• Toxicological Research
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• v.9 no.2
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• pp.217-224
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• 1993

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is a well-known dopamine neuron-specific toxin. But the involvement of oxidative damage in the pathogenesis of MPTP-induced parkinsonism is still uncertain. In this study, by using 2',7',-dichlorofluorescin diacetate(DCFH-DA) that detects intracellular oxidative processes, the effect of MPTP on dichlorofluorescein fluorescence in dissociated cells from fetal rat mesencephalon in culture was investigated. At 7th day in culture, cells were loaded with DCFH-DA, and exposed to 1 mM MPTP or MPP+. MPTP induced dichlorofluorescein-fluorescence which was peaked at 3 min and mostly faded away 30 min after MPTP treatment.

• Journal of Marine Bioscience and Biotechnology
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• v.1 no.2
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• pp.126-135
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• 2006

Phlorotannins which is oligomeric polyphenol of phloroglucinol unit were isolated from solvent fractions of methanolic extract of the brown alga, Ecklonia cava (EC). The inhibitory effects of phlorotannins from EC solvent fractions on oxidative stress were examined in human dermal fibroblasts (HDFs) related to wrinkle formation. Among the solvent fractions, phlorotannins from ethyl acetate fraction exerted the highest scavenging effect on DPPH radical, hydroxyl radical and alkyl radical analyzed by electron spin resonance (ESR) spectroscopy. The levels of intracellular reactive oxygen species (ROS) and lipid peroxidation were measured using 2',7'-dichlorofluorescin diacetate (DCFH-DA) and diphenyl-1-pyrenylphosphine (DPPP), respectively. Their levels were significantly decreased in the presence of phlorotannins from ethyl acetate fraction, compared with other fractions obtained from EC extract (P < 0.01). Furthermore, intracellular glutathione (GSH) level was significantly increased in a time dependent manner by the phlorotannins. Therefore, these results suggest that phlorotannins from EC extract could have a therapeutic potential for prevention and treatment of several diseases such as wrinkle formation related to oxidative stress.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.28 no.1
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• pp.42-57
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• 2002

본 연구는 널리 알려져 있는 항산화제들의 세포 수준에서의 anti-oxidative stress 효과 및 그 기작을 알아보기 위한 연구이다. 연구에 사용한 항산화제로는 지용성인 retinol, $\alpha$-tocopherol, propyl gallate(PG) 및 butylated hydroxy toluene(BHT)과 수용성인 ascorbic acid, $\alpha$-glucosyl rutin 및 green tea extract를 사용하였으며 이들 항산화제들의 시간별 세포 생존율을 NR assay 로 측정한 후 적정 농도에서 DCFH-DA(2', 7'-dichlorofluorescin-diacetate) 를 이용하여 항산화제들의 anti-oxidative stress 억제 효과를 시간별로 측정하였다. 또한 이들 항산화제의 항산화 기작을 알아보기 위하여 NBT(Nitro-blue-tetrazolium) 및 DPPH(Diphenyl-picry-hydrazl)도 병행하여 실시하였다. Anti-oxidative stress 실험에서 지용성 항산화제들은 전반적으로 수용성 항산화제에 비하여 세포에 대한 독성이 상대적으로 강하여 retinol 의 경우에는 0.01%에서 oxidative stress 억제 효과를 관할할 수 있었으며 1 시간경과 후 측정시 53.1%의 억제 효과를 보여 주었다. PG 의 경우에는 0.1%에서 2 시간 경과 후 측정시 50%의 oxidative stress 억제 효과를 보여주었다. 수용성 항산화제인 green tea extract 및 $\alpha$-glucosyl rutin의 경우에는 1%에서 1시간 경과 후 측정시 각 각 51.6% 및 69.7%의 oxidative stress 억제 효과를 관찰할 수 있었다. 또한 시료처리 후 자외선 조사시 oxidative stress 억제 효과의 경우 수용성 항산화제인 ascorbic acid, $\alpha$-glucosyl rutin 및 green tea extract 와 지용성 항산화제 중에서는 $\delta$-tocopherol 에서만 oxidative stress 억제 효과가 관찰되었으나 자외선을 조사 하지 않았을 때 보다 약 20%-40%까지 억제 효과가 감소되었다. 그리고 PG 및 retinol 의 경우에는 자외선 조사시 독성이 증가하여 oxidative stress 억제 효과를 측정할 수 없었다. NBT실험에서 $\alpha$-glucosyl rutin, $\alpha$-tocopherol 및 PG 1%에서 70%이상의 superoxide anion 생성 억제 효과를 보였으며 DPPH 실험에서는 ascorbic acid 와 PG 1%에서 98%의 hydroxyl radical 생성 억제 효과를 보여 주었다. 본 실험을 통하여 BHT 를 제외하고 전반적으로 세포 수준에서의 oxidative stress 에 대한 억제 효과를 확인해 볼 수 있었으며 특히 수용성 항산화제들에서 두드러진 효과를 보여 주었다.

