發(fā)布時(shí)間：2018-04-25 10:22

  本文選題：重組腺病毒 + 神經(jīng)細(xì)胞　； 參考：《遵義醫(yī)學(xué)院》2011年碩士論文

【摘要】：目的：用小鼠雌激素受體a(Estrogen receptorα, Erα)重組腺病毒感染神經(jīng)細(xì)胞及小鼠中樞神經(jīng)系統(tǒng)(central nervous system CNS),為研究Era在CNS發(fā)育中的作用及其與一些神經(jīng)系統(tǒng)疾病的關(guān)系奠定基礎(chǔ)。 方法：無血清培養(yǎng)C57BL/6小鼠神經(jīng)細(xì)胞,免疫組化方法鑒定神經(jīng)元的純度。腺病毒Adeno-GFP感染原代培養(yǎng)的神經(jīng)細(xì)胞,流式細(xì)胞儀(Flow cytometry, FCM)檢測細(xì)胞感染率及凋亡率,用于判斷腺病毒的最佳感染復(fù)數(shù)(Multiplicity of infection, MOI)。應(yīng)用最佳MOI重組腺病毒感染神經(jīng)細(xì)胞,實(shí)時(shí)定量PCR和western blot分別檢測受感染的神經(jīng)細(xì)胞中Era mRNA和蛋白的表達(dá)。應(yīng)用立體定位方法體內(nèi)感染CNS,觀察熒光表達(dá)。 結(jié)果：成功原代培養(yǎng)神經(jīng)細(xì)胞,神經(jīng)元純度達(dá)90%以上。MOI=100可作為Era重組腺病毒感染神經(jīng)細(xì)胞的最佳MOI, FCM檢測其感染效率為84.42±4.47%,凋亡率為3.62±0.72%。Era重組腺病毒可成功感染神經(jīng)細(xì)胞并能上調(diào)ErαmRNA及蛋白的表達(dá),亦可以成功感染CNS。 結(jié)論：Era重組腺病毒能成功感染神經(jīng)細(xì)胞及CNS并能上調(diào)ErαmRNA和蛋白表達(dá)。
[Abstract]:Aim: to study the role of a(Estrogen receptor 偽 (er 偽) in the development of CNS and its relationship with some nervous system diseases by recombinant adenovirus containing a(Estrogen receptor 偽 (er 偽) 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 鹵4.47.The apoptotic rate was 3.62 鹵0.72%.Era and the expression of er 偽 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 偽 mRNA and protein.
【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R363

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本文編號：1800919