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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。
Objective: To analyze the effect of exogenous wild-type α-synuclein on the proliferation of dopaminergic neurons and its molecular mechanism. METHODS: The experiment was performed at the Neurobiology Room of Beijing Institute of Geriatrics, 2005-04 / 11. Recombinant human wild-type α-synuclein was added to the culture medium of MES23.5 dopaminergic neurons and incubated for 48 hours, the distribution of α-synuclein in the cells was detected by immunofluorescence labeling and Western blot analysis, The growth curve was drawn by MTS method to observe the cell proliferation rate. The gene expression profile of α-synuclein-treated cells was observed by gene chip technique. Results: Recombinant human wild-type α-synuclein can enter into MES23.5 dopaminergic neurons and promote their proliferation. There was a significant change in the expression of 22 genes in α-synuclein-treated cells, with 3 genes related to endocytosis, 5 related to transcription, 3 related to protein synthesis, and 3 to cell growth and proliferation , 4 were related to differentiation, 1 was related to proliferation and differentiation, 2 were related to the formation of vesicles and the release of neurotransmitter, and 1 was related to the physiological cycle. CONCLUSION: Exogenous α-synuclein may enter into MES23.5 dopaminergic neurons through endocytosis to promote its proliferation; the effect of α-synuclein on cell proliferation may be related to its effect on the expression of certain genes.