• Title/Summary/Keyword: LLC-$PK_1$ renal epithelial cell

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Effect of Cisplatin on Sodium-Dependent Hexose Transport in LLC-$PK_1$ Renal Epithelial Cells

  • Lee, Suk-Kyu;Kim, Jee-Yeun;Yu, Tai-Hyun;Kim, Kyoung-Ryong;Kim, Kwang-Hyuk;Park, Yang-Saeng
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.1
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    • pp.35-43
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    • 1997
  • Cis-dichlorodiammine platin${\mu}M$II (Cisplatin), an effective chemotherapeutic agent, induces acute renal failure by unknown mechanisms. To investigate direct toxic effects of cisplatin on the renal proximal tubular transport system, LLC-$PK_1$ cell line was selected as a cell model and the sugar transport activity was evaluated during a course of cisplatin treatment. Cells grown to confluence were treated with cisplatin for 60 min, washed, and then incubated for up to 5 days. At appropriate intervals, cells were tested for sugar transport activity using ${\alpha}-methyl-D-[^{14}C]glucopyranoside$ (AMG) as a model substrate. In cells treated with 100 ${\mu}M$ cisplatin, the AMG uptake was progressively impaired after 3 days. The viability of cells was not substantially changed with cisplatin of less than 100 ${\mu}M$, but it decreased markedly with 150 and 200 ${\mu}M$. In cisplatin-treated cells, the $Na^+$ -dependent AMG uptake was drastically inhibited with no change in the $Na^+$ -independent uptake. Kinetic analysis indicated that Vmax was suppressed, but Km was not altered. The $Na^+$ -dependent phlorizin binding was also decreased in cisplatin-treated cells. However, the AMG efflux from preloaded cells was not apparently retarded by cisplatin treatment. These data indicate that the cisplatin treatment impairs $Na^+$ -hexose cotransporters in LLC-$PK_1$ cells and suggest strongly that defects in transporter function at the luminal plasma membrane of the proximal tubular cells constitute an important pathogenic mechanism of cisplatin nephrotoxicity.

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Antioxidative effects of hesperidin and hesperetin under cellular system (Hesperidin과 hesperetin의 cellular system에서의 항산화 효과)

  • Cho, Eun-Ju;Li, Li;Yamabe, Noriko;Kim, Hyun-Young
    • 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.

Protective Effects of Chungkookjang Extract on High Glucose Induced Oxidative Stress in LLC-PK1 Cells

  • Yi, Na-Ri;Seo, Kyoung-Chun;Choi, Ji-Myung;Cho, Eun-Ju;Song, Young-Ok;Han, Ji-Sook
    • Preventive Nutrition and Food Science
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    • v.13 no.2
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    • pp.84-89
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    • 2008
  • This study was designed to investigate the protective effect of a methanol extract of Chungkookjang (CKJ) on high glucose induced oxidative stress in LLC-$PK_1$ cells (renal tubular epithelial cells), which are susceptible to oxidative stress. Freeze dried CKJ powder was extracted with methanol, and the extract solution was concentrated, and then used in this study. To determine the protective effect of CKJ extract, oxidative stress was induced by exposing of LLC-$PK_1$ cells to high glucose (30 mM) or normal glucose (5 mM) for 24 hr. Exposure of LLC-$PK_1$ cells to high glucose for 24 hr resulted in a significant (p<0.05) decrease in cell viability, catalase, SOD and GSH-px activity and a significant (p<0.05) increase in intracellular ROS level and thiobarbituric acid reactive substances (TBARS) formation in comparison to the cells treated with 5 mM glucose. CKJ extract treatment decreased intracellular ROS level and TBARS formation, and increased cell viability and activities of antioxidant enzymes including catalase, SOD and GSH-px in high glucose pretreated LLC-$PK_1$ cells. These results suggest that CKJ extract may be able to protect LLC-$PK_1$ cells from high glucose-induced oxidative stress, partially through the antioxidative defense systems.

Free radical scavenging effect and protective activity from oxidative stress of broccoli flowers and sprouts (Broccoli flower와 Broccoli sprout의 라디칼 소거능 및 산화적 스트레스 개선 효과)

  • Kim, Hyun-Young;Lee, Young-A;Cho, Eun-Ju
    • 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.

