• 한국식품영양과학회지
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• 제43권5호
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• pp.668-674
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• 2014

채소즙이란 생채소를 마쇄하여 인체가 영양소를 흡수하기 쉬운 상태로 제조된 즙으로 생채소의 영양 섭취 효율을 높일 수 있는 식품이라 할 수 있다. 본 연구에서는 재배방법에 따라 생산된 유기농 및 일반농 케일을 착즙하여 다양한 실험을 통하여 항산화 활성을 비교하였다. DPPH radical, NO, $O_2{^-}$, ${\cdot}OH$ 라디칼 소거능에서 유기농 케일 착즙액은 일반농케일 착즙액보다 더 우수한 효과를 나타내었다. LLC-PK1 세포를 이용하여 산화적 스트레스 개선효과에서 유기농 케일 착즙액은 일반농 케일 착즙액에서보다 NO, $O_2{^-}$와 $ONOO^-$에 의해 유발된 산화적 스트레스에 대한 세포 생존율을 증가시키고, 지질과산화물을 억제시켜 유리라디칼에 대한 보호효과를 유의적으로 나타내었다. 위의 결과로 보아 유기농케일 착즙액은 일반농 케일 착즙액보다 산화적 스트레스에 대한 개선효과가 뛰어난 것으로 사료되며 재배방법의 차이에 따라 그 효과도 차이가 있었다.

• Food Science and Biotechnology
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• 제18권1호
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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.

• Toxicological Research
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• 제11권1호
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• pp.1-8
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• 1995

Fumonisins are a family of mycotoxins produced by the fungus Fusarium moniliforme which is a common contaminant in corn. Fumonisins are potent inhibitors of sphingosine and sphinganine N-acyltransferase (ceramide synthase), key enzymes in sphingolipid metabolism. The purpose of this study was to provide the evidence that the elevated levels of free sphingoid bases (primarily sphinganine) and depletion of complex sphingolipids were closely related to the inhibition of cell growth in LLC-$PK_1$ cells exposed to fumonisin $B_1$$(\leq 35 {\mu}M)$. Concentrations of fumonisin $B_1$ between 10 and $35 {\mu}M$ were known to inhibit cell growth without cytotoxicity in $LLC-PK_1$ cells (Yoo et al. Toxicol. Appl. Pharmacol. 114, 9-15, 1992). Cells exposed to 35$\mu M$ fumonisin B$_1$ for 48 and 72 hr developed a fibroblast-like (elongated and spindle-shaped) appearance and were less confluent than normal cells. At between 24 and 48 hr after exposure to fumonisin $B_1$ cells were beginning to show the inhibition of cell growth and at 72 hr the number of viable cells in fumonisin-treated cultures was about 50% of concurrent control cultures. During the 24 hr lag period preceding inhibition of cell growth, the free sphinganine levels in cells exposed to $35 {\mu}M$ fumonisin $B_1$ were highly elevated (approximately 230 fold higher than normal cells). The elevated levels of free sphinganine were $435\pm14$$pmoles/{10^6}$ cells at 48 hr and approximately TEX>$333\pm11$$pmoles/{10^6}$ cells in cells exposed to $35{\mu}M$ fumonisin$B_1$ at 72 hr, while the levels of free sphinganine in normal cells were less than 2$pmoles/{10^6}$ cells. Under the same condition, depletion of intracellular complex sphingolipids as a consequence of fumonisin inhibition of de novo sphingolipid biosynthesis and turnover pathway was appeared. Content of free sphingold bases in dividing cells was more elevated than in confluent cells at 24-48 hr after cells were exposed to $20{\mu}M$ fumonisin $B_1$. The dividing cells were showing the inhibition of cell growth at 48-72 hr and $20{\mu}M$ fumonisin $B_1$. The results of this study support the hypothesis that the inhibition of cell growth is very well related to the disruption of sphingolipid metabolism in $LLC-PK_1$ cells.

