摘要
目的 探讨丁苯酞（NBP）对β淀粉样蛋白（Aβ）诱导的PC12细胞凋亡的保护作用及其机制.方法 不同浓度的NBP（0.1、1.0、10、100 μmol/L）作用到经Aβ诱导或未经诱导的PC12细胞上,然后分别采用细胞存活率检测（MTT）及PI单染、透射电镜方法检测细胞凋亡,采用western印迹法检测Bcl-2和Cyt-C蛋白的表达,进而观察NBP对Aβ25-35诱导的PC12细胞凋亡的保护作用.结果 MTT显示不同浓度（0.1、1.0、10、100 μmol/L）NBP组细胞存活率分别为76.5%±1.1%、84.2%±1.3%、89.5%±1.3%、81.9%±1.9%,明显高于模型组（71.7%±1.4%）,其中10 μmol/LNBP组最明显,差异有统计学意义（P＜0.05） 流式细胞学结果显示10 μmol/L NBP组细胞凋亡率明显低于模型组（P＜0.05） 电镜下模型组细胞凋亡特征明显,10 μtmol/L NBP组细胞形态接近正常 Western印迹显示：10 μmol/L NBP组Bcl-2蛋白表达增加,而模型组Bcl-2表达明显减少（P＜0.05） 模型组胞质Cyt-C表达最强,而10μmol/L NBP组有较弱的Cyt-C表达（P＜0.05）.结论 NBP对Aβ诱导的PC12细胞凋亡有保护作用,其机制可能和NBP增加Bcl-2蛋白水平,抑制Cyt-C从线粒体的释放有关.
Objective To study the protective effects of butylphthalide （NBP） on the Aβ25-35-induced apoptosis in PC-12 cells.Methods The apoptosis of PC-12 cell was analyzed by MTT assay,transmission electron microscope and PI method at different concentrations of NBP （0.1,1.0,10,100μmol/L） with the addition of Aβ or not.The expressions of Bcl-2 and cytochrome C （Cyt-C） were detected by Western blot.Results As demonstrated by the MTT assay,the values of cell viability were 76.5% ±1.1%,84.2% ±1.3%,89.5% ±1.3% and 81.9% ±1.9% at various concentration （0.1,1.0,10,100 μmol/L） of NBP respectively.The model group was 71.7% ± 1.4%.It was revealed that the former could significantly prevent the cell viability under the induction of Aβ25-35 （P 〈 0.05）.A pretreatment with 10 μmol/L NBP could significantly inhibit the decrease of viability under the induction of Aβ25-35 （P 〈0.05）.PI showed that the apoptosis rate of the 10 μmol/L NBP treatment group was significantly lower than that of the model group.Under electron microscope,the characteristics of cell apoptosis were significant in the model group.And the cell morphology of the 10 μmol/L NBP treatment group was normal.The expression rate of Bcl-2 protein in the 10 μmol/L NBP treatment group was obviously higher than that in the model group.Cyt-C was weakly expressed in nerve cells of the normal and the 10 μmol/L NBP groups.But it had a strong expression in the model group.Conclusion NBP prevents the injury of PC12 cells by Aβ.And the mechanism may be related to the elevated level of Bcl-2 and the inhibited mitochondrial release of Cyt-C.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2010年第45期3235-3237,共3页
National Medical Journal of China
关键词
AΒ25-35
凋亡
丁苯酞
PC12细胞
Beta-amyloid peptide 25-35 Apoptosis 3-n-butylphthalide PC12 cell
