• 한국발생생물학회지:발생과생식
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• 제2권1호
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• pp.29-37
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• 1998

포유류 배아의 초기발생과정에서 수란관이 하는 역할을 알아보기 위하여 소의 수란관내액이 생쥐 2-세포기 배아의 체외발생에 미치는 영향을 조사하였다. 대조군의 경우 5%의 배아만이 체외배양과정 중 퇴화한데 비해 5% 혹은 그 이상의 소의 수란관내액 (bOF)이 함유된 배양액 내에서 배양된 배아는 48시간이 지났을 때 모두 퇴화하였다. bOF를 65 \circ C에서 30분간 가열한 후 배양액에 첨가한 결과 bOF의 독성은 변화가 없었으나 90 \circ C에서 30분간 가열하였을때에는 독성이 거의 사라져 95% 의 배아가 정상적으로 발생하였다. bOF를 chymotrypsin으로 1시간 혹은 24시간동안 처리한 후 배양액에 첨가한 경우에도 독성이 사라져 각각 95.5%의 배아가 상실배 혹은 포배로 발생을 진행하였다. 산화방지제인 10mM 농도의 glutathione (GSH)을 bOF와 함께 배양액에 첨가한 결과 마찬가지로 bOF의 독성이 사라져 91.0%의 배아가 상실배 이상으로 발생한 반면 산화 방지능력이 없는 산화형 GSH(GSSG)를 bOF와 함께 배양액에 처리한 경우에는 bOF의 독성은 전혀 제거되지 않았다. 또한 mercaptoethanol, dithiothreitol, cysteamine 등의 다른 산화방지제도 GSSG와 마찬가지로 bOF의 독성을 제거하지 못하였다. 이와 같은 실험 결과로 미루어 소의 수란관내액에는 체외에 노출될 경우 독성을 나타내는 단백질 성분으로 된 물질이 있으며 이 물질의 독성은 대기 중의 산소의 산화작용에 의해 활성화되는 것으로 여겨진다.

• Bulletin of the Korean Chemical Society
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• 제34권9호
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• pp.2671-2674
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• 2013

To elucidate the role of the evolutionarily conserved Tyr8 residue in rice Phi-class GSTF3, this amino acid was replaced with alanine and phenylalanine by site-directed mutagenesis, respectively. The replacement of Tyr8 with Ala significantly affected the catalytic activity and the kinetic parameters, whereas the substitutions of Tyr8 with Phe had almost no effect. The Y8A mutant resulted in approximately 90-100% decrease of the specific activity. Moreover, the Y8A mutant resulted approximately in 2-fold increase of $K_m$, approximately 60-80% decrease of $k_{cat}$, and approximately 6.5-fold decrease in $k_{cat}/K_m$. From the pH/log $k_{cat}/K_m$ plot, $pK_a$ values of the GSH in the wild-type enzyme-GSH complex, Y8A-GSH complex and Y8F-GSH complex were estimated to be approximately 6.8, 8.5 and 6.9, respectively. From these results, we suggest that the evolutionarily conserved Tyr8 residue in OsGSTF3 seems to influence the structural stability of the active site of OsGSTF3 rather than directly its catalytic activity.

• BMB Reports
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• 제28권5호
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• pp.420-426
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• 1995

This study was conducted to compare the effects of n-6 linoleic acid and n-3 linolenic acid on lipid peroxidation and the activities of enzymes defending against oxidation, which are involved in the tumor promotion, and histolOgical changes of hepatocarcinogen treated rat liver. In this study, weanling male Sprague-Dawley rats were fed one of three diets, containing 15% (w/w) of beef fat (BF), com oil (CO) or perilla oil (PO), for 11 weeks. During the 3rd week, experimental groups were injected with 2-AAF (50 mg/kg of BW) intraperitoneally 3 times. Findings show that the com oil diet group has greater liver MDA content than the beef fat and perilla oil diet groups. Also, it is observed that the perilla oil diet group has lower MDA content than beef fat and com oil diet groups, even though perilla oil is more desaturated than beef fat and com oil. In terms of activity, mixed-function oxidase activity is not Significantly affected by the different dietary fats and 2-AAF treatment. GSH-peroxidase, GSH-reductase and GSH-Stransferase activities are significantly higher in the CO+AAF group than those of the other groups. GST and GSH-Px are activated by 2-AAF treatment in the com oil diet group only. The hepatocytes of the BF+AAF group were the most severely degenerated, the second was the CO+AAF group and the least was the PO+AAF group. It was also found that dietary com oil increased lipid peroxidation and activated defense enzymes against oxidation in liver, but dietary perilla oil did not, or supressed defense enzymes. Therefore it is concluded that dietary n-3 linolenic acid in perilla oil inhibits lipid peroxidation and carcinoenesis in rat liver following 2-AAF treatment.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.78-78
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• 2003

