• Korean Journal of Environmental Biology
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• v.27 no.1
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• pp.66-72
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• 2009

Diabetic Retinopathy (DR) is a leading cause of blindness among adults in the western countries. Hyperglycemia is a condition, that induces apoptotic cell death in a variety of cell types in diabetes, but the mechanism remains unclear. The aim of the study is to understand the effects of high Glucose on Human Retinal Endothelial Cells. Retinal endothelial cells were cultured in Iscove's Modified Dulbecco's Medium (IMDM) containing 5, 25 and 50 mM Glucose, incubated for 24, 36 and 48 hours in humidified 5 % CO$_2$ incubator at 37$^{\circ}C$. Human Retinal Endothelial Cell Line (HREC) were characterized for morphology with different treatment by phase contrast microscopic analysis. Number of dead and viable cells was counted by trypan blue exclusion and supported by MTT assay. The intracellular Hydrogen peroxide (H$_2$O$_2$), a Reactive Oxygen Species (ROS) generation in high glucose conditions was assessed by FOX II assay and apoptosis by caspase-3 assay. The high glucose treated cells undergoing DNA fragmentation was witnessed by Agarose gel electrophoresis. We found that the cells incubated with 25 and 50 mM glucose containing medium for 48 hours altered the morphology of the cell, induced apoptosis and DNA fragmentation. The dead cell number were high in 25 and 50 mM when compared to the cells incubated with 5 mM glucose for 24, 36, and 48 hours. Also, the H$_2$O$_2$ levels and the activity of caspase-3 were increased in high glucose treated cells. Conclusions/interpretation: Our results demonstrated that elevated glucose induces apoptosis in cultured HREC. The hyperglycemia-induced increase in apoptosis may be dependent on caspase activation. The association between ROS generation and caspase-3 activation on high glucose treated cells is yet to be investigated.

• Journal of Nutrition and Health
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• v.50 no.5
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• pp.415-425
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• 2017

Purpose: Many studies have suggested that neuronal cells protect against oxidative stress-induced apoptotic cell death by polyphenolic compounds. We investigated the neuroprotective effects and the mechanism of action of Momordica charantia ethanol extract (MCE) against $H_2O_2-induced$ cell death of human neuroblastoma SK-N-MC cells. Methods: The antioxidant activity of MCE was measured by the quantity of total phenolic acid compounds (TPC), quantity of total flavonoid compounds (TFC), and 2,2-Diphenyl-1-pycrylhydrazyl (DPPH) radical scavenging activity. Cytotoxicity and cell viability were determined by CCK-8 assay. The formation of reactive oxygen species (ROS) was measured using 2,7-dichlorofluorescein diacetate (DCF-DA) assay. Antioxidant enzyme (SOD-1,2 and GPx-1) expression was determined by real-time PCR. Mitogen-activated protein kinases (MAPK) pathway and apoptosis signal expression was measured by Western blotting. Results: The TPC and TFC quantities of MCE were 28.51 mg gallic acid equivalents/extract g and 3.95 mg catechin equivalents/extract g, respectively. The $IC_{50}$ value for DPPH radical scavenging activity was $506.95{\mu}g/ml$ for MCE. Pre-treatment with MCE showed protective effects against $H_2O_2-induced$ cell death and inhibited ROS generation by oxidative stress. SOD-1,2 and GPx-1 mRNA expression was recovered by pre-treatment with MCE compared with the presence of $H_2O_2$. Pre-treatment with MCE inhibited phosphorylation of p38 and the JNK pathway and down-regulated cleaved caspase-3 and cleaved PARP by $H_2O_2$. Conclusion: The neuroprotective effects of MCE in terms of recovery of antioxidant enzyme gene expression, down-regulation of MAPK pathways, and inhibition apoptosis is associated with reduced oxidative stress in SK-N-MC cells.

