• Nutrition Research and Practice
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• v.1 no.2
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• pp.105-112
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• 2007

Ascorbic acid has been reported to extend replicative life span of human embryonic fibroblast (HEF). Since the detailed molecular mechanism of this phenomenon has not been investigated, we attempted to elucidate. Continuous treatment of HEF cells with ascorbic acid at ($200{\mu}M$) from 40 population doubling (PD) increased maximum PD numbers by 18% and lowered $SA-{\beta}-gal$ positive staining, an aging marker, by 2.3 folds, indicating that ascorbic acid extends replicative life span of HEF cells. Ascorbic acid treatment lowered DCFH by about 7 folds and Rho123 by about 70%, suggesting that ascorbic acid dramatically decreased ROS formation. Ascorbic acid also increased aconitase activity, a marker of mitochondrial aging, by 41%, indicating that ascorbic acid treatment restores age-related decline of mitochondrial function. Cell cycle analysis by flow cytometry revealed that ascorbic acid treatment decreased G1 population up to 12%. Further western blot analysis showed that ascorbic acid treatment decreased levels of p53, phospho-p53 at ser 15, and p21, indicating that ascorbic acid relieved senescence-related G1 arrest. Analysis of AP (apurinic/apyrimidinic) sites showed that ascorbic acid treatment decreased AP site formation by 35%. We also tested the effect of hydrogen peroxide treatment, as an additional oxidative stress. Continuous treatment of $20{\mu}M$ of hydrogen peroxide from PD 40 of HEF cells resulted in premature senescence due to increased ROS level, and increased AP sites. Taken together, the results suggest that ascorbic acid extends replicative life span of HEF cells by reducing mitochondrial and DNA damages through lowering cellular ROS.

• Nutrition Research and Practice
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• v.11 no.5
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• pp.365-372
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• 2017

BACKGROUND/OBJECTIVES: Owing to health concerns related to the consumption of traditional snacks high in sugars and fats, much effort has been made to develop functional snacks with low calorie content. In this study, a new recipe for Korean rice cookie, dasik, was developed and its antioxidative, lipid-lowering, and anti-inflammatory effects and related mechanisms were elucidated. The effects were compared with those of traditional rice cake dasik (RCD), the lipid-lowering effect of which is greater than that of traditional western-style cookies. MATERIALS/METHODS: Ginseng-added brown rice dasik (GBRD) was prepared with brown rice flour, fructooligosaccharide, red ginseng extract, and propolis. Mice were grouped (n = 7 per group) into those fed a normal AIN-76 diet, a high-fat diet (HFD), and HFD supplemented with RCD or GBRD. Dasik in the HFD accounted for 7% of the total calories. The lipid, reactive oxygen species, and peroxynitrite levels, and degree of lipid peroxidation in the plasma or liver were determined. The expression levels of proteins involved in lipid metabolism and inflammation, and those of antioxidant enzymes were determined by western blot analysis. RESULTS: The plasma and hepatic total cholesterol concentrations in the GBRD group were significantly decreased via downregulation of sterol regulatory element-binding protein-2 and 3-hydroxy-3-methylglutaryl-CoA reductase (P < 0.05). The hepatic peroxynitrite level was significantly lower, whereas glutathione was higher, in the GBRD group than in the RCD group. Among the antioxidant enzymes, catalase (CAT) and glutathione peroxidase (GPx) were significantly upregulated in the GBRD group (P < 0.05). In addition, nuclear factor-kappaB (NF-${\kappa}B$) expression in the GBRD group was significantly lower than that in the RCD group. CONCLUSIONS: GBRD decreases the plasma and hepatic cholesterol levels by downregulating cholesterol synthesis. This new dasik recipe also improves the antioxidative and anti-inflammatory status in HFD-fed mice via CAT and GPx upregulation and NF-${\kappa}B$ downregulation. These effects were significantly higher than those of RCD.

