Cell Biology Laboratory, School of Life Science, Beijing Institute of Technology, Beijing 100081, China.
Acta Biochim Biophys Sin (Shanghai). 2014 Jan;46(1):22-30. doi: 10.1093/abbs/gmt127. Epub 2013 Nov 18.
Neurotoxins and alterations in Ca2+ homeostasis have been associated with Parkinson's disease (PD), but the role of store-operated Ca2+ entry channels is not well understood. Previous studies have shown the neurotoxicity of salsolinol and 1-methyl-4-phenylpyridinium ion on SH-SY5Y cells and cytoprotection induced by transient receptor potential protein 1 (TRPC1). In the present study, N-methyl-(R)-salsolinol was tested for its cellular toxicity and effects on TRPC1 expression. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays, DAPI (4',6-diamidino-2-phenylindole), fluorescein isothiocyanate-Annexin-V/propidium iodide, western blot analysis, and JC-1 labeling revealed that the three indicated drugs could induce caspase-dependent, mitochondrial-mediated apoptosis. Exposure of SH-SY5Y cells to the indicated drugs resulted in a significant decrease in thapsigargin-mediated Ca2+ influx and TRPC1 expression. Immunocytochemistry experiments revealed that neurotoxins treatment induced TRPC1 translocation to the cytoplasm. Taken together, our results indicate that treatment with neurotoxins may alter Ca2+ homeostasis and induce mitochondrial-mediated caspase-dependent cytotoxicity, an important characteristic of PD.
神经毒素和钙稳态的改变与帕金森病（PD）有关，但储存操作的钙进入通道的作用尚不清楚。先前的研究表明，salsolinol 和 1-甲基-4-苯基吡啶离子对 SH-SY5Y 细胞具有神经毒性，瞬时受体电位蛋白 1（TRPC1）可诱导细胞保护作用。在本研究中，测试了 N-甲基-（R）-salsolinol 的细胞毒性及其对 TRPC1 表达的影响。MTT（3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐）测定法、DAPI（4'，6-二脒基-2-苯基吲哚）、荧光素异硫氰酸酯-Annexin-V/碘化丙啶、western blot 分析和 JC-1 标记显示，这三种药物均可诱导半胱天冬酶依赖性、线粒体介导的细胞凋亡。暴露于 SH-SY5Y 细胞的指示性药物导致 thapsigargin 介导的 Ca2+内流和 TRPC1 表达显著减少。免疫细胞化学实验表明，神经毒素处理诱导 TRPC1 向细胞质易位。总之，我们的结果表明，神经毒素的治疗可能会改变钙稳态并诱导线粒体介导的半胱天冬酶依赖性细胞毒性，这是 PD 的一个重要特征。
