• 대한바이러스학회지
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• 제26권1호
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• pp.1-8
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• 1996

Human cytomegalovirus (HCMV) replication and $Ca^{2+}$ response in human cell lines of neuronal origin were investigated. SK-N-SH (neuroblastoma cells) and A172 cells (glioblastoma cells) were used. SK-N-SH cells were permissive for HCMV multiplication with a delay of one day compared to virus multiplication in human embryo lung (HEL) cells. The delay of HCMV multiplication in SK-N-SH cells appeared to be correlated with a delay in the $Ca^{2+}$ response. The cytoplasmic free $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) began to increase at 12 h p.i. in HCMV-infected SK-N-SH cells, while $[Ca^{2+}]_i$ increase in HCMV-infected HEL cells was observed as early as 3 h p.i. On the whole, the level of the increase in $[Ca^{2+}]_i$ in SK-N-SH cells was about 30% of that in HEL cells. On the other hand, in A172 cells infected with HCMV, neither production of infectious virus nor detectable increase in $[Ca^{2+}]_i$ was observed. Treatment with TPA of HCMV-infected SK-N-SH cells resulted in $[Ca^{2+}]_i$ increase at 6 h p.i. The stimulatory effect of TPA on HCMV- induced $[Ca^{2+}]_i$ increase continued until 12 h p.i., but TPA failed to stimulate the $Ca^{2+}$ response in SK-N-SH cells at 24 h p.i., suggesting that the effect of TPA had disappeared in SK-N-SH cells at that time point. In conclusion, SK-N-SH cells are permissive for HCMV replication and the delay in $Ca^{2+}$ response may be a consequence of the lower responsiveness of SK-N-SH cells than HEL cells to HCMV infection.

• 대한한방내과학회지
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• 제25권3호
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• pp.482-491
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• 2004

Alzheimer's disease(AD) is a geriatric dementia that is widespread in old age. In the near future AD may be the biggest problem in public health service. Although a variety of oriental therapies in the study of Jimitang have been traditionally utilized for the treatment of AD, their pharmacological effects and active mechanisms have not been fully elucidated. This study in an investigation of effects of Jimitang on apoptotic cell death induced by CT105 overexpression in SK-N-SH neuroblastoma cell lines. DNA fragmentation, neurite outgrowth assay and LDH activity assay were examined. The regeneratory and inhibitory effects on Alzheimer's disease in pCT105-induced neuroblastoma cell lines by Jimitang water extract were examined. Findings from these experiments have shown that Jimitang inhibits the synthesis or activities of CT105, which has neurotoxicities and apoptotic activities in cell lines. In addition, pretreatment of $Jimitang(>50\;{\mu}g/mL\;for\;12\;hours)$ partially prevented CT(105)-induced cytotoxicity in SK-N-SH cell lines, and were inhibited by pretreatment. $Jimitang(>50\;{\mu}g/mL\;for\;12\;hours)$ repaired CT(105)-induced neurite outgrowth when SK-N-SH cell lines were transfected with CT(105). Results of this study show that. in the Jimitang group, the apoptosis in the nervous system in inhibited, the repair against the degerneration of neuroblastoma cells by CT105 expression is promoted. In addition, Jimitang was found to inhibit DNA fragmentation induced by CT105 overexpression, and promote neurite outgrowth. These findings suggest that Jimitang is beneficial for the treatment of AD.

• Asian Pacific Journal of Cancer Prevention
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• 제15권21호
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• pp.9445-9451
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• 2014

Neuroblastoma is the most common extracranial solid tumor in children. Approximately half of the affected patients are diagnosed with high-risk poor prognosis disease, and novel therapies are needed. Sanguinarine is a benzophenanthridine alkaloid which has anti-microbial, anti-oxidant and anti-inflammatory properties. The aim of this study is whether sanguinarine has in vitro apoptotic effects and which apoptotic genes might be affected in the human neuroblastoma cell lines SH-SY5Y (N-myc negative), Kelly (N-myc positive, ALK positive), and SK-N-BE(2). Cell viability was analysed with WST-1 and apoptotic cell death rates were determined using TUNEL. After RNA isolation and cDNA conversion, expression of 84 custom array genes of apoptosis was determined. Sanguinarine caused cell death in a dose dependent manner in all neuroblastoma cell lines except SK-N-BE(2) with rates of 18% in SH-SY5Y and 21% in Kelly human neuroblastoma cells. Cisplatin caused similar apoptotic cell death rates of 16% in SH-SY5Y and 23% in Kelly cells and sanguinarine-cisplatin combinations caused the same rates (18% and 20%). Sanguinarine treatment did not affect apoptototic gene expression but decreased levels of anti-apoptotic genes NOL3 and BCL2L2 in SH-SY5Y cells. Caspase and TNF related gene expression was affected by the sanguinarine-cisplatin combination in SH-SY5Y cells. The expression of regulation of apoptotic genes were increased with sanguinarine treatment in Kelly cells. From these results, we conclude that sanguinarine is a candidate agent against neuroblastoma.