Embryonic stem (ES) cells, derived from preimplantation embryo, are able to differentiate into various types of cells consisting the whole body, or pluripotency. In contrast, terminally differentiated cells do not usually alter their nature but frequently die or transform if they are exposed to inappropriate external stimulations. In addition to the plasticity, ES cells are expected to be different from terminally differentiated cells in very many ways, such as patterns of gene expressions, ability and response of the cells in confronting environmental stimulations, metabolism, and growth rate. As a model system to differentiate these two types of cells, human ES cells (MB03) and terminally differentiated cells (HeLa), we examined the ability of these two types of cells in confronting a severe oxidative insult, that is $H_2O$$_2$. Approximately 1$\times$10$^4$ cells were plated in 96 well plate and serum starved for overnight. The conditioned cells were exposed to a various concentration of $H_2O$$_2$ fur 24 hrs and loaded with neutral red (50$\mu\textrm{g}$/ml) for 4 hrs, washed with PBS for 2 min three times, and entrapped dye was dissolved out using acetic ethanol. Cytotoxicity was determined by reading the amount of dye in the medium using microplate reader. equipped with 575 nm filter. Relative amount of the dye entrapped within MB03 or HeLa were not significantly different when cells were exposed up to 0.4 mM $H_2O$$_2$. However, this sharply decreased down to 0.12% in HeLa cells when the cells were exposed to 0.8 mM $H_2O$$_2$, while it was approximately 54% in MB03 suggesting that this concentration of $H_2O$$_2$ is the defensive threshold for HeLa cells. The resistance to oxidative stimulation reversed, however, when cells were co-treated with BSO (L-buthionine- 〔S, R〕-sulfoximine) which chelates intracellular GSH. This result suggests that cellular GSH is the major defensive mechanism of human ES cells. Induction of enzymes involved in GSH metabolism and type of cell death is currently being studied.

• Asian-Australasian Journal of Animal Sciences
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• 제17권8호
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• pp.1162-1167
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• 2004

The present study was designed to define whether dietary conjugated linoleic acid (CLA) could affect antioxidant enzymes including superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), and glutathione S transferase (GST), and the level of malondialdehyde (MDA), a marker of lipid peroxidation, in the small intestine and liver from broiler chickens. A total of twenty-four 3 wk-old male broiler chickens were assigned to three dietary treatments (1.5% corn oil, 0.75% corn oil plus 0.75% CLA, and 1.5% CLA, isocalorically), and fed a grower-finisher diet from 22 to 35 days. In the small intestinal mucosae, the specific activities of SOD, GSH-Px, CAT, and GST, and the level of MDA were not substantially influenced by dietary CLA. In the liver, the specific activities of SOD, GSH-Px, and GST, and the level of MDA were also unaffected by dietary CLA at the level of either 0.75% or 1.5% compared with corn oil at the level of 1.5%. However, the broiler chickens fed the diet containing 1.5% CLA resulted in a significant increase in peroxisomal CAT activity and a marked decrease in total lipid and non-esterified fatty acids (NEFA) from liver tissues compared with those fed the diet containing 1.5% corn oil. In conclusion, ability of CLA to increase hepatic CAT activity suggest that dietary CLA may affect, at least in part, antioxidant defense system as well as lipid metabolism in the liver of broiler chickens.

• Journal of Ginseng Research
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• 제28권2호
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• pp.78-86
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• 2004

Panax ginseng occupies an important role in the folk medicine of China, Korea and Japan. The present study was undertaken to determine the radioprotective efficacy of ginseng root extract in the liver of Swiss albino mice. The animals were divided into 4 groups. Group I-Only vehicle was administered. Group II-The animals received 10 mg/kg body weight ginseng root extract i.p. for 4 consecutive days. Group III-Animals were irradiated with 8Gy gamma radiation at the dose rate of 1.69 Gy/min at the distance of 80 ems. Group IV-Animals were given by ginseng root extract (10 mg/kg body weight) continuously for 4 days and on 4th day they were irradiated with 8 Gy gamma radiation after 30 min. The animals from above groups were autopsied on 1,3,7,14 and 30 days. Biochemical estimations of phosphatases (acid ＆ alkaline), LDH (lactate dehydrogenase), LPO (lipid peroxidation) and GSH (reduced glutathione) in liver and SGOT (serum glutamate oxaloacetate transaminase), SGPT (serum glutamate pyruvate transaminase) and alkaline phosphatase in serum were done. In ginseng treated group acid phosphatase (ACP), alkaline phosphatase (ALP), LPO and LDH in liver and SGOT, SGPT and alkaline phosphatase in serum did not show any significant alteration. However, a significant increase in GSH content in liver was recorded. In irradiated group there was a significant increase in ACP, ALP and LPO content in liver and SGOT ＆ SGPT in serum was noted. Whereas, a significant decrease was recorded in GSH and LDH activity in liver and alkaline phosphatase activity in serum. Pretreatment of ginseng with radiation significantly alters the biochemical parameters in liver and serum. A significant decline in ACP, ALP activity and LPO content in liver and SGOT and SGPT activity in serum was observed. However, a significant increase in GSH content and LDH activity in liver and ALP activity in serum was estimated. The present study suggests that pretreatment of ginseng before irradiation significantly protects the liver and maintains the enzyme activity.

