摘要
目的探讨PDGFR-β在神经干细胞(NSC)多向分化过程中的作用机制。方法分离与培养出生后第1、28天(P1、P28)PDGFR-β基因敲除小鼠(PDGFR-β-/-)SVZ区NSC,利用实时定量PCR、细胞免疫荧光染色方法检验PDGFR-β在NSC神经元分化能力方面的作用机制。采用方差分析、Fisher精确检验进行统计学处理。结果在对NSC进行诱导分化的过程中,PDGFs和PDGFRs的表达量明显增加,且源自NSC的神经元和星形胶质细胞均可高表达磷酸化的PDGFR-β,在对照组细胞中,P28 NSC的神经元分化能力明显低于P1细胞(MAP2表达量:0.27±0.03 vs 1.00±0.15,t=4.215,P<0.01;MAP2+神经元数量:8.93±4.24 vs 29.7±6.69,t=3.991,P<0.01;Dcx+神经元数量:9.05±2.12vs 17.4±3.87,t=3.338,P<0.05),而胶质细胞分化能力明显高于P1细胞(GFAP表达量:4.82±1.21 vs 1±0.22,t=3.921,P<0.01;Olig2表达量:1.45±0.46 vs 1±0.51,t=4.337,P<0.05;GFAP+胶质细胞数量:73.2±17.9 vs 40.5±4.11,t=6.221,P<0.01;Olig2+少突胶质前驱细胞数量:72.1±14.2 vs 42.7±6.59,t=3.224,P<0.05;Sox10+少突胶质前驱细胞数量:61.8±12.5 vs 33.4±3.59,t=3.013,P<0.05)。同时,P1 PDGFR-β-/-NSC细胞的神经元分化能力明显低于P1对照组(MAP2表达量:0.59±0.06 vs 1±0.15,t=3.674,P<0.05;MAP2+神经元数量:22.1±4.03 vs 29.7±6.69,t=5.223,P<0.05;Dcx+神经前驱细胞数量:8.61±1.74 vs 17.4±3.87,t=4.997,P<0.05),而两组中的胶质细胞分化能力相当。结论本研究证明PDGFR-β信号系统在NSC多向分化方面起重要作用。因此,激活NSC中PDGFR-β信号途径,可能成为今后神经系统疾病细胞替代疗法又一新的治疗策略。
Objective To investigate the role of platelet-derived growth factor β-receptor(PDGFR-β) in multipotency of neural stem cells(NSC). Methods In this study, NSC of subventricular zone were isolated and cultured from PDGFR knockout(PDGFR-β-/-) mice on postnatal day 1(P1) and P28; the roles of PDGFR-β in maintaining multipotency of NSC were examined using RT-PCR and immunocytochemistry. Comparisons between two experimental groups were performed using one-way ANOVA, followed by Fisher's PLSD test for each group. Results The expressions of PDGFs and PDGFRs in wide-type NSCs increased after differentiation induction, and phosphorylated PDGFR-β was co-localized with neuronal and astrocyte differentiation markers. In the control, the neuronal differentiation was decreased(MAP2: 0.27 ± 0.03 vs 1 ± 0.15,t = 4.215,P < 0.01;MAP2+ NSC :8.93 ± 4.24 vs 29.7 ± 6.69,t = 3.991,P < 0.01;Dcx+ NSCs :9.05 ± 2.12 vs 17.4 ± 3.87,t = 3.338,P < 0.05), and the glial differentiation was increased from P1 to P28 NSC(GFAP: 4.82 ± 1.21 vs 1 ± 0.22,t = 3.921,P < 0.01;Olig2: 1.45 ± 0.46 vs 1 ± 0.51,t = 4.337,P < 0.05;GFAP+ NSCs :73.2 ± 17.9 vs 40.5 ± 4.11,t = 6.221,P < 0.01;Olig2+ NSCs :72.1 ± 14.2 vs 42.7 ± 6.59,t = 3.224,P < 0.05;Sox10+ NSCs :61.8 ± 12.5 vs 33.4 ± 3.59,t = 3.013,P < 0.05).Compared with P1 controls, neuronal differentiation was reduced in P1 PDGFR-β-/- NSC(MAP2: 0.59 ± 0.06 vs 1 ± 0.15,t = 3.674,P < 0.05;MAP2+ NSCs :22.1 ± 4.03 vs 29.7 ± 6.69,t = 5.223,P < 0.05;Dcx+ NSCs :8.61 ± 1.74 vs 17.4 ± 3.87,t = 4.997,P < 0.05), whereas glial differentiation was comparable between the two groups. Conclusion These results suggest that PDGFR-β signaling is important for the multipotency of NSC, particularly in neonatal NSC. Accordingly, the activation of PDGFR-β in NSC may be a novel therapeutic strategy of neurological diseases.
出处
《中华细胞与干细胞杂志（电子版）》
2015年第3期154-163,共10页
Chinese Journal of Cell and Stem Cell（Electronic Edition）
基金
国家自然科学基金(81360186)
关键词
血小板
受体
生长因子
神经干细胞
细胞分化
Blood platelets
receptors,growth factor
neural stem cell
cell differentiation