• 대한한의학회지
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• 제25권2호
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• pp.138-150
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• 2004

Objectives : Alzheimer's disease (AD) is a geriatric dementia that is widespread in old age. In the near future AD will be the biggest problem in public health service. Although a variety of oriental prescriptions, including Sesim-tang, have been traditionally utilized for the treatment of AD, their pharmacological effects and action mechanisms have not yet been fully elucidated. The present study investigated the effects of Sesim-tang on apoptotic cell death induced by CT105 (carboxy terminal 105 amino acid peptide fragment of APP) overexpression in SK-N-SH neuroblastoma cell lines. Methods: We studied the regenerative and inhibitory effects on Alzheimer's disease in CT105-induced SK-N-SH cell lines by Sesim-tang water extract. We examined for cell morphological pattern, DNA fragmentation, LDH activity assay, zymography assay, and immunohistochemistric analysis. Additionally, we investigated the association between the CT105 and neurite degeneration caused by CT105-induced apoptotic response in neurone cells. Results: Findings from our experiments have shown that Sesim-tang inhibits the synthesis or activities of CT105, which has neurotoxicities and apoptotic activities in the cell line. In addition, pretreatment with Sesim-tang ($>50\mu\textrm{g}/ml$ for 12 hours) partially prevented CT105-induced cytotoxicity in SK-N-SH cell lines. SK-N-SH cell lines overexpressed with CT105 exhibited remarkable apoptotic cell damage. Based on morphological observations by phase-contrast microscope and LDH activity measurements in the culture media, the CT105-induced cell death was significantly inhibited by Sesim-tang water extract. Sesim-tang was found to reduce the expression of APP and caspase-3 induced by CT105 in SK-N-SH cell lines and in rat hippocampus. Conclusions: As the result of this study, in the Sesim-tang group, apoptosis in the nervous system is inhibited, the repair against the degeneration of SK-N-SH cell lines by CT105 expression is promoted. Hence, Sesim-tang may be beneficial for the treatment of AD.

• 한국동물학회지
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• 제38권4호
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• pp.542-549
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• 1995

Extracellular matrix(ECM) 단백질이 SK-N-SH 및 IMR-32 세포주가 신경계 세포로 분화되는 데 미치는 영향을 조사하였다. Laminin과 collagen으로 도말한 배양기에서 7일간 배양했을 때 SK-N-SH세포는 잘 발달된 신경측색생장을 보였으나 IMR-32세포는 뚜렷한 형태변화를 나타내지 않았다. 왜 IMR-32세포가 ECM 단백질에 반응을 하지 않는가를 규명하기 위하여 ECM단백질에 의한 초기 신호전달기작을 두 세포주에서 분석하였다. ECM 단백질을 도말한 배양기에 세포를 깔았을 때 한시간 만에 tyrosine 인산화된 단백질이 두 세포 모두 증가함을 볼 수 있었다. 아울러 focal adhesion kinase(FAK)의 tyrosine 인산화도 두 세포주 모두에서 증가하였다. 이러한 결과는 두 세포주가 ECM 단백질에 의한 초기 신호전달체계가 정상임을 의미한다. 신경세포 분화과정에 증가한다고 알려진 Bcl-2 및 NSE의 량을 ECM 단백질 처리후 조사하였을 때 SK-N-SH 세포주는 두 단백질이 증가 했지만 IMR-32 세포주는 변화가 없었다. 이러한 결과는 IMR-32 세포주가 ECM 단백질에 반응하지 않는 것이 ECM 단백질에 의한 신호전달체계에 문제가 있다기 보다 신경계세포로 분화되는 데 필요한 유전인자의 발현조절에 문제가 있음을 시사한다.

• Journal of Acupuncture Research
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• 제25권3호
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• pp.53-69
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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-SH 세포에서 암세포성장의 억제 및 그 기전에 대하여 살펴보고자 하였다. 방법 : SVT를 처리한 후 SK-N-SH의 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-SH 세포에 SVT를 처리한 후, 신경아세포종 세포의 성장, Apoptosis의 유발 및 기전의 영향을 관찰하여 다음과 같은 결과를 얻었다. 1. SVT를 처리한 SK-N-SH 세포 관찰에서 세포독성은 농도의존적으로 증가하는 경향이 있는 반면 암세포성장의 유의한 억제는 $20{\mu}g/m{\ell}$ SVT에 의해서만 나타났다. 2. 세포자멸사 평가에서 SVT를 처리한 SK-N-SH 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-SH 세포의 세포주기, 세포 내 칼슘양 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-SH는 세포자멸사 관련 단백 발현에서 Bax에 대해 유의한 증가를 나타내지 않았으나 caspase-3 및 caspase-9의 유의한 증가와 Bcl-2의 유의한 감소를 나타내었다. 결론 : 이상의 결과는 SVT가 세포 내 활성산소를 증가시킴으로써 미토콘드리아의 세포막전위에 변화를 일으켜 인간 신경아세포종 세포주인 SK-N-SH의 세포박리와 유관한 세포자멸사를 유발함으로써 증식억제 효과가 있음을 입증한 것이다.

