硝普钠诱导体外培养的海马神经元凋亡的研究

Apoptosis of cultured rat hippocampal neurons induced by sodium nitroprusside

目的 :观察一氧化氮 (NO)供体硝普钠 (SNP)对体外培养的海马神经元凋亡的影响。方法 :用终浓度分别为 0、2 5、5 0、10 0、2 0 0、40 0、60 0 μmol/L的SNP处理海马神经元 2 4h ,用MTT比色法分析细胞存活率 ,倒置显微镜、Hoechst3 3 2 5 8荧光染色观察凋亡的形态学改变 ,DNA琼脂糖凝胶分析凋亡的生化特征。结果 :SNP可剂量依赖性的降低神经元的存活率 ,当SNP浓度为 5 0 μmol/L时 ,其存活率为 5 6.2 % ;倒置显微镜观察可见神经元胞体固缩 ,突起断裂 ,网络消失 ;荧光显微镜可见染为高亮蓝色的典型凋亡小体 ,其细胞核明显固缩、凝聚和断裂 ,且随SNP剂量的增加 ,出现凋亡小体的细胞明显增多 ;5 0、10 0、2 0 0 μmol/LSNP处理海马神经元 ,电泳图谱显示清晰的DNA梯度。 结论

Objective:To observe the effect of sodium nitroprusside(SNP)on apoptosis of the cultured rat hippocampal neurons.Methods:The cultured rat hippocampal neurons weretreated with different concentrations of SNP(0 600 μmol/L)for 24 hours.The viability of neurons was determined by MTT assay,and the morphological and biochemical changes were observed by invert microscope and fluorescent staining (Hoechst 33258)and by agarose gel electrophoresis,respectively.Results:SNP lowered the survival rate of neurons in a dose dependent manner,and there was 58.2% of survival rate at strength of 50 μ mol/L.When exposed to higher concentrations of SNP(50 200 μ mol/L),neurons exhibited the morphological and biochemical features of apoptosis,including cell shrinkage,neurite degeneration,network disappearance,condensation and aggregation of nuclear chromatin,and the formation of DNA ladder.There were more apoptotic bodies with the increase of SNP concentration.Conclusion:SNP could induce the apoptosis of cultured hippocampal neurons in rats.