• 제목/요약/키워드: LLC-PK1 cells

검색결과 51건 처리시간 0.024초

머위 에틸아세테이트 분획물의 LLC-PK1 세포에서의 Nrf-2 매개 항산화 효과 (Ethyl Acetate Fraction from Petasites japonicus Attenuates Oxidative Stress through Regulation of Nuclear Factor E2-Related Factor-2 Signal Pathway in LLC-PK1 Cells)

  • 김지현;이재민;이상현;조은주
    • 생약학회지
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    • 제47권1호
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    • pp.55-61
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    • 2016
  • Antioxidant effects and nuclear factor E2-related factor-2 (Nrf-2) signal pathway of methanol extract and 4 fractions [n-hexane, methylene chloride, ethyl acetate (EtOAc), and n-butanol fractions] from Petasites japonicus were investigated. The EtOAc fraction showed highest polyphenol and flavonoid contents among other fractions. In addition, EtOAc fraction showed stronger scavenging activity against superoxide anion radical than other fractions. Furthermore, we investigated antioxidants effects of the EtOAc fraction under cellular system using $LLC-PK_1$ cells. The EtOAc fraction dose-dependently increased the antioxidant protein expressions of heme oxygenase 1 (HO-1) and thioredoxin reductase 1 (TrxR1) known to be involved in oxidative stress, through activation of Nrf-2. The treatment of EtOAc fraction ($100{\mu}g/mL$) led to the elevation of the high expression of Nrf-2-dependent factor such as HO-1 and TrxR1. These results indicated that the EtOAc fraction of P. japonicus showed high antioxidant activity by regulation of Nrf-2 signaling pathway.

Phloroglucinol Attenuates Free Radical-induced Oxidative Stress

  • So, Mi Jung;Cho, Eun Ju
    • Preventive Nutrition and Food Science
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    • 제19권3호
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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.

Protective Effect of Chungkukjang from Sunchang Province against Cellular Oxidative Damage

  • Choi, Ji-Myung;Yi, Na-Ri;Seo, Kyoung-Chun;Han, Ji-Sook;Song, Young-Ok;Cho, Eun-Ju
    • Preventive Nutrition and Food Science
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    • 제13권2호
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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.

지골피(地骨皮)가 $H_{2}O_{2}$에 의한 $LLC-PK_1$ 세포의 Redox Status 및 $NF-{\kappa}B$ Signaling에 미치는 영향 (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)

  • 최규호;신현철
    • 대한한방내과학회지
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    • 제30권1호
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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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순무 (Brassica rapa) 뿌리 에탄올 추출물의 Cisplatin 유도에 의한 LLC-$PK_1$ 돼지 신장 세포주 및 흰쥐에서 신독성 보호효과 (Protective Effect of the Ethanol Extract of the Roots of Brassica rapa on Cisplatin-Induced Nephrotoxicity in LLC-$PK_1$ Cells and Rats)

  • 이경태;조웅;김양희;김용원;백남인;정선아;정해곤;정태숙;최명숙
    • 한국약용작물학회:학술대회논문집
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    • 한국약용작물학회 2008년도 춘계학술발표회
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    • pp.190-191
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    • 2008
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시스플라틴에 의한 $LLC-PK_1$의 알파-메틸글루코스 흡수 감소 기전 (Mechanism of Inhibition of ${\alpha}$-Methylglucose Uptake by Cisplatin in $LLC-PK_1$)

  • 서경원;김효정;정세영
    • 약학회지
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    • 제40권6호
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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.

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

  • 손종석;정지천
    • 동의생리병리학회지
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    • 제30권4호
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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.

