α-突触核蛋白对MES23.5多巴胺能神经细胞的促增殖作用

Effect of alpha-synuclein in promoting the proliferation of MES23.5 dopaminergic neuronal cells

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Westernblot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.