• International Journal of Oral Biology
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• v.40 no.2
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• pp.103-109
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• 2015

Growing evidence suggests that mitochondrial reactive oxygen species (ROS) are involved in various pain states. This study was performed to investigate whether ROS-induced changes in neuronal excitability in trigeminal subnucleus caudalis are related to ROS generation in mitochondria. Confocal scanning laser microscopy was used to measure ROS-induced fluorescence intensity in live rat trigeminal caudalis slices. The ROS level increased during the perfusion of malate, a mitochondrial substrate, after loading of 2',7'-dichlorofluorescin diacetate ($H_2DCF-DA$), an indicator of the intracellular ROS; the ROS level recovered to the control condition after washout. When pre-treated with phenyl N-tert-butylnitrone (PBN) and 4-hydroxy-2,2,6,6-tetramethylpiperidene-1-oxyl (TEMPOL), malate-induced increase of ROS level was suppressed. To identify the direct relation between elevated ROS levels and mitochondria, we applied the malate after double-loading of $H_2DCF-DA$ and chloromethyl-X-rosamine (CMXRos; MitoTracker Red), which is a mitochondria-specific fluorescent probe. As a result, increase of both intracellular ROS and mitochondrial ROS were observed simultaneously. This study demonstrated that elevated ROS in trigeminal subnucleus caudalis neuron can be induced through mitochondrial-ROS pathway, primarily by the leakage of ROS from the mitochondrial electron transport chain.

• Journal of Life Science
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• v.19 no.5
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• pp.598-606
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• 2009

Oxidative stress by free radicals is a major cause of neuronal cell death. Excitotoxicity in hypoxia/ischemia causes an increase in reactive oxygen species (ROS) and a loss of mitochondrial membrane potential (MMP), resulting in dysfunction of the mitochondria and cell death. Pinelliae Rhizoma (PR) is a traditional medicine for incipient stroke. We investigated the effects of PR Water-Extract on the modulation of ROS and MMP in a hypoxic model using cultured rat cortical cells. PR Water-Extract was added to the culture medium at various concentrations (0.25${\sim}$5, 5.0 ${\mu}g/ml$) on day in vitro 12(DIV12), given a hypoxic shock (2% $O_2$/5% $CO_2$, $37^{\circ}C$, 3 hr), and cell viability was assessed on DIV15 by Lactate Dehydrogenase Assay (LDH assays). PR Water-Extract showed a statistically significant effect on neuroprotection (10${\sim}$15% increase in viability; p<0.01) at 1.0 and 2.5 ${\mu}g/ml$ in normoxia and hypoxia. Measurement of ROS production by $H_2DCF-DA$ stainings showed that PR Water-Extract efficiently reduced the number and intensity of ROS-producing neurons, especially at 1 hr post shock and DIV15. When MMP was measured by JC-1 stainings, PR Water-Extract efficiently maintained high-energy charged mitochondria. These results indicate that PR Water-Extract protects neurons in hypoxia by preventing ROS production and preserving the cellular energy level.

• 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 Korean society of Dental Hygiene
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• v.14 no.5
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• pp.775-781
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• 2014

Objectives : The purpose of the study is to investigate the protective effect of coenzyme $Q_{10}$ on cytotoxicity effect of dental monomers in odontoblast(MDPC-23). Methods : MDPC-23 was incubated with the(co)monomers triethylene glycol dimethacrylate (TEGDMA) with and without addition of coenzyme $Q_{10}$. The cell proliferation and survival was determined using WST-1 assay. The level of reactive oxygen species(ROS) was measured by immunofluorescent staining for DCF-DA. Results : TEGDMA treatment decreased the cell proliferation by dose dependently(0.1, 1, 2.5, 5, 10 mM) on the growth of MDPC-23 cells. Coenzyme $Q_{10}$ showed cell proliferation from 5 to $500{\mu}M$ by WST-1 assay. Pre-treatment coenzyme $Q_{10}$ showed the antioxidant effect on proliferation and viability of MDPC-23 after 48h(p<0.05). The positive cells were observed in non-coenyme $Q_{10}$ treatment group(group 2) in comparison with coenyme $Q_{10}$ pre-treatment group(group 1) by DCF-DA. The fluorescence positive cells showed 14.715(group 1) and 19.788(group 2) using image J system. Conclusions : TEGDMA induced cytotoxicity. The MDPC-23 cell death was associated with the increasing ROS. Coenyme $Q_{10}$ showed the antioxidant effects by decreasing ROS. This effects may contribute to the treatment of periodontal disease induced by TEGDMA after operation.

