α-突触核蛋白可保护SH-SY5Y细胞免受多巴胺毒性的影响。
alpha-Synuclein protects SH-SY5Y cells from dopamine toxicity.
作者信息
Colapinto Monica, Mila Silvia, Giraudo Sabrina, Stefanazzi Paola, Molteni Monica, Rossetti Carlo, Bergamasco Bruno, Lopiano Leonardo, Fasano Mauro
机构信息
Department of Structural and Functional Biology, and Centre of Neuroscience, University of Insubria, Via Alberto da Giussano 12, 21052 Busto Arsizio, VA, Italy.
出版信息
Biochem Biophys Res Commun. 2006 Nov 3;349(4):1294-300. doi: 10.1016/j.bbrc.2006.08.163. Epub 2006 Sep 5.
Dopaminergic human neuroblastoma SH-SY5Y cells were stably transformed to increase expression of alpha-synuclein, a Parkinson's disease-related protein. Transformed cells were more resistant to oxidative insults, showing a cytoprotective role of alpha-synuclein. The expression of redox chaperonins (DJ-1, HSP70, and 14-3-3) was evaluated by Western blotting. Expression of alpha-synuclein reduced HSP70 levels even in the presence of dopamine, with a twofold increase of DJ-1 in the absence of oxidants. DJ-1 is significantly reduced by dopamine, and even more by dopamine and Cu(II). Increased alpha-synuclein expression did not affect 14-3-3, although dopamine increased its level by 60% in wild-type cells. alpha-Synuclein not only upregulated DJ-1, but also shifted all DJ-1 forms to a single spot at pI=5.7 not observed in wild-type cells. Dopamine gradually restored the distribution of DJ-1 forms to a situation similar to wild-type cells, with the form at pI=6.1 progressively enriched under oxidative conditions.
多巴胺能人类神经母细胞瘤 SH-SY5Y 细胞被稳定转化以增加与帕金森病相关的蛋白质 α-突触核蛋白的表达。转化后的细胞对氧化损伤更具抗性，显示出 α-突触核蛋白的细胞保护作用。通过蛋白质免疫印迹法评估氧化还原伴侣蛋白（DJ-1、HSP70 和 14-3-3）的表达。即使在多巴胺存在的情况下，α-突触核蛋白的表达也会降低 HSP70 的水平，在无氧化剂的情况下 DJ-1 增加两倍。多巴胺会显著降低 DJ-1 的水平，多巴胺和铜（II）共同作用时降低得更多。α-突触核蛋白表达增加并未影响 14-3-3 的水平，尽管多巴胺使野生型细胞中 14-3-3 的水平提高了 60%。α-突触核蛋白不仅上调 DJ-1 的表达，还将所有 DJ-1 形式转移到野生型细胞中未观察到的等电点为 5.7 的单个条带处。多巴胺逐渐将 DJ-1 形式的分布恢复到与野生型细胞相似的状态，在氧化条件下，等电点为 6.1 的条带逐渐富集。
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