• Preventive Nutrition and Food Science
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• v.13 no.2
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• pp.90-94
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• 2008

The protective effect of chungkukjang from Sunchang province against oxidative stress was evaluated in the cellular system using LLC-$PK_1$ renal epithelial cells. The LLC-$PK_1$ cells showed decrease in cell viability and elevation in lipid peroxidation by the treatment with the generators of nitric oxide (NO) and superoxide anion ($O_2^-$) produced by sodium nitrouprusside and pyrogallol, respectively. However, the methanol extract of chungkukjang significantly inhibited cellular loss and lipid peroxidation in a dose-dependent manner; in particular K chungkukjang (KC) exerted the strongest protective effect. In addition, the protective effect of chungkukjang from 3-morpholinosydnonimine, as a source of peroxynitrite, with simultaneous generations of NO and $O_2^-$, was also studied. Treatment with chungkukjangs significantly preserved the cell viability and inhibited lipid peroxidation caused by SIN-1 with dose-dependence. The present study suggests that chungkukjang from Sunchang province, especially KC, would have protective potential from oxidative stress induced by free radicals under cellular oxidative damage.

• The Journal of Internal Korean Medicine
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• v.31 no.1
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• pp.153-165
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• 2010

Objectives : The aims of this study were to investigate the cytoprotective, antioxidative and inflammation genes inhibitory effects of Patriniae Radix on the mouse LLC-$PK_1$ cells (renal epithelial cells). Methods : The cytoprotective effect of Patriniae Radix was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The antioxidative effect was measured in terms of generation amount of superoxide anion radical (${\cdot}{O_2}^-$) by 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), nitric oxide (NO) by 4,5-diaminofluorescein (DAF-2), peroxynitrite ($ONOO^-$) by dihyldrorhodamine 123 (DHR 123) and prostaglandin $E_2$ ($PGE_2$) by $PGE_2$ immunoassay on $H_2O_2$-treated LLC-$PK_1$ cells. For measuring of inflammation genes inhibitory effects, western blot was performed to detect IKK-$\alpha$, phospho-$I{\kappa}B-\alpha$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, IL-$1{\beta}$ and VCAM-1 protein level in cytosol fractions from LLC-$PK_1$ cells. Results : Patriniae Radix extract reduced the $H_2O_2$-induced cell death and inhibited the amount of $H_2O_2$-induced ${\cdot}{O_2}^-$, NO, $ONOO^-$, $PGE_2$ generation dose-dependently on the mouse LLC-$PK_1$ cells in vitro. Also Patriniae Radix extract inhibited the expression of IKK-$\alpha$, phospho-$I{\kappa}B-\alpha$, COX-2, iNOS, IL-$1\beta$ and VCAM-1 genes dose-dependently by means of decreasing activation of NF-${\kappa}B$. Conclusions : According to above results, it was identified that Patriniae Radix had the cytoprotective, antioxidative and inflammation genes inhibitory effects. So it was suggested that Patriniae Radix would be effective to the treatment for the inflammatory process and inflammation-related diseases.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.11
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• pp.1674-1680
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• 2014

The aim of this study was to investigate the protective effects of methanolic extracts of cooked mixed grain rice samples, including grain rice (sorghum, black bean, proso millet, and Job's tears) mixed with fermented brown rice (GR), GR added with 0.5% water extract of Sanghwang mushroom (GRS) or 0.1% curry (GRK), and traditional five grain mixed rice (TMR, Ohgokbap), on $H_2O_2$-induced oxidative injury in LLC-PK1 pig renal epithelial cells. White rice (WR) was used as a positive control. Cells were first exposed to $H_2O_2$ ($250{\mu}M$) for 4 hr, followed by treatment with $100{\mu}g/mL$ of different GR extracts for 24 hr. $H_2O_2$ significantly induced cell damage (P<0.05). Cellular levels of reactive oxygen species (ROS), lipid peroxidation, and antioxidant enzymes, including catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH-px), were measured. In addition, mRNA levels of antioxidant enzymes were determined by RT-PCR assay. Mixed grain rice, particularly GRS and GRK, were able to reduce cellular levels of ROS, decrease lipid peroxidation, and also increase mRNA expression of antioxidant enzymes compared to other samples. These results suggest that mixed grain rice, specifically GRS and GRK, have strong protective effects against $H_2O_2$-induced oxidative injury in LLC-PK1 cells through inhibition of lipid peroxidation, reduction of ROS levels, and elevation of antioxidant enzyme activities.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.6
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• pp.809-815
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• 2015

