HSP70蛋白在神经元样PC12细胞氧糖剥夺后的表达变化

发布时间：2018-01-02 02:06

本文关键词：HSP70蛋白在神经元样PC12细胞氧糖剥夺后的表达变化　出处：《中国老年学杂志》2013年12期 　论文类型：期刊论文

【摘要】：目的探讨神经细胞氧糖剥夺(OGD)造模后PC12细胞凋亡和热休克蛋白(HSP)70水平,评价OGD模型。方法利用PC12细胞,经NGF刺激诱导向神经细胞分化,利用OGD建立神经细胞OGD模型;利用MTT法、共聚焦显微镜鉴定PC12细胞转化为神经元样细胞及OGD模型的建立,Western印迹检测OGD后HSP70蛋白的表达。结果应用终浓度50 ng/ml NGF的DMEM完全培养基培养PC12细胞,随培养时间的增加细胞呈典型的神经元形态。微管相关蛋白(MAP)2免疫荧光细胞化学染色呈强阳性;建立OGD后神经元出现不同程度的损伤、坏死及凋亡,随OGD时间延长存活率逐渐降低、凋亡率升高、HSP70蛋白表达增加。结论 NGF有效地诱导PC12细胞转化为神经细胞,OGD可以有效诱导HSP70表达。
[Abstract]:Objective to investigate the apoptosis of PC12 cells and the level of heat shock protein (HSP70) in PC12 cells induced by oxygen glucose deprivation (OGD), and to evaluate the OGD model. Methods PC12 cells were used in this study. NGF was stimulated to induce neuronal differentiation and OGD was used to establish the neural OGD model. MTT method and confocal microscope were used to identify the transformation of PC12 cells into neuron-like cells and the establishment of OGD model. Western blotting was used to detect the expression of HSP70 protein after OGD. Results PC12 cells were cultured in 50 ng/ml NGF DMEM culture medium. With the increase of culture time, the cells showed typical neuronal morphology, and the microtubule-associated protein mitogen-activated protein (MAPF-2) was strongly positive by immunocytochemical staining. After the establishment of OGD, the neuronal damage, necrosis and apoptosis appeared in varying degrees. The survival rate decreased gradually and the apoptosis rate increased with the prolongation of OGD time. Conclusion NGF can effectively induce the transformation of PC12 cells into neuronal cells. NGF can effectively induce the expression of HSP70.
【作者单位】：吉林大学中日联谊医院神经内一科;吉林省人民医院;
【基金】：吉林省自然基金资助项目(No.201015181) 吉林省科技发展计划国际合作资助项目(No.20120723) 吉林省科技发展计划青年基金资助在项目(No.20130522024JH,201201016)
【分类号】：R363
【正文快照】： 缺血性脑损伤相关信号转导机制一直受到广泛关注〔1〕,在缺血性脑损伤研究中需建立可模拟脑缺血损伤的体外细胞模型。本实验采用神经细胞株PC12细胞以神经细胞生长因子(NGF)诱导分化为神经样细胞,进行氧糖剥夺(OGD)建立稳定神经细胞缺血缺氧模型,为进一步研究缺血性脑损伤相

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