• 생명과학회지
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• 제21권8호
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• pp.1190-1198
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• 2011

한약재의 추출방법에 따른 인체신경모세포 SK-N-SH에 대한 보호 효과를 연구하였다. 당귀, 건지황, 작약 및 천궁을 시료로 사용하였고, 열수추출(환류냉각, 5 시간), 증숙추출(100$^{\circ}C$ 및 120$^{\circ}C$, 90분 후 열수추출) 방법과 에탄올추출(환류냉각, 5 시간)방법을 비교하였다. 추출물을 농도별로 SK-N-SH 세포에 2 시간 처리한 후 $H_2O_2$로 250${\mu}M$로 2 시간 산화스트레스를 유발한 다음 세포독성 및 apoptosis와 caspase-3의 발현 정도를 측정하였다. 모든 약재의 열수추출물이 다른 추출물보다 세포 증식을 촉진하였고, apoptosis를 억제하였다. 한약재 열수추출물 1${\mu}g/{\mu}l}$ 농도까지는 세포증식을 촉진하였지만, 그 이상의 농도에서는 오히려 감소시켰다. 열수추출물은 다른 추출물보다 총페놀성 화합물이 많이 함유되어 있었고, 항산화능이 높았다. 또한, 이와 같은 효과는 당귀의 열수추출물이 다른 약재 열수추출물보다 우수하였다. 본 연구결과는 약재의 열수추출법이 인체신경모세포인 SK-N-SH의 증식과 세포사멸 억제를 위해 가장 우수한 방법이었고, 당귀 열수추출물이 가장 우수한 효과를 지니고 있어 기억력 보호나 상실억제제로 활용할 수 있을 것이다.

• Biomolecules & Therapeutics
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• 제8권4호
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• pp.285-292
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• 2000

The effects of the members of the same nuclear receptor superfamily (all-trans retinoic acid (RA), thyroid hormone(T3) or hydrocortisone) on proliferation and differentiation in the SK-N-SH neuroblastoma (NB) cell lines were studied. NB cells were treated with RA, T3, or hydrocortisone at concentration of 10$^{-6}$ M or 10$^{-8}$ M for 3 days or 7 days. RA induced concentration- and time-dependent morphologic differentiation(neurite outgrowth and microtubule-associated protein expression) and growth inhibition in NB cells. Treatment of 10$^{-7}$ M T3 for 7 days increased viability and differentiation of NB cells. Treatment of 10$^{-6}$ M hydrocortisone for 7 days increased viability of NB cells. Although these three effectors are members of the same receptor superfamily, the regulation of brain development may be carried out in a different manner.

• 대한본초학회지
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• 제28권2호
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• pp.45-53
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• 2013

Objectives : The purpose of the study is to determine the neuroprotective effects of the water and 80% EtOH extract of some herbal medicine plant on ischemia reperfusion-induced cell death in SK-N-SH human brain neuronal cells. Methods : SK-N-SH cells were treated with 3mM sodium azide and 10 mM 2-deoxy-D-glucose for 45 min, ptior to the addition of different concentrations of herbal medicine plant extract (0, 10, 25, 50, 100, 250, 500, 1000 ${\mu}g/ml$) for 2 hr and then reperfused with growth medium, incubated for 24 h. Cell viability was determined by WST-1 assay, and ATP/ADP levels were measured by ADP/ATP ratio assay kit. Results : Herbal medicine plant extract significantly inhibited decreasing the cell viability in ischemia-induced SK-N-SH cells. Also increased the ratio of ADP/ATP in ischemia-induced neuronal cells. Conclusions : Our results suggest that herbal medicine plant extract has a neuroprotective property via increasing the energy levels in neuronal cells, suggesting that extract may has a therapeutic potential in the treatment of ischemic brain injury. The exact component and mechanism remains for the future study.

• Journal of Ginseng Research
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• 제34권2호
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• pp.138-144
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• 2010

Ginseng (Panax ginseng C.A. Meyer) has been shown to have anti-stress effects in animal studies. However, most studies have only managed to detect altered levels of biomarkers or enzymes in blood or tissue, and the actual molecular mechanisms by which ginseng exerts these effects remain unknown. In this study, the anti-oxidative effect of Korean red ginseng (KRG) was examined in human SK-N-SH neuroblastoma cells. Incubation of SK-N-SH cells with the oxidative stressor hydrogen peroxide resulted in significant induction of cell death. In contrast, pre-treatment of cells with KRG decreased cell death significantly. To elucidate underlying mechanisms by which KRG inhibited cell death, the expression of apoptosis-related proteins was examined by Western blot analysis. KRG pre-treatment decreased the expression of the pro-apoptotic gene caspase-3, whereas it increased expression of the anti-apoptotic gene Bcl-2. Consistent with this, immunoblot analysis showed that pre-treatment of the SK-N-SH cells with KRG inhibited expression of the pro-inflammatory gene cyclooxygenase 2 (COX-2). RT-PCR analysis revealed that the repression of COX-2 expression by KRG pre-treatment occurred at the mRNA level. Taken together, our data indicate that KRG can protect against oxidative stress-induced neuronal cell death by repressing genes that mediate apoptosis and inflammation.