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目的探讨乙醇对胎鼠神经干细胞凋亡的影响。方法将妊娠14 d胎鼠脑新皮质来源的神经干细胞暴露于终浓度分别为0(对照)、25、50、100 mmol/L的乙醇培养基溶液3 d。采用Annexin V/PI法检测细胞早期和晚期凋亡情况,采用JC-1探针法检测线粒体膜电位的改变,并检测凋亡相关基因Bcl-2、Bax、Caspase-3 mRNA和蛋白的表达。结果与对照组比较,50、100 mmol/L乙醇染毒组神经干细胞的早期凋亡细胞率明显升高,差异有统计学意义(P<0.05);而各浓度乙醇染毒组神经干细胞的晚期凋亡率无明显改变。且随着乙醇染毒浓度的升高,神经干细胞的早期凋亡细胞率呈上升趋势。仅100 mmol/L乙醇染毒组神经干细胞线粒体膜电位低于对照组,差异有统计学意义(P<0.05)。与对照组比较,各浓度乙醇染毒组神经干细胞Bax mRNA的表达水平均上升,差异有统计学意义(P<0.05);而Bcl-2和Caspase-3 mRNA的表达水平均无明显改变。且随着乙醇染毒浓度的升高,Bax mRNA表达量呈上升趋势。与对照组比较,50、100 mmol/L乙醇染毒组神经干细胞Bax蛋白的表达水平均升高,而Bcl-2/Bax蛋白的比值均降低,差异有统计学意义(P<0.05);各浓度乙醇染毒组神经干细胞Bcl-2、Caspase-3蛋白的表达水平均无明显改变。结论乙醇可引起神经干细胞早期凋亡,其机制可能与线粒体损伤和Bax表达上调有关。
Objective To investigate the effect of ethanol on the apoptosis of neural stem cells in fetal rats. Methods The neural cortex cells derived from the cortex of 14-day gestation fetus were exposed to the final concentrations of 0 (control), 25, 50 and 100 mmol / L ethanol medium for 3 days, respectively. The apoptosis of early and late cells was detected by Annexin V / PI assay. The changes of mitochondrial membrane potential were detected by JC-1 probe, and the expressions of Bcl-2, Bax, Caspase-3 mRNA and protein were detected. Results Compared with the control group, the apoptotic rate of neural stem cells in 50,100 mmol / L ethanol-treated group was significantly higher than that in the control group (P <0.05). However, the late stage of neural stem cells in ethanol- No significant change in apoptosis rate. And with the increase of ethanol exposure, the rate of early apoptotic cells in neural stem cells showed an upward trend. The mitochondrial membrane potential of neural stem cells exposed to 100 mmol / L ethanol only was lower than that of the control group, the difference was statistically significant (P <0.05). Compared with the control group, the expression of Bax mRNA in neural stem cells of various concentrations of ethanol exposure group increased significantly (P <0.05), while the expressions of Bcl-2 and Caspase-3 mRNA had no significant changes. And with the increase of ethanol concentration, Bax mRNA expression increased. Compared with the control group, the expression of Bax protein in neural stem cells of 50,100 mmol / L ethanol-treated group increased, but the ratio of Bcl-2 / Bax protein decreased (P <0.05); The concentration of Bcl-2 and Caspase-3 in the neural stem cells of ethanol exposure group had no significant change. Conclusion Ethanol can induce early apoptosis of neural stem cells, which may be related to mitochondrial damage and up-regulation of Bax expression.