• Journal of the Korean Applied Science and Technology
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• v.36 no.3
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• pp.966-974
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• 2019

Arthrospira platensis has been reported to contain a variety of substances such as phycocyanin, ${\beta}$-carotene, vitamin E and other carotenoids. In this study, zebrafish were treated with indoor cultivation spirulina ethanol extracts(ICAE) to determine toxicity(coagulation rate, hatching rate, heart rate). We used the DCFH-DA staining method to detect the effect of reactive oxygen species(ROS) generation on lipopolysaccharide(LPS)-induced zebrafish embryos ROS various concentrations(0.01, 0.05, 0.1, 0.5mg/ml) of ICAE. Cell toxicity was measured by WST-1 assay on RAW 264.7 macrophage cell line. Also, measured the inhibitory effect of nitric oxide(NO) and prostaglandin $E_2$ ($PGE_2$) production in RAW264.7 macrophages induced by LPS at various concentrations of ICAE. The results of embryo coagulation rate, hatching rate, heart rate of zebrafish at various(0.01, 0.05, 0.1mg/ml) of ICAE was no toxicity. The ICAE treated group had an inhibitory effect on NO and $PGE_2$ production compared and decreased with concentration. The results of this study ethanol extract of Arthrospira platensis has an anti-inflammatory effect and suggest that is worthy of use cosmetics for skin protection.

• The Korean Journal of Food And Nutrition
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• v.11 no.1
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• pp.107-113
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• 1998

This experiment has conducted to evaluate whether single injection of red ginseng extract including 50% ethanol extract, crude saponin, and lipid soluble fraction can induce oxidative burst of mouse peritoneal macrophages with use of fluorescence spectrophotometer. To optimize conditions of fluorescent spectrophotometry, concentrations of DCFH-DA(2', 7' -dichlorofluorescin diacetate) was 1.6 ${\mu}{\textrm}{m}$ and control oxidative burst by Zymosan A and PMA(phorbol myristate acetate) were 100$\mu\textrm{g}$, 250ng, respectively. Though in vitro macrophages failed to induce increment of H2O2 production, but 50% ethanol extract group induced significant enhancement of H2O2 production when zymosan A triggered oxidative burst. On the other hand, lipid soluble fraction enhanced significantly H2O2 production than that of control group. These findings consisted with the other reports which showed ginsenosides inhibited nitric oxide production and lipid soluble fraction activated colony stimulating factor(granulocyte - monocyte) activity in bone marrow stem cells. As is well known, lipid soluble fraction contains phenol compound, polyacetylene compound and alkaloids. Further study would unravel which component of it can induce H2O2 production of macrophages. Key words : Red ginseng(Panax ginseng), H2O2 production, macrophages.

• Preventive Nutrition and Food Science
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• v.16 no.2
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• pp.127-134
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• 2011

The purpose of this study was to purify antioxidant substances from steamed squid extract (SSE). The yield of SSE was 8% by dry weight. The approximate compositions of SSE proteins, lipids, moisture, carbohydrate and ash were 64.95%, 1.69%, 7.23%, 4.44% and 21.69%, respectively. The major amino acids in SSE were taurine (29.17%), glycine (20.33%), alanine (12.51%), and glutamic acid (9.83%). Antioxidant activities were evaluated using 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging activity, which was measured as 24.7% at 1.0 mg/mL. Four SSE fractions were isolated by Sephadex G-25 gel chromatography; the F2 fraction showed the highest DPPH radical scavenging activity. The F2 fraction was separated by reverse-phase high performance liquid chromatography (HPLC) using an octadecylsilane (ODS) column, yielding a purified antioxidant substance with a DPPH radical scavenging activity of 64.41% at 1.0 mg/mL, representing a 2.64-fold increase in the scavenging activity of SSE purified by the 3-step procedure. The amino acid compositions showed that purified SSE was rich in taurine, glycine, glutamic acid and alanine. The purified SSE significantly elevated 2',7'-dichlorodihydrofluororescein diacetate (DCFH-DA) fluorescence probe, which confirms its effective radical scavenging potential in cellular ROS. In addition, the SSE significantly inhibited oxidative damage of purified genomic DNA. These results suggest that a purified antioxidant substance from SSE can be used as a potential natural compound-based antioxidant in the functional food and pharmaceutical industries.