• Journal of Ginseng Research
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• v.37 no.4
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• pp.442-450
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• 2013

The aim of this study was to determine and compare the preventive effect of Korean White Ginseng and Red Ginseng on oxidative stress in $H_2O_2$-treated HepG2 cells. The roots of ginseng were extracted with 70% methanol and partitioned with butanol to obtain saponin fractions, which have been known as bioactive constituents of ginseng. 2',7'-Dichlorofluorescein diacetate (DCF-DA) assay and malondialdehyde (MDA) content were measured for evaluating intracellular reactive oxygen species (ROS) generation. Also, mRNA expressions and activities of antioxidant enzymes were analyzed to determine the antioxidant activity of saponin or non-saponin fractions of ginsengs. According to DCF-DA assay, $H_2O_2$-induced MDA release and ROS generation were significantly reduced by treatment with saponin fractions of white and red ginseng roots. Also, saponin fractions increased effectively intracellular antioxidant enzyme activities including catalase, glutathione peroxidase and superoxide dismutase in $H_2O_2$-treated HepG2 hepatoma cells. In general, red ginseng was more effective than white ginseng for reducing oxidative stress. These results indicate that administration of red ginseng may certainly contribute relatively stronger than white ginseng to prevent from damaging liver function by oxidative stress.

• Biomedical Science Letters
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• v.28 no.4
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• pp.237-246
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• 2022

Ischemia-reperfusion results in excess reactive oxygen species (ROS) that affect myocardial cell damage. ROS production inhibition is effectively proposed in treating cardiovascular diseases including myocardial hypertrophy. Studies have shown that oxidizing cultured cells in in vitro experiments gradually decreases the permeability of mitochondrial membranes time- and concentration-dependent, resulting in increased mitochondrial membrane damage due to secondary ROS production and cardiolipin loss. However, recent studies have shown that 5-iodo-6-amino-1,2-benzopyrone (INH₂BP), an anticancer and antiviral drug, inhibited peroxynitrite-induced cell damage in in vitro and alleviated partial or overall inflammation in animal experiments. Therefore, in this paper, we studied the preventive effect of INH₂BP on H9c2 cells derived from mouse heart damaged by oxidative stress using 700 μM of hydrogen peroxide. As a result of oxidative stress to H9c2 cells by hydrogen peroxide whether the treatment of INH₂BP or not, hydrogen peroxide caused serious damage in H9c2 cells. These results were confirmed with cell viability and Hoechst 33342 assays. And this damage was through cell death. However, it was confirmed that H9c2 cells pretreated with INH₂BP significantly reduced cell death by hydrogen peroxide. In addition, measurements with DCF-DA assay to determine whether ROS is produced in H9c2 cells treated with only hydrogen peroxide produced ROS significantly, but H9c2 cells pretreated with INH₂BP significantly reduced ROS production by hydrogen peroxide. Taken together, it is believed that INH₂BP can be useful for the prevention and treatment of cardiovascular diseases induced through oxidative stress such as heart damage caused by ischemia/reperfusion.

