• Journal of Life Science
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• v.26 no.8
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• pp.948-954
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• 2016

We investigated the flavonoid and phenol contents and antioxidant effect of wine by-product extract. Antioxidant effects were measured with 1,1-diphenyl-2-picryhydrazyl (DPPH) and 2.2'-azino-bis(3-ethylbenothiazoline-6-sulfonic acid) diammonium salt radical cation (ABTS+) assays. Cellular reactive oxygen species (ROS) were measured with the dichlorofluorescein-diacetate (DCFH-DA) assay. The flavonoid and phenol contents of the methanol (MeOH) extract were greater than those of the acetone+methylene chloride (A+M) extract. Among fractions, the 85% aqueous methanol (85% aq. MeOH) fraction contained the highest flavonoid contents, while the n-BuOH fraction had more phenol contents. In the DPPH and ABTS assays, the MeOH extract showed a scavenging effect greater than that of the A+M extract (p<0.05). The n-BuOH fraction (0.5 mg/ml concentrations) showed scavenging effects of 72% and 92%, respectively, in the DPPH and ABTS assays (p<0.05). However, the 85% aq. MeOH fraction showed a 90% scavenging effect in the DPPH assay only. In 120 min ROS production assay, all tested fractions dose-dependently decreased cellular ROS production induced by H₂O₂ in comparison with that produced by exposure to the extract-free control. The MeOH extract showed a higher sinhibitory effect on cellular ROS producing than that of the A+M extract at all concentrations tested. Treatment with the n-BuOH fraction (0.1 mg/ml concentrations) inhibited cellular ROS production by 60%. These results indicate that the n-BuOH fraction of wine by-product extract inhibited cellular oxidation and may contain valuable bioactive compounds such as flavonoids and phenols.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.3
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• pp.671-678
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• 2007

Diabetes is a disease in which the body does not produce or properly use insulin. Etiological studies of diabetes and its complications showed that oxidative stress might play a major role. Therefore, many efforts have been tried to regulate free oxygen radicals for treating diabetes and its complications. Gamigukihwandong-hwan has been known to be effective for the treatment of diabetes. The present study was carried out to investigate the effect of Gamigukihwandong-hwan on renal function, peroxynitrite (ONOO$^-$) scavenging activity and polyol pathway in streptozotocin-induced diabetic rats. The crushed Gamigukihwandong-hwan was extracted 3 times, each time with 3 volumes of methyl alcohol at 60$^{\circ}C$ for 24 h. The extract was filtered and evaporated under a reduced pressure using a rotary evaporator to yield 74.95 g. Gamigukihwandong-hwan extract was oral-administered 100 mg per 1 kg of body weight for 20 days to the diabetic rats induced by streptozotocin (60 mg/kg). The effects of Gamigukihwandong-hwan extract on the streptozotocin-induced diabetic rats were observed by measuring the serum level of glucose, insulin, lipid components, creatinine and BUN, and also the kidney levels of superoxide anion radical (${\cdot}O_2^-$), nitric oxide (NO) and ONOO$^-$, and also the enzyme activities involved in polyol pathway. The Effects of Gamigukihwandong-hwan on the streptozotocin-induced diabetic rats with regards to body weight, blood glucose and insulin levels, creatinine and BUN levels, total cholesterol and triglyceride levels, and HDL-cholesterol levels were all shown to be good enough to cure and prevent the diabetes and its complications. Gamigukihwandong-hwan inhibited the generation of ${\cdot}O_2^-$, NO and ONOO$^-$ in the kidney of streptozotocin-induced diabetic rats. Renal aldose reductase and sorbitol dehydrogenase activities were increased in the streptozotocin-induced diabetic rats, whereas the ones in the Gamigukihwandong-hwan administered group among the streptozotocin-induced diabetic rats were reversed toward the natural activities. Gamigukihwandong-hwan might inhibit the development of diabetic nephropathy by scavenging reactive oxygen and nitrogen species, thereby by reducing oxidative stresses and also by regulating the activities of polyol pathway enzymes, all of which could help to recover the function of kidney.

