摘要
目的应用Aβ25~35孵育SH-SY5Y神经母细胞瘤株制作细胞损伤模型,以研究复智散对Aβ25~35细胞损伤模型凋亡相关指标的影响。方法将制备好的SH-SY5Y细胞分为正常对照组、单纯应用复智散孵育组、Aβ25~35细胞损伤模型组、复智散与Aβ25~35共同孵育SH-SY5Y细胞组。采用异硫氰酸荧光素(fluoresceinisothiocyanate,FITC)标记的AnnexinⅤ及碘化丙锭双染流式细胞技术检测各组细胞的凋亡率,并利用荧光探针JC-1测定线粒体膜电位,统计分析不同组间神经细胞凋亡率及线粒体膜电位的变化情况,以确认药物的保护作用。结果单纯应用复智散孵育组神经细胞凋亡率及线粒体膜电位的变化与正常对照组相比,差异无显著性意义(P>0.05);Aβ25~35细胞损伤模型组培养的SH-SY5Y细胞线粒体膜电位下降,凋亡率增高(P<0.05);复智散与Aβ25~35共同孵育组神经细胞线粒体膜电位上调,细胞凋亡率降低,提示复智散对培养的SH-SY5Y细胞具有保护作用。结论复智散可拮抗Aβ25~35降低培养神经细胞线粒体膜电位、增加凋亡率的不良效应,具有神经细胞保护功能。
Objective A nerve cell injury model established by incubating SH-SY5Y neuroblastoma cell line in the presence of Aβ 25-35 was adopted to study the effect of Fuzhisan on apoptosis of nerve cells.Meth ods The prepared SH-SY5Y cells were divided into four groups:control,Fuzhishan treated,Aβ 25-35 treated nerve cell injury model,incubated with Fuzhisan and Aβ 25-35 .FITC-labeled AnnexinⅤand propidium iodide were adopted to evaluate the rate of cell apoptosis in different groups by means of flow cytometry,fluo-rescence probe JC-1was used to measure mitochondria transmembrane potentials,the apoptotic rate and the change of mitochondria transmembrane potential in different groups were statistically analyzed for study the injury and protective tendency.Re sults The apoptotic rates and the changes of mitochondria transmembrane potentials in Fuzhisan incubated group were not significantly different from those in control group(P>0.05),cells incubated in the presence of Aβ 25-35 showed decreased mitochondria transmembrane potentials and in-creased apoptotic rate(P<0.05),but cells treated with Fuzhisan and Aβ 25-35 revealed increasing transmem-brane potential and decreasing apoptotic rate,it demonstrated that Fuzhisan had protective effect on cultured SH-SY5Y cell line.Con clu sion Fuzhisan can resist the adverse effects of Aβ 25-35 on decreasing the mem-brane potential of nerve cell mitochondria and increasing apoptotic rate,thus this drug possesses the protective function toward nerve cells.[
基金
黑龙江省"九五攻关"课题(Z99C20-24)
关键词
复智散
神经细胞
细胞凋亡
老年期痴呆
Drug therapy Pulvers Dementia Amyloid beta-protein Apoptosis Cells,cultured