• The Korean Journal of Physiology and Pharmacology
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• v.4 no.1
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• pp.47-54
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• 2000

Transition metal ions including $Se^{2+},\;Cd^{2+},\;Hg^{2+}\;or\;Mn^{2+}$ have been thought to disturb the bone metabolism directly. However, the mechanism for the bone lesion is unknown. In this study, we demonstrated that MC3T3E1 osteoblasts, exposed to various transition metal ions; selenium, cadmium, mercury or manganese, generated massive amounts of reactive oxygen species (ROS). The released ROS were completely quenched by free radical scavengers-N-acetyl cysteine (NAC), reduced glutathione (GSH), or superoxide dismutase (SOD). First, we have observed that selenium $(10\;{\mu}M),$ cadmium $(100\;{\mu}M),$ mercury $(100\;{\mu}M)$ or manganese (1 mM) treatment induced apoptotic phenomena like DNA fragmentation, chromatin condensation and caspase-3-like cysteine protease activation in MC3T3E1 osteoblasts. Concomitant treatment of antioxidant; N-acetyl-L-cysteine (NAC), reduced-form glutathione (GSH), or superoxide dismutase (SOD), prevented apoptosis induced by each of the transition metal ions. Catalase or dimethylsulfoxide (DMSO) has less potent inhibitory effect on the apoptosis, compared with NAC, GSH or SOD. In line with the results, nitroblue tetrazolium (NBT) stain shows that each of the transition metals is a potent source of free radicals in MC3T3E1 osteoblast. Our data show that oxidative damage is associated with the induction of apoptosis in MC3T3E1 osteoblasts following $Se^{2+},\;Cd^{2+},\;Hg^{2+}\;or\;Mn^{2+}$ treatment.

• The Korean Journal of Physiology and Pharmacology
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• v.19 no.1
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• pp.43-50
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• 2015

It has been shown that the extracts including eupatilin and quercetin-3-${\beta}$-D-glucuronopyranoside had mucoprotective effects on the esophagus and stomach through their antioxidant activities. This study was designed to investigate the anti-inflammatory effect of these flavonoid compounds in an animal model of inflammatory bowel disease induced by 2,4,6-trinitrobenzene sulfonic acid. Experimental colitis was induced by intracolonic administration of 2,4,6-trinitrobenzene sulfonic acid. Extracts including eupatilin or quercetin-3-${\beta}$-D-glucuronopyranoside were orally administered to animals 48, 24, and 1 h prior to the induction of colitis and then again 24 h later. The animals were sacrificed 48 h after by 2,4,6-trinitrobenzene sulfonic acid treatment and the macroscopic appearance of the colonic lesions was scored in a blinded manner on a scale of 1 to 10. The inflammatory response to colitis induction was assessed by measuring myeloperoxidase activity, nitric oxide production, tumor necrosis factor-${\alpha}$ expression, total glutathione levels, and malondialdehyde concentrations in the colon. The results indicated that extracts including eupatilin and extracts including quercetin-3-${\beta}$-D-glucuronopyranoside dose-dependently improved the morphology of the lesions induced by 2,4,6-trinitrobenzene sulfonic acid and reduced the ulcer index accordingly. In addition, rats receiving extracts including eupatilin and extracts including quercetin-3-${\beta}$-D-glucuronopyranoside showed significantly decreased levels of mucosal myeloperoxidase activity, nitric oxide production, tumor necrosis factor-${\alpha}$ expression, and malondialdehyde levels, and increased total glutathione levels. Extracts including eupatilin and extracts including quercetin-3-${\beta}$-D-glucuronopyranoside ameliorated the inflammatory response and colonic injury in acute colitis by decreasing oxidative stress and neutrophil activation. Extracts including eupatilin and extracts including quercetin-3-${\beta}$-D-glucuronopyranoside may inhibit acute colitis.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.4
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• pp.936-941
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• 2006

