjugation with glutathione (GSH). Such activation is an exception to the general mode of detoxification via GSH S-transferase action. The primary DNA adduct (>95) is S-[2-(N7-guanyl)ethyl] GSH and a minor adduct is S-[2-(N7-guanyl)ethyl]cysteine, which is excreted in the urine and may serve as a biomarker of damage.

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Effect of N-Acetylcysteine on the Supetoxide Release, Chemotaxis from the Neutrophils and Glutathione Level of Plasma and Neutrophils (N-Acetylcysteine이 호중구의 Superoxide, Chemotaxis 및 혈장과 호중구의 Glutathione에 미치는 영향)

  • Song, Jeong-Sup;Lee, Sook-Young;Moon, Hwa-Sik;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.5
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    • pp.475-483
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    • 1994
  • Background: N-acetylcysteine(ACE) is used both orally and intravenously in a variety of experimental pathologies resembling human disease states which exhibit endothelial toxicity as a result of oxidative stress, including acute pulmonary oxygen toxicity, septicemia and endotoxin shock. Despite these observations in vivo, it is not certain how this thiol drug produces its protective effects. ACE is a cysteine derivative which is able to direct1y react with oxygen radicals and may also act as a cysteine and glutathione(GSH) precursor following deacetylation. In this paper, we tried to know whether the therapeutic doses of ACE can modify the inflammatory function of the neutrophils and can increase the glutathione level of plasma in chronic obstructive pulmonary disease(COPD) patients. In addition, the effect of ACE to the purified neutrophil in terms of superoxide release and glutathione synthesis were observed. Method: Firstly, we gave 600mg of ACE for seven days and compare the release of superoxide, luminol-enhanced chemiluminescence from the neutrophils, neutrophil chemotaxis, and plasma GSH levels before and after ACE treatment in COPD patients. Secondly, we observed the dose dependent effect of ACE to the purified neutrophil's superoxide release and GSH levels in vitro. Results: 1) Usual oral therapeutic doses(600mg per day) of ACE for seven days did affect neither on the neutrophil's superoxide release, chemiluminescence, chemotaxis, nor on the plasma GSH concentration in the COPD patients. 2) ACE decreases the purified neutrophil's superoxide release and increase the GSH production in dose dependent fashion in vitro. Conclusion: Despite the fact that oral ACE treatment did not affect on the neutrophil's inflammatory function and plasma GSH concentration in COPD patients in usual therapeutic doses, it decreases the superoxide release and increases the GSH production from the isolated neutrophils in high molar concentrations. These findings suggest that to obtain an antioxidative effects of ACE, it might be needed to increase the daily dosage of ACE or therapeutic duration or change the route of adminisration in COPD patients.

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Glutathione Contents in Various Plant Cell Lines (다양한 식물배양세포주의 Glutathione 함량)

  • 이정은;안영옥;권석윤;이행순;김석원;박일현;곽상수
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.1
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    • pp.57-61
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    • 2000
  • We investigated the levels of glutathione (GSH) and its oxidized form (GSSG) in 24 cell lines derived from various plant species to understand the antioxidative mechanism in plant cell cultures. The total glutathione content was 98$\pm$27 $\mu$g/g cell fresh wt, showing a slight difference in plant species. The average contort of GSH and GSSG was 72$\pm$20 and 26$\pm$10 $\mu$g/g cell fresh wt, respectively. The average GSH content in plant cell lines occupies approximately 73% in total glutathione. During the suspension cultures of Scutellaria baicalensis, one of the plant species we tested, the GSH content decreased in proportion to the cell growth during the exponential growth stage, showing the low level at the stationary growth stage (84 $\mu$g/g cell fresh wt), whereas the GSSG content increased to the stationary growth stage (31 $\mu$g/g cell fresh wt). The results suggested that the ratio of GSH and GSSG should be involved in the cell growth and antioxidative mechanism in cultured cells.

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Comparison of Gangji-hwan-1, 2, 3, 4 and Combination of Gangji-hwan-1 and Gamisoche-hwan in the Reducing Effects of Body Weight in a High Fat Diet-Fed Obese Mice (고지방식이 비만마우스 모델에서 파키스탄산 및 중국산 마황으로 조성된 강지환(降脂丸)-1, 2, 3, 4와 강지환(降脂丸)-1합가미소체환(合加味消滯丸)의 체중감량효과 비교)

