• 대한한의학회지
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• 제31권3호
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• pp.55-65
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• 2010

Background: Nitric oxide (NO) is a reactive free radical gas and a messenger molecule. NO has many physiological functions, but excessive NO production induces neurotoxicity. Objective: The present study investigated whether the aqueous extract of Polygala tenuifolia Willdenow possesses a protective effect on NO-induced apoptosis in human neuroblastoma cell line SK-N-MC. Method: For this study, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, 4,6-diamidino-2-phenylindole (DAPI) staining, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and caspase-3 enzyme assay were performed. Result: Sodium nitroprusside (SNP) exposure significantly decreased the viability of cells. The cells treated with SNP exhibited several apoptotic features such as increasing of Bax expression, caspase-3 enzyme activity and inhibiting of Bcl-2 expression. On the other hand, the viability of cells pre-treated with the aqueous extract of Polygala tenuifolia Willdenow was increased dose-dependently. The cells pre-treated for 1 h with the aqueous extract of Polygala tenuifolia Willdenow followed by treatment with SNP showed a decreased occurrence of apoptotic features like decreasing Bax expressions, caspase-3 enzyme activity and increasing Bcl-2 expressions. The aqueous extract of Polygala tenuifolia Willdenow reduced apoptotic cell death in neuroblastoma cell line SK-N-MC through the inhibition of Bax-dependent caspase-3 activation and the increasing of Bcl-2 expression. Conclusion: Based on the present results, it is possible that Polygala tenuifolia Willdenow has therapeutic value for the treatment of a variety of NO-induced brain diseases.

• 약학회지
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• 제53권1호
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• pp.41-44
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• 2009

A series of 2-alkyl-2,3-dihydro-1H-2-azacyclopenta[b]anthracene-5,10-diones ($3a{\sim}h$) were synthesized and evaluated in vitro cytotoxicity against colon cancer cell lines (HCT116 and SW620) and nuroblastoma cell lines (SK-N-SH and SK-N-MC). Among them, compound 3f showed significant cytotoxic activity ($IC_{50}$ against SK-N-SH; $14.8{\mu}M$, $IC_{50}$ against SK-N-MC; $11.3{\mu}M$).

• 대한한의학회지
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• 제22권1호
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• pp.90-95
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• 2001

목적：인진 약침액이 SK-N-MC 신경아세포주에서 에탄올에 의해 유발된 아폽토시스에 미치는 영향을 조사하였다. 방법 : SK-N-MC cell line에서의 아폽토시스 변화를 관찰하기 위해서 MTT assay, DAPI staining 및 flow cytometric analysis 방법을 이용하였다. 결과： MTT assay를 이용하여 분석한 결과 농도에 따른 세포 독성의 효과가 에탄올 투여로부터 관찰되었다. 또한 인진 약침액으로 전처치하고 에탄올을 처치하였을 때 세포 독성이 크게 감소되었다. DAPI staining에서 인진 약침액 투여군은 에탄올 투여군에 비해서 fragmentation이 억제되었다. Flow cytometry를 통하여 인진 약침액 투여군은 에탄올 투여군에 비하여 세포주기 중 sub $G_1$ 분획의 증가가 억제되었다. 결론 : SK-N-MC 신경아세포주에서 에탄올에 의해서 유발된 아폽토시스는 전형적인 세포사별 형태를 나타내었다. 또한 인진 약침액은 에탄올에 의해서 유발된 아폽토시스에서 세포보호 효과가 있음이 확인되었다.

• 대한한의학회지
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• 제26권4호
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• pp.74-86
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• 2005

Background: There are increasing neuro-degenerative disorders with aging. Paeoniae Radix(PR) possesses various pharmacological effects such as sedative, analgesic, anti-inflammatory, anti-stress and neuro-protective actions. Also antiaging and anti-cancer effects of PR were reported. Our purpose was to investigate whether PR is useful on the treatment of Parkinson's disease, one of the neuro-degenerative disorders. Objective: We investigated whether Paeonia Radix possesses a protective effect against 1-methyl-4 phenylpyridine(MPP+)-induced cytotoxicity in neuronal cells. Methods: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay, flow cytometry, DNA fragmentation assay, reverse transcription-polymerase chain reaction(RT-PCR), and Western blotting were performed on SK-N-MC neuroblastoma cells. Results: Cells treated with MPP+ exhibited several apoptotic features, while cells pre-treated with Paeonia Radix prior to MPP+ exposure showed s decrease in the occurrence of apoptotic features. Conclusions: These results suggest that Paeonia Radix may exert a protective effect against MPP+-induced apoptosis in SK-N-MC neuroblastoma cells.

