Caspase-2 and caspase-8 trigger caspase-3 activation following 6-OHDA-induced stress in human dopaminergic neurons differentiated from ReNVM stem cells
Caspase-2 and caspase-8 trigger caspase-3 activation following 6-OHDA-induced stress in human dopaminergic neurons differentiated from ReNVM stem cells
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DOI：
10.1179/1743132812y.0000000135
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发表时间：
2013-05-01
影响因子：
1.9
通讯作者：
Ahmed, Bushra Y.
中科院分区：
文献类型：
--
作者：
Chaudhry, Zohara L.;Ahmed, Bushra Y.
目的：本研究的目的是建立一个合适的模型来研究分化的多巴胺能神经元（dDCN）中帕金森病（PD）的发病机制。具体目的是证明 caspase 家族的参与，并鉴定由 6-羟基多巴胺 (6OHD) 处理激活导致 dDCN 死亡的特定 caspase。 方法：将 ReNcell VM 细胞分化为 dDCN，并暴露于 6OHD 以诱导应激。在未处理和 6OHD 处理的 dDCN 中进行了 caspase-2、-3 和 -8 的蛋白质印迹 (WB) 和双重免疫荧光分析。 zVADfmk 用于确定它是否可以在 6OHD 介导的应激后抑制 dDCN 中 caspase-2、-3 和 -8 的激活。结果：我们的免疫荧光和 WB 数据显示，6OHD 触发 caspase-2 和 -8 激活，进而激活 caspase-3 导致 dDCN 死亡。此外，WB 分析显示，在 6OHD 处理的细胞中，zVADfmk 减少了 caspase-2、-3 和 -8 的激活。讨论：该研究表明，6OHD 诱导的毒性触发了 caspase 介导的 dDCN 死亡。这一发现支持了之前使用不同 PD 模型的研究，表明 6OHD 可以通过凋亡途径诱导 caspase-2 和 -3 激活，并且这两种 caspase 都可以激活 PD 中的 caspase-3。此外，我们的结果表明 caspase-2 可能通过间接 NF kappaB 途径导致细胞死亡。这项研究建立了帕金森病模型，可以在生化和分子水平上更好地了解帕金森病的发病机制，从而更好地了解帕金森病和新治疗的潜力。
Objectives: The purpose of the study was to establish a suitable model to study Parkinson's disease (PD) pathogenesis in differentiated dopaminergic neurons (dDCN). The specific aim was to demonstrate the involvement of the caspase family and to identify specific caspases which are activated by 6-hydroxydopamine (6OHD) treatment leading to death of dDCN.Methods: ReNcell VM cells were differentiated into dDCN and were exposed to 6OHD to induce stress. Western blot (WB) and double immunofluorescent analyses of caspases-2, -3, and -8 were carried out in untreated and 6OHD-treated dDCN. zVADfmk was used to determine if it could inhibit activation of caspases-2, -3, and -8 in dDCN following 6OHD-mediated stress.Results: Our immunofluorescent and WB data showed that 6OHD triggered caspases-2 and -8 activation which in turn activated caspase-3 leading to death of dDCN. Additionally, WB analysis revealed that caspases-2, -3, and -8 activation was reduced by zVADfmk in 6OHD-treated cells.Discussion: The study showed that 6OHD-induced toxicity triggered caspase mediated death of dDCN. This finding is in support of previous studies using different PD model showing that 6OHD can induce caspases-2 and -3 activation through apoptotic pathway and that both caspases can activate caspase-3 in PD. In addition, our results suggest that caspase-2 cause's cell death might be via an indirect NF kappaB route. This study has established a PD model which can provide better insight to PD pathogenesis on a biochemical and molecular level, leading to a better understanding of PD and potential for new treatments.