• 동의신경정신과학회지
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• 제21권2호
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• pp.29-44
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• 2010

Objectives : The purpose of this study was to investigate the effect of Gentianae Radix on neurogenesis and apoptosis in ethanol- induced newborn rats hippocampus dentate gyrus. Methods : In vivo, laboratory animals were divided into three groups; Normal group(N), Control group(C) and Treated group (TG)(n=7 for each group). N were treated saline daily for five days. C were treated 1.5 g/kg ethanol and saline daily for five days. TG were treated 1.5 g/kg ethanol and 300 mg/kg Gentianae Radix daily for five days. BrdU(5-bromo-2-deoxyuridine) assay was used to test neurogenesis in the dentate gyrus. And TUNEL(Terminal deoxynucleotidyl transferase dUTP nick end labeling) assay was used to test apoptosis in the dentate gyrus. Three groups were measured body weight, serum ethanol concentration, BrdU-positive cells and TUNEL-positive cells in the dentate gyrus. In vitro, MTT(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used to test viability in SK-N-MC cells. BrdU assay was used to test neurogenesis in SK-N-MC cells. DNA fragmentation and caspase-3 enzyme activity assay were used to test apoptosis in SK-N-MC cells. And treated ethanol and Gentianae Radix of all in vitro tests were made various concentration. Results : In vivo, Gentianae Radix modulated ethanol-induced neurogenesis and apoptosis in newborn rats hippocampus dentate gyrus. In vitro, TG 100 ${\mu}g/ml$ have significantly modulated ethanol-induced neurogenesis and apoptosis in SK-N-MC cells. And only TG 100 ${\mu}g/ml$ have significantly protected SK-N-MC cells from ethanol-induced cytotoxicity. Conclusions : Gentianae Radix may have the effect that modulated ethanol-induced neurogenesis and apoptosis in SK-N-MC cells.

• 동의생리병리학회지
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• 제16권1호
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• pp.172-180
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• 2002

Polychlorinated biphenyls(PCBs) are large scale industrial chemicals which are using in diverse applications. The goal of this study was to determine if exposure to 2,2',5,5'-tetrachlorobiphenyl (PCB 52) leads to an increase in the production of active oxidants, and subsequently promotes apoptosis of neuronal SK-N-MC cells. Reactive oxygen species (ROS) formation was examined in SK-N-MC cells after treatment of PCB 52 by concentrations and incubation times, respectively. It showed that the rate of ROS production in the cells was increased in a does-dependent manner to 45 min, followed by a return towards control levels after 120 min treatment. We also examined the association of PCB-induced apoptosis with the modulation of biomakers of oxidative damage to lipids (malondialdehyde [MDA]) in SK-N-MC cells. Increased MDA was observed in a dose-dependent manner in groups treated with 10, 15, and 20 figJ me of PCB 52 for 24 h. After treatment of PCB 52, the cells did not show any significant change in the rate of Cu/Zn-superoxide dismutase (Cu/Zn-SOD) activity. Whereas, the cells had a two-fold greater rate of change in catalase activity at 20 ㎍/㎖ of PCB 52 for 24 h when compared to control group. Korean Ginseng is one of the most important crude drugs which has been used as a traditional Oriental medicine. We next investigated protective effect of extracts of ginseng on cytotoxicity induced by PCB 52 in SK-N-MC cells. Pretreatment of SK-N-MC cells with 25-200 μg/ml of ginseng were reduced cell death in a dose-dependent manner in PCB 52-treated cells. To examine the sensitivity of beta-catenin to ginseng, the protective effect of a range of ginseng concentrations was examined in SK-N-MC cells treated with PCB 52. The result demonstrated that ginseng efficiently blocked PCB 52 inducible beta-catenin proteolysis in a concentration dependent manner. The ROS formation was also measured in the presences of extract of ginseng and superoxide dismutase (inhibitor of oxygen free radical production). The both SOD (400 U/ml) and ginseng (200 μg/ml) significantly inhibited RDS generation in PCB 52-treated group.