  • Yoo, Jae Sang;Ku, Ja Ryong;Yoon, Ki Hyeon;Jo, Ju Heum;Jang, Du Hyon;Jung, Yang Sam;Kim, Jong Hoon;Kim, Byeong Chul;Seok, Hoa Jun;Yoon, Michung;Roh, Jong Seong;Shin, Soon Shik
    • Journal of Korean Medicine for Obesity Research
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    • v.15 no.1
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    • pp.9-23
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    • 2015
  • Objectives: This study was investigated the improvement effects of Pakistani Ephedra herba-containing Gangji-hwan-1, 2, 3 (Di-fatty; DF-1, 2, 3), Chinese Ephedra herba-containing Gangjihwan-4 (DF-4) and combination of DF-1 and Gamisoche-hwan (GSH) on obesity in a high fat diet-fed obese mouse model. Methods: Eight-week-old C57BL/6N mice were divided into seven groups: a normal lean group given a standard diet, an obese control group given a high fat diet, and DF-1, 2, 3, 4, and DF-1+GSH groups given a high fat diet with DF-1, 2, 3, 4 (40, 80, 160, 80 mg/kg), and DF-1+ GSH (80 mg/kg), respectively. After 8 weeks of treatment, body weight gain, feeding efficiency ratio (FER), blood lipid markers, liver histology, and fat weight and histology were examined. Results: Body weight gain was significantly decreased in DF and DF-1+GSH groups compared with control. The extent of decreases was eminent in DF-1+GSH group. FER and circulating concentration of leptin were decreased in DF and DF-1+GSH groups compared with control. Circulating concentrations of triglyceride, glucose and insulin were decreased in DF and DF-1+GSH groups compared with control. The size of adipocytes were decreased by DF and DF-1+GSH groups compared with control, whereas the adipocyte number per unit area was increased by them, suggesting that DF and DF-1+GSH groups decreased the number of large adipocytes. Conclusions: In conclusion, these results suggest that DF and DF-1+GSH groups decrease FER, plasma leptin concentration, blood anti-obesity biomarkers and fat mass, improves body weight gain. In addition, these effects were more effective in DF-1+GSH combination group than in DF-1, 2, 3, 4 groups.

Effect of glutathione on tetraploid embryo development in the pigs

  • Kim, Hwa-Young;Lee, Sang-Hee;Hwangbo, Yong;Lee, Seung Tae;Lee, Eunsong;Park, Choon-Keun
    • Journal of Embryo Transfer
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    • v.31 no.3
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    • pp.207-213
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    • 2016
  • The objective of this study was to investigate to influence of glutathione (GSH) on development and antioxidant enzyme activity in tetraploid porcine embryos. Tetraploid embryos were produced using parthenogenetic 2-cell embryo by electrofusion method. Tetraploid embryo development was observed every 24 hours and intracellular antioxidant enzyme activity was measured at 120 hours after electrofusion. The 4-cell to 16-cell stage tetraploid embryos was increased in 100 and $500{\mu}M$ GSH-treated groups compared control group at 48 hours (P < 0.05) but cleavage rates were not significantly different among the GSH treatment groups at 48, 72, 96, and 120 hours. Blastocyst formation was significantly increased by 300 and $500{\mu}M$ GSH at 120 hours in tetraploid embryos (P < 0.05). But blastocyst cell number were not significantly different among the GSH treatment groups ($16.4{\pm}0.8$, $16.8{\pm}2.6$, $18.5{\pm}2.8$ and $17.5{\pm}1.8$). The intracellular antioxidant enzyme level was increased in $500{\mu}M$ GSH compared to 0 and $100{\mu}M$ GSH (P < 0.05). We suggest that GSH may be improve development of tetraploid embryo in pigs.

Functional analysis of Tyr7 residue in human glutathione S-transferase P1-1 (Human glutathione S-transferase 중 tyrosine 7 잔기의 기능 분석)

  • Kong, Kwang-Hoon;Park, Hee-Joong;Yoon, Suck-Young;Cho, Sung-Hee
    • Analytical Science and Technology
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    • v.10 no.5
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    • pp.378-385
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    • 1997
  • In order to clarify the functional role of Tyr7 in human glutathione S-transferase P1-1, we extensively investigated the effect of mutation of Tyr7 on the substrate specificity and inhibition characteristics. The mutational replacement of Tyr7 with phenylalanine lowered the specific activities with 1,2-dichloro-4-nitrobenzene and 1,2-epoxy-3-(p-nitrophenoxy) propane for GSH-conjugation reaction to 3~5% of the values for the wild-type enzyme. The pKa of the thiol group of GSH bound in Y7F was about 2.4 pK units higher than that in the wild-type enzyme. The $I_{50}$ of hematin for Y7F was similar to that for the wild-type enzyme and those of benastatin A and S-(2,4-dinitrophenyl)glutathione were only moderately decreased. These results suggest that Tyr7 is considered to be important the catalytic activities not only for GSH-chloronitrobenzene derivatives but also for GSH-epoxide conjugation reaction, rather than to binding of the substrates.

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