小鼠雌激素受体α重组腺病毒感染神经细胞及小鼠中枢神经系统的鉴定

发布时间：2018-04-25 10:22

本文选题：重组腺病毒 + 神经细胞　；参考：《遵义医学院》2011年硕士论文

【摘要】：目的：用小鼠雌激素受体a(Estrogen receptorα, Erα)重组腺病毒感染神经细胞及小鼠中枢神经系统(central nervous system CNS),为研究Era在CNS发育中的作用及其与一些神经系统疾病的关系奠定基础。方法：无血清培养C57BL/6小鼠神经细胞,免疫组化方法鉴定神经元的纯度。腺病毒Adeno-GFP感染原代培养的神经细胞,流式细胞仪(Flow cytometry, FCM)检测细胞感染率及凋亡率,用于判断腺病毒的最佳感染复数(Multiplicity of infection, MOI)。应用最佳MOI重组腺病毒感染神经细胞,实时定量PCR和western blot分别检测受感染的神经细胞中Era mRNA和蛋白的表达。应用立体定位方法体内感染CNS,观察荧光表达。结果：成功原代培养神经细胞,神经元纯度达90%以上。MOI=100可作为Era重组腺病毒感染神经细胞的最佳MOI, FCM检测其感染效率为84.42±4.47%,凋亡率为3.62±0.72%。Era重组腺病毒可成功感染神经细胞并能上调ErαmRNA及蛋白的表达,亦可以成功感染CNS。结论：Era重组腺病毒能成功感染神经细胞及CNS并能上调ErαmRNA和蛋白表达。
[Abstract]:Aim: to study the role of a(Estrogen receptor 伪 (er 伪) in the development of CNS and its relationship with some nervous system diseases by recombinant adenovirus containing a(Estrogen receptor 伪 (er 伪) from mouse central nervous system (CNS) and central nervous system (CNS) of mouse central nervous system (CNS). Methods: C57BL/6 mouse nerve cells were cultured without serum, and the purity of neurons was identified by immunohistochemical method. The infection rate and apoptosis rate of primary cultured nerve cells were detected by flow cytometry (FCM) with adenovirus Adeno-GFP, which was used to determine the optimal infection number of adenovirus, multiplicity of infecting, moi. The optimal MOI recombinant adenovirus was used to infect nerve cells. The expression of Era mRNA and protein in infected neurons was detected by real-time quantitative PCR and western blot, respectively. The expression of CNS in vivo was observed by stereotactic method. Results: nerve cells were successfully cultured in primary culture. The purity of neurons was more than 90%. MOI could be used as the best moi for Era recombinant adenovirus infection. The infection efficiency of FCM was 84.42 卤4.47.The apoptotic rate was 3.62 卤0.72%.Era and the expression of er 伪 mRNA and protein could be upregulated. CNS can also be infected successfully. Conclusion the recombinant adenovirus can infect nerve cells and CNS successfully and upregulate the expression of er 伪 mRNA and protein.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

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