摘要
目的探讨线粒体ATP敏感钾离子通道开放剂二氮嗪、线粒体膜渗透性转换孔抑制剂环孢菌素A,以及两者协同对α-突触核蛋白片段的非淀粉样成分(NAC)致PC12多巴胺能细胞凋亡的分子机制。方法采用25μmol/L的NAC对PC12细胞进行诱导建立帕金森病细胞模型,分为对照组、NAC处理组(NAC组)、NAC+二氮嗪干预组(干预1组)、NAC+环孢菌素A干预组(干预2组)、NAC+二氮嗪+环孢菌素A干预组(干预3组)。二氮嗪、环孢菌素A及两者协同对其干预48h,用流式细胞术检测细胞凋亡率,Western blot检测细胞色素C、Bcl-2、Bax、Bcl-2/Bax比值情况。结果与对照组比较,NAC组细胞凋亡率、细胞色素C明显升高,Bcl-2、Bcl-2/Bax比值明显降低(P<0.01),Bax未见明显变化(P>0.05);与NAC组比较,干预1组、干预2组、干预3组细胞凋亡率、细胞色素C明显降低,Bcl-2、Bcl-2/Bax比值明显升高(P<0.01),Bax未见明显变化(P>0.05)。结论二氮嗪、环孢菌素A及两者协同作用通过抑制线粒体凋亡通路相关蛋白的表达来拮抗NAC的细胞凋亡作用。
Objective To study the molecular mechanism of combined diazoxide andcyclosporine A in preventing synuclein fragment-induced apoptosis of PC12 cells. Methods A Parkinson's disease model was established with 25 μmol/L synuclein fragment. PC12 cells were divided into control group, synuclein fragment treatment group, synuclein fragment+ diazoxide intervention group (intervention group 1),synuclein fragment+ cyclosporine A intervention group (intervention group 2),and synuclein fragment+ diazoxide+ cyclosporine A intervention group (intervention group 3). Apoptosis of PC12 cells was detected by flowcytometry. Expression levels of cytochrome C, Bcl-2, Bax,and Bcl-2/Bax ratio were measured by Western blot 48 h after diazoxide+ cyclosporine A intervention and their combination intervention. Results The apoptosis rate and expression level of cytochrome C were significantly higher whereas the expression level of Bcl-2 and the Bcl-2/Bax ratio were significantly lower in synuclein fragment treatment group than in control group (P〈0.01). The apoptosis rate and expression level of cytochrome C were significantly lower whereas the expression level of Bcl-2 and the Bcl-2/Bax ratio were significantly higher in inter-vention groups 1-3 than in synuclein fragment treatment group (P〈0.01) with no significant difference in expression level of Bax (P〉0.05). Conclusion Diazoxide and cyclosporine A and their combination can prevent synuclein fragment-induced apoptosis of PC12 cells by inhibiting mitochondrial apoptotic pathyway-related protein expression.
出处
《中华老年心脑血管病杂志》
CAS
北大核心
2013年第9期976-978,共3页
Chinese Journal of Geriatric Heart,Brain and Vessel Diseases
基金
烟台市科学技术发展计划项目(2010148-8)