• Korean Journal of Microbiology
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• v.52 no.1
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• pp.1-9
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• 2016

A unique Schizosaccharomyces pombe $TrxR^+$ gene encoding thioredoxin reductase (TrxR) was found to be positively regulated by stress-inducing agents through the stress-responsive transcription factor Pap1. In the present study, the protective roles of S. pombe TrxR were evaluated using the TrxR-overexpressing recombinant plasmid pHSM10. In the presence of hydrogen peroxide ($H_2O_2$) and superoxide anion-generating menadione (MD), S. pombe TrxR increased cellular growth and the total glutathione (GSH) level, while it reduced levels of intracellular reactive oxygen species (ROS). The nitric oxide (NO) levels of the TrxR-overexpressing cells, in the presence of $H_2O_2$ and MD, were maintained to be similar to those of the corresponding non-treated cells. Although S. pombe TrxR was able to scavenge NO generated by sodium nitroprusside (SNP), it had no significant modulating effects on cellular growth, ROS levels, or the total GSH level of SNP-exposed yeast cells, compared with the differences in those of the two non-treated cell cultures. TrxR increased the cellular growth and total GSH level, which were diminished by nitrogen starvation. It also scavenged ROS and NO produced during nitrogen starvation. Taken together, the S. pombe TrxR protects against oxidative, nitrosative, and nutritional stresses.

• Journal of Nutrition and Health
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• v.33 no.7
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• pp.745-754
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• 2000

Effects of oral taurine supplementation (6g/day)for 2-4 weeks on activities of red blood cell(RBC)total superoxide dismutase (SOD) and plasma glutathione peroxidase(GSH-Px) and the level of malondialdehyde(MDA) were evaluated in healthy female adults (23.6$\pm$0.3 years old). Compared to the value for 0 week plasma GSH-Px activity of the subjects was significantly lower after 2 weeks of taurine supplementation(p<0.05) and recovered to the value similar to 0 week after 4 weeks of taurine supplementation. RBC total SOD activity tended to be decreased after 2 weeks of taurine supplmentation compared to the values for 0 week although the difference between the means of the two group was not statistically significant. Plasma MDA level was not significantly decreased by taurine supplementation most probably due to the fact that the subjects participated in the present study were healthy and their antioxidant defense system had been in the 'normal' range. Plasma MDA concentration was negatively correlated with plasma taurine concentration(r=-0.2003m p<0.05) but tended to be positively correlated with plasma cholesterol concentration(r=0.2465, p=0.0645) as expected Plasma GSH-Px activity was positively associated with the percentage of 22:0 (r=0.2892, p<0.05) or 20:4w6(r=0.2939, p<0.05). On the other hand plasma MDA concentration was positively correlated with the percentage of 20:5w3 in plasma total lipids(r=0.2635 p<0.05) and negatively correlated with $\Delta$5 desaturation index of w6 fatty acids(20:3w6⇒20:4w6) in plamsa total lipids(r=-0.2714, p<0.05) as well as in phospholipids(r=-0.2864, p<0.05). From these results protective effect of taurine supplementation against lipid peroxidation and antioxidant defense system in humans appears to be minimal when the subjects are in a relatively healthy state. Further studies concerning the antioxidant efficacy of taurine should be conducted in human subjects under various disease states related to oxidative stress such as diabetes and artheroxclerosis.

