乙醇对胎鼠脑新皮质神经干细胞凋亡的影响

发布时间：2018-10-23 14:15

【摘要】：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(P0.05);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(P0.05)。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(P0.05);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(P0.05);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。
[Abstract]:Objective to investigate the effect of ethanol on apoptosis of neural stem cells in fetal rats. Methods the neural stem cells derived from the neocortex of the fetal brain on the 14th day of gestation were exposed to ethanol medium with final concentration of 0 (control) and 250 mmol/L for 3 days. The changes of mitochondrial membrane potential and the expression of apoptosis-related gene Bcl-2,Bax,Caspase-3 mRNA and protein were detected by Annexin V/PI assay and JC-1 probe method. Results compared with the control group, the early apoptotic rate of neural stem cells in the 50100 mmol/L ethanol group was significantly higher than that in the control group (P0.05), but the late apoptotic rate of the neural stem cells in different concentration ethanol groups had no significant change. With the increase of ethanol concentration, the early apoptotic cell rate of neural stem cells increased. The mitochondrial membrane potential of neural stem cells in the 100 mmol/L ethanol group was significantly lower than that in the control group (P0.05). Compared with the control group, the Bax mRNA expression level of neural stem cells increased in all concentration ethanol groups, the difference was statistically significant (P0.05), while the expression level of Bcl-2 and Caspase-3 mRNA did not change significantly. The expression of, Bax mRNA increased with the increase of ethanol concentration. Compared with the control group, the expression of Bax protein in neural stem cells was increased and the ratio of Bcl-2/Bax protein was decreased in 50100 mmol/L ethanol group. The difference was statistically significant (P0.05); the expression level of Bcl-2,Caspase-3 protein of neural stem cells in all concentration ethanol groups were not significantly changed. Conclusion ethanol can induce early apoptosis of neural stem cells, which may be related to mitochondrial damage and up-regulation of Bax expression.
【作者单位】：复旦大学公共卫生学院环境卫生学教研室公共卫生安全教育部重点实验室健康风险预警治理协同创新中心;
【基金】：国家自然科学基金(30800893) 高等学校博士学科点基金(200802461122)
【分类号】：R114

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