• Journal of Applied Biological Chemistry
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• v.66
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• pp.250-257
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• 2023

Glutamate is an excitatory neurotransmitter distributed in the central nervous system of mammals. However, high concentrations of glutamate are known to cause neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and stroke by causing nerve cell death. In this study, the antioxidant activity and neuroprotective effect of subtropical natural products were analyzed. Among 11 subtropical plant extracts mainly tested, Sallacca wallichiana extract (SE) showed the greatest free radical scavenging activity. Then, we confirmed through WST-1 assay that SE protected HT22 cells against glutamate-induced cell death in a concentration-dependent manner. The protective effects of SE against glutamate-induced apoptosis in HT22 cells were also confirmed by flow cytometry analysis using Annexin V/PI double staining. We also confirmed using H₂DCF-DA single staining that SE inhibits glutamate-induced intracellular reactive oxygen species. And we were confirmed through that SE inhibited glutamate-induced phosphorylation of Mitogen-activated Protein kinases. Consequently, our results propose that SE may contribute to the development of therapeutics to prevent neurodegenerative diseases.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.9
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• pp.1249-1256
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• 2016

Black ginseng (BG) obtained by a 9-fold steaming process of Panax ginseng has been reported to have anti-oxidative, anti-obesity, and anti-diabetes effects. The current study evaluated the protective effect of BG by steaming time in an HCl/ethanol-induced acute gastritis model. BG was divided into four samples according to steaming-drying processing (Gin1, Gin3, Gin6, and BG). High performance liquid chromatography analysis, free radical scavenging activity, and total phenol and flavonoid contents were examined in ginseng and four BG samples. Compared with ginseng, BG showed a stronger radical scavenging effect and higher contents of total phenol and flavonoids. To evaluate the anti-gastritic effect of BG, mice were distributed into five groups: normal mice (N), acute gastritic mice with distilled water (CON), acute gastritic mice with 100 mg/kg of ginseng (Gin0), acute gastritic mice with 100 mg/kg of BG (BG), and acute gastritic mice with 10 mg/kg of sucralfate (SC). After 1 hour of pre-treatment with water, extracts (Gin0 and BG), or drug (SC), experimental groups except for N were orally administered 0.5 mL of 150 mM HCl/60% ethanol (v/v) mixture. Blood was collected 1 hour later from the heart, and gastric tissue was harvested. Reactive oxygen species (ROS) levels were measured in serum, and related protein expression was examined by Western blot assay. In HCl/ethanol-induced acute gastritic mice, treatment with ginseng or BG improved mucosal damage in the histological evaluation. The serum ROS level significantly decreased in the BG-treated group compared with the CON group. Furthermore, expression of inflammatory cytokines significantly decreased in the BG-treated group compared with the CON group. Based on these results, antioxidant and anti-gastritic activities of ginseng were enhanced by streaming-drying processing, in part due to an increase in biological active compounds.

• Korean Journal of Food Science and Technology
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• v.47 no.2
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• pp.233-239
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• 2015

This study determined the optimum extraction conditions based on five response variables (yield, total phenolics, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radical scavanging activity, oxygen radical absorbance capacity (ORAC), and ${\beta}$-1,3-glucan content) in chaga mushroom (Inonotus obliquus) using the response surface methodology, where three independent variables (ethanol concentration, extraction temperature, and extraction time) were optimized using a central composite design. The optimum ethanol concentration, extraction temperature, and extraction time were 50% (w/w), $88.7^{\circ}C$, and 14.5 h; 9.2%, $92.7^{\circ}C$, and 14.5 h; 50.8%, $92.7^{\circ}C$, and 14.5 h; 9.2%, $92.7^{\circ}C$, and 1.5 h; and 90.8%, $92.7^{\circ}C$, and 1.5 h for yield, total phenolics, ABTS, ORAC, and ${\beta}$-1,3-glucan content, respectively. The predicted values of the response variables were compared with those of the extracts under the optimal extraction conditions to verify the models. The optimum extraction condition for the five response variables was predicted to be 81.4% ethanol at $92.7^{\circ}C$ for 14.5 h.

• Journal of Chest Surgery
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• v.29 no.8
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• pp.850-860
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• 1996

Oxygen free radicals and their metabolites have been implicated as possible causes of reperrusion injury In animal models. Their role in the clinical setting is still controversial. The aim of this study was to evaluate the degree of tissue damage, oxidative stress. and changes in the antioxidant enzyme system in patients undergoing cor nary artery bypass graft operations(CABG) with myocardial protection by cold blood cardioplegia. In patients undergoing CABG(n:10). the levels of lactate dehydrogenate(LDH), creatine phosphokinase MB fraction(CK-MB), and malondialdehyde(M DA) were measured In the coronary sinus effluent before aortic cross clamping and 20 minutes after reperfusion. At the same time, the myocardial tissue activities of superoxide dismutase(SOD). catalase(CAT), glutathione peroxiddse(GSHPX), glutathione reductase (GSSGRd), and glucose 6-phosphate dehydrogenate(GfPDH ) were determined in the right atrial auricle excised before aortic cross clamping and in the left atrial auricle excised 20 minutes after reperfuslon. The levels of increased significantly after reperrusion(p< U.05). There were no significant changes in CAT and CfPDH levels. Western blot analysis was performed to study the induction of antioxidant enzyme and demonstrated increased amount of Cu,Zn-SOD.

