The establishment of a reliable cytotoxic system with SK-N-SH neuroblastoma cell culture
The establishment of a reliable cytotoxic system with SK-N-SH neuroblastoma cell culture
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DOI：
10.1016/s0165-0270(02)00324-2
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发表时间：
2003-02-15
影响因子：
3
通讯作者：
Benishin, CG
中科院分区：
文献类型：
--
作者：
Ba, F;Pang, PKT;Benishin, CG
一个可靠的体外细胞毒性系统是神经细胞毒性和神经保护研究的基础。本研究检测了SK-N-SH人神经母细胞瘤细胞系的四种细胞毒性损伤。这些是β-淀粉样蛋白（Abeta），1-甲基-4-苯基-1，2，3，6-四氢吡啶（MPTP），高密度培养，和血清剥夺诱导的神经元死亡。培养96 h后，这些损伤以浓度依赖性方式诱导细胞数量显著减少。在所有的侮辱，MPTP，血清剥夺，和高密度培养诱导细胞凋亡后96小时，而Abeta推测诱导坏死性神经元死亡，因为细胞凋亡是不可检测的。p38 MAP激酶抑制剂SB 203580（1 μ M）和PKC抑制剂白屈菜红碱（5 μ M）分别成功地抑制了由Abeta和高密度培养物引起的活力丧失。其他激酶抑制剂，包括非特异性蛋白激酶抑制剂H7、PKA抑制剂14-22 Amide、PKG抑制剂KT 5823和蛋白酪氨酸激酶抑制剂AG 18对四种细胞毒性模型中的任何一种都没有影响。该系统允许在可以在一个细胞系统内考虑神经元的不同途径和机制的条件下研究神经保护，从而消除可能由于所研究的不同细胞类型而导致的变化。目前的研究描述了一个有效的模型系统筛选潜在的神经保护剂。(C)2002 Elsevier Science B. V.保留所有权利。
A reliable in vitro cytotoxic system is essential in neurocytotoxic and neuroprotective research. The present study examined four cytotoxic insults with the SK-N-SH human neuroblastoma cell line. These were beta-amyloid protein (Abeta), 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), high density culture, and serum deprivation induced neuronal death. These insults induced significant reduction in cell numbers after 96 h culture, in a concentration dependent manner. Among all the insults, MPTP, serum deprivation, and high density culture induced apoptosis after 96 h, while Abeta presumably induced necrotic neuronal death since apoptosis was not detectable. The p38 MAP kinase inhibitor, SB203580 (1 muM), and the PKC inhibitor, chelerythrine (5 muM) successfully inhibited the loss in viability caused by Abeta and the high density culture, respectively. Other kinase inhibitors, including the non-specific protein kinase inhibitor, H7, the PKA inhibitor 14-22 Amide, the PKG inhibitor, KT5823, and the protein tyrosine kinase inhibitor, AG18 had no effect on any of the four cytotoxic models. This system allows the study of neuroprotection under conditions where the different pathways and mechanisms of the neurons can be considered within one cellular system, removing variations which may be due to different cell type studied. The present studies describe an effective model system for screening potential neuroprotective ;agents. (C) 2002 Elsevier Science B.V. All rights reserved.