• The Korean Journal of Physiology and Pharmacology
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• 제13권3호
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• pp.139-145
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• 2009

The purpose of this study was to evaluate the effects of the water extract of Samultang (SMT), a Chinese herb, on apoptotic cell death by $H_2O_2$-induced oxidative stress in SK-N-M C cells. A nuclear fragmentation was observed via fluorescence imaging 12 h after exposure to 30 ${\mu}M$ $H_2O_2$ and DNA laddering was detected via agarose electrophoresis gel. In addition, increases in sub-G1 phase and cleavage of the PARP protein were observed. However, treatment with SMT for 2 h prior to $H_2O_2$ exposure significantly reduced apoptotic cell death induced by incubation with 30 ${\mu}M$ $H_2O_2$ in SK-N-MC cells. Pre-incubation with water extract of SMT for 2 h prevented the $H_2O_2$-induced decrease in mitochondrial transmembrane potential. SMT also attenuated the increase in caspase-3 activity and the breakdown of PARP protein caused by $H_2O_2$-induced oxidative stress. These results suggest that the water extract of SMT provides inhibition of apoptotic cell death against oxidative injury in SK-N-MC cells.

• 한국식품영양학회지
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• 제29권6호
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• pp.923-929
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• 2016

The purpose of this study was to evaluate the protective effect of $PineXol^{(R)}$ on $H_2O_2$-induced cell death in SK-N-MC cells, and in early stage focal ischemia rodent model. SK-N-MC cells were pre-treated with $200{\mu}M$ $H_2O_2$ or various concentrations of $PineXol^{(R)}$ (10, 30, and 50 pg/mL) for 24 h, and then exposed to $H_2O_2$ for 3 h. Cell death was assessed by the CCK-8 assay, reactive oxygen species (ROS) assay, and lactate and dehydrogenase (LDH) release assay. Superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) expressions were also analyzed by western blotting. Focal ischemia rodent model was used as the in vivo model, and different concentrations of $PineXol^{(R)}$ (1, 10, and 100 mg/kg) were administered. One week after administration, reduction of infarct volume was analyzed by TTC staining. Cell viability of $H_2O_2$-treated SK-N-MC cells significantly increased by pre-treatment of $PineXol^{(R)}$ (p<0.05). $PineXol^{(R)}$ pre-treatment also induced significant decrease of ROS and LDH expressions. However, $PineXol^{(R)}$ did not affect the infarct volume. These results suggest that $PineXol^{(R)}$ has significant neuroprotective effect in vitro, but statistical significance was not confirmed in the in vivo focal ischemia model.

• Journal of Acupuncture Research
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• 제25권2호
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• pp.41-57
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• 2008