• Preventive Nutrition and Food Science
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• 제20권3호
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• pp.198-203
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• 2015

The present study was conducted to assess the status of vitamin C among healthy young adults in relation to serum antioxidant parameters [glutathione (GSH), thiols, and total antioxidant capacity, (TAC)], and oxidative stress markers [malondialdehyde (MDA), and nitrites plus nitrates (NN)]. A prospective study included 200 young adults, and their dietary intake was assessed by using food diaries. Fasting plasma vitamin C, serum levels of GSH, thiols, TAC, MDA, and NN were measured using biochemical assays. It was observed that 38% of the enrolled subjects, n=76, had an adequate dietary intake of vitamin C (ADI group). Meanwhile, 62%, n=124, had a low dietary intake of vitamin C (LDI group) as compared to the recommended dietary allowances. The fasting plasma level of vitamin C was significantly higher in the ADI group as compared to the LDI group. Oxidative stress in the sera of the LDI group was evidenced by depletion of GSH, low thiols levels, impairment of TAC, an elevation of MDA, and increased NN. In the ADI group, positive correlations were found between plasma vitamin C and serum antioxidant parameters (GSH, thiols, and TAC). Meanwhile, the plasma vitamin C was negatively correlated with serum MDA and NN levels. This study reveals a significant increase of oxidative stress status and reduced antioxidant capacity in sera from healthy young adults with low intake of the dietary antioxidant, vitamin C.

• Safety and Health at Work
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• 제12권1호
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• pp.127-132
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• 2021

Background: Herbicides such as glyphosate, paraquat, and 2,4-dichlorophenoxyacetic acid have been reported to cause adverse side effects through production of reactive oxygen species. However, there were no data representing the adverse effects of a mixture herbicide usage in farmers, especially the changes in oxidative marker and antioxidant defense. This study aimed to determine the urinary malondialdehyde (MDA) and glutathione (GSH) level in farmers using mixed herbicides. Methods: Ninety-three farmers were recruited, and two spot urine samples (before and after work) were collected. The urinary MDA level was evaluated by thiobarbituric acid reactive substance assay, and the urinary GSH level was determined using the enzymatic recycling method. Results: Sixty-two percent of the participants were men, and 59% of the participants worked in a farm for 20-40 years. The common combinations of herbicide usage were glyphosate with 2,4-dichlorophenoxyacetic acid (36.5%). There was no significant difference between pre- and post-work urinary MDA and GSH levels among the 3 groups of herbicides. However, the urinary MDA levels in farmers using the combination of glyphosate and paraquat were significantly higher than those found in farmers using glyphosate alone. The associated factors with changes in MDA levels found that the exposure intensity index (B = 0.154), the cumulative exposure intensity index (B = 0.023), and wearing gloves while working (B = -2.347) were found to be significantly associated with MDA level. Conclusion: The results suggest that the combined use of glyphosate and paraquat caused a significant increase in urinary MDA levels. Moreover, intensity of exposure to herbicide and wearing gloves were associated with the level of MDA.

• 한국식품위생안전성학회지
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• 제26권3호
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• pp.260-265
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• 2011