• Journal of Nutrition and Health
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• v.37 no.10
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• pp.872-880
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• 2004

This study was performed to investigate the effect of Puerariae radix-ethanol extracts rich in isoflavone on the antio-xidative system of rats. For this purpose, first, Puerariae radix was extracted with ethanol, and its total isoflavone and puerarin contents were analysed. Second, female Sprague Dawley rats were fed for 6 weeks with four diets which were based on AIN96G diet and supplemented with Puerariae radix-ethanol extracts to contain isoflavone. The isoflavone contents of four experimental diets were 0 mg, 500 mg, 1,000 mg, 2,000 mg per kg diet, respectively (control, P0.05%,P0.1%, P0.2%). Liver and erythrocyte activities of antioxidative enzyme such as superoxide dismutase (SOD), catalase,glutathione peroxidase (GSHpx) were measured. Also, plasma and liver malondialdehyde (MDA) concentrations, liver glutathione (GSH) and oxidized glutathione (GSSG) concentrations were measured. The total isoflavone content of Puerariae radix-ethanol extract was 3067.6 mg per 100 g extract and the content of puerarin was 2557.4 mg per 100 g extract. The erythrocyte activities of GSH-Px and catalase were higher in group P0.1% and P0.2%. But SOD activity of erythocyte did not show any difference by the Puerariae radix-ethanol extract supplementation in diet. The activity of SOD in liver increased significantly by the supplementation of extract, showing highest level in P0.1% group. The liver GSH concentration increased significantly in group of P0.05%, P0.1%, and P0.2% compared with control group (p <0.05). The GSSG concentration in liver showed no difference by the supplementation of Puerariae radix extract from the control group, except P0.2% group. The plasma MDA concentration did not show any significant differences by the extract supplementation. But the liver MDA concentration decreased by the extract supplementation, showing the lowest level in P0.1 % diet group. These results suggest that the supplementation of Puerariae radix-ethanol extract can inhibit lipid peroxidation in liver and enhance the antioxidative defense competence of rats.

• The Journal of Korean Medicine
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• v.38 no.3
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• pp.43-58
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• 2017

Objectives: We performed this study to evaluate the antioxidative and hypolipidemic effect of Artemisia iwayomogi (韓茵蔯), Curcuma longa L. (鬱金) and Raphanus sativus L. (蘿?子) (ACR). Method: We enriched Artemisiae Capillaris, Curcumae Longae and Raphani Semen compound with alcohol. ACR extract is treated to HepG2 cell. Cell groups are devided into 3 groups: normal, control and ACR treated group. We measured polyphenol, flavonoids, DPPH and ABTS radical scavenging activity, ROS, glutathione, GSH peroxidase, GSH reductase, SOD, catalase, free fatty acid, lipid peroxidation and suppression of ACAT1 and HMG-CoA reductase expression on mRNA level. Results: 1. ACR contained polyphenol and flavonoids and increased GSH significantly in HepG2 cell. 2. ACR increased GPx, GR, and catalase activity significantly in HepG2 cell. 3. ACR increased DPPH and ABTS radical scavenging activity significantly in HepG2 cell and decreased ROS. 4. ACR decreased free fatty acid and MDA significantly in HepG2 cell. 5. ACR suppressed ACAT1 and HMG-CoA reductase expression on mRNA level in HepG2 cell. Conclusion: This study suggests that ACR has antioxidative and hypolipidemic effect and might be effective in prevention and treatment of dyslipidemia.

• Environmental Analysis Health and Toxicology
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• v.20 no.1
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• pp.75-85
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• 2005

This study was aimed to know the effect of cadmium chloride (CdCl₂) on glucose transport in L6 myotube and its action mechanism. CdCl₂ increased the 2-deoxy- (l-3H)-D-glucose (2-DOG) uptake 1.9 and 2.4 fold at 10 and 25 μM respectively. To investigate the stimulating-mechanism of glucose transport induced by CdCl₂, the wortmannin and PD98059 were used as PI3K (phosphatidylinositol 3-kinase) inhibitor and MAPK inhibitor respectively, which did not affect 2-DOG uptake. This fact suggests that CdCl₂ induced 2-DOG uptake may not be concerned to the insulin signalling pathway. Whereas nifedipine, a calcium channel blocker, and trifluoperazine, a calmodulin inhibitor, were found to inhibit the 2-DOG uptake stimulted by CdCl₂. In addition, we also measured the ROS (reactive oxygen species) production and GSH level in L6 myotube to investigate the correlation between the glucose uptake and ROS. CdCl₂(25 μM) increased ROS generation approximately 1.5 fold and changed the cellular GSH level, but GSSG/GSH ratio remained unchanged. CdCl₂ stimulated 2-DOG uptake and ROS generation were inhibited by N-acetylcystein. And BSO pretreatment, a potent inhibitor of γ-GCS, resulted in the dramatic decrease of 2-DOG uptake and also the increase of the sensitivity to cadmium cytotoxicity. The obtained results suggest that CdCl₂-stimulated glucose uptake might be based on the activation of HMP shunt as an antioxidant defense mechanism of the cells.