In this study, we confirmed biological compounds from methanol (MeOH) extract of processed Polygoni Multiflori Radix (PPMR), and the radical scavenging effect and oxidative stress protective activity of MeOH extract of PPMR were investigated under in vitro conditions using LLC-$PK_1$ renal epithelial cells. In HPLC analysis, MeOH extract of PPMR contained four species of biological compounds named 2,3,5,4'-tetrahydroxystilbene 2-O-${\beta}$-D-glucoside, emodin, chrysophanol, and rhein. 2,3,5,4'-Tetrahydroxystilbene 2-O-${\beta}$-D-glucoside was detected as the main compound in PPMR as 115.02 mg/kg. MeOH extract of PPMR showed 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) diammonium salt (ABTS), and hydroxyl radical scavenging activities in a concentration- dependent manner. In particular, upon $50{\mu}g/mL$ of PPMR extract treatment, DPPH, ABTS, and hydroxyl radical scavenging activities were approximately 48.4%, 57.9%, and 81.2%, respectively. LLC-$PK_1$ cell viability declined in response to oxidative stress induced by pyrogallol, sodium nitroprusside (SNP), and morpholinosydnonimine (SIN-1) generators of NO, $O_2{^-}$, and $ONOO^-$, respectively. However, MeOH extract of PPMR significantly and dose-dependently inhibited oxidative-stressed LLC-$PK_1$ cell cytotoxicity. In fact, upon $50{\mu}g/mL$ of PPMR extract treatment, LLC-$PK_1$ cell viabilities were approximately 82.1%, 89.1%, and 77.6% compared to stress levels induced by pyrogallol, SNP, and SIN-1, respectively.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.8
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• pp.1151-1156
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• 2005

The comparison on antioxidative activity of 13 kinds of vegetables that showed efficient oxygen radical absorbance capacity was carried out under in uitro and cellular model using LLC-$ PK_{1}$ renal epithelial cell, and also the total Phenol contents were analyzed. Beets, eggplant, and kale exerted the strong 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging effect and also these vegetables showed high contents of total phenol, indicating the positive relationship between DPPH radical scavenging effect and total phenol content. In addition, the treatment of 1 mM 2,2'-azobis (2-amidinopropane) dihydrochloride for 24 hrs to LLC-$PK_{1}$ cell susceptible to oxidative stress led to the decline of cell viability to 68.1$\%$, whereas the cellular oxidative damage was ameliorated by vegetables, especially eggplant and cauliflower, resulting in the elevation of cell viability to higher than 90$\%$ at the concentration of 5 $\mu$g/mL. This study suggests that 13 kinds of vegetables exert antioxidative activity under in uitro and cellular oxidative damage model, in particular among them eggplant showed the most effective antioxidative activity with higher total phenol content.

• Korean Journal of Food Science and Technology
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• v.47 no.4
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• pp.517-524
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• 2015

The objectives of the present study were to investigate the protective effects of Angelica keiskei juice and Oenanthe javanica juice against oxidative damage in LLC-$PK_1$ renal epithelial cells and to evaluate their free radical-scavenging activities. Both A. keiskei juice and O. javanica juice showed a strong in vitro antioxidant activities such as ${\alpha}$, ${\alpha}$-diphenyl-${\beta}$-picrylhydrazyl (DPPH), nitric oxide (NO), $O_2{^-}$, and ${\cdot}OH$ radical-scavenging activities. The LLC-$PK_1$ cells showed significant lipid peroxidation and cell death due to oxidative stress when it was induced by 2, 2'-azobis (2-amidinopropane) dihydrochloride (AAPH), sodium nitroprusside (SNP), pyrogallol, and 3-morpholinosydnonimine (SIN-1). Treatment with both A. keiskei juice and O. javanica juice significantly increased cell viability and inhibited lipid peroxidation. These results suggest that A. keiskei juice and O. javanica juice are potential natural antioxidants.

• Korean Journal of Agricultural Science
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• v.39 no.4
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• pp.467-471
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• 2012

In this study, we investigated the antioxidative and antiaging activity of Perilla frutescens using LLC-$PK_1$ porcine renal epithelial cell and WI-38 human diploid fibroblast cell. The extract from Perilla frutescens showed strong protective effect against nitric oxide (NO) and superoxide ($O_2{^-}$)-induced oxidative stress generated by sodium nitroprusside (SNP) and pyrogallol, respectively. The result showed that P. frutescens increased the cell viability and showed scavenging activity of NO and $O_2{^-}$. In addition, the extract of P. frutescens exerted the protective effect against peroxynitrite ($ONOO^-$) induced by 3-morpholinosydnonimine. It suggests that P. frutescens would have the protective role against $ONOO^-$ itself and its precursors, NO and $O_2{^-}$. Furthermore, the aging model of hydrogen peroxide ($H_2O_2$)-treated WI-38 human diploid fibroblast was employed to investigate the anti-aging effect of P. frutescens. $H_2O_2$-treated WI-38 cells showed the loss of cell viability, however before-treatment with P. frutescens to WI-38 cells under premature senescence could delay the cellular aging process. The present study suggests the antioxidative and antiaging potential against free radical-induced oxidative damage of P. frutescens.