The Effects of Lycium Chinense Milie on the $H_{2}O_{2}$-treated $LLC-PK_1$ Cell's Redox Status and $NF-{\kappa}B$Signaling (지골피(地骨皮)가 $H_{2}O_{2}$에 의한 $LLC-PK_1$ 세포의 Redox Status 및 $NF-{\kappa}B$ Signaling에 미치는 영향)

  • Choi, Gyu-Ho;Shin, Hyeon-Cheol
    • The Journal of Internal Korean Medicine
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    • v.30 no.1
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    • pp.36-50
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    • 2009
  • Objectives : This study was aimed to verify the cytoprotective function, antioxidative effect and inflammation genes inhibitory effects of Lycium chinense Milie. Therefore the generation of superoxide anion radical ( $O_2\;^-$), peroxynitrite ($ONOO^-$), nitric oxide (NO) and prostaglandin $E_2$ $(PGE_2)$ was investigated in the renal epithelial cells of mouse. Effects of Lycium chinense Milie on the expression of inflammation-related proteins, $IKK-{\alpha}$. $p-IKK-\alpha\beta$, $p-I{\kappa}B-\alpha$, $NF-{\kappa}B$ (p50, p65), COX-2 and iNOS, were examined by western blotting. Methods : For this study, the fluorescent probes were used, namely dihydrorhodamine 123 (DHR 123), 4.5-diaminofluorescein (DAF-2) and 2',7'-dichlorodihydrofluorescein diacetate (DCFDA). Western blotting was performed using anti-$IKK-\alpha$, anti-phospho $IKK-\alpha\beta$, anti-phospho $I{\kappa}B-\alpha$, anti-$NF-{\kappa}B$ (p50, p65), anti-COX-2 and anti-iNOS, respectively. Results : Lyciutn chinense Milie reduced $H_{2}O_{2}$-induced cell death dose-dependently. It inhibited the generation of $O_2\;^-$, $ONOO^-$, NO and $PGE_2$ in the $H_{2}O_{2}$-treated renal epithelial cells of mouse in vitro. Lycium chinense Milie inhibited the expression of $IKK-\alpha$, $p-IKK-\alpha\beta,\;p-I{\kappa}B-\alpha$, COX-2 and iNOS genes by means of decreasing activation of $NF-{\kappa}B$. Conclusions : According to above results. Lycium chinense Milie recommended to be applied in treatment for the inflammatory process and inflammation-related diseases.

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Effects of Polygoni Cuspidati Radix on the $H_2O_2$-treated LLC-$PK_1$ Cell's Redox Status and NF-${\kappa}B$ Signaling (호장근(虎杖根)이 $H_2O_2$에 노출된 LLC-$PK_1$ 세포의 Redox Status 및 NF-${\kappa}B$ Signaling에 미치는 영향)

  • Kim, Sol-Ri;Jeong, Ji-Cheon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.26 no.4
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    • pp.483-490
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    • 2012
  • This study was designed to identify the effects of Polygoni cuspidati Radix(PCR) on the generation of superoxide anion radicals (${\cdot}O_2{^-}$), nitric oxide (NO), peroxynitrite ($ONOO^-$) in the renal epithelial cells of mouse(LLC-$PK_1$). The effects of PCR on the expression of inflammation-related proteins, IKK-${\alpha}$, phospho-$I{\kappa}B-{\alpha}$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, IL-$1{\beta}$, VCAM-1, were examined by western blotting. For this study, the fluorescent probes, namely dihydrorhodamine 123 (DHR 123), 2',7'-dichloro dihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) were used. Protein expression levels of IKK-${\alpha}$, phospho-$I{\kappa}B-{\alpha}$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, IL-$1{\beta}$, VCAM-1 were assayed by western blot. PCR reduced $H_2O_2$-induced cell death dose-dependently. It inhibited the generation of ${\cdot}O_2{^-}$, NO, $ONOO^-$ and $PGE^2$ in the $H_2O_2$-treated LLC-PK1 cells in vitro. PCR inhibited the espression of IKK-${\alpha}$, phospho-$I{\kappa}B-{\alpha}$, COX-2, iNOS, IL-$1{\beta}$ and VCAM-1 genes by means of decreasing the NF-${\kappa}B$ activation. These results suggest that PCR is an effective NO, ${\cdot}O_2{^-}$, $ONOO^-$ scavenger, and this substance recommended to be applied in treatment for the inflammatory process and inflammation-related disease.

Protective effect of ginsenosides Rk3 and Rh4 on cisplatin-induced acute kidney injury in vitro and in vivo