국내산 법제 하수오의 라디칼 소거능 및 산화적 스트레스 개선 효과 (Free Radical Scavenging Effect and Oxidative Stress Protective Activity of Domestic Processed Polygoni Multiflori Radix)

  • 김현영;김준영;조은주;최지명;황정은;이희율;안민주;이진환;김윤근;고건희;구영민;오경렬;조계만
    • 한국식품영양과학회지
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    • 제44권6호
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    • pp.809-815
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    • 2015
  • 본 연구에서는 국내산 법제 하수오 메탄올 추출물로부터 생리활성물질을 확인하였고, 추출물의 라디칼 소거능과 LLC-$PK_1$ cell을 이용한 산화적 스트레스 개선 효과를 살펴보았다. HPLC 분석 결과 4종의 생리활성물질인 2,3,5,4'-tetrahydroxystilbene 2-O-${\beta}$-D-glucoside, emodin, chrysophanol 및 rhein을 확인하였다. 특히 주요 화합물인 2,3, 5,4'-tetrahydroxystilbene 2-O-${\beta}$-D-glucoside는 115.02 mg/kg이었다. 법제 하수오 추출물의 DPPH, ABTS, hydroxyl 라디칼 소거능은 농도 의존적으로 증가하였다. 추출물의 $50{\mu}g/mL$ 처리 시 DPPH 라디칼 소거능은 48.4%, ABTS 라디칼 소거능은 57.9% 및 hydroxyl 라디칼 소거능은 81.2%로 나타내었다. 한편 LLC-$PK_1$ cell에서 각각의 NO, $O_2{^-}$, 및 $ONOO^-$ 생성물질인 pyrogallol, sodium nitroprusside(SNP) 및 morpholinosydnonimine(SIN-1) 처리에 의해 유도된 산화적 스트레스 상에서 세포 생존율은 감소하였다. 그러나 법제 하수오 메탄올 추출물은 농도 의존적으로 세포 독성을 저해하였다. 추출물의 $50{\mu}g/mL$ 처리 시 세포 생존율은 각각 82.1%(pyrogallol), 89.1%(SNP) 및 77.6%(SIN-1)였다.

The altered $Na^+,\;K^+$-pump activity following the fumonisin exposure to LLC-PKl cells

  • Choi, Heon-Kyo;Yoo, Jae-Myung;Tudev, Munkhtsetseg;Lee, Yong-Moon;Yun, Yeo-Pyo;Yoo, Hwan-Soo
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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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)

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Blockade of P-Glycoprotein Decreased the Disposition of Phenformin and Increased Plasma Lactate Level

  • Choi, Min-Koo;Song, Im-Sook
    • Biomolecules & Therapeutics
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    • 제24권2호
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    • pp.199-205
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    • 2016
  • This study aimed to investigate the in vivo relevance of P-glycoprotein (P-gp) in the pharmacokinetics and adverse effect of phenformin. To investigate the involvement of P-gp in the transport of phenformin, a bi-directional transport of phenformin was carried out in LLC-PK1 cells overexpressing P-gp, LLC-PK1-Pgp. Basal to apical transport of phenformin was 3.9-fold greater than apical to basal transport and became saturated with increasing phenformin concentration ($2-75{\mu}M$) in LLC-PK1-Pgp, suggesting the involvement of P-gp in phenformin transport. Intrinsic clearance mediated by P-gp was $1.9{\mu}L/min$ while passive diffusion clearance was $0.31{\mu}L/min$. Thus, P-gp contributed more to phenformin transport than passive diffusion. To investigate the contribution of P-gp on the pharmacokinetics and adverse effect of phenformin, the effects of verapamil, a P-gp inhibitor, on the pharmacokinetics of phenformin were also examined in rats. The plasma concentrations of phenformin were increased following oral administration of phenformin and intravenous verapamil infusion compared with those administerd phenformin alone. Pharmacokinetic parameters such as $C_{max}$ and AUC of phenformin increased and CL/F and Vss/F decreased as a consequence of verapamil treatment. These results suggested that P-gp blockade by verapamil may decrease the phenformin disposition and increase plasma phenformin concentrations. P-gp inhibition by verapamil treatment also increased plasma lactate concentration, which is a crucial adverse event of phenformin. In conclusion, P-gp may play an important role in phenformin transport process and, therefore, contribute to the modulation of pharmacokinetics of phenformin and onset of plasma lactate level.