目的:观察常氧和低氧环境下大鼠大脑皮层神经干细胞(NSCs)体外增殖情况,并比较P13K/Akt、JNK和Notch3条信号通路在不同氧环境下NSCs增殖时所发挥作用的异同。方法:在常氧和4%02浓度下悬浮培养新生SD大鼠大脑皮层NSCs,在培养细胞24、36和48h时使用图像分析软件测量NSCs增殖后所形成的神经球直径。结果:在培养细胞24、36和48h后,在不同时间段低氧组神经球直径都大于常氧对照组;在常氧浓度环境下分别使用0.5%DMSO和10μmol/LLY294002(P13K/Akt抑制剂)、SP600125(JNK抑制剂)和DAPT(Notch抑制剂)培养细胞,在培养细胞36h和48h时,3个抑制剂组神经球直径都小于常氧DMSO对照组。在低氧浓度环境下培养细胞36h和48h时,LY294002和SP600125组神经球直径与低氧DMSO组相比均无差异,而DAPT组神经球直径明显小于低氧DMSO对照组。结论:P13K/Akt、JNK和Notch3条信号通路均介导了常氧浓度环境下NSCs增殖,但低氧浓度环境诱导的NSCs增殖主要由Notch信号通路介导。
Objective: To observe the cell proliferation of the cerebral cortex in newborn SD rats under normoxia and hypoxia, and to compare the roles of three signaling pathways including PI3K/AKT, JNK and Notch in the proliferation under normoxia and hypoxia. Methods: The neural stem cells (NSCs) of the cerebral cortex in newborn SD rats were isolated and cultured under normal and 4% oxygen concentration. Each group was mixed with DMSO/LY294002/SP600125/DAPT, and then the diameters of neurospheres were measured at 24 h/36 h/48 h using the image analysis software. Results: The diameters of neurospheres under hypoxia were greater than those of normorxia at 24 h/36 h/48 h. The diameters of neurospheres treated with 0. 5%DMSO, LY294002 (inhibitor of PI3K/Akt), SP600125 (inhibitor of JNK), DAPT(inhibitor of Notch) under normal oxygen were smaller than those of DMSO group in normal oxygen. The diameters of neurospheres treated with LY294002/SP600125 had no significance difference compared to the DMSO group, but the diameters of neurospheres treated with DAPT were much smaller than those of the DMSO group. Conclusion: PI3-K/Akt, JNK and Notch signaling pathway all play important roles in the proliferation of NSCs, but Notch pathway mainly regulates the proli{eration of NSCs induced by hypoxia.