Fe-NTA는 전립선에 산화 스트레스를 일으킨다고 보고되어 있으며, 본 연구자는 랫드에 항산화 효과가 있다고 알려진 차전초잎 추출물(P. asiatica leaf extract, PLE)올 1, 2 또는 4 g/kg body weight (b.w.) 1주일간 경구 투여하고 ferric nitrilotriacetate (Fe-NTA)의 복강 주사로 전립선에 손상을 일으킨 후 차전초가 전립선에서 산화 스트레스를 얼마나 억제하여 주는지를 항산화 바이오 마커들인 reduced glutathione (GSH), glutathione-S-transferase (GST), glutathione reductase (GR) 및 지질과산화물의 척도인 malondialdehyde (MDA)를 통해서 측정하였다. 전립선 기능 이상 시 증가되는 GSH 및 GR은 아무것도 처리하지 않은 대조군(193.84${\pm}$37.78 mmol/g tissue, 31.32${\pm}$3.85)에 비해 Fe-NTA로 산화스트레스를 유발한 그룹에서 101.89${\pm}$24.31, 15.74${\pm}$2.92mmol/g tissue 48%의 감소를 나타낸다. 반면 PLE 1 g/kg b.w.을 투여한 그룹에서는 GSH 및 GR 수치가 119.01${\pm}$1.23 mmol/g tissue와 19.24${\pm}$0.53 mmol/g tissue, 2 g/kg b.w.을 투여한 그룹에서는 150.80${\pm}$34.11 mmol/g tissue와 19.17${\pm}$3.31 mmol/g tissue 마지막으로 4 g/kg b.w. 투여 그룹에서는 182.99${\pm}$10.89와 26.88${\pm}$4.40 mmol/g tissue로 Fe-NTA만 처리한 그룹에 비해 농도 의존적으로 GSH와 GR 함량이 회복되는 경향을 보이며, 이는 GSH 함량이 농도에 따라 약 l.2, 1.5, 1.8배 증가, GR 함량이 약 1.2, 1.22, 1.7배 증가한 것이다. 이렇듯 정자 보호와 전립선 조직 세포 보호에 관련이 있는 GSH와 GR값이 유의적 차이로 회복되는 결과에 따라 PLE의 전립선 보호 효과를 확인하였다. 또한, 항산화의 또 다른 바이오 마커인 GST 값은 아무것도 처리하지 않은 그룹 96.11${\pm}$6.23 mmol/g tissue에 비해 Fe-NTA 만 처리한 그룹 53.29${\pm}$11.45 mmol/g tissue이 약 45% 정도 감소하였지만, PLE 1, 2 또는 4 g/kg b.w.을 투여 한 그룹에서 각각 65.74${\pm}$9.79 mmol/g tissue, 76.54${\pm}$4.44 mmol/g tissue, 91.66${\pm}$5.53 mmol/g tissue의 값을 나타내며 Fe-NTA에 의해 생성된 산화 스트레스의 자유라디칼을 효과적으로 억제함을 확인하였다. 또한, 전립선에서 철 이온에 의해 유도된 산화스트레스에 의해 발생된 지질과산화물을 측정하였을 때 Fe-NTA에 의해 산화 스트레스만 유발한 그룹은 192.74${\pm}$33.20mmol/g tissue로 대조군 그룹의 75.66${\pm}$14.90 mmol/g tissue 보다 2.55배 높은 MDA를 생성한 것으로 지질과산화가 많이 일어난 것을 확인하였으나, PLE 1, 2 또는 4 g/kg을 투여한 그룹의 MDA의 생성량은 137.84${\pm}$23.29, 125.16${\pm}$16.69, 85.98${\pm}$5.12 mmol/g tissue로 약 29%, 35%, 55% 감소한 것을 확인하였다. 랫드의 전립선에서, 독성을 유발하는 Fe-NTA를 투여 후 PLE를 투여하였을 때 전립선 인지질 막의 손상지표인 MDA의 농도 의존적 감소와 항산화 및 정자 보호의 전립선 기능의 지표인 GSH, GR의 값이 증가하였다. 위의 결과들을 종합하였을 때, 우리는 Fe-NTA가 전립선에서 산회스트레스를 유발하고, 전립선 인지질 막의 손상을 줄 수 있으며, PLE는 이러한 전립선 손상을 항산화 효과를 기본으로 하여 전립선 보호 효과가 있음을 확인하였다.

• 한국식품위생안전성학회지
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• 제26권2호
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• pp.107-113
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• 2011

