蛇床子素对神经干细胞体外分化的影响

Effects of Osthole on Differentiation of Neural Stem Cells in vitro

目的探讨蛇床子素(Ost)对神经干细胞(NSCs)分化的影响及机制。方法体外分离并培养新生小鼠脑源NSCs,免疫细胞化学法鉴定;取第5代NSCs置于含不同浓度(0,10,50,100μmol·L-1)Ost的分化培养液中,免疫荧光细胞化学法检测NSCs分化为神经元、星形胶质细胞和少突胶质细胞情况;RT-PCR检测Notch 1基因及其靶基因Mash1和Neurogenin 2(Ngn2)表达情况。结果免疫荧光细胞化学法显示神经球显Nestin/Sox2阳性,即所培养的细胞为NSCs;Ost可促进NSCs更多地向神经元(P<0.01)和少突胶质细胞(P<0.05)分化,而非星形胶质细胞。Ost可减少Notch 1(P<0.01)并增加Ngn 2(P<0.01)mRNA表达,其中以Ost 100μmol·L-1组最明显。结论 Ost可促使NSCs更多地向神经元和少突胶质细胞分化,其机制可能与Ost抑制Notch信号通路有关。

Objective To investigate the effects of osthole on neural stem cells （ NSCs） differentiation and explore the potential mechanism. Methods Brain-derived NSCs from newborn mice were isolated and cultured in vitro and determined by immunofluorescence. The P5 generations of NSCs were placed in culture solution with osthole at concentrations of （0,10,50, 100 μmol·L-1 ） . The neuron, astrocyte and oligodendroglia cell differentiation were determined by immunofluorescence. The mRNA expression of Notch 1 and its target genes Mash 1 and Neurogenin 2 were assessed by RT-PCR. Results The neurosphere displayed Nestin and Sox 2-postive by immunofluorescence, suggesting that the cultured cells were NSCs. Osthole promoted NSCs differentiating into more neuron（P〈0. 01） and oligodendrocyte（P〈0. 05）, but not astrocyte. Meanwhile, osthole significantly reduced the mRNA expression of Notch 1（P〈0. 01） and increased Ngn 2（P〈0. 01）at the dose of 100 μmol·L-1. Conclusion Osthole enhances NSCs differentiating into more neuron and oligodendrocyte via probablly inhibiting Notch signal pathway.