载神经生长因子纳米粒诱导神经干细胞分化为神经元及PI3K/Akt通路的影响  

Influence of NGF-PEG-PLGA-NPs on neuronal differentiation of neural stem cells and PI3K/Akt signaling pathway

作　　者：陈艳[1] 包国庆[2] 刘菲菲[2] 张君度[2] 潘翠环[1] 龙大宏[2] 

机构地区：[1]广州医科大学附属第二医院康复医学科,广东省广州市510260 [2]广州医科大学解剖教研室,广东省广州市510182

出　　处：《中国组织工程研究》2015年第28期4576-4581,共6页Chinese Journal of Tissue Engineering Research

基　　金：广州医科大学青年基金项目(2011A17);项目名称:NGF与b-FGF联合诱导神经干细胞分化为胆碱能神经元~~

摘　　要：背景:课题组前期研究证实NGF-PEG-PLGA-NPs在体外有良好的缓释效能及生物活性,可以诱导PC12细胞向神经元样细胞分化。目的:探讨NGF-PEG-PLGA-NPs诱导胎鼠大脑隔区来源神经干细胞分化为神经元的可行性及其对PI3K/Akt信号通路的影响。方法:采用优化处方,复乳化溶剂扩散法制备NGF-PEG-PLGA-NPs。设对照组、NGF组、NGF-PEG-PLGA-NPs组、LY294002组、LY294002+NGF组、LY294002+NGF-PEG-PLGA-NPs组,诱导神经干细胞分化为神经元,细胞免疫荧光染色对神经元进行鉴定,Werstern-blotting检测PI3K/Akt信号通路Akt磷酸化水平。结果与结论:对照组、NGF组、NGF-PEG-PLGA-NPs组、LY294002组、LY294002+NGF组、LY294002+NGF-PEG-PLGA-NPs组β-TubulinⅢ阳性神经元分化率分别为(22.80±2.58)%,(35.80±3.98)%,(35.40±5.77)%,(26.60±3.87)%,(21.20±2.59)%,(25.80±7.22)%。NGF组与NGF-PEG-PLGA-NPs组神经元分化率差异无显著性意义(P>0.05),但两组神经元分化率均高于其他各组(P<0.05)。Western blotting检测结果显示:NGF组及NGF-PEG-PLGA-NPs组Akt磷酸化水平差异无显著性意义(P>0.05),但均高于其他各组(P<0.05);LY294002+NGF组及LY294002+NGF-PEG-PLGA-NPs组Akt磷酸化水平与对照组比较差异无显著性意义(P>0.05),但均高于LY294002组(P<0.05)。结果表明NGF-PEG-PLGA-NPs可促进神经干细胞分化为神经元,其可能机制与促进PI3K/Akt信号通路Akt的磷酸化有关。BACKGROUND：Our previous studies confirmed that NGF-PEG-PLGA-NPs has good sustained release effect and biological activity in vitro, and can induce the differentiation of PC12 cel s into neuron-like cel s. OBJECTIVE：To investigate the feasibility of neuronal differentiation of neural stem cel s from septal area of fetal brain induced by NGF-PEG-PLGA-NPs and its influence on PI3K/Akt signaling pathway. METHODS：According to optimization prescription, NGF-PEG-PLGA-NPs were prepared by multiple emulsion solvent diffusion method. Neural stem cel s were induced to neuronal differentiation in six groups, including control group, NGF group, NGF-PEG-PLGA-NPs group, LY294002 group, LY294002＋NGF group and LY294002＋NGF-PEG-PLGA-NPs group. Neurons were identified by immunofluorescence, while phosphorylation levels of Akt in PI3K/Akt signaling pathway were detected by western blotting. RESULTS AND CONCLUSION：The proportions ofβ-Tubulin III-positive neurons in control group, NGF group, NGF-PEG-PLGA-NPs group, LY294002 group, LY294002＋NGF group and LY294002＋NGF-PEG-PLGA-NPs group were （22.80±2.58）%, （35.80±3.98）%, （35.40±5.77）%, （26.60±3.87）%, （21.20±2.59）%and （25.80±7.22）%, respectively. There were no statistical differences in neuronal differentiation between NGF group and NGF-PEG-PLGA-NPs group （P〉0.05）, but the ratios of neural differentiation in the two groups were both higher than that in the other four groups （P〈0.05）. Western blotting results revealed that there were no statistical differences in Akt phosphorylation levels between NGF group and NGF-PEG-PLGA-NPs group （P〉0.05）, but the phosphorylation levels of Akt were both higher than other four groups （P〈0.05）. There were also no significant differences between LY294002＋NGF and LY294002＋NGF-PEG-PLGA-NPs groups and control group （P〉0.05）, but the phosphorylation levels of Akt were higher than LY294002 group （P〈0.05）. Results suggest that NGF-PEG-PLGA-NPs promoted neural differentiat

关 键 词：干细胞 分化 神经生长因子 纳米粒 神经干细胞 神经元 信号通路 

分 类 号：R394.2[医药卫生—医学遗传学]