목적 : 이 연구는 Vipera lebetina turanica의 사독약침액(蛇毒藥鍼液)(Snake venom toxin, SVT)이 인간 신경아세포종의 암세포주인 SK-N-MC 세포에서 암세포성장의 억제 및 그 기전에 대하여 살펴보고자 하였다. 방법 : SVT를 처리한 후 SK-N-MC의 성장억제를 관찰하기 위해 CCK-8 assay와 LDH assay를 시행하였고, apoptosis 평가에는 세포형태의 관찰과 DAPI, TUNEL, Annexin V-PI double staining assay 및 cell detachment assay를 시행하였다. 세포자멸사 관련 세포기전을 보기 위하여 세포주기, 세포내 칼슘량, 세포내 활성산소량 및 미토콘드리아의 세포막전위 변화를 측정하였고, DNA fragmentation assay를 시행하였으며, 세포자멸사 조절 단백인 Bax, Bcl-2, caspase-3, -9의 발현 변화 관찰에는 western blot analysis를 시행하였다. 결과 : SK-N-MC 세포에 SVT를 처리한 후, 신경아세포종 세포의 성장, Apoptosis의 유발 및 기전에 영향을 관찰하여 다음과 같은 결과를 얻었다. 1. SVT를 처리한 SK-N-MC 세포 관찰에서 $20{\mu}g/m{\ell}$ SVT 처리가 암세포성장의 유의한 억제를 나타내었다. 세포독성 관찰에서 SVT처리는 처리하지 않은 것에 비하여 증가를 나타내었다. 2. 세포자멸사 평가에서 SVT를 처리한 SK-N-MC 세포는 세포자멸사의 특징적 형태를 나타내었다. TUNEL assay에서는 세포자멸사 활성세포가 미약하게 나타난 반면 cell detachment assay와 Annexin V-PI double staining에서는 각각 세포박리와 세포자멸사 활성세포의 유의한 증가를 나타내었다. 3. 세포자멸사 관련 세포기전연구에서 SVT를 처리한 SK-N-MC 세포의 세포주기, 세포내 칼슘량 및 DNA fragmentation에는 유의한 변화가 관찰되지 않은 반면 세포내 활성산소 양은 유의한 증가를 나타내었고, 그에 따른 미토콘드리아 세포막 전위의 유의한 변동이 관찰되었다. 4. SVT를 처리한 SK-N-MC는 세포자멸사 관련 단백 발현에서 caspase-9에 대해 유의한 증가를 나타내지 않았으나 Bax 및 caspase-3의 유의한 증가와 Bcl-2의 유의한 감소를 나타내었다. 결론 : 이상의 결과는 SVT가 세포내 활성산소를 증가시키므로 미토콘드리아의 세포막전위에 변화를 일으켜 인간 신경아세포종 세포주인 SK-N-MC의 세포박리와 유관한 세포자멸사를 유발하므로 증식억제 효과가 있음을 입증한 것이다.

• 대한한의학회지
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• 제28권1호통권69호
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• pp.198-210
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• 2007

Objectives : Nitric oxide(NO) is a reactive free radical and a messenger molecule in many physiological functions. However, excessive release NO of induces neurotoxicity. We investigated whether a mixture of red ginseng and paeonia radix prossesses a protective effect against sodium nitroprusside(SNP)-induced apoptosis in the human neuroblastoma cell line SK-N-MC. Methods : We performed 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay, 4,6-diamidino-2-phenylindole(DAPD) staining, terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick end labeling(TUNEL)assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction(RT-PCR), Western blot analysis, and caspase-3 enzyme activity assay in SK-N-HFC cells. Result : MTT assay showed that SNP treatment significantly reduced the viabilities of cells and that pre-treatment with the red ginseng and paeonia radix mixture alleviated SNP-induced cytotoxicity. The cells treated with SNP exhibited several apoptotic features, while those pre-treated fir 1 h with the mixture of red ginseng and paeonia radix 1 h prior to SNP expose showed reduced apoptotic features. In addition, the cells pre-treated with the red ginseng and paeonia radix mixture for 1 h prior to SNP expose increased bel-2 expressions, decreased Bax expressions, and decreased caspase-3 enzyme activity. Conclusions : These results show that the red ginseng and paeonia radix mixture exerts a protective effect against SNP-induced apoptosis in SK-N-MC cells.

• 동의생리병리학회지
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• 제22권2호
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• pp.328-332
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• 2008

The purpose of this study was to identify the protective effect of Codonopis pilosula extract on cell death induced by $H_2O_2$ in SK-N-MC neuroblastoma cells. We measured the antioxidant effect by DPPH radical scavenging analysis, BSA analyssis and examined the cell viability by crystal violet and cytochrome C, Bax, Bcl-2, p53, p21 by using Western blot analysis. Codonopis pilosula extract scavenged DPPH radical in a dose-dependent manner and shown direct free radical scavenging effect, suggested that Codonopis pilosula extract have antioxidant effect in vitro. Treatment of cells with hydrogen peroxide, a reactive oxygen species, was to induce cell death and pretreatment with Codonopis pilosula extract attenuated the occurrence of $H_2O_2-induced$ cell death. To elucidate the protective mechanisms of action of Codonopis pilosula extract, Western blot analyses for Bcl-2 and Bax expression and cytochrome c release were carried out. Pretreatment with Codonopis pilosula extract induced the expression of Bcl-2 and suppressed the release of cytochrome c and Bax into the cytosol, thereby arresting $H_2O_2-induced$ apoptotic cell death. Especially p21 and p53 were decreased prior to $H_2O_2$ treatment. These results suggest that Codonopis pilosula extract is associated with the cell cycle and anti-apoptotic cell death.