• The Korean Journal of Physiology and Pharmacology
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• v.5 no.3
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• pp.223-230
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• 2001

Melatonin, a pineal gland hormone, is believed to act as an antioxidant via the stimulation of radical detoxifying enzymes and scavenging of free radicals. In this study, effects of in vitro and in vivo treatments of melatonin on the cisplatin-induced lipid peroxidation, LDH release and plasma creatinine were determined in rabbit renal cortical cells. The level of malondialdehyde (MDA) was assayed as an index of lipid peroxidation and the level of LDH release as an indicator of cellular damage. In in vitro studies, cisplatin increased the levels of MDA and LDH release in a concentration-and time-dependent manner. Melatonin inhibited the cisplatin-induced lipid peroxidation and LDH release in a concentration-dependent manner. The minimal effective concentration of melatonin that significantly reduced the $300\;{\mu}M$ cisplatin-induced lipid peroxidation and LDH release was 1 mM. In in vivo studies, the levels of lipid peroxidation and LDH release in renal cortical cells increased significantly 24 or 48 hours after a single injection of cisplatin (6 mg/kg). When the cisplatin-injected rabbits were pretreated with 10 mg/kg of melatonin, a significant reduction in both lipid peroxidation and LDH release was observed. The plasma creatinine level increased from $0.87{\pm}0.07$ mg/dl in control to $6.33{\pm}0.54$ mg/dl in cisplatin-injected rabbits (P＜0.05). Melatonin partially prevented the increase in serum creatinine level $(1.98{\pm}0.11\;mg/dl)$ by cisplatin (P＜0.05). In the proximal tubules from cisplatin-treated group, tubular cells had microvilli of variable heights. Necrotic debris was seen in tubular lumens. In most of cells, the mitochondria and lysosomes were increased in frequency. The endocytic vacuoles were not prominent and distribution of the brush border was irregular and shortened. These cisplatin-induced morphological changes were moderate in the melatonin-pretreated group. These results suggest that the toxicity of cisplatin is associated with the generation of reactive oxygen free radicals and that melatonin is a powerful antioxidant, which prevents some of the adverse effects of cisplatin.

• Journal of Applied Biological Chemistry
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• v.57 no.3
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• pp.205-210
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• 2014

Persicaria orientalis (L.) Spach (Po) and Potentilla fragarioides var. major Maxim (Pf) extracts were analyzed to investigate anti-inflammation through their suppressing effects on free radicals such as reactive oxygen species (ROS). In addition, with regard to Po and Pf, an analysis was conducted of their inhibitory effect on nitric oxide, which is produced in lipopolysaccharide (LPS)-treated murine macrophage RAW 264.7 cells, and their inhibitory effect on the translocation of the nucleus of nuclear factor-kappa B (NF-${\kappa}B$). The $IC_{50}$ value of ROS, which was induced by $50{\mu}M$ 3-morpholinosydnonimine hydrochloride (SIN-1), was found to be $23.35{\pm}1.27{\mu}g/mL$ due to the effect of the Po extract, and $8.46{\pm}1.22{\mu}g/mL$ due to the effect of the Pf extract. In addition, the $IC_{50}$ value of peroxynitrite treated with the Po extract was $2.19{\pm}0.04{\mu}g/mL$, whereas that of peroxynitrite treated with the Pf extract was $0.80{\pm}0.02{\mu}g/mL$. ROS and peroxynitrite were induced by $50{\mu}M$ 3-morpholinosydnonimine hydrochloride. There was an increase in the amount of nitric oxide in the RAW 264.7 cells treated with LPS ($1{\mu}g/mL$), whereas the level of NO was observed to significantly and dose-dependently decrease in the cells treated with Po and Pf. The amount of nitric oxide produced by the group treated with $10{\mu}g/mL$ of the Pf extract was $11.45{\pm}0.57{\mu}M$. Furthermore, the Po extracts inhibited the translocation of the nucleus of NF-${\kappa}B$ in LPS-treated RAW 264.7 cells. Therefore, it is highly possible that Po and Pf have anti-inflammatory properties.

