• Korean Journal of Pharmacognosy
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• v.47 no.1
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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.

• 한국약용작물학회:학술대회논문집
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• 2008.05a
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• pp.190-191
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• 2008

• 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.

• Biomolecules & Therapeutics
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• v.24 no.2
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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.

• 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.

• 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.

• 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.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.9 no.6
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• pp.1787-1794
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• 2008

Coenzyme $Q_{10}$ and six derivatives of coenzyme $Q_n$ were synthesized and tested for their antioxidative effects occurred in proximal tubular epithelial cell (LLC-PK1 cell) and cytotoxicities using in NIH/3T3 cell. As the result, synthetic coenzyme $Q_n$ derivatives showed a potent antioxidative effect compared to coenzyme $Q_{10}$. Among these synthetic compounds, coenzyme $Q_3$-C at ranged 0.04 to 0.4 mmol showed the $107.7{\sim}135.9%$ of cell viability in LLC-PK1 cell. In the test of NIH/3T3, all synthesized coenzyme $Q_n$ derivatives showed the similar effect compared with coenzyme $Q_{10}$. A correlation between isoprene unit number of coenzyme $Q_n$ derivatives and its biological effects, we suggest reduction of isoprene unit number of $Q_n$ derivatives may be related to the increase of antioxidants effects and the reduction of cytotoxicities.

• 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.

• Natural Product Sciences
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• v.18 no.4
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• pp.261-265
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• 2012

Xanthohumol was isolated from hops (Humulus lupulus L.), and then investigated anti-oxidant effect by AAPH-induced LLC-PK1 cell and oxygen radical absorbance capacity (ORCA) assays and MMP-12 inhibitory effect by direct MMP-12 inhibition assay. The treatment of xanthohumol protected LLC-PK1 cells from AAPH-induced cell damage such as cell viability, SOD and GSH-px reduction in a dose dependant manner (0.1, 1, and $5{\mu}M$), the SOD value was 2.98, 4.51, and 5.77 U/mg protein, and GSH-px value was 30.12, 49.32, and 60.11 U/mg protein. ORAC value of xanthohumol was showed as 4320, 12004, and $14209{\mu}M$ TE/g at the concentration 0.1, 1, and $5{\mu}M$, respectively. The change of SOD and GSH-px values was significantly correlated with the results of ORAC assay, that is, AAPH-induced cell and ORCA assays. In addition, inhibition of MMP-12 that is known to play an important role in skin aging was 14%, 37%, 46%, and 79% at the concentration of 0.01, 0.1, 1, and $5{\mu}M$, respectively. On the basis of these results, xanthohumol from hops (Humulus lupulus L.) showed interesting biological and pharmacological activity such as anti-oxidant effect and anti-aging.