• Title, Summary, Keyword: LLC-PK1 cells

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Beneficial effects of fermented black ginseng and its ginsenoside 20(S)-Rg3 against cisplatin-induced nephrotoxicity in LLC-PK1 cells

  • Han, Myoung-Sik;Han, Im-Ho;Lee, Dahae;An, Jun Min;Kim, Su-Nam;Shin, Myoung-Sook;Yamabe, Noriko;Hwang, Gwi Seo;Yoo, Hye Hyun;Choi, Suk-Jung;Kang, Ki Sung;Jang, Hyuk-Jai
    • 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.

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.

Protective Effects of Phellinus linteus and Curry-Added Cooked Mixed Grain Rice Extracts on Oxidative Stress-Induced LLC-PK1 Cell Damage (상황버섯 및 카레를 첨가한 잡곡밥 추출물의 LLC-PK1 세포에서의 산화적 스트레스 보호 효과)

  • Lee, Jung-Sook;Song, Jia-Le;Kil, Jeung-Ha;Jeong, Byung-Jin;Jeong, Jong-Sung;Huh, Tae-Gon;Park, Kun-Young
    • 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.

Effect of Nephrotoxicants on $\alpha$-Methylglucose Uptake in LLC-$PK_1$ (LLC-$PK_1$을 이용한 신독성 물질들의 $\alpha$-methyl glucose uptake에 미치는 영향의 평가)

  • Seo, Kyung-Won;Kim, Hyo-Jung;Chung, Se-Young
    • Environmental Analysis Health and Toxicology
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    • v.9 no.1_2
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    • pp.25-35
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    • 1994
  • Many nephrotoxic agents exert their effect primarily on the cells of the proximal tubules. We used the LLC-$PK_1$, kidney epithelial cell line as a model system for studies on nephrotoxicity and investigated whether the uptake of $\alpha$-methylglucose($\alpha$-MG) could serve as a parameter to assess effects of nephrotoxicants on the functional integrity of the cells at an early time of toxicity. The enzyme leakage test which has been used to be as a conventional cytotoxic parameter in vitro, was conducted to compare with $\alpha$-MG uptake. Treatment with cisplatin for 24 and 48 hours significantly increased activities of lactate dehydrogenase and $\gamma$-glutamyltransferase in culture medium at a concentration of 50$\mu$M. However, above 100$\mu$M of concentration, activities of these enzymes in media were dramatically decreased by cisplatin. These observations indicate that cisplatin has direct inhibitory effect on the activities of these enzymes and make it doutful to use enzyme leakage test to demonstrate damage of kidney cells by chemicals such as cisplatin over the appropriate range of concentration. Cisplatin inhibited $\alpha$-MG uptake at a low concentration which enzymes were not leaked. Also cadmium chloride and mercuric chloride which are acutely nephrotoxic in vivo, significantly inhibited $\alpha$-MG uptake at a low concentration. These results indicate that the uptake of $\alpha$-methylglucose in LLC-$PK_1$cell line is a useful biomarker for the study of nephrotoxicity.

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Antioxidative Effects of Common and Organic Kale Juices (유기농 및 일반농 케일 착즙액의 항산화 활성)

  • Kim, Jong-Dai;Lee, Ok-Hwan;Lee, Jong Seok;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.5
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    • pp.668-674
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    • 2014
  • The objective of the present study was to investigate the protective and free radical scavenging effects of conventionally and organically cultivated kale juices against oxidative damage in $LLC-PK_1$ cells. The DPPH, NO, $O_2{^-}$, and ${\cdot}OH$ radical scavenging activities of organically cultivated kale were higher than those of conventionally cultivated kale juice. Oxidative damage induced by AAPH (2,2'-azobis(2-amidinopropane) dihydrochloride), SNP (sodium nitroprusside), pyrogallol, and SIN-1 (3-morpholinosydnonimine) led to loss of cell viability and increased lipid peroxidation in LLC-PK1 cells, whereas treatment with vegetable juices, especially organically cultivated kale juices, significantly increased cell viability and inhibited lipid peroxidation in a dose-dependent manner (P<0.05). These results suggest that organically cultivated kale juices have protective roles against oxidative stress induced by free radicals.

