神经干细胞在缺氧后的分化情况及相关信号通路初探  

A preliminary study on the differentiation of neural stem cells after hypoxia and related signaling pathways

作　　者：王雨晴 陈志刚[2] 张斯佳 王荣亮 罗玉敏 赵海苹 WANG Yuqing;CHEN Zhigang;ZHANG Sijia;WANG Rongliang;LUO Yumin;ZHAO Haiping(Cerebrovascular Diseases Research Institute,Xuanwu Hospital of Capital Medical University,Beijing 100053,China;Department of Encephalopathy,Dongfang Hospital of Bejing University of Traditional Chinese Medicine,Beijing 100078)

机构地区：[1]首都医科大学宣武医院脑血管病研究室,北京100053 [2]北京中医药大学东方医院脑病一科,北京100078

出　　处：《中国比较医学杂志》2021年第1期7-15,共9页Chinese Journal of Comparative Medicine

基　　金：国家自然科学基金项目(81771412,81801149)。

摘　　要：目的观察缺氧缺糖后神经干细胞在不同时间点的分化趋势,以及与信号通路促红细胞生成素受体(EPOR和βCR)和PDZ连接激酶(PBK)的相关性。方法分离小鼠原代神经干细胞,制备氧糖剥夺(OGD)模型,缺氧3 h,复氧1 h、2 h、4 h、6 h、8 h,观察神经干细胞OGD的分化趋势,采用RT-PCR技术检测少突胶质细胞标志物CNP和MBP、星型胶质细胞标志物GFAP和S100B、神经元标志物RBFOX3、MAP2和TUBB3以及信号分子EPOR、CSF2RB、PBK的基因表达,并进行了Pearson相关性分析。结果 (1) OGD处理促进神经干细胞向少突胶质细胞方向分化,Cnp和Mbp表达呈升高趋势,复氧4 h Cnp显著升高。(2) OGD处理促进神经干细胞向星形胶质细胞方向分化,复氧2-8 h S100b显著升高,复氧2 h和6 h Gfap表达显著降低,复氧8 h表达显著增加。(3) OGD处理减少神经干细胞向神经元方向分化,Rbfox3以及Tubb3表达显著降低,复氧2 h和6 h最为显著。(4) Pbk在复氧1 h和8 h显著降低,Epor、Pbk与Cnp的表达呈正相关,Pbk与Mbp的表达呈正相关。结论缺氧缺糖损伤显著促进神经干细胞向少突胶质细胞和星形胶质细胞新生,同时抑制向神经元方向的分化。EPOR以及PBK可能参与到脑缺血后神经干细胞向少突胶质细胞分化的调控过程,为进一步研究相关信号通路在缺血后神经再生中的调控机制提供了基础。Objective To observe the trend of differentiation of neural stem cells(NSCS)at different time points after oxygen glucose deprivation/reoxygenation(OGD/R)and its correlation with Erythropoietin Receptor(EPOR),β-Common Receptor(βCR)and PDZ-binding-kinase(PBK).Methods The primary neural stem cells of mice were isolated and the OGD model was made.The OGD was hypoxia for 3 hours,reoxygenation for 1 hour,2 hours,4 hours,6 hours and8 hours.The differentiation trend of neural stem cells was observed.CNP and MBP,GFAP and S100 B,RBFOX3,MAP2 and TUBB3,EPOR,CSF2 RB and PBK were detected by RT-PCR and Pearson correlation analysis was carried out.Results 1,OGD treatment promoted the differentiation of neural stem cells to oligodendrocytes,and the expressions of Cnp and Mbp were increased obviously at 4 h after OGD.2,OGD treatment promoted the differentiation of neural stem cells to astrocytes,and the expression of Gfap was decreased at 2~6 h after OGD,while the expression of Gfap was increased at1 h and 8 h after OGD.The expression of S100 b was significantly increased at 2~8 h after OGD.3,OGD treatment reduced neuronal differentiation of neural stem cells,and the expression of Rbfox3 and Tubb3 were decreased after OGD significantly at 2 h and 6 h after OGD.4,Pbk was reduced at 1 h and 8 h after OGD.Epor and Pbk were positively correlated with the expression of Cnp,and Pbk was positively correlated with the expression of Mbp.Conclusions Hypoxia injury significantly promoted the formation of oligodendrocytes and astrocytes and inhibited the neuronal differentiation of neural stem cells.EPOR and PBK may be involved in the differentiation of neural stem cells to oligodendrocytes.These result may provide a basis for the further research on the related mechanism of nerve regeneration after ischemia.

关 键 词：神经干细胞分化 缺氧 促红细胞生成素受体 T-LAK细胞源蛋白激酶 

分 类 号：R-33[医药卫生]