• Biomolecules & Therapeutics
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• 제14권3호
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• pp.137-142
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• 2006

Apigenin, a natural flavonoid found in a variety of vegetables and fruits, has been shown to possess many biological functions. In this study we investigated the role of apigenin in the production of reactive oxygen species (ROS) through the modulation of activity of $K^+-Cl^-$-cotransport (KCC) in IMR-32 human neuroblastoma cells. Apigenin induced $Cl^-$-dependent $K^+$ efflux, a hallmark of KCC activity, which was markedly prevented by different kinds of KCC inhibitors (calyculin-A, genistein and $BaCl_2$). These results indicate that KCC is functionally present, and activated by apigenin in the IMR-32 cells. Treatment with apigenin also induced a sustained increase in the level of intracellular ROS. The KCC inhibitors also significantly inhibited the apigenin-induced ROS generation. Taken together, these results suggest that apigenin can modulate ROS generation through the activation of a membrane ion transporter, KCC. These results further suggest that the alteration of KCC activity may play a role in the mechanism of degenerative diseases and/or carcinogenesis in neuronal tissues through the regulation of ROS production.

• Molecular & Cellular Toxicology
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• 제2권1호
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• pp.60-66
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• 2006

Methylmercury (MeHg), one of the heavy metal compounds, can cause severe damage to the central nervous system in humans. Many reports have shown that MeHg is poisonous to human body through contaminated foods and has released into the environment. Despite many studies on the pathogenesis of MeHg-induced central neuropathy, no useful mechanism of toxicity has been established so far. This study, using of suppression subtractive hybridization (SSH) method, was peformed to identify differentially expressed genes by MeHg in SH-SY5Y human neuroblastoma cell line. We prepared to total RNA from SH-SY5Y cells treated with solvent (DMSO) and $6.25\;{\mu}M\;(IC_{50})$ MeHg and performed forward and reverse SSH. Differentially expressed cDNA clones were screened by dot blot, sequenced and confirmed that individual clones indeed represent differentially expressed genes with real time RT-PCR. These sequences were identified by BLAST homology search to known genes or expressed sequence tags (ESTs). Analysis of these sequences may provide an insight into the biological effects of MeHg in the pathogenesis of neurodegenerative disease and a possibility to develop more efficient and exact monitoring system of heavy metals as ubiquitous environmental pollutants.

• 대한한의학회지
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• 제21권4호
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• pp.183-192
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• 2000

Objectives : The present study was carried out to investigate the effects of Danchun-hwan(DCH) on the peroxynitrite-induced neural cell death in human neuroblastoma cell line, SH-SY5Y. Methods : The cultured cells were pretreated with DCH and exposed to 3-morpholinosydnonimine(SIN-1) that simultaneously generates NO and superoxide, thus possibly forming peroxynitrite. The cell damage was assessed by using MTT assay and crystal violet staining. Results : Exposure of the cells to SIN-1 for 24hr induced 75% apoptotic cell death, as evaluated by the occurrence of morphological nuclear changes characteristic of apoptosis using 4', 6-diamidino-2-phenylinole(DAPI). However, pretreatment of SH-SY5Y with the water extracts of DCH, inhibited the apoptotic cell death in a dose-dependent manner. DCH also inhibited SIN-1-induced apoptotic caspase 3-like protease activity in a dose-dependent manner. DCH recovered the depleted glutathione levels by SIN-1. Conclusions : Taken together, it is suggested that DCH protected human neuroblastoma cell line, SH-SY5Y, from the free radical injury mediated by peroxynitrite by a mechanism of elevating antioxidant, GSH.

• Archives of Pharmacal Research
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• 제18권3호
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• pp.159-163
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• 1995

Previously, we have reported that methylene blue (MB) induces cytotoxicity in human brain tumor cells through the generation of free radicals. In this study the effect of deferoxamine (DFO), an iron chelator, on MB-induced cytotoxicity was investigated using SK-N-MC human neuroblastoma and U-373 MG human astrocytoma cells as model cellular systems. The cytotoxic effect of MB was potentiated by DFO. The potentiation effect of DFO was significantly blocked by either stoichiometric amounts of ferric ion, various antioxidants, hydroxyl radical scavengers or intracellular $Ca^{2+}$ release blockers. These results suggest that hydroxyl radical and intracellular $Ca^{2+}$ may act as important mediators of the enhanced cytotoxicity by MB and DFO. These results further suggest that the combined treatment with MB and DFO may be useful for the therapeutical applications of human brain tumors.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2020년도 춘계학술대회
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• pp.76-76
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• 2020

