• Title/Summary/Keyword: Neuronal cell culture

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Anti-neuronal Injury Effect of Evodiae Fructus Water Extract in Sodium Cyanide-induced SK-N-SH Cell Lines (Sodium Cyanide로 유도된 신경아세포종 세포주에서 오수유의 신경상해 보호효과)

  • Jang Woo-Seok;Lee So-Yeon;Yoon Hyeon-Deok;Shin Oh-chul;Park Chang-Gook;Park Chi-Sang
    • The Journal of Korean Medicine
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    • v.26 no.3 s.63
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    • pp.135-145
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    • 2005
  • Objectives : This study investigated effect of Evodiae fructus water extract (EVOR) on apoptotic cell death induced by NaCN in SK-N-SH neuroblastoma cell lines. NaCN stimulates glutamate release which can activate glutamate receptors to initiate excitotoxic processes. This study examines the role of EVOR in mediating NaCN-induced cytotoxicity. Methods & Results : Cytotoxicity was assessed by measuring lactate dehydrogenase (LDH) in the culture media. NaCN(0.1mM) produced cytotoxicity following 12hrs of incubation. NaCN-induced cytotoxicity was partially blocked by EVOR. The treatment of EVOR in simultaneous exposure of cultures to NaCN provided complete protection against cytotoxicity. NaCN-induced cytotoxicity was found to inhibit DNA fragmentation, repaired by cell cycle and simultaneous exposure to NaCN, regenerated with neurite outgrowh by EVOR. These results indicate thaf damage by NaCN in neumnal cell cultures was repaired by EVOR, whereas NaCN-induced cytotoxicity is blocked Primarily by activation of anti-apoptosis. Conclusions : These results suggest that EVOR may be beneficial for the treatment of dementia and other degenerative problems of the central nervous system.

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Transduced Human Copper Chaperone for Cu,Zn-SOD (PEP-1-CCS) Protects Against Neuronal Cell Death

  • Choi, Soo Hyun;Kim, Dae Won;Kim, So Young;An, Jae Jin;Lee, Sun Hwa;Choi, Hee Soon;Sohn, Eun Jung;Hwang, Seok-Il;Won, Moo Ho;Kang, Tae-Cheon;Kwon, Hyung Joo;Kang, Jung Hoon;Cho, Sung-Woo;Park, Jinseu;Eum, Won Sik;Choi, Soo Young
    • Molecules and Cells
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    • v.20 no.3
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    • pp.401-408
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    • 2005
  • Reactive oxygen species (ROS) contribute to the development of various human diseases. Cu,Zn-superoxide dismutase (SOD) is one of the major means by which cells counteract the deleterious effects of ROS. SOD activity is dependent upon bound copper ions supplied by its partner metallochaperone protein, copper chaperone for SOD (CCS). In the present study, we investigated the protective effects of PEP-1-CCS against neuronal cell death and ischemic insults. When PEP-1-CCS was added to the culture medium of neuronal cells, it rapidly entered the cells and protected them against paraquat-induced cell death. Moreover, transduced PEP-1-CCS markedly increased endogenous SOD activity in the cells. Immunohistochemical analysis revealed that it prevented neuronal cell death in the hippocampus in response to transient forebrain ischemia. These results suggest that CCS is essential to activate SOD, and that transduction of PEP-1-CCS provides a potential strategy for therapeutic delivery in various human diseases including stroke related to SOD or ROS.

Effects of Water-extract Mixture of Scutellariae baicalensis GEORGI, Acarus gramineus SOLAND and Gastrodia elata BLUME on Cultured Rat Cortical Neurons and Enhancement of Learning and Memory Power (황금(黃芩), 석창포(石菖蒲), 천마(天麻) 물추출액 혼합물이 배양한 흰쥐 대뇌신경세포의 활성과 학습능력 증진 효능)