• Archives of Pharmacal Research
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• 제24권2호
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• pp.136-143
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• 2001

Fumonisins are specific inhibitors of ceramide synthase in sphingolipid metabolism. An alteration in sphingolipid metabolism as a result of fumonisin exposure is related to cell death (Yoo et al., 1992). The objective of this study was to investigate whether elevated free sphinganine levels are related to the sensitivity of cultured cells to fumonisin exposure. Fumonisin $B_1$ elevated the intracellular free sphinganine concentraions in both LLC-$PK_1$ and Chinese hamster ovary (CHO) cells. However, CHO cells are resistant to fumonisin cytotoxicity at 50${u}m$, while LLC-$PK_1$ cells are sensitive at concentrations greater than 357M. The intracellular concentration of free sphinganine in LLC-$PK_1$ cells treated at 50${u}m$ fumonisin $B_1$ for 72 h was approximately 1450 pmol/mg protein relative to the 37 pmol observed in the control culture. Under the same conditions, the population of apoptotic cells in the 50${u}m$ fumonisin $B_1$-treated culture was approximately 37% of the total compared to 12% in the control. The caspase III-like activity after 72 h in the 50${\mu}$M fumonisin $B_1$-exposed culture Increased to approximately 50 $pmol/mg$ protein/hr compared to 6 $pmol/mg$ protein/hr in the control. L-cycloserine, a serine palmitoyltransferase inhibitory reduced the fumonisin $B_1$-stimulated caspase III-like activity down to the control level. Under the same culture conditions, the intracellular concentration of free sphinganine after-cycloserine plus fumonisin $B_1$ treatment was 140 pmol/mg protein compared to 1450 $pmol/mg$ protein in fumonisin $B_1$ alone. The intracellular concentration of free sphinganine in CHO cells treated with 50${u}m$ fumonisin $B_1$ for 72 h was al)proximately 460 pmol/mg protein, indicating that the mass amount of elevated free sphinganine in the CHO cells was about 32% of that in LLC-$PK_1$ cells. Adding exogenous sphinganine to the CHO cells along with 50${u}m$ fumonisin $B_1$ treatment for 72 h caused both necrosis and apoptosis. In conclusion, the elevated endogenous sphinganine acts as a contributing factor to the fumonisin-induced cell death.

• Journal of Ginseng Research
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• 제41권2호
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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.

• 대한한방내과학회지
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• 제31권1호
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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 Ginseng Research
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• 제41권3호
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• pp.284-289
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• 2017

Background: Compound FK506 is an immunosuppressant agent that is frequently used to prevent rejection of solid organs upon transplant. However, nephrotoxicity due to apoptosis and inflammatory response mediated by FK506 limit its usefulness. In this study, the protective effect of Korean Red Ginseng (KRG) against FK506-induced damage in LLC-PK1 pig kidney epithelial cells was investigated. Methods: LLC-PK1 cells were exposed to FK506 with KRG and cell viability was measured. Western blotting and RT-PCR analyses evaluated protein expression of MAPKs, caspase-3, and KIM-1. TLR-4 gene expression was assessed. Caspase-3 activities were also determined. The number of apoptotic cells was measured using an image-based cytometric assay. Results: The reduction in LLC-PK1 cell viability by $60{\mu}M$ FK506 was recovered by KRG cotreatment in a dose-dependent manner. The phosphorylation of p38, p44/42 MAPKs (ERK), KIM-1, cleaved caspase-3, and TLR-4 mRNA expression was increased markedly in LLC-PK1 cells treated with $60{\mu}M$ FK506. However, with the exception of p-ERK, elevated levels of p-p38, KIM-1, cleaved caspase-3, and TLR-4 mRNA expression were significantly decreased after cotreatment with KRG. Activity level of caspase-3 was also attenuated by KRG cotreatment. Moreover, image-based cytometric assay showed that apoptotic cell death was increased by $60{\mu}M$ FK506 treatment, whereas it was decreased after cotreatment with KRG. Conclusion: Taken together, these results suggest that the molecular mechanism of KRG in the FK506-induced nephrotoxicity may lead to the development of an adjuvant for the inhibition of adverse effect FK506 in the kidney.

• 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.

• 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.

• 대한약학회:학술대회논문집
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• 대한약학회 2000년도 NEW STRATEGY FOR DRUG DEVELOPMENT IN POST-GENOMIC ERA(대한약학회)
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• pp.206.1-206.1
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• 2000