• Restorative Dentistry and Endodontics
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• v.34 no.2
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• pp.95-102
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• 2009

The aim of this study was to evaluate the influence of light energy on the tooth whitening effect of bleaching agent in vitro..Extracted human mandibular molars were sectioned to 2 fragments(mesial. distal) and lingual portions of crown were used in this study. All specimens were stained using a red wine for 24 hours and immersed in artificial saliva. Specimens divided into four groups, group 1 and 2 light-activated by LumaCool (LED, LumaLite, Inc., Spring Valley, USA), group 3 and 4 light-activated by FlipoWhite2 (Plasma acr lamp, Lokki. Australia). Group 1 and 3 bleached with Luma White (LumaLite, Inc., Spring Valley, USA), group 2 and 4 bleached with Polaoffice(SDI, Victoria, Australia). Bleaching treatment performed during 10 minutes every 24 hours and repeated 6 times. During bleaching treatment, distal fragments was light-activated (L) but mesial fragments was not(NL). Shade assessment employed before and after bleaching treatment using spectrophotometer. The results of the change in shade was compared and analysed between NL and L by using paired-sample T test with 95 % level of confidence. There were no significant differences between NL and L with a few exceptions. In group 2, $a^*$ value more change in L, in group 3, $b^*$ value more change in L, in group 4, $a^*$ value less change in L. After bleaching, $L^*$ value and ${\Delta}E$ increased in all groups and the value of $a^*$ and $b^*$ decreased in all groups. Within the limitation of this test conditions, the results of this study indicate that the light energy has no obvious improving impact on the tooth whitening effect of a bleaching agent.

• Journal of Preventive Medicine and Public Health
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• v.39 no.2
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• pp.130-134
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• 2006

Objectives : Oxidative DNA damage is a known risk factor of lung cancer. The glutathione peroxidase (GPX) antioxidant enzyme that reduces hydrogen peroxide and lipid peroxides plays a significant role in protecting cells from the oxidative stress induced by reactive oxygen species. The aim of this case-control study was to investigate effects of oxidative stress and genetic polymorphisms of the GPX1 genes and the interaction between them in the carcinogenesis of lung cancer. Methods : Two hundreds patients with lung cancer and 200 age- and sex-matched controls were enrolled in this study. Every subject was asked to complete a questionnaire concerning their smoking habits and their environmental exposure to PAHs. The genotypes of the GPX1 and 8-oxoguanine glycosylase 1 (hOGG1) genes were examined and the concentrations of urinary hydroxypyrene (1-OHP), 2-naphthol and 8-hydroxydeoxyguanosine (8-OH-dG) were measured. Results : Cigarette smoking was a significant risk factor for lung cancer. The levels of urinary 8-OH-dG were higher in the patients (p<0.001), whereas the urinary 1-OHP and 2-naphthol levels were higher in the controls. The GPX1 codon 198 polymorphism was associated with an increased risk of lung cancer. Individuals carrying the Pro/Leu or Leu/Leu genotype of GPX1 were at a higher risk for lung cancer (adjusted OR=2.29). In addition, these individuals were shown to have high urinary 8-OH-dG concentrations compared to the individuals with the GPX1 Pro/Pro genotype. On the other hand, the polymorphism of the hOGG1 gene did not affect the lung cancer risk and the oxidative DNA damage. Conclusions : These results lead to a conclusion that individuals with the GPX1 Pro/Leu or Leu/Leu genotype would be more susceptible to the lung cancer induced by oxidative stress than those individuals with the Pro/Pro genotype.

• Korean Journal of Food Science and Technology
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• v.46 no.3
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• pp.384-389
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• 2014

The objective of the present study was to investigate the chemical composition of anthocyanin-enriched extract of radiation-induced blackberry (Rubus fruticosus L.) mutant (${\gamma}$-B201) as well as the protective effect of ${\gamma}$-B201 against oxidative stress in vitro. The cytotoxicity, reactive oxygen species (ROS) scavenging capacity, and DNA damage were assessed by WST-1 assay, flow cytometry, and comet assay, respectively. Lactate dehydrogenase, superoxide dismutase, and catalase activities were determined by using a commercial kit. The in vitro results showed that ${\gamma}$-B201 increased the cell viability, reduction of lactate dehydrogenase release, and intracellular ROS scavenging capacity in hydrogen peroxide ($H_2O_2$)-treated HepG2 cells. Furthermore, treatment with ${\gamma}$-B201 attenuated DNA damage in $H_2O_2$-treated HepG2 cells and treatment with ${\gamma}$-B201 restored the activity of superoxide dismutase and catalase in $H_2O_2$-treated HepG2 cells. In conclusion, the present study suggests that ${\gamma}$-B201 blackberry extract can exert a significant cytoprotective effect against $H_2O_2$-induced cell damage.