• Nutrition Research and Practice
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• v.14 no.4
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• pp.334-351
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• 2020

BACKGROUND/OBJECTIVES: This study was designed to investigate the improvement effect of white ginseng extract (GS-KG9) on D-galactosamine (Ga1N)-induced oxidative stress and liver injury. SUBJECTS/METHODS: Sixty Sprague-Dawley rats were divided into 6 groups. Rats were orally administrated with GS-KG9 (300, 500, or 700 mg/kg) or silymarin (25 mg/kg) for 2 weeks. The rats of the GS-KG9- and silymarin-treated groups and a control group were then intraperitoneally injected Ga1N at a concentration of 650 mg/kg for 4 days. To investigate the protective effect of GS-KG9 against GalN-induced liver injury, blood liver function indicators, anti-oxidative stress indicators, and histopathological features were analyzed. RESULTS: Serum biochemical analysis indicated that GS-KG9 ameliorated the elevation of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) in GalN-treated rats. The hepatoprotective effects of GS-KG9 involved enhancing components of the hepatic antioxidant defense system, including glutathione, glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT). In addition, GS-KG9 treatment inhibited reactive oxygen species (ROS) production induced by GalN treatment in hepatocytes and significantly increased the expression levels of nuclear factor erythroid-2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) proteins, which are antioxidant proteins. In particular, by histological analyses bases on hematoxylin and eosin, Masson's trichrome, α-smooth muscle actin, and transforming growth factor-β1 staining, we determined that the administration of 500 mg/kg GS-KG9 inhibited hepatic inflammation and fibrosis due to the excessive accumulation of collagen. CONCLUSIONS: These findings demonstrate that GS-KG9 improves GalN-induced liver inflammation, necrosis, and fibrosis by attenuating oxidative stress. Therefore, GS-KG9 may be considered a useful candidate in the development of a natural preventive agent against liver injury.

• Journal of the Korean Applied Science and Technology
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• v.28 no.4
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• pp.482-490
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• 2011

In this study, the antioxidative effects and inhibitory effects on tyrosinase and elastase of Sorbus commixta (S. commixta) twig extracts were investigated. The aglycone fraction of S. commixta twig extract showed the prominent free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity($FSC_{50}$, $13{\mu}g/mL$). Reactive oxygen species (ROS) scavenging activities of S. commixta twig extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated by the luminol-dependent chemiluminescence assay. The 50 % ethanol extract among extracts showed the most prominent ROS scavenging activity ($OSC_{50}$, $0.189{\mu}g/mL$). The cellular protective effects of extract/fractions of S. commixta twig on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The 50 % ethanol extract and ethyl acetate fraction showed the cellular protective effects against ROS in a concentration dependent manner ($5{\sim}50{\mu}g/mL$). The inhibitory effect of S. commixta twig extract on tyrosinase was investigated to assess the whitening efficacy. The ethyl acetate ($IC_{50}$, $113.2{\mu}g/mL$) and aglycone fraction($IC_{50}$, $105.3{\mu}g/mL$) on tyrosinase showed more remarkable inhibitory effect than arbutin($IC_{50}$, $226.88{\mu}g/mL$), known as the whitening agent. The inhibitory effect of aglycone fraction ($IC_{50}$, $6.9{\mu}g/mL$) on elastase was simillar to quercetin($IC_{50}$, $6.1{\mu}g/mL$), flavonoid known as reference compound. These results indicate that extract/fractions of S. commixta twig can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. S. commixta twig extracts can be applicable to new functional cosmetics for anti-aging products.