• Biomedical Science Letters
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• v.15 no.2
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• pp.153-160
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• 2009

Photodynamic therapy(PDT) is a treatment utilizing the generation of singlet oxygen and other reactive oxygen species(ROS), which selectively accumulate in target cells. The aim of present work is to investigate the photodynamic therapy mechanism of 9-HpbD-a-mediated PDT in HeLa cell lines. We studied the general reactive oxygen species(G-ROS) activation after 9-HpbD-a PDT using fluorescence stain with $H_2DCF-DA$. G-ROS activation observed after 9-HpbD-a PDT and higher activation condition was 1 hour after PDT at 0.5 ${\mu}g/ml$ 9-HpbD-a concentration. Sodium azide and reduced glutathione(the singlet oxygen quencher) could protect HeLa cells from cell death induced by 9-HpbD-a PDT. But D-mannitol(the hydroxyl radical scavenger) could not protect cell death. Singlet oxygen played a decisive role in 9-HpbD-a PDT induced HeLa cell death. Type II reaction was the main type of ROS formation at 9-HpbD-a PDT.

• KSBB Journal
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• v.29 no.3
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• pp.199-204
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• 2014

To investigate the effect of Plantago asiatica L. Root extract on skin care, we measured anti-oxidant activity and whitening action. As a result of measuring DPPH radical scavenging activity to examine independent anti-oxidation of PRE, there was slight scavenging activity. Fluorescent materials DHE, DCF-DA and DHR were each used to measure superoxide anion, hydrogen peroxide, and hydroperoxide created in RAW 264.7 cells, all concentrations were found to dependently prevent ROS production. Tyrosinase activation was found to be blocked dose-dependant. Melanin production was also prevented dose-dependant, but the effects were slight. Therefore, it is expected to be used effectively in development of functional cosmetic materials.

• KSBB Journal
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• v.31 no.1
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• pp.14-19
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• 2016

To investigate the effect of peanut sprout extract on skin care, we measured anti-oxidant activity and whitening action. As a result of measuring DPPH radical scavenging activity to examine independent anti-oxidation of peanut sprout extract, there was strongly scavenging activity. Fluorescent material DCF-DA was used to measure hydrogen peroxide created in RAW 264.7 cells, and all concentration dependently decreased ROS production. As a result of measuring nitric oxide to examine anti-inflammation of peanut sprout extract, there was strongly inhibited nitric oxide production in RAW 264.7 cells. Tyrosinase activation was found to inhibited dose-dependant. Melanin production was also prevented dose-dependant. Therefore, it is expected to be used effectively in development of functional cosmetic materials.

• Tuberculosis and Respiratory Diseases
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• v.65 no.6
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• pp.504-511
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• 2008

Background: Particulate matter may be toxic to human tissue. Ambient air particulate matter ${\leq}10{\mu}m$ in aerodynamic size ($PM_{10}$), which changes under different environmental conditions, is a complex mixture of organic and inorganic compounds. The Asian dust event caused by meteorological phenomena can also spread unique particulate matter in affected areas. We evaluated production of ROS, $TGF-{\beta}$, fibronectin, and $NF{\kappa}B$ by exposing normal epithelial cells to Asian dust particulate matter. Methods: Bronchial epithelial cells were exposed to 0, 50, ${\leq}100{\mu}g/ml$ of a suspension of $PM_{10}$ for 24 h. ROS were detected by measurement of DCF release from DCF-DA by FACScan. $TGF-{\beta}$, fibronectin, and $NF{\kappa}B$ were detected by western blotting. Results: $PM_{10}$ exposure increased the expression of $TGF-{\beta}$, fibronectin, and $NF{\kappa}B$. ROS production and $TGF-{\beta}$ levels were significantly higher with 50 or ${\leq}100{\mu}g/ml$ $PM_{10}$. Fibronectin and $NF{\kappa}B$ production were significantly higher after ${\leq}100{\mu}g/ml$ of $PM_{10}$. Conclusion: $PM_{10}$ from Asian dust particles might have fibrotic potential in bronchial epithelial cells via ROS induction after $PM_{10}$ exposure.