• 동의생리병리학회지
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• 제16권3호
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• pp.602-608
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• 2002

To investigate whether Puerariae radix (PR) possesses protective effect against ethanol (EtOH)-inducecl apoptosis in the central nervous system, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, flow cytometric analysis, DNA fragmentation assay. and reverse transcription-polymerase chain reaction (RT-PCR) were performed on human neuroblastoma cell line SK-N-MC. Morphological and biochemical analyses demonstrated that SK-N-MC cells treated with EtOH exhibit classical apoptotic features. On the other hand, cells pre-treated with PR prior to EtOH exposure showed decreased occurrence of classical apoptotic features. In addition, it was shown that PR pre-treatment inhibits EtOH-induced increases in the levels of mRNA expression of bax and caspase-3, while it further enhances the level of bcl-2 expression. These results suggest that PR may exert protective effects against EtOH-induced apoptosis in human neuroblastoma cells.

• Biomolecules & Therapeutics
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• 제10권3호
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• pp.142-151
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• 2002

The present study was attempted to investigate the effect of apamin on catecholamine (CA) secretion evoked by ACh, high $K^+$, DMPP, McN-A-343, cyclopiazonic acid and Bay-K-8644 from the isolated perfused rat adrenal gland and to establish the mechanism of its action. The perfusion of apamin (1 nM) into an adrenal vein for 20 min produced greatly potentiation in CA secretion evoked by ACh (5.32 $ imes$ $10^{-3}$ M), high $K^+$, (5.6 $ imes$ $10^{-2}$), DMPP ($10^{-4}$ M for 2 min), McN-A-343 ($10^{-4}$ M for 2 min), cyclopiazonic acid ($10^{-5}$ M for 4 min) and Bay-K-8644 ($10^{-5}$ M for 4 min). However, apamin itself did fail to affect basal catecholamine output. Furthermore, in adrenal glands preloaded with apamin (1 nM) under the presence of glibenclamide ($10^{-6}$ M), an antidiabetic sulfonylurea that has been shown to be a specific blocker of ATP-regulated potassium channels (for 20 min), CA secretion evoked by DMPP and McN-A-343 was not affected. However, the perfusion of high concentration of apamin (100 nM) into an adrenal vein for 20 min rather inhibited significantly CA secretory responses evoked by ACh, high $K^+$, DMPP, McN-A-343, cyclopiazonic acid and Bay-K-8644. Taken together, these results suggest that the low concentration of apamin causes greatly the enhancement of CA secretion evoked by stimulation of cholinergic (both nicotinic and muscarinic) receptors as well as by membrane depolarization. These findings suggests that apamin-sensitive SK ($Ca^{2+}$) channels located in rat adrenal medullary chromaffin cells may play an inhibitory role in the release of catecholamines mediated by stimulation of cholinergic nicotinic and muscarinic receptors as well as membrane depolarization. However, it is thought that high concentration of apamin cause the inhibitory responses in catecholamine secretion evoked by stimulation of cholinergic receptors as well as by membrane depolarization from the rat adrenal gland without relevance with the SK channel blockade.

• 생약학회지
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• 제27권4호
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• pp.350-353
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• 1996

The effects of acid hydrolysis product of Panax ginseng and MeOH extract of Euphorbia humifusa on the growth of human brain tumor cells were evaluated using U-373 MG human astrocytoma and SK-N-MC human neuroblastoma cells as model cellular systems. These plant extracts induced cytotoxicity in both cells in a dose-dependent manner. These cytotoxic effects were significantly inhibited by GSH, an antioxidant, in both cells. BAPTA/AM, an intracellular $Ca^{2+}$ chelator, significantly blocked the cytotoxic effects of these extracts in U-373 cells, but enhanced these effects in SK-N-MC cells. These results suggest that the plant extracts may be a valuable choice for the studies on the treatment of human brain tumors.