To prevent human body injury from oxidative stress, antioxidants are very important and many research about antioxidants are generally being conducted. Hydrogen peroxide$(H_20_2)$ that is one of vitality oxygen species has been seen that cause various diseases, DNA damage and gene change. We have already known that the inhibition effect of Zizania latifolia Radix, Rhizoma on apoptosis induced by $H_2O_2$ in Neuro2A cell. And the purpose of this study was that we made a comparative study on the inhibition effect of apoptosis in Neuro2A cell on the region of Zizania latifolia(Radix, Rhizoma, Herba). Neuro2A cells were cultivated in RPMI(GibcoBRL) with 5% FBS and treated with $H_2O_2$ and Zizania latifolia(Radix, Rhizoma, Herba). Separately we measured the cell viability and analyzed DNA fragmentation. Activity of PARP, Cytochrome C, caspase-9, caspase-3, p53, p21, Bax and Bcl-2 in the cell was examined by using western blot. The results obtained were as Follows: The cell viability in all of Zizania latifolia (Radix, Rhizoma, Herba) treatment (60ug/m1<) decreased significantly compared with that of none treatment(p<0.001). Zizania latifolia Radix increased cell viability was most effective of three regions. But we had no significant difference among three regions. All of Zizania latifolia (Radix, Rhizoma, Herba) increased cell viability about twice as much as that being injury by $H_2O_2$,(Zizania Latifolia (Radix, nhizoma, Herba) 20ug/m1, $H_2O_2$ 200uM, p<0.001). DNA fragmentation developed by $H_2O_2$, but was not developed in all of Zizania latifolia (Radix, Rhizoma, Herba) treatment. PARP, Cytochrome C, caspase-9 and caspase-3 activated all by $H_2O_2$ but were not activated in all of Zizania latifolia (Radix, Rhizoma, Herba) treatment. P53, P2l and Bax activated by $H_2O_2$, and Bcl-2 got into inactivation. But the opposite results appeared in all of Zizania latifolia (Radix, Rhizoma, Herba) treatment. In conclusion, these results suggest that all of Zizania latifolia (Radix, Rhizoma, Herba) inhibit the development of DNA fragmentation and apoptosis by $H_2O_2$and the antioridant action of all of Zizania latifolia (Radix, Rhizoma, Herba) is effective.

• Biomolecules & Therapeutics
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• v.28 no.5
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• pp.443-455
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• 2020

The thioredoxin (Trx) system plays critical roles in regulating intracellular redox levels and defending organisms against oxidative stress. Recent studies indicated that Trx reductase (TrxR) was overexpressed in various types of human cancer cells indicating that the Trx-TrxR system may be a potential target for anti-cancer drug development. This study investigated the synergistic effect of auranofin, a TrxR-specific inhibitor, on sulforaphane-mediated apoptotic cell death using Hep3B cells. The results showed that sulforaphane significantly enhanced auranofin-induced apoptosis by inhibiting TrxR activity and cell proliferation compared to either single treatment. The synergistic effect of sulforaphane and auranofin on apoptosis was evidenced by an increased annexin-V-positive cells and Sub-G1 cells. The induction of apoptosis by the combined treatment caused the loss of mitochondrial membrane potential (ΔΨm) and upregulation of Bax. In addition, the proteolytic activities of caspases (-3, -8, and -9) and the degradation of poly (ADP-ribose) polymerase, a substrate protein of activated caspase-3, were also higher in the combined treatment. Moreover, combined treatment induced excessive generation of reactive oxygen species (ROS). However, treatment with N-acetyl-L-cysteine, a ROS scavenger, reduced combined treatment-induced ROS production and apoptosis. Thereby, these results deduce that ROS played a pivotal role in apoptosis induced by auranofin and sulforaphane. Furthermore, apoptosis induced by auranofin and sulforaphane was significantly increased through inhibition of the phosphoinositide 3-kinase (PI3K)/Akt pathway. Taken together, the present study demonstrated that down-regulation of TrxR activity contributed to the synergistic effect of auranofin and sulforaphane on apoptosis through ROS production and inhibition of PI3K/Akt signaling pathway.

• Journal of Life Science
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• v.12 no.3
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• pp.349-356
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• 2002

In order to observe the bioactivity of ovariectomized rats, nonovariertoized (sham) group, ovariectomized (Ovx) group, ovariectomized ginseng total saponin (GTS)-treated (Ovx+ GTS) group and ovariectomized ginseng water extract (GW)-treated (Ovx+CW) group were made. We measured AST (L-aspartate aminotransferase) and ALT (L-alanin aminotransferase) in sera, and MDA (malondialdehyde:lipid peroxidation), SOD (superoxide dismutase), catalase, total-glutathione (GSH + GSSG) and GPx (glutathione peroxidase) in liver tissue total homogenates of rat. AST activity of serum in Ovx group was 2.11 times increased, but ALT activity was not changed compared to Sham group. In AST activity, they tend to decrease significantly in each substance such as GTS and GW administered group. Lipidperoxides of each fraction in Ovx group were highly increased compared to Sham group. Extracts of ginseng-treated group markedly inhibited lipid peroxidation by 62% ∼72%. And as the result of the measurements of SOD, catalase, total-glutathione and GPx which are antioxidant enzyme, antioxidant enzymes in Ovx group much lower than in Sham group. But they were significantly increased in each substance such as GTS and GW, administered group. Based on the results, it is supposed that more produced free radicals decreased antioxidant enzyme. And it is also thought that extracts of ginseng can inhibit aging by reducing antioxidant enzyme.

