本文選題：LINGO- + 脊髓神經(jīng)干細胞　； 參考：《中國脊柱脊髓雜志》2017年09期

【摘要】：目的 :觀察LINGO-1在大鼠脊髓神經(jīng)干細胞(spinal cord derived neural stem cells,SpNSCs)分化過程中的表達特征及其對SpNSCs分化的影響。方法:分別在體外分離培養(yǎng)3只10周齡Fischer 344大鼠SpNSCs,培養(yǎng)至第3代對其進行干性Nestin表達鑒定和觀察在其分化過程中LINGO-1的表達量(分化第0、1、3、5天)。隨后用包裝好的LINGO-1 shRNA慢病毒及Scramble shRNA慢病毒感染SpNSCs,觀察感染效率及對比感染后下調(diào)LINGO-1蛋白表達的效果。在確定LINGO-1 shRNA的下調(diào)效果后,將第4代SpNSCs分為干擾組及對照組,干擾組感染LINGO-1 shRNA慢病毒,對照組感染Scramble-sh RNA慢病毒,于分化5d后使用免疫熒光染色檢測神經(jīng)元及星形膠質(zhì)細胞的比例。結(jié)果:體外分離培養(yǎng)的SpNSCs表達Nestin陽性。LINGO-1蛋白的積分光密度(integrated option density,IOD)在第0、1、3、5天分別為514±45、715±68、1017±92、654±61,LINGO-1從SpNSCs分化第1天起表達量上升,為第0天表達量的1.39倍,至分化第3天為第0天表達量的1.98倍,隨后表達下降,至第5天為第0天表達量的1.36倍,以上4個時間點LINGO-1蛋白表達量有統(tǒng)計學(xué)差異(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蛋白表達量為感染Scramble-sh RNA慢病毒Sp NSCs的26%。干擾組神經(jīng)元分化比例為對照組的3.85倍,星形膠質(zhì)細胞分化比例為對照組的0.65倍,兩組神經(jīng)元分化比例及星形膠質(zhì)細胞分化比例的差異均有統(tǒng)計學(xué)意義(P0.05)。結(jié)論 :LINGO-1在大鼠SpNSCs分化初期表達上升,抑制LINGO-1表達可促進Sp NSCs向神經(jīng)細胞分化,減少其向星形膠質(zhì)細胞分化。
[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 鹵45715 鹵681017 鹵92654 鹵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.
【作者單位】： 空軍軍醫(yī)大學(xué)第一附屬醫(yī)院康復(fù)醫(yī)學(xué)科;新疆軍區(qū)總醫(yī)院全軍骨科中心;
【基金】：國家自然科學(xué)基金(編號:81100932) 中國博士后科研基金(編號:20100481518)
【分類號】：R329.2

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