目的:分析外源性野生型α-突触核蛋白对多巴胺能神经细胞增殖的影响及其分子机制。 方法:实验于2005—04/11在北京老年病研究所神经生物室完成。向MES23.5多巴胺能神经细胞的培养基中加入重组人野生型α-突触核蛋白,孵育48h后用细胞免疫荧光标记法和Western blot分析法检测α-突触核蛋白在细胞内的分布,用MTS法绘制生长曲线观察细胞增殖率,用基因芯片分技术观察α-突触核蛋白处理细胞的基因表达谱变化。 结果:重组人野生型α-突触核蛋白可以进入MES23.5多巴胺能神经细胞,并促进其增殖。在α-突触核蛋白处理的细胞。有22个基因的表达发生明显变化,有3个基因与内吞相关,5个与转录有关,3个与蛋白质合成有关,3个与细胞生长和增殖有关,4个与分化有关,1个与增殖及分化均有关,2个与小泡生成和神经递质释放有关,1个与生理周期有关。 结论:外源性α-突触核蛋白可能通过内吞作用进入MES23.5多巴胺能神经细胞。从而促进其增殖;α-突触核蛋白的促细胞增殖作用可能与其影响某些基因表达有关。
AIM: To analyze the influence of the exogenous wild type alpha-synuclein (α-synuclein) on the proliferation of MES23.5 dopaminergic neuronal cells and the molecular mechanism. METHODS: The experiment was carried out in the Beijing Institute of Geriatrics between April and November 2005. Recombinant wild type human α-synuclein was added to the medium of MES23.5 dopaminergic neuronal cells. Immunofluorescent labeling and Western blot were used to detect the distribution of α-synuclein in the cells after 48-hour incubation. The growth curve was drawn With the MTS method to observe the proliferative rate ceils. Gene chips technique was introduced to analyze the gene expression of the α- synuclein-treated cells. RESULTS: Recombinant a-synuclein could enter into ME523.5 cells after addition to the culture medium and promote the proliferation of the cells, Of the α-synuclein-treated ceils, 22 genes were found to he changed obviously, including 3 genes were correlated with endocytosis, 5 with transcription, 3 with protein synthesis, 3 with cell growth mid proliferation, 4 with differentiation, 1 with proliferation and differentiation, 2 with generation Of vesicle and neurotransmitter release, and 1 with cell cycle. CONCLUSION; Exogenous wild type α-synuclein can enter into MES23.5 dopaminergic neuronal cells by endocytosis, and then promote its proliferation, which may be correlated with its influence on the gene expression.