• Microbiology and Biotechnology Letters
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• v.41 no.4
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• pp.433-441
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• 2013

In this study, the anti-oxidative, anti-inflammatory, anti-melanogenic activities of Endlicheria anomala (Nees) Mez methanol extract (EAME) were evaluated by use of in vitro assays and cell culture model systems. The results revealed that EAME scavenges various radicals such as 1,1-diphenyl-2-picryl hydrazyl hydrogen peroxide induced reactive oxygen species, and lipopolysaccharide induced nitric oxide. Furthermore, EAME induced the expression of anti-oxidative enzymes such as heme oxygenase 1, thioredoxin reductase 1, NAD(P)H dehydrogenase 1, and their upstream transcription factor, nuclear factor-E2-related factor 2. Moreover, EAME inhibited in vitro DOPA oxidation and 3-isobutyl-1-methylxanthine induced melanogenesis in B16F10 cells. Its anti-melanogenic activity will have originated from the inhibition of tyrosinase enzyme activity and melanogenesis related protein expression. Taken together, these results provide the important new insight that E. anomala possesses various biological activities such as anti-oxidative, anti-inflammatory, and anti-melanogenic. Therefore, it might be utilized as a promising material in the fields of nutraceuticals and cosmetics.

• The Korean Journal of Physiology and Pharmacology
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• v.8 no.6
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• pp.339-344
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• 2004

Recent data have shown the importance of oxidative stresses in the pathogenesis of inflammatory bowel disease, crohn's disease and ulcerative colitis. $H_2O_2$, reactive oxygen species (ROS) donor, has been reported to act as a signaling molecule involved in a variety of cellular functions such as apo/ptosis and proliferation. In the present study, we investigated viability of cultured ileal smooth muscle cells (ISMC) after stimulation with $H_2O_2$. Trypan blue method revealed that the cell viability of ISMC treated with 1 mM $H_2O_2$ was not different from that of controls at up to 2 h time point, while treatment of ISMC with 1 mM $H_2O_2$ for 48 h finally induced significant decrease in the cell viability. Therefore, we evaluated whether $H_2O_2$ was capable of ERKs activation in ISMC for the short-term exposure and examined whether tyrosine kinase was involved in the process of ERK activation by $H_2O_2$ in ISMC. We also investigated the effects of $H_2O_2$ on activation of SAPK/JNK and p38 MAP kinase in ISMC. Thus, ISMC were cultured and exposed to $H_2O_2$, and western blot analysis was performed with phosphospecific MAP kinase antibodies. Robust activation of ERK occurred within 30 min of 1 mM $H_2O_2$ treatment. $H_2O_2-induced$ ERK activation was attenuated by a tyrosine kinase inhibitor, genistein, indicating that tyrosine kinase was probably involved in the ERK activation by $H_2O_2$. $H_2O_2$ was a moderate activator of SAPK/JNK, while p38 MAP kinase was not activated by $H_2O_2$. We suggest that ERK activation induced by short-term $H_2O_2$ treatment plays a critical role in cellular protection in the early stage of response to oxidative stress. The present study suggests the necessity of identification of MAPK signaling pathways affected by ROS, since it could ultimately elucidate cellular consequences involved in initiation and perpetuation of intestinal tissue damage in the diseases such as crohn's disease and ulcerative colitis, resulted from excessive ROS.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.32 no.3 s.58
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• pp.161-171
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• 2006

In this study, we investigated the whitening and anti-oxidant activity of 37 Jeju island native plants. The active ingredients of the plants were prepared by methanol extraction. Whitening activity of plant extracts was examined from the inhibitory effect of tyrosinase and the inhibition of melanin synthesis of the B16-F1 cell line. Their anti-oxidant activity was measured by electron donating ability of DPPH (1,1-diphenyl-2-picrylhydrazyl) and ROS (reactive oxygen species) scavenging activity in V79-4 lung fibroblast cells using DCF-DA (dichlorofluorescin diacetate). Cytotoxicity of the extracts on cell s based experiments was investigated by MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) assay. Also, the toxicity test using a rabbit and the human skin patch test were carried out for examining the safety of the extracts which showed the high whitening activity. It is interesting that the extracts of Lespedeza cuneata, Ligustrum lucidum (stem), Morus bombycis (stem) and Prunella vulgaris var. lilacina showed both potent whitening and anti-oxidant activities.