  • Bae, Chul-Hwan;Jung, Hyun-Jung;Jung, Seung-Hyun;Moon, Il-Soo;Lee, Won-Chul;Sin, Gil-Jo
    • Journal of Life Science
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    • v.19 no.6
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    • pp.756-764
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    • 2009
  • Scutellariae baicalensis GEORGt Acorus gramineus SOLAND and Gastrodia elata BLUME are traditional medicines used in the treatment of incipient stoke. In this study we investigated their effects on various aspects of neuronal differentiation in single or composite forms. Water-extracts of these medicines showed neuroprotective effects on cultured rat cortical neurons in normoxia and hypoxia. To understand the mechanism for neuroprotection we carried out various cell biological assays. They stimulated initial differentiation of neuronal development (transition from stage 1 to 2), and increased the number of spines and the length and number of dendritic processes. These effects were best manifested in the experimental group, which were given a mixture of the three kinds of extracts (p<0.01). To assess improvement of brain functions we carried out Morris water-maze tests for the mice that were fed on these extracts instead of water for 4 weeks. The experimental groups, especially those which were given the mixture of the three kinds of extract, showed significant (p<0.01) enhancement in memory as early as one day after the learning trial. These results indicate that these three kinds of extracts have synergistic effects on neuronal protection and improvement of brain functions.

Myelination by co-culture of neurons and schwann cells and demyelination by virus infection (뉴런세포와 슈반세포의 공동배양에 의한 수초화와 바이러스 감염에 의한 탈수초화)

  • Sa, Young-Hee;Kweon, Tae Dong;Kim, Ji-Young;Kim, Hyun Joo;Lee, Bae Hwan;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2018.10a
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    • pp.448-451
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    • 2018
  • The purpose of this study was to investigate the developmental process of myelination by neuron and Schwann cell cultures and the development of demyelination by herpes simplex virus-1 infection by electron microscopy and molecular biological analysis. The dorsal root ganglion (DRG) was isolated from the mouse embryo and Schwann cells and neuronal cells were cultured in vitro. Neuronal cells treated with mitotic inhibitors and purified Schwann cells were co-cultured together to induce myelination. The herpes simplex virus-1 was infected with the co-cultured cells, and the demyelination was induced. The myelin protein zero (MPZ) antibody, which means the presence of myelin formation, was used and electron microscopy was used to observe the development of myelin and dehydration.

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Effects of Taurine on Glutamate-induced Neurotoxicity and Interleukin-6 mRNA Expression in Astrocytes

  • Yang, Seong-Chil;Baek, Su-Yeon;Choe, In-Pyo;Lee, Chang-Jung
    • Animal cells and systems
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    • v.1 no.3
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    • pp.467-473
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    • 1997
  • Taurine (2-aminoethanesulfonic acid), one of bioactive amino acid in the mammalian brain, is known to exert inhibitory effects on neurons via GABA receptor. In the present study, we examined effects of taurine on glutamateinduced neurotoxicity on hippocampal neuron cell culture using cell counting method and lactate dehydrogenase (LDH) assay. After 10 d of culture, cells were stimulated with appropriate drugs. Only 43% of cultured neuronal cells survived at one day after stimulation with 500 uM L-glutamate for 10 min. Survival rate was enhanced by 82% in the presence of 10 mM taurine. LDH activity from the culture supernatant incubated with a combination of L-glutamate and taurine was less than half of that with L-glutamate alone. In the next series of experiments, interleukin-6 (IL-6) mRNA expression in cultured astrocytes was investigated using reverse tanscription-PCR (RT-PCR). IL-6 mRNA was detected in the astrocytes stimulated with L-glutamate in a dose-dependent manner, while not detected in the unstimulated control astrocytes. The expression of IL-6 mRNA caused by 10 mM glutamate was inhibited by taurine, but not by GABA. These findings demonstrated a neuroprotective action of taurine against glutamate-induced toxicity.

