• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.4
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• pp.996-1001
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• 2003

It is well known that oxidative stress of reactive oxygen species(ROS) may be a causative factor in the pathogenesis of bone disorder. The purpose of this study was to evaluate the cytotoxicity of oxidative stress. Cell viability by MTS assay or INT assay, activity of glutathione peroxidase(GPx), lipid peroxidation(LPO) activity and cell viablity. And also protctive effect of glutamate receptors against ROS-induced osteotoxicity was examined by protein synthesis, alkaline phosphatase (ALP) activity and lactate dehydrogenase (LDH) activity in cultured rat osteoblasts and osteoclasts. XO/HX decreased cell viability and GPx activity, protein synthesis and ALP activity, but increased LPO activity and LDH activity. In the protective effect, N-methyl-D-aspartate (NMDA) receptor antagonists or AMPA/kainate receptor antagonists such as D-2-amino-5-phosphonovaleric acid (APV), 7-chlorokynurenic acid (CKA), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 6,7-dinitroquinoxaline-2,3-dione (DNQX), NMDA receptor antagonists but AMPA/kainate receptor antagonists showed protective effect on xanthine oxidase (XO) and hypoxanthine (HX) in these cultures by the increse of protein synthesis, ALP activity.

• Herbal Formula Science
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• v.13 no.1
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• pp.9-33
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• 2005

Acetaminophen, which causes acute liver min in humans and animals, has made useful inducer of hepatoxicity for studying hepatopreventive drugs. Injinhotang is known as one of the hepatopreventive drugs. However, its mechanism of recovery of hepatoxicity treated with acetaminophen is poorly understood. this study was performed to observe the antioxidative effect of injinhotang extract and its several combination groups. The results were obtained as follows:1. In the study on free radical scavenging effect in vitro(the suppressing effect on peroxidation of linoleic acid on concentration, the scavenging effect of DPPH radical, inhibitory effect of superoxide in xanthine-xanthine oxidase system and the inhibitory effect on lipid peroxidation reaction by hydroxy radical in H2O2-Fe2+system, injinhotang have more effect than its components groups relatively. 2. In the study on antioxidants system in vivo(the level of serum LPO, the level of hepatic LPO, catalase, GSH, GST), only injnhotang has a significant effect. 3. In the study on hepatotoxicity(GOT, GPT, $\gamma$-GTP, ALP, LDH, b ilirubin), only injinhotang has a significant effect. These results suggest that injinhotang has the protective effect on acetaminophen-induced hepatoxicity. The mechanisms of these are supposed to be involved in antioxidant and three drugs' cooperative synergy effect.

• Herbal Formula Science
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• v.10 no.2
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• pp.159-188
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• 2002

The purpose of this study was to reseach the effect of Daehwanggamchoeumja(大黃甘草飮子) and its component groups on diabetes, free radicals, and antioxidants system in Alloxan-induced diabetic rats. The experimental group was divided into three groups: Daehwanggamchoeumja(DG), and its components groups, Gamdutang (Gamcho&Daedu; DG-I) and Daehwanggamchotang(DG-2). The results were obtained as follows: 1. In the study of effect on diabetic metabolic dysfunction(Glucose, Triglyceride, Total Cholesterol, HDL Cholesterol, Total Protein, Albumin, Creatine, BUN), only DG has a significant effect. 2. In the study on free radical scavenging effect in vitro(the suppressing effect on peroxidation of linoleic acid on concentration, the scavenging effect of DPPH radical, inhibitory effect of superoxide in xanthine-xanthine oxidase system, inhibitory effect on lipid peroxidation reaction by hydroxy radical in $H_2O_2Fe^{2-}$system, and the effect on Nitrate reductase activity), DG and DG-2 have more effect than DG-l relatively. 3. In the study on antioxidants system in vivo(The level of serum LPO, The level of hepatic LPO, Catalase, GSH, GST), only DG has a significant effect. These results suggest that Daehwanggamchoeumja(大黃甘草飮子) has an effect on diabetes, peroxidative damage by free radical, so it seems to be useful to prevent and treat diabetes. The mechanisms of these are supposed to be involved in antioxidant and three drugs' cooperative synergy effect.