Protective effect of ginsenoside Rh3 against anticancer drug-induced apoptosis in LLC-PK1 kidney cells

  • Lee, Hye Lim;Kang, Ki Sung
    • Journal of Ginseng Research
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    • v.41 no.2
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    • pp.227-231
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    • 2017
  • Background: Ginsenosides are active components of Panax ginseng that exert various health benefits including kidney protection effect. The medicinal activity of ginsenosides can be enhanced by modulating their stereospecificity by heat processing. Ginsenosides Rk2 and Rh3 represent positional isomers of the double bond at C-20(21) or C-20(22). Methods: The present study investigated the kidney-protective effects of ginsenosides Rk2 and Rh3 against cisplatin, a platinum based anticancer drug, induced apoptotic damage in renal proximal LLC-PK1 cells. Results: As a result, ginsenoside Rh3 shows a stronger protective effect than that shown by Rk2. Cisplatin-induced elevated protein levels of phosphorylated c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), p38, and cleaved caspase-3 decreased after cotreatment with ginsenoside Rh3. The increase in the percentage of apoptotic LLC-PK1 cells induced by cisplatin treatment also significantly reduced after cotreatment with ginsenoside Rh3. Conclusion: These results demonstrate that inhibition of the JNK and ERK mitogen-activated protein kinase signaling cascade plays a critical role in mediating the renoprotective effect of ginsenoside Rh3.

The Effect of Patriniae Radix on the Oxidative Stress and the NF-${\kappa}B$ Signaling in Mouse LLC-$PK_1$ Cell (Mouse의 신장상피세포에서 패장(敗醬)추출물이 산화 스트레스 및 NF-${\kappa}B$ signaling에 미치는 영향)

  • Kim, Hyun-Young;Jang, Soo-Young;Choi, Gyu-Ho;Shin, Hyeon-Cheol
    • 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.

Antioxidative Effects of Delphinidin under in vitro and Cellular System

  • Noh, Jeong-Sook;Cho, Yun-Ju;Kim, Boh-Kyung;Park, Kun-Young;Cho, Eun-Ju
    • Food Science and Biotechnology
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    • v.18 no.1
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    • pp.167-171
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    • 2009
  • This study examined the antioxidative activity of delphinidin, a kind of anthocyanidin from eggplant. Cellular protective potential from oxidative damage by nitric oxide (NO), superoxide anion ($O_2^-$), and peroxynitrite ($ONOO^-$) using epithelial cell line LLC-PK1 cell as well as in vitro radical scavenging effects were investigated. Delphinidin showed strong in vitro radical scavenging effects against NO, $O_2^-$, and hydroxyl radical (${\cdot}OH$) in dose-dependent manners. In addition, delphinidin increased cell viability in LLC-PK1 cells in a concentration-dependent manner when viability was reduced by $ONOO^-$-induced oxidative damage. To elucidate the protective mechanisms of delphinidin from $ONOO^-$, sodium nitroprusside (SNP), and pyrogallol were also employed to generate NO and $O_2^-$, respectively. The treatment of delphinidin recovered reductions in cell viability caused by SNP and pyrogallol, indicating that delphinidin can attenuate oxidative stress induced by NO and $O_2^-$. The present study suggests that delphinidin is a promising anti oxidative agent.

In Vitro Cytotoxicity of Novel Platinum(II) Coordination Complexes Containing Diaminocyclohexane and Diphenylphosphines

  • Jung, Jee-Chang;Kim, Young-Kyu;Park, Seung-Joon;Chung, Joo-Ho;Chang, Sung-Goo;Lee, Kyung-Tae;Baek, Min-Son;Park, Jong-Jip;Rho, Young-Soo
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.3
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    • pp.395-401
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    • 1998
  • We have synthesized new platinum(II) analogs containing 1,2-diaminocyclohexane (dach) as a carrier ligand, 1,3-bis(diphenylphosphino) propane (DPPP) /1,2-bis(diphenylphosphino)ethane (DPPE) as a leaving group and nitrates to improve solubility. In the present study, the cytotoxicity of $[Pt(trans-l-dach)(DPPP)]\;2NO_3$ (KHPC-001) and $[Pt(trans-l-dach)(DPPE)]\;2NO_3$ (KHPC-002) was evaluated and compared on various P-388 cancer cell lines and porcine kidney cell line ($LLC-PK_1$). The new platinum complexes demonstrated high efficacy on P-388 mouse leukemia cell line as well as cisplatin-resistant (P-388/CDDP) and adriamycin-resistant (P-388/ADR) P-388 cell lines. The intracellular platinum content was measured by a flame atomic absorption spectrophotometer (FAAS), and it was comparable to the results of $IC_{50}$ of the three complexes on $LLC-PK_1$ and P-388/S cells, while only DPPE compound was accumulated in high volume in P-388/ADR and P-388/CDDP cells. While the DNA-interstrand cross-links of KHPC-001, KHPC-002 and cisplatin were similar on P-388/S leukemia cells, these new platinum complexes were much less DNA cross-linking to a kidney derived cell line, $LLC-PK_1$. These results indicate that KHPC-001 and KHPC-002 are a third-generation platinum complexes with potent antitumor activity and low nephrotoxicity.

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