Stachys sieboldii Miq. (chinese artichoke), which has been extensively used in oriental traditional medicine to treat of ischemic stroke; however, the role of Stachys sieboldii Miq. (SSM) in cerebral ischemia/reperfusion (I/R) injury is not yet fully understood. In the current study, the neuroblastoma cell line (SH-SY5Y) were subjected to oxygen-glucose deprivation/reperfusion (OGD/R) to simulate I/R injury in vitro model. The results showed that SSM improved OGD/R-induced inhibitory effect on cell viability of SH-SY5Y Cells. SSM displayed anti-oxidative activity as proved by the decreased levels of reactive oxygen species (ROS) and malondialdehyde (MDA), and increased activities of superoxide dismutase (SOD) and glutathione peroxidase (GPx) in OGD/R-induced SH-SY5Y Cells. In addition, cell apoptosis was markedly decreased after SSM treatment in OGD/R-induced SH-SY5Y Cells. The up-regulation of Bcl-2 and down-regulation of Bax, thus reducing the Bax/Bcl-2 ratio that in turn protected the activation of caspase-9 and -3, and inhibition of poly (ADP-ribose) polymerase cleavage, which was associated with the blocking of cytochrome c release to the cytoplasm. Collectively, SSM protected human neuroblastoma SH-SY5Y cells from OGD/R-induced injury via preventing mitochondrial-dependent pathway through scavenging excessive ROS, suggesting that SSM might be a potential agent for the ischemic stroke therapy.

• Bulletin of the Korean Chemical Society
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• 제34권5호
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• pp.1503-1507
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• 2013

The two major symptoms characterizing Alzheimer's disease are the formation of amyloid-${\beta}$ extracellular deposits in the form of senile plaques and intracellular neurofibrillary tangles (NFTs) that consist of pathological hyperphosphorylated tau protein aggregated into insoluble paired helical filaments (PHFs). Neurons of the central nervous system have appreciable amounts of tau protein, a microtubule-associated protein. To maintain an optimal operation of nerves, the microtubules are stabilized, which is necessary to support cell structure and cellular processes. When the modified tau protein becomes dysfunctional, the cells containing misfolded tau cannot maintain cell structure. One of the pathological hallmarks of Alzheimer's disease is hyperphosphorylated tau protein. This paper shows that the small heat shock protein from humans (Hsp27) reduces hyperphosphorylated tau and prevents hyperphosphorylated tau-induced cell death of the human neuroblastoma cell line SH-SY5Y.

• BMB Reports
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• 제46권5호
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• pp.276-281
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• 2013

In this study, we aimed to compare the morphogenetic and neuronal characteristics between monolayer cells and spheroids. For this purpose, we established spheroid formation by growing SH-SY5Y cells on the hydrophobic surfaces of thermally-collapsed elastin-like polypeptide. After 4 days of culture, the relative proliferation of the cells within spheroids was approximately 92% of the values for monolayer cultures. As measured by quantitative assays for mRNA and protein expressions, the production of synaptophysin and neuronspecific enolase (NSE) as well as the contents of cell adhesion molecules (CAMs) and extracellular matrix (ECM) proteins are much higher in spheroids than in monolayer cells. Under the all-trans-retinoic acid (RA)-induced differentiation condition, spheroids extended neurites and further up-regulated the expression of synaptophysin, NSE, CAMs, and ECM proteins. Our data indicate that RA-differentiated SH-SY5Y neurospheroids are functionally matured neuronal architectures.

• 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.

• IMMUNE NETWORK
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• 제1권1호
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• pp.53-60
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• 2001

Background: The human mic2 gene is a pseudoautosomal gene that encodes a cell surface antigen, CD99. High levels of CD99 constitute a tumor marker in Ewing s sarcoma (ES). We have recently demonstrated that CD99-induced apoptosis occurs only in undifferentiated ES cells, not in differentiated ES cells, raising the possibility of the involvement of CD99 in neural ontogeny. Methods: To elucidate the relations between the expression of CD99 and the differentiation of neural cells and the mechanism by which the expression of CD99 is regulated, we analyzed the differential patterns of CD99 expression in SH-SY5Y by treatment of 12-O-tetradecanoyl-13-phorbol acetate (TPA) and retinoic acid. In addition, to explore the transcriptional activity of CD 99 during neural cell differentiation, SH-SY5Y cells were transiently transfected with a CD99 promoter-driven luciferase construct, and treated with the inducers. Results: In immunoblotting and flow cytometry, the expression level of CD99 was increased on differentiated SH-SY5Y cells induced by TPA and retinoic acid. The luciferase activity was elevated by the treatment with TPA, known to mature SH-SY5Y cells toward a sympathetic neuronal lineage, whereas retinoic acid inducing a sympathetic chromaffin lineage displayed little effect. Conclusion: The result indicates that CD99 might be expressed only on cells maturing toward a neuronal lineage among differentiating primitive neuronal cells. In addition, the expression of CD99 seems to be regulated at the transcriptional level during the differentiation.

• 한국환경독성학회:학술대회논문집
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• 한국환경독성학회 2000년도 추계심포지움 및 학술발표회:바이오모니터링 기법을 이용한 환경위해성 평가
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• pp.48-48
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• 2000