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Pineal Photoreceptor and Ganglion Cells in River Lamprey, Lampetra japonica -Two Types of Pineal Ganglion Cell-

  • Tamotsu, Satoshi;Kinugawa, Yoshimi;Kawano, Emi;Watanabe, Mai;Samejima, Michikazu;Oishi, Tadashi
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.21-24
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    • 2002
  • Pineal organs of poikilotherm vertebrates transform the environmental light information into a humoral message and a neuronal activity. The former is melatonin, and the latter is modulation of the impulse in ganglion cells. The ganglion cells are physiologically classified into luminosity (achromatic) type and chromatic one, as the neural activity is modulated in two ways. We attempted to classify the pineal ganglion cells with morphological characteristics by means of the three- dimensional reconstruction method. In the pineal ganglion cells of river lamprey, there are two different features, oval and spherical. For comparison of their projection region in the brain, the tracing investigation was also carried out. The application of the neural tracer near mesencephalic tegmentum showed that only oval-shaped ganglion cells were labeled in the pineal organ. These results suggest that the oval-shaped ganglion cell is functionally different from the spherical one.

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Protection of spontaneous and glutamate-induced neuronal damages by Soeumin Sibjeundaibo-tang and Soyangin Sibimijihwang-tang in cultured mice cerebrocortical cells

  • Lee, Mi-Young;Ma, Jin-Yeul;Choo, Young-Kug;Jung, Kyu-Yong
    • Advances in Traditional Medicine
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    • v.1 no.1
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    • pp.55-63
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    • 2000
  • Soeumin Sibjeundaibo-tang (SJDBT) and Soyangin Sibimijihwang-tang (SMJHT) have been used traditionally to improve the systemic blood circulation and biological energy production in the patients with circulatory and neuronal diseases. The object of this study is to determine the protective effects of SJDBT and SMJHT extracts on the spontaneous and glutamate-induced neuronal damages in cultured cells derived from mice cerebral cortex. At 14 days after beginning the cultures, the activity of lactate dehydrogenase released into the culture media was significantly decreased by treatment of cerebroneuronal cells with SJDBT and SMJHT (0.1 mg/ml) for 7 days. By comparison with the normal cells, cerebroneuronal morphology was dramatically changed by treatment of glutamate (1 mM) for 12 hrs, and this was conspicuously recovered by pretreatment of cerebroneural cells with SJDBT and SMJHT (0.1-1.0 mg/ml) for 2 days. Moreover, glutamated-induced DNA fragmentation was also protected by pretreatment of cerebroneuronal cells with those extracts. These results suggest that naturally occurring and glutamate-induced degeneration of cultured cerebrocortical cells may be related, in part, to the process of apoptotic cell death. The pharmacological properties of SJDBT and SMJHT extracts to improve cerebroneuronal degeneration may be considered as one of useful medicines that can prevent cerebrocortical impairments resulted from age-dependent and excitotoxicity-induced neuronal degeneration in human brain.

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Ginsenosides Inhibit NMDA Receptor-Mediated Epileptic Discharges in Cultured Hippocampal Neurons

  • Kim, Sun-Oh;Rhim, Hye-Whon
    • Archives of Pharmacal Research
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    • v.27 no.5
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    • pp.524-530
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    • 2004
  • Epilepsy or the occurrence of spontaneous recurrent epileptiform discharges (SREDs, seizures) is one of the most common neurological disorders. Shift in the balance of brain between excitatory and inhibitory functions due to different types of structural or functional alterations may cause epileptiform discharges. N-Methyl-D-aspartate (NMDA) receptor dysfunctions have been implicated in modulating seizure activities. Seizures and epilepsy are clearly dependent on elevated intracellular calcium concentration ([C $a^{2+}$]$_{i}$ ) by NMDA receptor activation and can be prevented by NMDA antagonists. This perturbed [C $a^{2+}$]$_{i}$ levels is forerunner of neuronal death. However, therapeutic tools of elevated [C $a^{2+}$]$_{i}$ level during status epilepticus (SE) and SREDs have not been discovered yet. Our previous study showed fast inhibition of ginseng total saponins and ginsenoside R $g_3$ on NMDA receptor-mediated [C $a^{2+}$]$_{i}$ in cultured hippocampal neurons. We, therefore, examined the direct modulation of ginseng on hippocampal neuronal culture model of epilepsy using fura-2-based digital $Ca^{2+}$ imaging and neuronal viability assays. We found that ginseng total saponins and ginsenoside R $g_3$ inhibited $Mg^{2+}$ free-induced increase of [C $a^{2+}$]$_{i}$ and spontaneous [C $a^{2+}$]$_{i}$ oscillations in cultured rat hippocampal neurons. These results suggest that ginseng may playa neuroprotective role in perturbed homeostasis of [C $a^{2+}$]$_{i}$ and neuronal cell death via the inhibition of NMDA receptor-induced SE or SREDs.d SE or SREDs..

