文章摘要
张〓俊,刘锦波,张志坚,龚爱华,刘志元.嗅鞘细胞条件培养液对PC12细胞神经丝蛋白?突触蛋白?突触素蛋白表达的影响[J].南京医科大学学报,2010,(3):328~
嗅鞘细胞条件培养液对PC12细胞神经丝蛋白?突触蛋白?突触素蛋白表达的影响
Effect of olfactory ensheathing cell conditioned medium on NF,synapsin and synaptophysin expression in PC12 cells
投稿时间:2009-10-14  
DOI:10.7655
中文关键词: 嗅鞘细胞  条件培养液  PC12细胞  神经丝蛋白  synapsin  synaptophysin
英文关键词: olfactory ensheathing cells  conditioned medium  PC12 cells  NF  synapsin  synaptophysin
基金项目:江苏省自然基金项目(BK2009101),国家自然基金项目(30571878)资助
作者单位
张〓俊 南京医科大学附属常州二院中心实验室,江苏 常州〓213003 
刘锦波 常州第一人民医院骨科,江苏 常州〓213003 
张志坚 江苏大学基础医学与医学技术学院,江苏 镇江〓212013 
龚爱华 江苏大学基础医学与医学技术学院,江苏 镇江〓212013 
刘志元 南京医科大学附属常州二院中心实验室,江苏 常州〓213003 
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中文摘要:
      目的:观察嗅鞘细胞(olfactory ensheathing cells,OECs)条件培养液对PC12细胞神经丝蛋白(neurofilament protein,NF)?突触蛋白(synapsin)和突触素蛋白(synaptophysin)表达的影响,探讨嗅鞘细胞条件培养液促神经细胞分化生长的作用机制?方法:原代培养并纯化OECs,收集培养上清制备成嗅鞘细胞培养液(OEC culture medium,OECCM),与PC12细胞培养24?48?72 h,观察细胞形态学变化;通过免疫荧光法检测PC12细胞NF的表达及分布;蛋白质印迹法检测NF?突触蛋白和突触素的相对含量?结果:OECCM培养的PC12细胞长有突起,并随着培养时间的延长,细胞形态酷似神经元;免疫荧光染色显示NF起初在核周分布,OECCM作用后NF的分布范围逐渐增加,其分布面积与β-tubulin分布面积的比值显著增高(P < 0.05)?蛋白质印迹法显示OECCM组NF?突触蛋白(synapsin)和突触素蛋白(synaptophysin)的表达水平明显高于对照组(P < 0.05)?结论:嗅鞘细胞能分泌一些促PC12细胞分化的因子,这些因子能促进PC12细胞NF骨架的组装,并诱导神经元功能蛋白的表达,使该细胞在结构和功能上向神经元分化?
英文摘要:
      Objective:To observe the effect of olfactory ensheathing cells (OECs) conditioned medium on NF,synapsin and synaptophysin expression in PC12 cells,and investigate the mechanism of olfactory ensheathing cell conditioned medium to promote growth of nerve cell differentiation. Methods:Primarily cultured OECs were prepared and purified,and the purified cell conditioned medium was collected into the olfactory ensheathing cell culture medium,then used to culture PC12 cells in 24 h,48 h,72 h. In vitro the morphological changes of the PC12 cells were observed under a phase contrast microscope;The expression and distribution of NF was determined by immunofluorescence methed;The relative content of NF,synapsin and synaptophysin proteins was detected by Western blot. Results:PC12 cells have long processes cultured in OECCM,and with the extension of the culture time,the cell morphology resembles neurons. Immunofluorescence staining showed the distribution of NF initially in the perinuclear. After the effects of OECCM the distribution of NF,increased and the ratio to β-tubulin was significantly high(P < 0.05).Western blotting showed the expression level of NF synapsin,and synaptophysin was significantly higher in OECCM group than in control group(P < 0.05). Conclusion:Olfactory ensheathing cells can secrete a number of cytokines to promote PC12 cell differentiation. These cytokines can promote the assembly of NF skeleton in PC12 cells,and induce neuronal function protein expression. So the cells can differentiate to the neurons in the structure and function.
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