异氟醚抑制 Notch-1通路诱导小鼠神经干细胞凋亡

摘要

目的：探讨异氟醚对小鼠神经干细胞的影响及其可能的机制。方法：分离小鼠神经干细胞并培养，分为对照组、磷酸盐缓冲液（PBS ）组和异氟醚组。对照组无任何干预，异氟醚组给予异氟醚，PBS组给予等量的PBS ，培养24 h。采用CCK‐8法检测细胞增殖率；实时定量–聚合酶链反应法检测凋亡相关基因的 mRNA水平；蛋白质印迹法检测凋亡相关蛋白的表达；Annexin V‐FITC／PI法检测细胞凋亡。结果：与对照组和PBS组比较，异氟醚组细胞增殖率明显降低，同时Notch‐1、CBF‐1、Hes‐1基因的mRNA及相应蛋白表达水平也显著下降。异氟醚组细胞凋亡数目与对照组、PBS组比较，差异有统计学意义。结论：异氟醚能够诱导小鼠神经干细胞的凋亡，此作用与Notch‐1信号通路的抑制密切相关。%Objective:To investigate the influence of isoflurane on neural stem cells and its possible mechanism .Methods:Mouse neural stem cells ,isolated and cultured in vitro ,were divided into the control group ,the phosphate‐buffered saline (PBS) group and the isoflurane group .The control group was not intervened .The isoflurane group exposed to isoflurane for 24 h ,while the PBS group exposed to the same amount of PBS .Then CCK‐8 assay was used to detect the cell proliferation rate . Real‐time quantitative polymerase chaln reaction was used to detect the mRNA levels of apoptosis‐related genes .Expression of apoptosis‐related protein was measured with Western blotting .Annexin V‐FITC/PI kit was used to assess cell apoptosis . Results:Compared with those in the control group and the PBS group ,cell proliferation rate ,as well as the mRNA and protein levels of Notch‐1 ,CBF‐1 and Hes‐1 ,in the isoflurane group ,decreased significantly .There were more apoptotic cells in the isoflurane group than in the control group and the PBS group .Conclusions:Isoflurane can induce the apoptosis of mouse neural stem cells ,and the process is closely related to the inhibition of Notch‐1 pathway .