Ceramide induces neuronal apoptosis through the caspase-9/caspase-3 pathway
Ceramide induces neuronal apoptosis through the caspase-9/caspase-3 pathway
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DOI：
10.1016/s0006-291x(02)02593-7
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发表时间：
2002-11-29
影响因子：
3.1
通讯作者：
Faden, AI
中科院分区：
文献类型：
--
作者：
Movsesyan, VA;Yakovlev, AG;Faden, AI
神经酰胺的细胞渗透性类似物c -2-神经酰胺引起培养的大鼠皮质神经元细胞死亡。c- 2神经酰胺诱导的神经元损失伴随着caspase-3活性的上调，通过其荧光底物Ac-DEVD-AMC的切割来测量。当皮层神经元培养物用鞘磷脂酶（一种在细胞中负责神经酰胺形成的酶）处理时，也得到了类似的结果。Hoechst 33258染色显示，c -神经酰胺处理的皮质神经元形态学表现为核凝聚和断裂。选择性caspase-3抑制剂z-DEVD-fmk或caspase-9抑制剂z-LEHD-fmk联合施用可显著降低c -2神经酰胺诱导的细胞死亡，而caspase-8抑制剂z-IETD-fmk联合施用可显著降低c -2神经酰胺诱导的细胞死亡。没有效果。对c -2神经酰胺处理的皮质神经元培养物的蛋白质提取物进行免疫印迹分析，发现活性caspase-9和caspase-3蛋白水平上调，而在该系统中未检测到活性caspase-8的免疫反应性。对SH-SY5Y人神经母细胞瘤细胞给予c -神经酰胺也可引起细胞凋亡。此外。在caspase-9显性阴性的SH-SY5Y细胞中，神经酰胺诱导的细胞死亡明显减少。而caspase-8显性阴性培养和模拟转染的细胞在c -神经酰胺处理后均显示出同样高水平的细胞死亡。综上所述，这些数据表明，神经酰胺诱导的神经元死亡可能与caspase-9/caspase-3调控的细胞凋亡内在途径有关。(C) 2002 Elsevier Science (USA)，版权所有。
C-2-ceramide, a cell-permeable analog of ceramide, caused cell death in cultured rat cortical neuronal cells. C-2-ceramide-induced neuronal loss was accompanied by upregulation of caspase-3 activity, measured by cleavage of its fluorogenic substrate Ac-DEVD-AMC. Similar results were obtained when cortical neuronal cultures were treated with sphingomyelinase, an enzyme responsible for ceramide formation in the cell. Morphological evaluation of C-ceramide-treated cortical neurons showed nuclear condensation and fragmentation as visualized by Hoechst 33258 staining. Co-administration of the selective caspase-3 inhibitor z-DEVD-fmk or caspase-9 inhibitor z-LEHD-fmk significantly reduced C-2-ceramide-induced cell death, while co-application of the caspase-8, inhibitor z-IETD-fmk. was without effect. Immunoblot analysis of protein extracts from C-2-ceramide-treated cortical neuronal cultures revealed upregulation of active caspase-9 and caspase-3 protein levels, whereas presence of active caspase-8 immunoreactivity was undetectable in this system. Administration of C-ceramide to SH-SY5Y human neuroblastoma cells also caused apoptotic cell death. Moreover. ceramide-induced cell death was significantly decreased in caspase-9 dominant-negative SH-SY5Y cells. while both caspase-8 dominant-negative cultures and mock-transfected cells showed equally high levels of cell death following C-ceramide treatment. Taken together, these data suggest that neuronal death induced by ceramide may be linked to the caspase-9/caspase-3 regulated intrinsic pathway of cellular apoptosis. (C) 2002 Elsevier Science (USA), All rights reserved.