• Korean Journal of Agricultural Science
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• v.38 no.4
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• pp.717-722
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• 2011

In this study, we investigated the antioxidant activity of hesperidin and hesperetin, which are the active compounds from Citrus junos, in the cellular system. Under cellular model of oxidative damage using LLC-$PK_1$ renal epithelial cell, the oxidative damage induced by 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH) led to the loss of cell viability, while treatment of hesperidin and hesperetin increased significantly the cell viability as dose-dependent manner. In addition, NO-induced cellular oxidative damage by sodium nitroprusside were significantly recovered by the treatment of hesperidin and hesperetin, showing the increase of cell viability. But hesperidin and hesperetin showed no significant protective effect on $O_2{^-}$-induced cellular oxidative damage. The present study indicates that hesperidin and hesperetin protect against free radical, especially AAPH-induced peroxyl radical. In particular, hesperetin has stronger protective effect against oxidative stress than hesperidin.

• 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.

• Preventive Nutrition and Food Science
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• v.14 no.1
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• pp.37-42
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• 2009

The protective activity of seolitae chungkukjang added with green tea against oxidative stress was investigated under the cellular systems using LLC-$PK_1$ cells. The treatment of 2,2'-azobis(2-aminopropane) dihydrochloride (AAPH) showed increase in lipid peroxidation, and decrease in endogenous antioxidant enzymes activity and cell viability. However, the methanol extract of seolitae chungkukjang inhibited lipid peroxidation by 58.3%, and increased cell viability up to more than 60%. In addition, it enhanced superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities. Seolitae chungkukjang improved oxidative stress-induced cellular injury through the radical scavenging activities. In particular, the addition of green tea in seolitae chungkukjang showed stronger effect against oxidative stress induced by AAPH. The more addition of green tea resulted in the greater antioxidative effect through elevation in activities of SOD and GSH-Px, and inhibition of lipid peroxidation, eventually leading to increase in cell viability. Theses results suggested that seolitae chungkukjang added with green tea have protective effects from cellular oxidative damage and could be considered as an application for the development of chungkukjang with functionality.

• YAKHAK HOEJI
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• v.40 no.6
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• pp.705-712
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• 1996

We have previously shown that determination of glucose uptake using ${\alpha}$-methylglucose(${\alpha}$-MG) is very sensitive and rapid parameter for the assessment of loss of cellular fu nction in renal cell line($LLC-PK_1$). The present study was designed to elucidate the mechanism of inhibition of ${\alpha}$-MG uptake and the intracellular site of toxic action of cisplatin(CIS). $LLC-PK_1$ cells were exposed to various concentrations(5 ${\mu}$M-l00 ${\mu}$M) of CIS for 5 hrs or 24 hrs and ${\alpha}$-MG uptake was determined. Mitochondrial function was evaluated by measuring intracellular ATP content and MTT reduction. The activities of marker enzymes for the basolateral membrane(Na$^+$-K$^+$ ATPase) and brush border membrane (alkaline phosphatase: ALP) were also measured. CIS treatment significantly inhibited the ${\alpha}$-MG uptake in a time- and dose-dependent manner above 25 ${\mu}$M for 5 hrs. Intracellular ATP content and MTT reduction were affected by 24 hr-treatment of 50 ${\mu}$M CIS. The activities of Na$^+$-K$^+$ ATPase and ALP were significantly decreased at 10 ${\mu}$M and 5 ${\mu}$M of CIS for 24 hrs, respectively. The incubation with CIS for 5 hrs had no effects on the intracellular ATP content, MTT reduction and the activities of marker enzymes up to 100 ${\mu}$M. These results partly indicate that inhibition of ${\alpha}$-MG uptake by CIS may not be attributed to the disturbance of mitochondrial function or inhibition of the activity of Na$^+$-K$^+$ ATPase and can be resulted from direct effect of CIS on the Na$^+$/glucose cotransporter in brush border membrane. This study shows that additional mechanistic information, indicating the intracellular site of nephrotoxic action, can be gained by coupling the ${\alpha}$-MG uptake and ATP content or the activity of Na$^+$-K$^+$ ATPase.

• Preventive Nutrition and Food Science
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• v.19 no.3
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• pp.129-135
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• 2014

The protective role of phloroglucinol against oxidative stress and stress-induced premature senescence (SIPS) was investigated in vitro and in cell culture. Phloroglucinol had strong and concentration-dependent radical scavenging effects against nitric oxide (NO), superoxide anions ($O_2{^-}$), and hydroxyl radicals. In this study, free radical generators were used to induce oxidative stress in LLC-PK1 renal epithelial cells. Treatment with phloroglucinol attenuated the oxidative stress induced by peroxyl radicals, NO, $O_2{^-}$, and peroxynitrite. Phloroglucinol also increased cell viability and decreased lipid peroxidation in a concentration-dependent manner. WI-38 human diploid fibroblast cells were used to investigate the protective effect of phloroglucinol against hydrogen peroxide ($H_2O_2$)-induced SIPS. Phloroglucinol treatment attenuated $H_2O_2$-induced SIPS by increasing cell viability and inhibited lipid peroxidation, suggesting that treatment with phloroglucinol should delay the aging process. The present study supports the promising role of phloroglucinol as an antioxidative agent against free radical-induced oxidative stress and SIPS.

