目的 研究藁本内酯对神经干/祖细胞（neural stem/progenitor cells，NS/PCs）的增殖作用及机制。方法 孕14 d Wistar大鼠脱颈椎处死，分离胎鼠脑皮层部位NS/PCs，采用悬浮法原代培养7 d。采用WST-1法检测藁本内酯对NS/PCs存活率的影响；采用Western blotting法检测藁本内酯对NS/PCs增殖相关蛋白磷酸化蛋白激酶B（phosphorylated protein kinase B，p-Akt）、Akt、磷酸化糖原合成酶激酶（phosphorylated glycose synthase kinase 3β，p-GSK3β）、GSK3β和active-β-catenin表达；给予藁本内酯第1～3天，观察NS/PCs神经球生长状态并测量其直径；分化培养基诱导NS/PCs分化5 d后，采用免疫荧光法检测藁本内酯对NS/PCs分化类型的影响，并采用WST-1法检测藁本内酯对分化细胞存活率的影响。结果 藁本内酯（25μmol/L）显著促进NS/PCs存活率（P<0.05），显著上调增殖相关蛋白p-Akt/Akt和active-β-catenin表达（P<0.05）；给予藁本内酯第1～3天，可增加NS/PCs神经球直径，但无统计学差异；藁本内酯对神经元、星型胶质细胞方向分化无显著影响，但分化后细胞存活率显著升高（P<0.05）。结论 藁本内酯可促进NS/PCs增殖，其作用机制与Akt/β-catenin通路有关。藁本内酯可能对神经元、星型胶质细胞以外的其他分化类型细胞有干预作用。

Objective To study the effect and mechanism of ligustilide on proliferation of neural stem/progenitor cells (NS/PCs). Methods Wistar rats were sacrificed on 14th day of pregnancy, NS/PCs in cerebral cortex of fetal rats were isolated and cultured in suspension for 7 d. WST-1 method was used to detect the effect of ligustilide on survival rate of NS/PCs; Western blotting was used to detect the effect of ligustilide on proliferation-related protein, including phosphorylated protein kinase B (p-Akt), Akt, phosphorylated glycose synthase kinase 3β (p-GSK3β), GSK3β and active-β-catenin expressions in NS/PCs; Ligustilide was given to cells for 1st to 3rd day, growth status of neurospheres in NS/PCs was observed and their diameter was measured; After 5 d of differentiation of NS/PCs induced by differentiation medium, immunofluorescence method was used to detect the effect of ligustilide on differentiated type of NS/PCs, and WST-1 method was used to detect the survival rate of differentiated cells. Results Ligustilide (25 μmol/L) significantly promoted the survival rate of NS/PCs (P < 0.05), and up-regulated the expressions of proliferation-related proteins p-Akt/Akt and active-β-catenin (P < 0.05); Administration of ligustilide on 1st to 3rd day, diameter of neurospheres was increased in NS/PCs, but there was no statistical difference; Ligustilide had no significant effect on differentiation of neurons and astrocytes, but the survival rate of cells after differentiation was significant increased (P < 0.05). Conclusion Ligustilide can promote the proliferation of NS/PCs, and its mechanism is related to Akt/β-catenin pathway. Ligustilide may has an intervening effect on other differentiated cells rather than neurons and astrocytes.

R285.5

国家自然科学基金资助项目（81873041）；国家自然科学基金资助项目（82030124）