• Tuberculosis and Respiratory Diseases
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• v.41 no.3
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• pp.222-230
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• 1994

Background: Paraquat, a widely used herbicide, is extremely toxic, causing multiple organ failure in humans. Paraquat especially leads to irreversible progressive pulmonary fibrosis, which is related to oxygen free radicals. However, its biochemical mechanism is not clear. Natural mechanisms that prevent damage from oxygen free radicals include changes in glutathione level, G6PDH, superoxide dismutase(SOD), catalase, and glutathione peroxidase. The authors think catalase is closely related to paraquat toxicity in the lungs Method: The effects of 3-amino-1,2,4-triazole(aminotriazole), a catalase inhibitor, on mice administered with paraquat were investigated. We studied the effects of aminotriazole on the survival of mice administered with paraquat, by comparing life spans between the group to which paraquat had been administered and the group to which a combination of paraquat and aminotriazole had been administered. We measured glutathion level, glucose 6-phosphate dehydrogenase(G6PDH), superoxide dismutase(SOD), catalase, and glutathione peroxidase(GPx) in the lung tissue of 4 groups of mice: the control group, group A(aminotriazole injected), group B(paraquat administered), group C(paraquat and aminotriazole administered). Results: The mortality of mice administered with paraquat which were treated with aminotriazole was significantly increased compared with those of mice not treated with aminotriazole. Glutathione level in group B was decreased by 20%, a significant decrease compared with the control group. However, this level was not changed by the administration of aminotriazole(group C). The activity of G6PDH in all groups was not significantly changed compared with the control group. The activities of SOD, catalase, and glutathione peroxidase(GPx) in the lung tissue were significantly decreased by paraquat administration(group B); catalase showed the largest decrease. Catalase and GPX were significantly decreased by aminotriazole treatment in mice administered with paraquat but change in SOD activity was not significant(group C). Conclusion: Decrease in catalase activity by paraquat suggests that paraquat toxicity in the lungs is closely related to catalase activity. Paraquat toxicity in mice is enhanced by aminotriazole administration, and its result is related to the decrease of catalase activity rather than glutathione level in the lungs. Production of hydroxyl radicals, the most reactive oxygen metabolite, is accelerated due to increased hydrogen peroxide by catalase inhibition and the lung damage probably results from nonspecific tissue injury of hydroxyl radicals.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.35 no.4
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• pp.287-292
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• 2009

Human skin is constantly exposed to environmental irritants such as smoke, chemicals and ultraviolet (UV). Free radicals and reactive oxygen species (ROS) caused by these environmental irritants play critical roles in cellular damage. In this study, to investigate the skin cell protective effect of Ophioglossum vulgatum extract, we investigated its effects on intercellular antioxidative activity and UVA-induced MMP expression in human dermal fibroblasts (HDFs). The dried O. vulgatum was extracted in a mixture of ethanol and water (1 : 1) for 24 h at room temperature. The extract was filtered and concentrated in vacuo and lyophilized. For testing intracellular ROS scavenging activity the cultured HDFs were analyzed by increase in DCF fluorescence upon exposure to UVB $20\;mJ/cm^2$. After treatment of O. vulgatum extracts, intracellular ROS levels were measured by luminescence spectrophotometer. Enzyme linked immuno sorbent assay (ELISA), and RT-PCR techniques were used for evaluating the effects of O. vulgatumon on MMP protein and mRNA expression in UVA irradiated HDFs. O. vulgatum extract was found to have ROS scavenging activity with the $IC_{50}$ values of $18.2\;{\mu}g/mL$ against superoxide radicals in the xanthine/xanthine oxidase system. After treatment of O. vulgatum extracts, the oxidation of CM-DCFDA was inhibited effectively and O. vulgatum extracts showed a potent free radical scavenging activity by 30.4 % at $100\;{\mu}g/mL$ in UVB-irradiated HDFs. UVA induced MMP protein expression was reduced 37.7 % by treatment with O. vulgatum extract, and MMP-1 mRNA expression was reduced in a dose-dependent manner. Taken together, these results suggest that O. vulgatum extract prevents the skin cell damage induced by UV irradiation, and implies that O. vulgatum extract may be useful as a new ingredient for anti-aging cosmetics.