• Journal of Food Hygiene and Safety
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• v.32 no.6
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• pp.507-512
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• 2017

There has been increasing concern regarding misuse of disinfectants and sanitizers such as ethanol, sodium hypochlorite, and hydrogen peroxide for food contact surfaces in the food industry. Examining the efficacy of the concentration of currently used disinfectants and sanitizers is urgently required in the Korean society. This study aimed to develop predictive reduction models for Escherichia coli and Staphylococcus aureus in suspension, as a function of $ClO_2$ (chlorine dioxide) and contact time using response surface methodology. E. coli ATCC 10536 and S. aureus ATCC 6538 (initial inoculum, 8-9 log CFU/mL) in tryptic soy broth were treated with different concentrations of $ClO_2$ (5, 20, and 35 ppm) for different contact times (1, 3, and 5 min) following a central composite design. The polynomial reduction models for $ClO_2$ on E. coli and S. aureus were developed under the clean condition. E. coli reduction by 35 ppm $ClO_2$ for 1, 3, and 5 min was 2.49, 2.70, and 3.65 log CFU/mL, respectively. Also, S. aureus reduction by 35 ppm $ClO_2$ for 1, 3, and 5 min was 4.59, 5.25, and 5.81 log CFU/mL, respectively. The predictive response polynomial models developed were $R=0.43231-0.056492^*X_1-0.097771^*X_2+9.24167E-003^*X_1^*X_2+3.06333E-003^*X_1{^2}$ ($R^2=0.98$) on E. coli and $R=1.10542-0.20896^*X_1-0.046062^*X_2+8.30000E-003^*X_1^*X_2+8.73300E-003^*X_1{^2}$ ($R^2=0.99$) on S. aureus, where R was the bacterial reduction (log CFU/mL), $X_1$ was the concentration and $X_2$ was the contact time. Our predictive reduction models should be validated in developing the optimal concentration and contact time of $ClO_2$ for inhibiting E. coli and S. aureus on food contact surfaces.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.9
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• pp.1139-1145
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• 2006

In this study, antioxidant activities of enzymatic and methanolic extracts from E. cava stem and leave were evaluated by measuring the scavenging activities on 1,1 diphenyl 2 picrylhydrazyl (DPPH), hydroxyl radical, hydrogen peroxide and the inhibitory effects on DNA damage induced by oxidative stress of cells. Enzymatic extracts were prepared by enzymatic hydrolysis of both stem and leave using food grade five different carbohydrases (Viscozyme, Celluclast, AMG, Termamyl, Ultraflo) and five proteases (Protamex, Kojizyme, Neutrase, Flavourzyme, Alcalase). The enzymatic extracts were lower than methanolic extracts in polyphenol contents, but higher in extraction yield by approximately 30%. The enzymatic extracts were superior to methanolic extracts in DPPH and H2O2 scavenging activities and DNA damage protective effect. There were no significant antioxidant activity difference between stem and leave, but the extracts of leave were relatively better than those of stem. In this study it is suggested that E. cava stem as well as its leave would be a good raw materials for antioxidants compound extraction and enzymatic hydrolysis would be a good strategy to prepare antioxidant extracts from seaweeds.