• Environmental Analysis Health and Toxicology
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• v.20 no.1
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• pp.87-95
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• 2005

Cadmium is well known as a toxic metal and has insulin mimicking effects in rat adipose tissue. This study was undertaken to investigate the effect of CdCl₂ on glucose transport and its mechanism in 3T3 - L1 adipocytes. CdCl₂ exhibits respectively 2.2 and 2.8 fold increases in the 2-deoxyglucose uptake when exposed to 10 and 25 μM of CdCl₂ for 12 hr. To investigate the stimulating mechanism of glucose transport induced by CdCl₂. Wortmannin and PD98059 were used respectively as PI3K inhibitor and MAPK inhibitor, which did not affect 2-DOG uptake. This results suggest that induced 2-deoxy-(l-3H)-D-glucose (2-DOG) uptake by CdCl₂ may not be concerned with the insulin signalling pathway. Whereas nifedipine, a calcium channel blocker inhibited the 2- DOG uptake stimulated by CdCl₂. In addition, we also measured the increased production of Reactive oxygen substances (ROS) and glutathione (GSH) level in 3T3-L1 adipocytes to investigate correlation between the glucose uptake and increased production of ROS with H2DCFDA. CdCl₂ increased production of ROS. Induced 2-DOG uptake and increased production of ROS by CdCl₂ were decreased by N-acetylcystein (NAC). And L-buthionine sulfoximine (BSO) a potent inhibitor of γ-GCS, decreased of 2-DOG uptake. Also NAC and BSO changed the cellular GSH level, but GSH/GSSG ratio remained unchanged at 10, 25 μM of CdCl₂.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.4
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• pp.357-365
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• 2020

The aim of present study was to investigate regulatory mechanism of alpha-linolenic acid (ALA) during in vitro maturation (IVM) on nuclear and cytoplasmic maturation of porcine oocytes. Basically, immature cumulus-oocyte complexes (COCs) were incubated for 22 h in IVM-I to which hormone was added, and then further incubated for 22 h in IVM-II without hormone. As a result, relative cumulus expansion was increased at 22 h after IVM and it was enhanced by treatment of ALA compared with control group (p < 0.05). During IVM process within 22 h, cAMP level in oocytes was decreased at 6 h (p < 0.05) and it was recovered at 12 h in ALA-treated group, while oocytes in control group recovered cAMP level at 22 h. In cumulus cells, it was reduced in all time point (p < 0.05) and ALA did not affect. Treatment of ALA enhanced metaphase-I (MI) and MII population of oocytes compared with oocytes in control group at 22 and 44 h, respectively (p < 0.05). Intracellular GSH levels in ALA group was increased at 22 and 44 h after IVM (p < 0.05), whereas it was increased in control group at 44 h after IVM (p < 0.05). In particular, the GSH in ALA-treated oocytes during 22 h of IVM was higher than control group at 22 h (p < 0.05). Lipid amount in oocytes from ALA group was higher than control group (p < 0.05). Treatment of ALA did not influence to absorption of glucose from medium. Cleavage and blastocyst formation of ALA-treated oocytes were enhanced compared with control group (p < 0.05). These findings suggest that supplementation of ALA could improve oocyte maturation and development competence through increasing GSH synthesis, lipid storage, and regulation of cAMP accumulation during early 22 h of IVM, and these might be mediated by cumulus expansion.