• Journal of the Korean Applied Science and Technology
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• v.35 no.2
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• pp.423-432
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• 2018

Arthrospira platensis is one of the oldest algae in the world and has been reported to have anti-aging properties, including phycocyanin, tocopherol and beta-carotene. In this study, we tried to search protective activities against UVB-induced reactive oxygen species(ROS) of Arthrospira platensis under indoor cultivation ethanol extracts(ICAE) and outdoor cultivation ethanol extracts(OCAE). The anti-oxidative capacities were evaluated by DPPH radical scavenging activity and SOD-like activities at various concentrations(0.1, 0.5, $1mg/m{\ell}$) of ICAE and OCAE. Zebrafish embryos and HaCaT cells were exposed to UVB radiation and treated with various concentrations(0, 0.01, 0.05, 0.1, 0.5, $1mg/m{\ell}$) of ICAE and OCAE. ROS levels of zebrafish and HaCaT cells were generated by UVB radiation. ROS levels were detected using a fluorescent microscope after DCFH-DA staining. The DPPH radical scavenging activity of ascorbic acid was 73% and SOD-like activity was 86% in the positive control group. ICAE and OCAE at $1mg/m{\ell}$ concentration showed 43, 57% DPPH radical scavenging activity and 20, 19% SOD-like activity. Anti-oxidative of ICAE and OCAE had lower effects than the positive control ascorbic acid but significant results. ROS of UVB-induced zebrafish embryos and HaCaT cells were higher than negative control. ICAE and OCAE treated group decreased ROS concentration dependently than UVB-induced positive control group. These results suggest that Arthrospira platensis ethanol extract may have usability value as a cosmetic material for skin protection.

• Journal of Life Science
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• v.20 no.9
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• pp.1365-1372
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• 2010

We investigated the inhibitory effects of solvent extracts from dried yam on $H_2O_2$-induced oxidative stress and growth of cancer cell lines (HT1080 human fibrosarcoma and HT-29 human colon cancer cells). Yam (Dioscoreacea) has been recognized as a healthy food due to its various biological activities, such as anti-obesity, anti-constipation, anti-proliferation, and anti-mutagenic activities, as well as its ability to decrease blood glucose and cholesterol levels. In order to determine the protective effect on $H_2O_2$-induced oxidative stress, DCFH-DA (dichlorodihydrofluorescin diacetate) assay was conducted. Acetone with methylene chloride (A+M) extract of dried yam appeared to reduce the levels of intracellular reactive oxygen species (ROS) with dose responses. Among the fractions, 85% aq. methanol fraction showed the highest protective effect on production of lipid peroxides. Inhibitory effects of A+M and methanol (MeOH) extracts on the growth of HT1080 and HT-29 cancer cells increased in a dose dependent manner. The treatments of n-hexane, 85% aq. methanol and n-butanol fractions (${\geqq}0.5$ mg/ml concentrations) significantly inhibited the growth of both cancer cells (p<0.05). From these results, 85% aq. methanol fraction showed inhibitory effects on cellular oxidation and growth of human cancer cells, suggesting that this fraction may contain active compounds of dried yam.

• Journal of Korean Ophthalmic Optics Society
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• v.13 no.3
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• pp.95-101
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• 2008

Purpose: Hydrogen peroxide which is one of the reactive oxygen species has been seen to cause various diseases, various cellular disinfections, gene transformation and cell death. The goals of this study were to determine the protective effect of EGCG against $H_2O_2$-induced apoptotic death in conjunctival cell lines. Methods: We measured cell viability by MTT assay and analyzed DNA fragmentation to check up a distinctive feature in cell death and measured the removal ability of free radicals by DPPH free radical scavenging assay and evaluated the oxygen free radical's quantity in the cell by DCFH-DA assay. The mRNA expression in the cell were examined by RT-PCR. Results: Cell viability and free radical scavening activites was significantly increased in dose dependently after cell was exposed to EGCG. And DNA fragmentation and intracellular ROS was decreased. It was showed the mRNA expression which increase of bcl-2, bcl-xL expression and decrease of bax expression. Conclusions: From these results, it suggests that EGCG has an antioxidant effect and protects conjunctival cell lines from the $H_2O_2$-mediated apoptosis through the modulation of the mRNA expression.

• Journal of Life Science
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• v.17 no.11
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• pp.1533-1538
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• 2007

This study was carried out to determine the inhibitory effects of solvent extracts from Sargassum hemiphyllum on growth of cancer cell lines (AGS human gastric adenocarcinoma and HT-29 human colon cancer cells) and production of lipid peroxides. Inhibitory effects of acetone with methylene chloride extract from S. hemiphyllum on the growth of AGS and HT-29 cancer cells were increased as dose dependent patterns (p<0.05). The methanol extract was more effective on inhibition of growth of AGS. The treatments of hexane, 85% aq. methanol, butanol and water fractions significantly inhibited the growth of cancer cells (p<0.05) and the inhibitory effect was stronger in HT-29. In DCFH-DA (dichlorodihydrofluorescin diacetate) assay, acetone with methylene chloride and methanol extracts showed a stronger inhibitory effect on the production of cellular lipid peroxides (p<0.05) compared with the butanol and hexane fractions. These results indicate that the consumption of S. hemiphyllum may be recommended as a potent functional food for preventing cellular oxidation and cancer.