叠氮钠对SH—SY5Y人神经母细胞瘤细胞线粒体跨膜电位 …

目的 观察线粒体呼吸链复合体ＩＶ（即细胞色素Ｃ氧化酶）抑制剂叠氮钠（ＮａＮ３）对人神经母细胞瘤细胞能量代身的影响，探讨线粒体缺陷在神经退行性疾病发病中的作用。方法 将叠氮钠与培养的人神经母细胞瘤细胞系ＳＨ－ＳＹ５Ｙ细胞共同孵育，用微测量法测定线粒体复合体ＩＶ活性，噻唑兰（ＭＴＴ）法测定细胞存活率，激光共聚焦显微镜和流式细胞仪检测线粒体跨膜电位变化。结果 １６￣６４ｍｍｏｌ／Ｌ叠氮钠与培养的ＳＨ－Ｓ

Effect of sodium azide on mitochondrial membrane potential in SH-SY5Y human neuroblastoma cells

OBJECTIVE: To study the role of mitochondrial deficiency in the pathogenesis of neurodegenerative disease by investigating the energy metabolism in a sodium azide inhibited cytochrome-c oxidase SH-SY5Y Cell model. METHODS: Human neuroblastoma SH-SY5Y Cells were exposed to sodium azide, then mitochondrial complex IV activity was assayed by microassay method; cell viability was measured by Thiazolyl blue(MTT) method; mitochondrial membrane potential (MMP) was detected by confocal microscopy and flow cytometry. RESULTS: Cultured SH-SY5Y cells were exposed to 16-64 mmol/L sodium azide for 1 hour, the mitochondrial complex IV activity decreased dose-dependently. MTT absorbance decreased does- and time-dependently in cultured nerve cells treated by 16-128 mmol/L sodium azide for 1-8 hours. After the treatment of 16 mmol/L sodium azide for 1 hour, both the fluorescence intensity of MMP and normal cell events reduced. Decrease of MMP was significant especially in cell processes. CONCLUSION: Sodium azide induced the impairment of mitochondrial energy synthesis in the cultured nerve cells which is an important cause in cell death.