• Korean Journal of Agricultural Science
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• v.39 no.1
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• pp.81-86
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• 2012

In this study, the antioxidative effect and protective potential against oxidative damage of extract and fractions from broccoli were investigated under in vitro and cellular system. The methanol (MeOH) extracts of broccoli flowers and sprouts were partitioned as dichloromethane, n-butanol (BuOH) and aqueous fractions. The comparison of antioxidative effect of broccoli flowers and sprouts showed that broccoli sprouts exerted the more effective protective activity from 2,2'-azobis (2-aminopropane) dihydrochloride (AAPH)-induced oxidative stress in LLC-$PK_1$ porcine renal epithelial cell. In addition, the extract and fractions from broccoli sprouts showed strong scavenging effect of 1,1-diphenyl-2-picrylhydrazyl radical and the BuOH fraction exerted the strongest activity. Therefore, the BuOH fraction was evaluated as the most active fraction with strong radical scavenging activity among the fractions of broccoli flowers and sprouts. The present study suggests the antioxidative potential against free radical-induced oxidative damage of flowers and sprouts of broccoli. In addition, the BuOH fraction of broccoli is considered as the active fraction with antioxidative effect.

• 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.

• Journal of Ginseng Research
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• v.40 no.2
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• pp.135-140
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• 2016

Background: Nephrotoxicity is a common side effect of medications. Panax ginseng is one of the best-known herbal medicines, and its individual constituents enhance renal function. Identification of its efficacy and mechanisms of action against drug-induced nephrotoxicity, as well as the specific constituents mediating this effect, have recently emerged as an interesting research area focusing on the kidney protective efficacy of P. ginseng. Methods: The present study investigated the kidney protective effect of fermented black ginseng (FBG) and its active component ginsenoside 20(S)-Rg3 against cisplatin (chemotherapy drug)-induced damage in pig kidney (LLC-PK1) cells. It focused on assessing the role of mitogen-activated protein kinases as important mechanistic elements in kidney protection. Results: The reduced cell viability induced by cisplatin was significantly recovered with FBG extract and ginsenoside 20(S)-Rg3 dose-dependently. The cisplatin-induced elevated protein levels of phosphorylated c-Jun N-terminal kinase (JNK), p53, and cleaved caspase-3 were decreased after cotreatment with FBG extract or ginsenoside 20(S)-Rg3. The elevated percentage of apoptotic LLC-PK1 cells induced by cisplatin treatment was significantly abrogated by cotreatment with FBG and the ginsenoside 20(S)-Rg3. Conclusion: FBG and its major ginsenoside 20(S)-Rg3, ameliorated cisplatin-induced nephrotoxicity in LLC-PK1 cells by blocking the JNKep53ecaspase-3 signaling cascade.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.8
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• pp.1179-1186
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• 2010

The antiradical property of hot water extract from dried radish (DR) or dried radish roasted with pressure (DRRP) was investigated in vitro and in LLC-PK1 cell system. The contents of total free amino acid and reducing sugar in DR were decreased by 72.86% and 3.17%, respectively, after pressurized roasting. In vitro test, $IC_{50}$ for DR and DRRP for DPPH radical scavenging activity were 646.70 and $135.45\;{\mu}g/mL$, 896.10 and $566.98\;{\mu}g/mL$ for superoxide anion radical, and 722.26 and $531.84\;{\mu}g/mL$ for hydroxy radical, respectively. The radical scavenging effects of DRRP was significantly greater than those for DR (p<0.001). These radical scavenging effects of DR and DRRP were confirmed in LLC-$PK_1$ at which oxidative stresses were induced by superoxide, nitric oxide and peroxynitrite generated in the treatment of pyrogallol, SNP, and SIN-1, respectively. Cell viability was increased in the presence of DR or DRRP, dose dependently (p<0.05), and TBARS formation was decreased. The protective effects of DRRP against oxidative damage in LLC-$PK_1$ were greater than those of DR at the same concentration tested (p<0.05). This superior antiradical activity of DRRP might be due to the products produced during the pressurized roasting in addition to the antioxidative compounds originally present in the radish. 5-hydroxyl methyl furfural (5-HMF) known as an intermediate product of the maillard reaction was detected in DRRP (0.57 mg/g), but not from DR. In conclusion, daily consumption of DRRP may prevent oxidative damage by retarding oxidative stress.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.110.1-110.1
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• 2003

Fumonisins are specific inhibitors of ceramide synthase in sphingolipid metabolism. Sphingolipids are biologically active lipid mediators in cellular physiology and involved in cell signaling, growth, transformation, angiogenesis and differentiation. The objective of this study was to determine the effect of fumonisin B1 on $Na^+, \;K^+$-pump activity when fumonisin B1 was exposed to LLC-PK1 cells. Fumonisin B1 elevated free sphingoid bases and their 1-phosphates, while total complex sphingolipids were depleted at 20$\mu$M fumonisin B1 during the 3 day exposure. (omitted)