• Journal of Nutrition and Health
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• v.52 no.1
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• pp.26-35
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• 2019

Purpose: The aim of this study was to estimate the antioxidant activities of 50%, 70%, and 100% ethanol extracts of Lycium barbarum leaf and chlorophyll removal extract. Methods: The antioxidant activities were estimated by measuring total polyphenol content and by assays of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfate) (ABTS) radical scavenging activities and ferric reducing antioxidant power (FRAP). In addition, reactive oxygen species (ROS) production, DNA fragmentation, and antioxidant enzyme (superoxide dismutase and catalase) activities of the extracts were measured in hydrogen peroxide ($H_2O_2$)-stressed HepG2 cells. Results: The total polyphenol content, DPPH and ABTS radical scavenging activities, and FRAP value of the extracts increased in an ethanol concentration-dependent manner. The antioxidant activities of the chlorophyll-removal extracts were much higher than those of the chlorophyll-containing extracts. Cytotoxicity was not observed in HepG2 cells with extracts up to $1,000{\mu}g/mL$. All extracts inhibited ROS production in a concentration-dependent manner from $31.3{\mu}g/mL$ and inhibited DNA damage at $250{\mu}g/mL$. The SOD and catalase activities of cell lines treated with the extracts and $H_2O_2$ were similar to those of normal cells, indicating a strong protective effect. Conclusion: Lycium barbarum leaf extracts had high antioxidant activities and protected $H_2O_2$-stressed HepG2 cells. Since the chlorophyll-removal extract exhibited higher antioxidant activities than the chlorophyll-containing ones and the cytoprotective effect was similar, chlorophyll removal extract of Lycium barbarum leaf could be developed as ingredients of functional food and cosmetics.

• Journal of Life Science
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• v.31 no.2
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• pp.126-136
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• 2021

Oxidative stress causes injury to and degeneration of retinal pigment epithelial (RPE) cells. It is involved in several retinal disorders and leads to vision loss. In the present study, we investigated the effect of 14 kinds of natural compounds and two kinds of synthetic compounds on oxidative stress-induced cellular damage in human PRE cell lines (ARPE-19). From among them, we selected five kinds of compounds, including auranofin, FK-509, hemistepsin A, honokiol, and spermidine, which have inhibitory effects against hydrogen peroxide (H2O2)-mediated cytotoxicity. In addition, we found that four kinds of compounds (excluding auranofin) have protective effects on H2O2-induced mitochondrial dysfunction. Furthermore, the expression of phosphorylation of histone H2AX, a sensitive marker of DNA damage, was markedly up-regulated by H2O2, whereas it was notably down-regulated by FK-506, honokiol, and spermidine treatment. Meanwhile, five kinds of candidate compounds had no effect on H2O2-induced intracellular reactive oxygen species (ROS) levels, suggesting that the five candidate compounds have protective effects on oxidative stress-induced cellular damage through the ROS-independent pathway. Taken together, according to the results of H2O2-mediated cellular damage―such as cytotoxicity, apoptosis, mitochondrial dysfunction, and DNA damage―spermidine and FK-506 are the natural and synthetic compounds with the most protective effects against oxidative stress in RPE. Although further studies on the identification of the mechanism responsible are required, the results of the present study suggest the possibility of using spermidine and FK-506 to suppress the risk of retinal disorders.

• Journal of Life Science
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• v.31 no.5
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• pp.488-495
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• 2021

The aim of this study was to investigate the possible antioxidant effect of Spirodela polyrhiza (SP) on rats fed a high fat and high cholesterol diet supplemented with either 5% (SPA group) or 10% (SPB group) SP for 4 weeks. The hepatic SOD activity of the HF group significantly decreased compared to that of the N group, but that of the SPA and SPB groups significantly increased. The GPx activity of the SPA and SPB groups in the liver was significantly greater than that of the HF group, and the hepatic catalase activity of the SPA and SPB groups significantly increased compared to the HF group. The hepatic superoxide radical content of the mitochondria and microsomes of the HF group significantly increased compared to that of the N group, but the contents were reduced in the group that took SP powder. The hepatic hydrogen peroxide content in the cytosol and mitochondria of the SP powder group was lower than in the HF group. The carbonyl content in the mitochondria and microsomes of the SPA and SPB groups was significantly lower than in the HF group. The TBARS values in the liver significantly decreased in the SPA and SPB groups. Spirodela polyrhiza was thus effective in reducing oxidative stress by regulating the hepatic antioxidant enzymes and the free radicals in rats fed high fat and high cholesterol diets.