• Molecules and Cells
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• 제26권1호
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• pp.12-17
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• 2008

The TrkA tyrosine kinase is activated by autophosphorylation in response to NGF, and plays an important role in cell survival, differentiation, and apoptosis. To investigate its role in cell fate determination, we produced stable TrkA-inducible SK-N-MC and U2OS cell lines using the Tet-On system. Interestingly, TrkA overexpression induced substantial cell death even in the absence of NGF, by stimulating ERK phosphorylation and caspase-7 activation leading to PARP cleavage. TrkA-mediated cell death was shown by the annexin-V binding assay to be, at least in part, apoptotic in both SK-N-MC and U2OS cells. Furthermore, the truncated form (p18) of Bax accumulated in the TrkA-induced cells, suggesting that TrkA induces mitochondria-mediated apoptosis. NGF treatment augmented the cell death induced by TrkA overexpression. This TrkA-induced cell death was blocked by the tyrosine kinase inhibitors, K-252a and GW441756. Moreover, TrkA overexpression inhibited long-term proliferation of both the neuronal SK-N-MC cells and the non-neuronal U2OS cells, suggesting a potential role of TrkA as a tumor suppressor.

• 한국식품과학회지
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• 제34권4호
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• pp.688-693
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• 2002

천연물을 이용하여 생리활성이 있는 성분을 추출 분리하여 그 유효 성분을 탐색함으로써 질병 예방 및 치료제로 이용하고자 하는 연구가 최근 활발히 진행되고 있다. 본 연구에서는 예부터 혈액 순환을 돕고, 지혈 작용에 효과가 있는 것으로 알려져 있는 식탁에 상용되는 야채인 부추를 이용하여 4종의 인체 암세포주인 HepG2, HeLa, MCF7 및 SK-N-MC를 이용하여 암세포 증식억제 및 암예방 QR 유도활성 효과를 측정하였다. MTT assay를 이용한 암세포 증식억제 실험 결과, 4종의 암세포주 모두 ethylether층인 ATMEE에서 아주 높은 농도 의존적이 암세포 증식억제 효과를 보였다. 즉 본 실험에 사용된 인체 암세포주 4종에서 시료의 농도가 $150\;{\mu}g/mL$일 때 각 세포주 모두 약 90% 이상의 높은 암세포 증식억제 효과가 나타났으며, 특히 신경아종세포주인 SK-N-MC 에서는 ethylether층의 시료 농도를 다른 층을 첨가시의 농도보다 적은 $100\;{\mu}g/mL$ 첨가에서도 이미 99.8%의 아주 높은 암세포 증식억제 효과를 나타내었다. 또 신경아종세포인 SK-N-MC에서는 ethylacetate층인 ATMEA의 경우에서도 최종 첨가 시료농도를 $200\;{\mu}g/mL$ 첨가했을 때 이미 91%의 높은 암세포 증식억제 효과가 나타났다. 암예방 quinone reductase(QR) 유도활성을 측정한 결과, 시료의 butanol 분획물 ATMB를 $50\;{\mu}g/mL$ 첨가했을 때 시료무처리구인 대조군을 1.0으로 한 경우 암예방 QR유도 효과가 아주 높은 3.92배의 활성이 나타났다. 본 연구 결과, 인체암세포 4종류에 대한 암세포 증식억제(cytotoxicity) 효과는 부추의 ethylether 분배층(ATMEE)에서 제일 뚜렷하였고, 암예방 QR 유도활성 효과는 butanol 분배층(ATMB)에서 가장 유도효과가 좋았다. 나아가 단계적인 생리활성 물질의 분리 동정이 계속 이루어져야 할 것으로 사료된다.

• 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.