• The Journal of Internal Korean Medicine
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• v.23 no.1
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• pp.57-64
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• 2002

Objective : This study was designed to investigate the anti-oxidative effects of Oldenlandiae Diffusae Herba Water extract (ODHW) on lipid peroxidation by free radicals oxidative hepatic injury. Methods : In order to evaluate anti-oxidative activities of ODHW in the liver cell, cultured normal rat liver cells(Ac2F) were incubated with or without ODHW. After 16 hours to 18 hours of experiment, cells were placed in DMEM medium without serum, and then incubated with 1mM tert-butyl hydro-peroxide(t-BHP) for two hours. Viable cells were detected by MTT assay. The levels of LPO induced by hydroxyl radical derived from H2O2-Fe2+ system in rat liver homogenate were determined by means of TBA. Inhibitory effect of ODHW on superoxide generation was measured by xanthine-xanthine oxidase system. Results : In the linoleic acid autoxidation system, ODHW exhibited antioxidant activity, which inhibited 85% of linoleic acid peroxidation. These effects were similar to those of dl-a-tocopherol. ODHW showed scavenging effects on DPPH radical, inhibited superoxide generation in xanthine-xanthine oxidase system, and also inhibited lipid peroxidation of rat liver tissue with hydroxyl radical derived from $H_2O_2-Fe^{2+}$ system. In addition, ODHW protected the cell death induced by t-BHP and it significantly increased cell viability in a normal rat liver cell(Ac2F)

• Korean Journal of Animal Reproduction
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• v.26 no.3
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• pp.275-289
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• 2002

The objective of this study was to develop an in vitro assessment of sperm fertilizing capacity of bulls and investigate the factors influencing sperm function and characteristics of frozen-thawed bovine spermatozoa. in vitro fertilization (IVF), the evaluation of motility and normal morphology, HOST (hypoosmotic swelling test), Ca-ionophore induced acrosome reaction, luminol and lucigenin-dependent chemiluminescence for the measurement of reactive oxygen species (ROS), the measurement of malondialdehyde formation for the analysis of lipid peroxidation (LPO), and the evaluation of DNA fragmentation using the method of 747-mediated nick end labelling (TUNEL) by flow cytometry were performed in frozen-thawed bovine spermatozoa. Correlations between the rates of fertilization, blastocyst formation after IVF and the values of respective assays were investigated. 1. IVF rate and blastocyst formation rate averaged 64.4% and 34.3% for spermatozoa from high -fertility bull group and averaged 18.5% and 6.2% for spermatozoa from low-fertility bull group, respectively. There were significantly different between two bull groups. Sperm motility and percentage acrosome reaction averaged 79.0% and 66.2% for spermatozoa from high-fertility bull group and averaged 40.7% and 22.9% for spermatozoa from low-fertility bull group, respectivitely. There were not different between two bull groups. 2. Luminol depenent chemiluminescence, LPO and DNA fragementation averaged 6.4, 2.0 nmol and 2.6% from spermatozoa from high-fertility bull group and averaged 6.5, 3.1 nmol and 7.4% for spermatozoa from low-fertility bull group, respectively. There were significantly different between two bull groups. There was no significant difference in lucigenin dependent chemiluminescence between two bull groups. 3. Fertilization rate was positively correlated with motility and the rate of Ca-ionophore induced acrosome reaction, but negatively correlated with the frequency of luminol-dependent chemiluminescence, the rate of LPO, and the percentage of sperm with DNA fragmentation. There was no correlation between fertilization rate and the percentage of swollen spermatozoa, normal morphology, and the frequency of lucigenin-dependent chemiluminescence. 4. Blastocyst formation rate was positively correlated with the rate of Ca-ionophore induced acrosome reaction, but negatively correlated with the frequency of luminol-dependent chemiluminescence, the rate of LPO, and the percentage of sperm with DNA fragmentation. There was no correlation between blastocyst formation rate and motility, the percentage of swollen spermatozoa, normal morphology, and the frequency of lucigenin-dependent chemiluminescence. In conclusion, these data suggest that ROS significantly impact semen quality. The assays of this study may provide a basis fur improving in vitro assessment of sperm fertilizing capacity.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.1
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• pp.81-86
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• 2005

The purpose of this study is to examine the antioxidant activity in the germ cells of the extract of Psoraleae fructus. The extract was studied for dipheny-picryl-hydrazyl (DPPH) radical scavenging activity, GC-1 cell viability by a modified MTT assay, the effects on $H_2O_2$-induced cytotoxicity by MTT assay and lipid perixidation by malondialdehyde (MDA) formation, respectively. The results showed that the extract scavenged DPPH radical with the IC50 being 0.427 mg/mL. The extract was dose-dependent in growth of GC-1 cell. $H_2O_2$-induced cytotoxicity (67.7 %) was blocked by the extract concentration- dependently. Furthermore, the extract also displayed a dose-dependent reduction of MDA formation on $H_2O_2$-induced lipid peroxidation. In conclusion, the extract of Psoraleae fructus has potent antioxidant activity.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.6
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• pp.1541-1545
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• 2005

The purpose of this study is to examine the antioxidant activity in the germ cells of the extract of Corm fructus. The extract was studied for diphenyl-picryl-hydrazyl (DPPH) radical scavenging activity, GC-1 cell viability by a modified MTT assay, the effects on $H_2O_2$-induced cytotoxicity by MTT assay and lipid perixidation by malondialdehyde (MDA) formation, respectively. The results showed that the extract scavenged DPPH radical with the IC50 being $200{\mu}g/mL$. The extract at concentrations of $10-500{\mu}g/ml$ showed dose-dependent in growth of GC-1 cell. $H_2O_2$-induced cytotoxicity (63.0%) was blocked by the extract (10, 50, 100, 250 and $500{\mu}g/ml$) concentration-dependently. Furthermore, the extract (50, 100 and $250{\mu}g/ml$) also displayed a dose-dependent reduction of MDA formation on $H_2O_2$-induced lipid peroxidation. In conclusion, the extract of Corm fructus has potent antioxidant activity.