• Biomolecules & Therapeutics
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• 제19권3호
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• pp.288-295
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• 2011

Amyloid beta ($A{\beta}$)-induced neurotoxicity is a major pathological mechanism of Alzheimer's disease (AD). In this study, we investigated the inhibitory effect of L-theanine, a component of green tea (Camellia sinensis) on $A{\beta}_{1-42}$-induced neurotoxicity and oxidative damages of macromolecules. L-theanine inhibited $A{\beta}_{1-42}$-induced generation of reactive oxygen species, and activation of extracellular signal-regulated kinase and p38 mitogenic activated protein kinase as well as the activity of nuclear factor kappa-B. L-theanine also signifi cantly reduced oxidative protein and lipid damage, and elevated glutathione level. Consistent with the reduced neurotoxic signals, L-theanine (10-50 ${\mu}g$/ml) concomitantly attenuated $A{\beta}_{1-42}$ (5 ${\mu}M$)-induced neurotoxicity in SK-N-MC and SK-N-SH human neuroblastoma cells. These data indicate that L-theanine on $A{\beta}$-induced neurotoxicity prevented oxidative damages of neuronal cells, and may be useful in the prevention and treatment of neurodegenerative disease like AD.

• Journal of Acupuncture Research
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• 제18권3호
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• pp.134-142
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• 2001

목적 : 인진(茵蔯)이 SK-N-MC 신경아세포주에서 에탄올에 의해 유발된 아폽토시스에 대한 보호작용의 기전을 연구하였다. 방법 : SK-N-MC cell line에서의 세포 보호 기전을 알아보기 위하여 reverse transcription polymerase chain reaction (RT-PCR) 기법을 이용하여 bcl-2, bax 및 caspase-3의 변화를 관찰하였다. 결과 : RT-PCR을 이용하여 분석한 결과 SK-N-MC neuroblastoma에서 에탄올 처치는 bax, bcl-2 및 caspase-3 mRNA의 발현을 증가시켰다. 인진액의 전처치후 에탄올 처치한 신경 아세포에서는 에탄올에 의해서 증가된 bax와 caspase-3 mRNA 발현이 억제되었으나, bcl-2의 발현에는 유의한 증가를 나타내지 않았다. 결론 : 이상의 결과를 통하여 에탄올에 의해서 유발된 신경아세포의 아폽토시스에서 인진이 세포보호 효과가 있음이 확인되었고 그 기전은 bax와 caspase-3의 억제에 기인할 가능성을 시사한다고 할 수 있다.

• Nutrition Research and Practice
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• 제14권1호
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• pp.3-11
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• 2020

BACKGROUND/OBJECTIVES: Oxidative stress causes cell damage and death, which contribute to the pathogenesis of neurodegenerative diseases. Urolithin A (UA), a gut microbial-derived metabolite of ellagitannins and ellagic acid, has high bioavailability and various health benefits such as antioxidant and anti-inflammatory effects. However, it is unknown whether it has protective effects against oxidative stress-induced cell death. We investigated whether UA ameliorates H₂O₂-induced neuronal cell death. MATERIALS/METHODS: We induced oxidative damage with 300 μM H₂O₂ after UA pretreatment at concentrations of 1.25, 2.5, and 5 μM in SK-N-MC cells. Cytotoxicity and cell viability were determined using the CCK-8 assay. The formation of reactive oxygen species (ROS) was measured using a 2,7-dichlorofluorescein diacetate assay. Hoechst 33342 staining was used to characterize morphological changes in apoptotic cells. The expressions of apoptosis proteins were measured using Western blotting. RESULTS: UA significantly increased cell viability and decreased intracellular ROS production in a dose-dependent manner in SK-N-MC cells. It also decreased the Bax/Bcl-2 ratio and the expressions of cytochrome c, cleaved caspase-9, cleaved caspase-3, and cleaved PARP. In addition, it suppressed the phosphorylation of the p38 mitogen-activated protein kinase (MAPK) pathway. CONCLUSIONS: UA attenuates oxidative stress-induced apoptosis via inhibiting the mitochondrial-related apoptosis pathway and modulating the p38 MAPK pathway, suggesting that it may be an effective neuroprotective agent.