LINGO-1在脊髓神经干细胞分化中的表达及影响

发布时间：2018-05-01 13:09

本文选题：LINGO- + 脊髓神经干细胞　；参考：《中国脊柱脊髓杂志》2017年09期

【摘要】：目的 :观察LINGO-1在大鼠脊髓神经干细胞(spinal cord derived neural stem cells,SpNSCs)分化过程中的表达特征及其对SpNSCs分化的影响。方法:分别在体外分离培养3只10周龄Fischer 344大鼠SpNSCs,培养至第3代对其进行干性Nestin表达鉴定和观察在其分化过程中LINGO-1的表达量(分化第0、1、3、5天)。随后用包装好的LINGO-1 shRNA慢病毒及Scramble shRNA慢病毒感染SpNSCs,观察感染效率及对比感染后下调LINGO-1蛋白表达的效果。在确定LINGO-1 shRNA的下调效果后,将第4代SpNSCs分为干扰组及对照组,干扰组感染LINGO-1 shRNA慢病毒,对照组感染Scramble-sh RNA慢病毒,于分化5d后使用免疫荧光染色检测神经元及星形胶质细胞的比例。结果:体外分离培养的SpNSCs表达Nestin阳性。LINGO-1蛋白的积分光密度(integrated option density,IOD)在第0、1、3、5天分别为514±45、715±68、1017±92、654±61,LINGO-1从SpNSCs分化第1天起表达量上升,为第0天表达量的1.39倍,至分化第3天为第0天表达量的1.98倍,随后表达下降,至第5天为第0天表达量的1.36倍,以上4个时间点LINGO-1蛋白表达量有统计学差异(P0.05)。LINGO-1 shRNA慢病毒感染效率90%,感染LINGO-1 shRNA慢病毒的SpNSCs的LINGO-1蛋白表达量为感染Scramble-sh RNA慢病毒Sp NSCs的26%。干扰组神经元分化比例为对照组的3.85倍,星形胶质细胞分化比例为对照组的0.65倍,两组神经元分化比例及星形胶质细胞分化比例的差异均有统计学意义(P0.05)。结论 :LINGO-1在大鼠SpNSCs分化初期表达上升,抑制LINGO-1表达可促进Sp NSCs向神经细胞分化,减少其向星形胶质细胞分化。
[Abstract]:Aim: to investigate the expression of LINGO-1 in the differentiation of spinal cord derived neural stem cells and its effect on SpNSCs differentiation in rats. Methods: SpNSCs of 3 10-week-old Fischer rats were isolated and cultured in vitro. The dry Nestin expression of SpNSCs was identified in the third generation and the expression of LINGO-1 was observed during the differentiation. Then LINGO-1 shRNA lentivirus and Scramble shRNA lentivirus were used to infect SpNSCs. The infection efficiency was observed and the effect of down-regulating LINGO-1 protein expression was compared. After determining the down-regulation effect of LINGO-1 shRNA, the fourth generation of SpNSCs was divided into interference group and control group. The interfering group was infected with LINGO-1 shRNA lentivirus, and the control group was infected with Scramble-sh RNA lentivirus. The proportion of neurons and astrocytes was detected by immunofluorescence staining 5 days after differentiation. Results: the integrated optical density of SpNSCs expressing Nestin positive. LINGO-1 protein in vitro was 514 卤45715 卤681017 卤92654 卤61LINGO-1 on the 5th day of SpNSCs differentiation, which was 1.39 times higher than that on day 0. On the third day of differentiation, the expression level was 1.98 times of that on day 0, then decreased, and on the fifth day it was 1.36 times of that on day 0. There was a significant difference in the expression of LINGO-1 protein among the four time points. The infection efficiency of LINGO-1 shRNA lentivirus was 90. The expression of LINGO-1 protein in SpNSCs infected with LINGO-1 shRNA lentivirus was 26% of that of Scramble-sh RNA lentivirus spp NSCs. The proportion of neuron differentiation in interference group was 3.85 times of that in control group, and the proportion of astrocyte differentiation in interference group was 0.65 times of that in control group. There were significant differences between the two groups in the proportion of neuron differentiation and astrocyte differentiation. Conclusion the expression of 1% LINGO-1 increased at the early stage of SpNSCs differentiation in rats. Inhibiting the expression of LINGO-1 could promote the differentiation of SP NSCs into neural cells and decrease the differentiation of Sp NSCs into astrocytes.
【作者单位】：空军军医大学第一附属医院康复医学科;新疆军区总医院全军骨科中心;
【基金】：国家自然科学基金(编号:81100932) 中国博士后科研基金(编号:20100481518)
【分类号】：R329.2

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