• Journal of Veterinary Clinics
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• v.28 no.1
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• pp.13-19
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• 2011

Semen cryopreservation induces the formation of reactive oxygen species (ROS), and the ROS cause sperm damage. We aimed to investigate the effects of the antioxidative enzyme catalase (CAT) on sperm quality and ROS during cryopreservation. Sperm rich fractions collected from five Duroc boars were cryopreserved in freezing extender with (200 or 400 U/mL) or without CAT (control). After thawing, sperm motility, viability, normal morphology, plasma membrane integrity, mitochondrial function and intracellular ROS were evaluated. CAT significantly improved total sperm motility at a concentration of 400 U/mL (P < 0.05), but didn't improve progressive sperm motility, viability, morphological defects, plasma membrane integrity and mitochondrial function in frozen-thawed boar sperm. In evaluation of ROS, CAT had no effect on reduction in ${\cdot}O_2$, but scavenged $H_2O_2$ in viable frozen-thawed boar sperm at concentrations of 200 and 400 U/mL (P < 0.05). In conclusion, CAT was not enough to improve quality of frozen-thawed sperm, but can reduce $H_2O_2$ generation in viable boar sperm during cryopreservation.

• Korean Journal of Veterinary Research
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• v.48 no.3
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• pp.337-345
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• 2008

There are accumulating evidences that high levels of dietary iron may play a role in colon carcinogenesis. Elevated iron status has been associated with oxidative stress. Phytic acid (PA) functions as an antioxidant by chelating divalent cations and prevents formation of reactive oxygen species responsible for cell injury and carcinogenesis. The protective effect of PA was investigated on formation of aberrant crypt foci (ACF) induced by azoxymethane (AOM) in iron-overloaded male F344 rats. After acclimation with AIN-93G purified diet (35 ppm Fe, normal control diet) for one week, animals were fed iron-overloaded diet (350 ppm Fe) and PA (0.5% or 2% PA in water) for 8 weeks. Animals received two (1st and 2nd week) injections of AOM (15 mg/kg b.w.) to induce colonic ACF. The colonic mucosa was examined for the total numbers of aberrant crypt (AC) and ACF after staining with methylene blue. The blood and serum were analyzed with a blood cell differential counter and an automatic serum analyzer. Iron-overloaded diet increased the concentration of iron in liver of the rats. But iron-related parameters in blood were not changed among experimental groups. The numbers of ACF per colon and AC per colon were $178.8{\pm}33.2$ and $448.4{\pm}110.2$ in the iron-overloaded F344 rats. The total AC was significantly increased, compared with normal-diet AOM control group (p < 0.05). The treatments of PA at the dose of 0.5% slightly decreased the number of ACF and AC per colon to $153.6{\pm}29.5$ and $396.3{\pm}107.5$. However, there were no significant differences in the total numbers of ACF and AC between the AOM control group and PA (0.5% or 2%)-treated groups. These results suggest that PA may not affect the formation of ACF or AC induced by AOM in ironoverloaded F344 rats.

• Korean Journal of Plant Resources
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• v.31 no.1
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• pp.1-9
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• 2018

As a part of an infrastructure project on medicinal herb-based remedies, we conducted a phytohemical investigation of the 100% MeOH extract from the aerial part of Boehmeria quelpaertense; our findings resulted in the isolation of flavonoids (1-2), isoquercitrin (1) and hyperoside (2). The identification and structural elucidation of these compounds were based on $^1H$-, $^{13}C-NMR$, and LC ESI IT-TOF MS data. All the compounds isolated from this plant were reported for the first time. In this study, we examined the antioxidant activity of the 1 and 2 on the hydrogen peroxide ($H_2O_2$)-induced oxidative stress in a Rat Cardiomyoblast cell line (H9c2). The pretreatment of the flavonoids showed that it protects against $H_2O_2$-mediated cell death in the H9c2 cell line. Also, it decreases the intracellular reactive oxygen species (ROS) levels by the flavonoids in the $H_2O_2$-treated H9c2 cell line. These results showed that the 1 and 2 are a source of antioxidants. As a result, they might be helpful in preventing the progress of various oxidative stress mediated diseases, including myocardial infarction.