• Development and Reproduction
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• v.23 no.1
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• pp.11-19
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• 2019

The study was conducted to investigate the effects of alpha-linolenic acid (ALA) combined with bovine serum albumin (BSA) or methyl-beta-cyclodextrin (MBCD) on plasma and acrosomal membrane damages, mitochondrial activity, morphological abnormality, motility, and oxidative stress in frozen-thawed boar sperm. In previous our study, 3 ng/mL ALA had been shown protective effect during freezing process of boar sperm. Therefore, we used 3 ng/mL ALA in present study and ALA was combined with same molar ratio of BSA or MBCD (ALA+BSA and ALA+MBCD, respectively). To confirm the effect of two carrier proteins, same volume of BSA and MBCD without ALA were added during cryopreservation. Membrane damage, mitochondrial activity, reactive oxygen species (ROS) and lipid peroxidation (LPO) levels were measured using flow cytometry, and movement of sperm tail as motility parameter and morphological abnormality were observed under light microscope. In results, all of sperm parameters were enhanced by ALA combined with BSA or MBCD compared to control groups (p<0.05). Mitochondrial activity, morphological abnormality, ROS and LPO levels in ALA+BSA or MBCD groups were no significant difference compared with ALA, BSA and MBCD treatment groups. On the other hand, plasma and acrosomal membrane intact, and sperm motility in ALA+MBCD group were higher than single treatment groups (p<0.05), whereas ALA+BSA did not differ. Our findings indicate that carrier proteins such as BSA and MBCD could improve the effect of ALA during cryopreservation of boar sperm, and treatment of ALA with carrier proteins enhance membrane integrity, mitochondrial activity through reduction of ROS-induced LPO.

• Journal of the Korean Society of Food Science and Nutrition
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• v.20 no.2
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• pp.111-120
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• 1991

In order to investigate the cellular peroxidative damage due to heated oil intake and the preventive effect of vitamin E on it rats were fed heated corn oil with acid value of 4.02 at the level of 10 Cal% and three different levels of vitamin E that were 0, 40 and 200 mg/kg diet. Control group was fed fresh corn oil and 40mg/kg diet of vitamin E. After ech feeding period of 0, 3 and 6 weeks, liver superoxide dismutase (SOD), glutathione peroxidase (GPX) activities and microsomal content of vitamin E and lipid peroxide (LPO) were measured as well as cellular morphology was examined. SOD activities and LPO contents were higher, while GPX activities and vitamin e contents were lower in heated oil groups than control group. Electromicroscopic observation revealed the loss of inner mitochondrial membrane and cristae and irregular arrangement of nuclear membrane and chromatin in heated oil groups. As dietary vitamin e level was increased, SOD activity and LPO content were decreased, but GPX activity and vitamin E content in the liver increased and cellular peroxidative damage reduced progressively. This phenomena was more remarkable in 6 weeks of feeding than 3 weeks.

• Bulletin of Food Technology
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• v.11 no.4
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• pp.49-60
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• 1998

Ascorbate-$Fe^3+$-EDTA에 의해 생성된 hydroxyl radical (HO.)을 이용하여 단백질과 지질의 산화적 손상을 유도하였으며, 이 손상에 대한 멜라토닌(melatonin), glutathione, $\alpha$-tocopherol의 방어효과를 분석하였다. 단백질로는 bovine serum albumin (BSA), 지질로는 phosphatidyl-choline (PC) liposome을 이용하였다. 그리고 BSA와 PC liposome의 혼합계에서도 각 항산화제의 효능을 분석하였다. 단백질의 산화적 손상은 carbonyl기의 생성 및 단백질 절단을, 지질산화는 lipid peroxidation (LPO, malondialdehyde 와 4-hydroxyalkenal) 분석을 통하여 조사하였다. 단백질의 산화는 멜라토닌과 glutathione이, 지질산화는 멜라토닌과 $\alpha$-tocopherol이 효과적으로 억제하였다. 그러나 glutathione은 지질, $\alpha$-tocopherol은 단백질의 산화를 억제하지 못하였다. BSA와 PC liposome이 동시에 존재하는 혼합물에서는 수용성과 지용성 특성을 모두 지닌 멜라토닌은 단백질, 지질 모두에 대해 산화방지효과가 있으나 수용성인 glutathione은 단백질에, 지용성인 $\alpha$-tocopherol은 지질에만 주로 효과가 있는 것으로 조사되었다.