The specific Na+/Ca2+ exchange inhibitor SEA0400 prevents nitric oxide-induced cytotoxicity in SH-SY5Y cells
The specific Na+/Ca2+ exchange inhibitor SEA0400 prevents nitric oxide-induced cytotoxicity in SH-SY5Y cells
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DOI：
10.1016/j.neuint.2011.03.026
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发表时间：
2011-08-01
影响因子：
4.2
通讯作者：
Matsuda, Toshio
中科院分区：
文献类型：
--
作者：
Nashida, Tetsuaki;Takuma, Kazuhiro;Matsuda, Toshio
Na+/Ca 2+交换器（NCX）在细胞内Ca 2+水平的调节中起作用，并且一氧化氮（NO）参与包括神经退行性疾病在内的许多病理状况。我们以前已经发现，硝普钠（SNP），一个NO供体，通过NCX介导的途径在培养的神经胶质细胞中引起神经胶质样细胞死亡，并且NO诱导的细胞毒性的机制是细胞类型依赖性的。本研究使用特异性NCX抑制剂2-[4-[（2，5-二氟苯基）甲氧基]苯氧基]-5-乙氧基苯胺（SEA 0400）检测NCX是否参与NO诱导的培养神经细胞损伤。SNP可诱导神经母细胞瘤SH-SY 5 Y细胞凋亡，其作用可被丝裂原活化蛋白（MAP）/细胞外信号调节激酶（ERK）激酶抑制剂U 0126和p38 MAP激酶（MAPK）抑制剂SB 203580阻断，但不能被c-Jun N末端激酶（INK）抑制剂SP 60012阻断。SNP增加细胞内Ca ~（2+）内流和细胞内Ca ~（2+）水平。此外，SNP增加ERK和p38 MAPK磷酸化，并以细胞外Ca 2+依赖的方式产生活性氧（ROS）。SNP的这些作用被SEA 0400所阻止。SNP诱导的细胞毒性不受Ca 2+，Na+和存储操作/容量通道的抑制剂的影响。此外，SNP诱导的细胞内Ca 2+水平增加，ROS产生和细胞活力下降被cGMP依赖性蛋白激酶（PKG）抑制剂阻断。上述结果提示，NO通过鸟苷酸环化酶/PKG途径激活NCX，并通过NCX的反向Ca ~（2+）内流参与NO诱导神经元凋亡的级联反应。(C)2011 Elsevier BY. All rights reserved.
The Na+/Ca2+ exchanger (NCX) plays a role in the regulation of intracellular Ca2+ levels, and nitric oxide (NO) is involved in many pathological conditions including neurodegenerative disorders. We have previously found that sodium nitroprusside (SNP), an NO donor, causes apoptotic-like cell death in cultured glial cells via NCX-mediated pathways and the mechanism for NO-induced cytotoxicity is cell type-dependent. The present study examined using the specific NCX inhibitor 2-[4-[(2,5-difluorophenyl)methoxy]phenoxy]-5-ethoxyaniline (SEA0400) whether NCX is involved in NO-induced injury in cultured neuronal cells. The treatment of neuroblastoma SH-SY5Y cells with SNP resulted in apoptosis and the cytotoxicity was blocked by the mitogen-activated protein (MAP)/extracellular signal-regulated kinase (ERK) kinase inhibitor U0126 and the p38 MAP kinase (MAPK) inhibitor SB203580, but not by the c-Jun N-terminal kinase (INK) inhibitor SP60012. SNP increased Ca2+ influx and intracellular Ca2+ levels. In addition, SNP increased ERK and p38 MAPK phosphorylation, and production of reactive oxygen species (ROS) in an extracellular Ca2+-dependent manner. These effects of SNP were prevented by SEA0400. SNP-induced cytotoxicity was not affected by inhibitors of the Ca2+, Na+ and store-operated/capacitative channels. Moreover, SNP-induced increase in intracellular Ca2+ levels, ROS production and decrease in cell viability were blocked by a cGMP-dependent protein kinase (PKG) inhibitor. These results suggest that Ca2+ influx via the reverse of NCX is involved in the cascade of NO-induced neuronal apoptosis and NO activates the NCX through guanylate cyclase/PKG pathway. (C) 2011 Elsevier BY. All rights reserved.