현대사회는 생활습관, 공해, 오염, 독성물질 등 여러 종류의 산화스트레스를 받는 환경에 노출되어 있으며, 이런 산화스트레스들은 인간에게 있어 여러 질병을 야기한다는 많은 증거 들이 나오고 있다. Ferric Nitrilotriacetate (Fe-NTA)는 iron-induced oxidative Stress로 인해 신장독성에 관여되어 있다고 보고된 물질이다. 이에 본 연구진은 랫드에 항산화 효과가 있는 차전초(Plantago asiatica) 추출물을 투여하고 Fe-NTA를 복강 주사하여 선장에서 iron-induced oxidative stress를 유발한 후, 산화스트레스를 얼마나 억제하여 주는 지를 혈액에서 신장기능 지표인 blood urea nitrogen (BUN)과 creatinine (Cr)을 측정하고, 신장조직에서는 항산화 바이오마커들인 reduced glutathione (GSH), glutathione-S-transferase (GST), glutathione reductase (GR) 및 지질과산화물 (lipid peroxide)인 malondialdehyde (MDA)를 측정 하였다. 신장기능 이상시 증가되는 BUN 및 Cr은 Fe-NTA에 의해 산화스트레스만 유발한 그룹에서는 $116.82{\pm}5.20$ mg/dL과 $2.70{\pm}0.20$ mg/dL로 아무것도 처리하지 않은 대조군의 $18.54{\pm}1.29$ mg/dL, $0.52{\pm}0.04$ mg/dL에 비해 약 6.3배 및 5.2배의 증가를 나타냈다. 반면 차전초를 투여한 1g/kg b.w. 군에 서 BUN 및 Cr의 활성 이 각각 $95.76{\pm}7.89$ mg/dL과 $2.16{\pm}0.340$ mg/dL, 2g/kg b.w. 군에서는 $89.34{\pm}26.93$ mg/dL 과 $2.20{\pm}0.51$ mg/dL, 4 g/kg b.w. 군에서는 $39.54{\pm}21.93$ mg/dL과 $1.16{\pm}0.55$ mg/dL으로 농도 의존적으로 유의적 (p < 0.05) 차이를 보이며 감소하는 결과를 보였다. 항산화의 바이오마커로 작용하는 GSH, GST 및 GR은 Fe-NTA에 의해 산화스트레스만 유발한 그룹에서는 각각 $59.45{\pm}12.32$ mmol GSH/g tissue, $49.88{\pm}4.55$ Units/g tissue, $56.70{\pm}5.40$ Units/g tissue로 아무것도 처리하지 않은 정상그룹의 $142.82{\pm}16.51$ mmol GSH/g tissue, $124.69{\pm}13.073$ Units/g tissue, $107.31{\pm}8.70$ Units/g tissue에 비해 58.4%, 60.0%, 47.2%로 현저하게 감소하였다. 반면 차전초 추출물을 1, 2, 4 g/kg b.w. 으로 투여한 그룹에서의 GSH는 $77.86{\pm}12.62$, $123.11{\pm}12.72$, $147.97{\pm}26.27$ mmol GSH/g tissue로 Fe-NTA만 처리한 그룹에 비해 약 1.3배, 2.1배, 2.5배 증가하였으며, GST는 $66.59{\pm}5.01$, $83.25{\pm}8.38$, $124.68{\pm}13.67$ Units/g tissue.로1.3배,1.7배, 2.5배 증가, GR은 $67.37{\pm}8.66$, $80.34{\pm}6.06$, $98.67{\pm}10.11$ Units/g tissue로 1.2배, 1.4배, 1.7배 증가하였다. 산화스트레스로 유발되는 지질과산화물을 신장에서 측정 하였을 때 Fe-NTA에 의해 산화스트레스만 유발한 그룹은 $215.70{\pm}49.73\;{\mu}mol$/g tissue로 대조군 그룹의 $46.20{\pm}10.65\;{\mu}mol$/g tissue보다 지질과산화를 많이 일으켜 4.7배 많은 MDA를 생성한 것을 나타냈지만, 차전초 추출물 1, 2,4 g/kg b.w. 투여한 그룹은 MDA의 생성량이 $141.74{\pm}10.79$, $116.11{\pm}9.19$, $86.52{\pm}22.30\;{\mu}mol$/g tissue로 약 34.3%, 46.2%, 59.9% 감소한 것을 확인하였다. 신장에서 iron-induced oxidative stress에 의해 독성을 유발하는 Fe-NTA를 처리 하였을시 신장손상 지표인 BUN과 Cr은 증가하고, 항산화 지표인 GSH와 GST, GR은 감소함과 동시에 지질과산 화물인 MDA는 증가하였다. 그러나 차전초 추출물을 농도를 달리하여 투여한 후 Fe-NTA로 산화스트레스를 유발한 그룹은 농도 의존적으로 BUN과 Cr은 감소하고, 항산화 지표들은 증가함과 동시에 MDA는 감소하는 경향을 보였으며 Fe-NTA만 처리한 그룹과 유의적 차이를 보였다. 위 결과들을 종합하면 Fe-NTA는 산화스트레스에 의해 신장에 심각한 손상을 줄 수 있으며, 차전초 추출물은 항산화 효과를 바탕으로 하여 Fe-NTA에 의한 신장 손상에 대한 보호 또는 개선 효과가 있음을 확인 하였다.