  • Baek, Seung-Hoon;Shin, Byong-kyu;Kim, Nam Jae;Chang, Sun-Young;Park, Jeong Hill
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.233-239
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    • 2017
  • Background: Nephrotoxicity is the major side effect in cisplatin chemotherapy. Previously, we reported that the ginsenosides Rk3 and Rh4 reduced cisplatin toxicity on porcine renal proximal epithelial tubular cells (LLC-PK1). Here, we aimed to evaluate the protective effect of ginsenosides Rk3 and Rh4 on kidney function and elucidate their antioxidant effect using in vitro and in vivo models of cisplatin-induced acute renal failure. Methods: An enriched mixture of ginsenosides Rk3 and Rh4 (KG-KH; 49.3% and 43.1%, respectively) was purified from sun ginseng (heat processed Panax ginseng). Cytotoxicity was induced by treatment of $20{\mu}M$ cisplatin to LLC-PK1 cells and rat model of acute renal failure was generated by single intraperitoneal injection of 5 mg/kg cisplatin. Protective effects were assessed by determining cell viability, reactive oxygen species generation, blood urea nitrogen, serum creatinine, antioxidant enzyme activity, and histopathological examination. Results: The in vitro assay demonstrated that KG-KH ($50{\mu}g/mL$) significantly increased cell viability (4.6-fold), superoxide dismutase activity (2.8-fold), and glutathione reductase activity (1.5-fold), but reduced reactive oxygen species generation (56%) compared to cisplatin control cells. KG-KH (6 mg/kg, per os) also significantly inhibited renal edema (87% kidney index) and dysfunction (71.4% blood urea nitrogen, 67.4% creatinine) compared to cisplatin control rats. Of note, KG-KH significantly recovered the kidney levels of catalase (1.2-fold) and superoxide dismutase (1.5-fold). Conclusion: Considering the oxidative injury as an early trigger of cisplatin nephrotoxicity, our findings suggest that ginsenosides Rk3 and Rh4 protect the kidney from cisplatin-induced oxidative injury and help to recover renal function by restoring intrinsic antioxidant defenses.

The Butanol Fraction of Bitter Melon (Momordica charantia) Scavenges Free Radicals and Attenuates Oxidative Stress

  • Kim, Hyun Young;Sin, Seung Mi;Lee, Sanghyun;Cho, Kye Man;Cho, Eun Ju
    • Preventive Nutrition and Food Science
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    • v.18 no.1
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    • pp.18-22
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    • 2013
  • To investigate radical scavenging effects and protective activities of bitter melon (Momordica charantia) against oxidative stress, in vitro and a cellular system using LLC-$PK_1$ renal epithelial cells were used in this study. The butanol (BuOH) fraction of bitter melon scavenged 63.4% and 87.1% of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals at concentrations of 250 and $500{\mu}g/mL$, respectively. In addition, the BuOH fraction of bitter melon effectively scavenged hydroxyl radicals (${\cdot}OH$). At all concentrations tested, the scavenging activity of the BuOH fraction was more potent than that of the positive control, ascorbic acid. Furthermore, under the LLC-$PK_1$ cellular model, the cells showed a decline in viability and an increase in lipid peroxidation through oxidative stress induced by pyrogallol, a generator of superoxide anion ($O_2{^-}$). However, the BuOH fraction of bitter melon significantly and dose-dependently inhibited cytotoxicity. In addition, 3-morpholinosydnonimine (SIN-1), a generator of peroxynitrite ($ONOO^-$) formed by simultaneous releases of nitric oxide and $O_2{^-}$, caused cytotoxicity in the LLC-$PK_1$ cells while the BuOH fraction of bitter melon ameliorated oxidative damage induced by $ONOO^-$. These results indicate that BuOH fraction of bitter melon has protective activities against oxidative damage induced by free radicals.

Phloroglucinol Attenuates Free Radical-induced Oxidative Stress

  • So, Mi Jung;Cho, Eun Ju
    • 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.

Effects of Prunellae Herba on the H2O2-Treated LLC-PK Cell's Redox Status and NF-κB Signaling (H2O2 처리된 LLC-PK₁세포에서 Redox Status 및 NF-κB Signaling에 대한 하고초(夏枯草)의 효과)

  • Son, Jong Seok;Jeong, Ji Cheon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.30 no.4
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    • pp.242-249
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    • 2016
  • This study was to investigate the anti-inflammatory effects of Prunellae Herba(PH). The generation of superoxide anion radical (․O2-), nitric oxide (NO), peroxynitrite (ONOO-) and Prostaglandin E₂(PGE2) were measured in the H2O2-Treated renal epithelial cells(LLC-PK1 cell) of mouse. And the effects of Prunellae Spica on the expression of NF-κB (p50, p65), IKK-α, phospho-IκB-α and inflammation-related proteins, COX-2, iNOS, IL-1β and VCAM-1, were examined by western blot. The fluorescent probes, 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) and dihyldrorhodamine 123 (DHR 123) were used to estimate the scavenging effect of Prunellae Spica on ․O2-, NO, ONOO-. Western blot was conducted to assess the protein expression levels of NF-κB (p50, p65), IKK-α, phospho-IκB-α, inflammation-related proteins, COX-2, iNOS, IL-1β, VCAM-1. PH inhibited H2O2-treated cell death dose-dependently. It reduced the generation of ·O2-, NO, ONOO- and PGE₂ in the H2O2-treated renal epitheial cells(LLC-PK1 cell) of mouse in vitro. PH reduced the expression of NF-κB, IKK-α, phospho-IκB-α, COX-2, iNOS, IL-1β and VCAM-1 genes through means of decreasing activation of NF-κB signaling as well. According to these results, PH has an antioxidative activity and anti-inflammatory effect by regulating the NF-κB pathway. This suggest that PH is expected to be used to regulating inflammatory process and treating inflammation-related disease.