• Nutrition Research and Practice
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• v.4 no.2
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• pp.114-120
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• 2010

The effects of grape seeds extract and grape peels extract prepared from grape pomace on the activity of antioxidant enzymes, degree of lipid peroxidation in serum and liver tissue were investigated in rabbits fed on high cholesterol diet. New Zealand white rabbits were divided as follows ; 1) NOR (normal group); 2) CHOL (cholesterol group); 3) GSH (cholesterol + grape seed extract group); 4) GPE (cholesterol + grape peel extract); 5) GSP (cholesterol + grape seed powder); 6) GPP (cholesterol + grape peel powder); 7) GE (cholesterol + grape seed and peel extract); 8) GP (cholesterol + grape seed and peel powder). Eight groups of rabbits were studied for 8 weeks. At the end of the experimental period, rabbits were sacrificed and the liver tissue were removed. Then, GSH, GPx, GST, CAT and MDA in the liver were measured. In liver tissues, total glutathione contents (GSH), glutathione peroxidase (GPx) and catalase (CAT) activity, which was significantly higher by grape seed extract supplementation. The level of malondialdehyde (MDA) was lower in the serum of rabbits fed grape seed extract or grape peel powder plus cholesterol than in the serum of rabbits fed cholesterol alone. It is therefore likely that grape seed extract prepared from grape pomace functioned as antioxidants in vivo, negating the effects of the oxidative stress induced by 1% cholesterol diet. The grape seed extract was found effective in converting the oxidized glutathione into reduced glutathione, and in removing $H_2O_2$ that is created by oxidative stress. The grape peel powder was found to have small influence on reduced glutathione content, CAT and GPX activity, but it increased GST activity in liver tissues, resulting in promoting the combination of lipid peroxide and glutathione (GSH), and further, lowering the formation of lipid peroxide in the serum. Therefore, grape pomace (grape seed extract and grape peel powder) supplementation is considered to activate the antioxidant enzyme system and prevent damage with hypercholesterolemia.

• 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.

• Herbal Formula Science
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• v.9 no.1
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• pp.355-366
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• 2001

Objectives : Oldenlandiae Diffusae herba has been used as a natural drug for tumor, inflammation and liver disease in traditional medicine. This study was performed in order to investigate the antioxidative effects of Oldenlandiae Diffusae herba methanol extract(ODHM) on acetaminophen induced acute liver injury in mice. Methods : In order to investigate the protective effect of ODHM on acute hepatic injury in vivo, ICR mice were pretreated with ODHM, and then treated with acetaminophen(500mg/kg). And the levels of LPO and glutathione(GSH), antioxidative enzyme activities were measured. The levels of LPO were measured by TBA method. And catalase activity was measured as the decrease in hydrogen peroxide absorbance at 240nm on spectrophotometer using 30mM hydrogen peroxide. Superoxide dismutase(SOD) was assayed by recording the inhibition of nitro blue tetrazolium reduction with xanthine and xanthine oxidase. Glutathione peroxidase(GPX) activity was determined by the modified coupled assay developed by Paglia and Lawrence. The reaction was started by addition of 2.2mM hydrogen peroxide as substrate. The change in absorbance at 340nm was measured for 1min on spectrophotometer. Glutathione-S-transferase(GST) activity was assayed with CDNB as substrate and enzyme activity of GST towards the glutathione conjugation of CDNB. And Total SH and GSH levels were measured. Results : In vivo study, LPO levels of acetaminophen treatment group were significantly higher than other groups. This increased level was significantly reduced by ODHM pretreatment. The acetaminophen treatment resulted in a decrease of catalase, GPX, SOD and GST activities. By contrast, ODHM pretreatment markedly increased compare to those of untreated groups. Total SH and GSH levels were reduced by of acetaminophen treatment, and ODHM pretreatment significantly increased GSH levels.