• Korean Journal of Food Science and Technology
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• v.46 no.3
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• pp.369-374
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• 2014

The physiological characteristics of kiwi (Actinidia delicosa) fruit were analyzed, which inclued its nutritional composition, in vitro-antioxidant activities, and neuronal cell protective effects. The most abundant components of mineral, amino acid, and fatty acid were found to be potassium (K), glutamic acid, and a-linolenic acid, respectively. The major free sugars were fructose, glucose, and sucrose. In addition, ${\beta}$-carotene and vitamin C contents were $1.35{\mu}g/100mL$ and 29.21 mg/100 g, respectively. The 2,2'-azinobis-(3-ethylbenothiazline-6-sulfonic acid) (ABTS) radical-scavenging activity of the n-hexane fraction obtained from the kiwi extract was 10.52% at a concentration of $1000{\mu}g/mL$. The malondialdehyde (MDA) inhibition of the n-hexane fraction was found to increased in a dose-dependent manner. The intracellular reactive oxygen species (ROS) accumulation after hydrogen peroxide ($H_2O_2$) treatment of PC12 cells was significantly reduced in the presence of the n-hexane fraction compared to PC12 cells treated with $H_2O_2$ only. Moreover, in the a MTT assay, the n-hexane fraction showed in vitro-protective effects against $H_2O_2$-induced neurotoxicity.

• The Korean Journal of Food And Nutrition
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• v.32 no.5
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• pp.462-472
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• 2019

The purpose of this study was to demonstrate the quality characteristics of Brassica juncea cultivated in Jeongseon (BJJ), South Korea. We analyzed the nutritional components and antioxidant activity of BJJ. As a result of the free sugar analysis, the contents of glucose and fructose in BJJ were $0.29{\pm}0.02g/100g$ and $0.10{\pm}0.00g/100g$, respectively. The major fatty acids were palmitic acid, octadecenoic acid and stearic acid. The palmitic acid was the highest at 31.22% of all fatty acids. The major minerals were identified as Ca, P, K, Mg and Na. The contents of vitamin $B_1$, vitamin $B_2$, vitamin $B_6$, vitamin C and vitamin E in BJJ were $0.02{\pm}0.00mg/100g$, $0.087{\pm}0.01mg/100g$, $0.02{\pm}0.00mg/100g$, $0.56{\pm}0.06mg/100g$ and $0.20{\pm}0.03mg\;{\alpha}-TE/100g$, respectively. As a result of the free amino acid analysis, total amino acid contents in BJJ were $2,801.21{\pm}115.38mg/100g$. L-proline content was the highest ($744.30{\pm}119.06mg/100g$) in BJJ. BJJ extract inhibits reactive oxygen species production in 3T3-L1 adipocytes. Also, BJJ extract exhibits a protective effect on oxidative stress in $H_2O_2$-induced human dermal fibroblast. These results indicate that BJJ comprises various valuable nutrients which can be used as functional food ingredients.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.1 s.49
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• pp.121-125
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• 2005

Most of the cells, specially in the skin, free radicals and reactive oxygen species induced aging, accompanying with diseases. The pinaceae family is very familiar plant with Korean spirit. However, those plants were not estimated for natural products in manufactural process. We employed 4 different plants, Pinus koraienis S et Z. Pinus banksiana LAMBERT. Pinus rigida MILL. Pinus densiflora S. et Z. in pinceae family. The ethanol extracts were prepared for three different parts of plant, wood, bark and leaf, respectively. In this study, we carried out free radical and superoxide radical scavenging assay to investigate the anti-oxidative activity The bark and leaf part of plants showed similar anti-oxidant effect. L-ascorbic acid and $\alpha$-tocopherol at $10{\mu}g/mL$. Also we tried to investigate inhibitory effects of elastase activity in in vitro experiments on pinaceae plant extract as a matters for functional cosmetics. Among those plants, the bark of Pinus rigida MILL. and Pinus densiflora S. et Z. showed inhibitory effect. The cell viability was evaluated with MTT assay. The potential relationship was shown between the cell viability and anti-oxidant effect because the anti-oxidant effects were positively correlated wvith the cell growth in MTT assay. As the results in our experiments, we expect the potential activities of pinaceae as a material of functional cosmetics.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.4 s.54
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• pp.329-335
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• 2005

