Yuan Yuhe, Jin Jin, Yang Bo, Zhang Wei, Hu Jinfeng, Zhang Yun, Chen Nai-Hong
Department of Pharmacology, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Key Laboratory of Bioactive Substances and Resources Utilization, Ministry of Education, Beijing, 100050, PR China.
Cell Mol Neurobiol. 2008 Jan;28(1):21-33. doi: 10.1007/s10571-007-9185-6. Epub 2007 Aug 22.
Alpha-synuclein is a presynaptic protein which is implicated in some neurodegenerative disorders including Parkinson's disease, dementia with Lewy bodies, multiple systems atrophy, and Hallervorden-Spatz disease, and its overexpression contributes to the loss of dopaminergic neurons. Although the role of alpha-synuclein in these paradigms has been widely documented, its exact function is still elusive. And the dysfunction of the transcription factor nuclear factor (NF-kappaB) also exists in many neurodegenerative diseases. In this reason the purpose of this study was to investigate the molecular mechanism of alpha-synuclein's toxicity and its association with NF-kappaB by MTT assay, Western blot method, and luciferase assay. Results showed that overexpressed alpha-synuclein and 1-methyl-4-phenylpyridinium (MPP(+)) suppressed the SH-SY5Y cell viability and attenuate NF-kappaB-mediated luciferase expression significantly. Moreover, the impairment function was enhanced with the increase of alpha-synuclein protein level. We also found that overexpressed alpha-synuclein localized both in the cytoplasms and nuclei, down-regulated the anti-apoptotic Bcl-2 expression and up-regulated the pro-apoptotic glycogen synthase kinase 3beta (GSK3beta) protein level. In conclusion, all these findings mentioned above suggested that alpha-synuclein shared some toxic functional homology with neurotoxin MPP(+), and the proapoptotic effects of alpha-synuclein might be mediated at least in part by the impairment of NF-kappaB signaling pathway which involves GSK3beta.
α-突触核蛋白是一种突触前蛋白,与包括帕金森病、路易体痴呆、多系统萎缩和哈勒沃登-施帕茨病在内的一些神经退行性疾病有关,其过度表达会导致多巴胺能神经元的丧失。尽管α-突触核蛋白在这些疾病中的作用已被广泛记录,但其确切功能仍不清楚。而且转录因子核因子κB(NF-κB)的功能障碍也存在于许多神经退行性疾病中。因此,本研究的目的是通过MTT法、蛋白质印迹法和荧光素酶测定法研究α-突触核蛋白毒性的分子机制及其与NF-κB的关系。结果表明,过表达的α-突触核蛋白和1-甲基-4-苯基吡啶鎓(MPP(+))显著抑制SH-SY5Y细胞活力并减弱NF-κB介导的荧光素酶表达。此外,随着α-突触核蛋白蛋白水平的增加,损伤功能增强。我们还发现,过表达的α-突触核蛋白定位于细胞质和细胞核中,下调抗凋亡蛋白Bcl-2的表达并上调促凋亡糖原合酶激酶3β(GSK3β)的蛋白水平。总之,上述所有发现表明,α-突触核蛋白与神经毒素MPP(+)具有一些毒性功能同源性,并且α-突触核蛋白的促凋亡作用可能至少部分是由涉及GSK3β的NF-κB信号通路受损介导的。