Induction of Demyelination of Neuronal cells by Sindbis Virus (Sindbis Virus에 의한 뉴런세포의 탈수초의 유도)

  • Sa, Young-Hee;Kim, Hyun Joo;Kweon, Tae Dong;Kim, Ji-Young;Lee, Bae Hwan;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2018.05a
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    • pp.584-587
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    • 2018
  • Many viruses including mouse hepatitis virus, corona, measles, and sidbis viruses are known as causative virus of inducing demyelination which means destruction of myelination in nervous system of mice. The purpose of this study is to investigate processing of myelination by co-culture of Schwann cells and neuronal cells and demyelination induced by infection of sindbis virusin rat. Schwann cells and neuronal cells from dorsal root ganglion (DRG) in embryos (E16) of rat were cultured in vitro respectively. The purified neuronal cells with anti-mitotic agents and purified Schwann cells were co-cultured. After that, infection of sindbis virus into this myelinated co-culture system was performed. Myelination and demyelination process were observed using antibody of myelin basic protein meaning presence of myelination.We identified myelination and demyelination processing using antibody of peripheral myelin protein 22 (PMP 22) meaning presence of myelinated neuron. This study was supported by the Basic Research Program through the National Research Foundation (NRF) funded by the Ministry of Science, ICT & Future Planning (NRF-2015R1C1A1A01053484 and 2017R1A2B3005753).

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Effects of Water Extract of Liriope platyphylla on the mRNA Expression and Protein Secretion of Nerve Growth Factors (맥문동 유수추출물이 NGF의 mRNA발현과 단백질 분비에 미치는 영향)

  • Choi, Sun-Il;Park, Jee-Hee;Her, Youn-Kyng;Lee, Yoen-Kyung;Kim, Ji-Eun;Nam, So-Hee;Goo, Jun-Seo;Jang, Min-Ju;Lee, Hee-Sup;Son, Hong-Ju;Lee, Chung-Yeol;Hwang, Dae-Youn
    • Korean Journal of Medicinal Crop Science
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    • v.18 no.5
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    • pp.291-297
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    • 2010
  • Liriope platyphylla has been though as an useful medical plant to improve the cough, sputum, neurodegenerative disorders, obesity, and diabetes in Korea and China from old times. In order to investigate the effects of Liriope platyphylla on expression and secretion of nerve growth factor (NGF), the mRNA expression and protein secretion were detected in the neuronal cell (B35) and neuroglial cell (C6) cultured with three differences concentration (5%, 10%, 15%) of Liriope platyphylla. In MTT assay and FACS anslysis, the some death of some B35 and C6 cells were observed in 15% extract-treated group, while other groups did not induce the death. Also, the mRNA expression of NGF were significantly increased in 5% and 10% extracts treated-group. Furthermore, the NGF protein concentration in supernatant collected from cultured cells showed the very similar pattern with mRNA expression. In order to verify the activity of secreted NGF, the culture supernatant collected from B35 and C6 cells cultured with Liriope platyphylla extracts for 24 hrs were treated into undifferentiated PC12 cells, and the differentiation level of PC12 cell were also observed with microscopes. The differentiation level of PC12 cell were significantly increased depend on the dose of extract. Therefore, these results suggested that the water extracts of Liriope platyphylla may contribute the regulation of NGF expression and secretion in the neuronal cell and be considered as an excellent candidate for a neurodegenerative disease-therapeutic drug.