In order to investigate the effects of Saussurea involucrata on (relationship between) aging (and Saussurea involucrata), we examined the activities of antioxidation, in vitro MMP inhibition and UVA-induced MMP-1 expression in human dermal fibroblasts. S. involucrata showed scavenging activities radicals and reactive oxygen species (ROS) with the $IC_{50}$ values of $3.89{\mu}g/mL$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and $67.29{\mu}g/mL$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. At the concentration of $1000{\mu}g/mL$, S. involucrata showed 93.27% inhibition on lipid peroxidation of linoleic acid. S. involucrata inhibited the activities of MMP-1 in a does-dependent manner and the $IC_{50}$ value calculated from semi-log plots was $97.18{\mu}g/mL$. Also, UVA induced MMP expression in human dermal fibroblasts was reduced 42.86% by treatment with S. involucrata, and MMP-1 mRNA expression was reduced in a dose-dependent manner. Therefore S. involucrata was able to significantly inhibit MMP expression in protein and mRNA level. All these results suggested that S. involucrata might act as an anti-aging agent by antioxidation and reducing UVA-induced MMP-1 production.

• Animal Bioscience
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• v.34 no.10
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• pp.1590-1599
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• 2021

Objective: This study investigates the expression patterns of toll-like receptors (TLRs) and intracellular mediators in horse muscle cells after exercise, and the relationship between TLRS expression in stressed horse muscle cells and immune cell migration toward them. Methods: The expression patterns of the TLRs (TLR2, TLR4, and TLR8) and downstream signaling pathway-related genes (myeloid differentiation primary response 88 [MYD88]; activating transcription factor 3 [ATF3]) are examined in horse tissues, and horse peripheral blood mononuclear cells (PBMCs), polymorphonuclear cells (PMNs) and muscles in response to exercise, using the quantitative reverse transcription-polymerase chain reaction (qPCR). Expressions of chemokine receptor genes, i.e., C-X-C motif chemokine receptor 2 (CXCR2) and C-C motif chemokine receptor 5 (CCR5), are studied in PBMCs and PMNs. A horse muscle cell line is developed by transfecting SV-T antigen into fetal muscle cells, followed by examination of muscle-specific genes. Horse muscle cells are treated with stressors, i.e., cortisol, hydrogen peroxide (H₂O₂), and heat, to mimic stress conditions in vitro, and the expression of TLR4 and TLR8 are examined in stressed muscle cells, in addition to migration activity of PBMCs toward stressed muscle cells. Results: The qPCR revealed that TLR4 message was expressed in cerebrum, cerebellum, thymus, lung, liver, kidney, and muscle, whereas TLR8 expressed in thymus, lung, and kidney, while TLR2 expressed in thymus, lung, and kidney. Expressions of TLRs, i.e., TLR4 and TLR8, and mediators, i.e., MYD88 and ATF3, were upregulated in muscle, PBMCs and PMNs in response to exercise. Expressions of CXCR2 and CCR5 were also upregulated in PBMCs and PMNs after exercise. In the muscle cell line, TLR4 and TLR8 expressions were upregulated when cells were treated with stressors such as cortisol, H₂O₂, and heat. Migration of PBMCs toward stressed muscle cells was increased by exercise and oxidative stresses, and combinations of these. Treatment with methylsulfonylmethane (MSM), an antioxidant on stressed muscle cells, reduced migration of PBMCs toward stressed muscle cells. Conclusion: In this study, we have successfully cultured horse skeletal muscle cells, isolated horse PBMCs, and established an in vitro system for studying stress-related gene expressions and function. Expression of TLR4, TLR8, CXCR2, and CCR5 in horse muscle cells was higher in response to stressors such as cortisol, H₂O₂, and heat, or combinations of these. In addition, migration of PBMCs toward muscle cells was increased when muscle cells were under stress, but inhibition of reactive oxygen species by MSM modulated migratory activity of PBMCs to stressed muscle cells. Further study is necessary to investigate the biological function(s) of the TLR gene family in horse muscle cells.