摘要
目的探讨毛蕊异黄酮对缺氧缺糖/复氧复糖PC12细胞线粒体凋亡通路的影响。方法将PC12细胞随机分为4组:正常组(control)、模型组(model)、毛蕊异黄酮组(calycosin)和尼莫地平组(nimodipine,阳性对照药组)。除control组外,其余各组进行缺氧缺糖2 h,复氧复糖24 h制作模型处理。Calycosin组和nimodipine组在复氧复糖的同时分别给予含有毛蕊异黄酮(0.07μmol/L)和尼莫地平(5.00μmol/L)的含药培养基处理。用CCK-8法检测各组细胞活性,流式细胞术检测各组细胞凋亡率,细胞免疫荧光法检测各组细胞Bax/Bcl-2比值,Western blotting法检测线粒体凋亡通路中关键蛋白细胞色素C(Cyt-C)、凋亡酶激活因子1(Apaf-1)和Caspase-3表达。结果与control组相比,model组细胞的活性明显降低(P<0.05),细胞凋亡率明显升高(P<0.05),Bax/Bcl-2比值明显升高(P<0.05),线粒体凋亡通路中关键蛋白Cyt-C、Apaf-1和Caspase-3的表达均明显升高(P<0.05);与model组相比,calycosin组和nimodipine组的细胞活性均明显提高(P<0.05),细胞凋亡率明显降低(P<0.05),Bax/Bcl-2比值明显降低(P<0.05),线粒体凋亡通路中关键蛋白Cyt-C、Apaf-1和Caspase-3的表达均明显降低(P<0.05),差异有统计学意义。结论毛蕊异黄酮可显著提高缺氧缺糖/复氧复糖PC12细胞活性,抑制细胞凋亡,其作用机制与毛蕊异黄酮抑制线粒体凋亡通路中关键蛋白Cyt-C、Apaf-1和Caspase-3表达密切相关。
Objective To investigate the effect of calycosin on mitochondrial apoptotic pathway in oxygen-glucose deprivation/reoxygenation PC12 cells.Methods PC12 cells were randomly divided into four groups:control group,model group,calycosin group and nimodipine group.Except for the control group,the other groups were treated with oxygen and glucose deprivation for 2 hours and compound oxygen and glucose for 24 hours.Calycosin group and nimodipine group were treated with drug-containing medium containing calycosin(0.07μmol/L)and nimodipine(5.00μmol/L)simultaneously with reoxygenation.CCK-8 method was used to detect cell survival rate,flow cytometry was used to detect cell apoptosis rate,immunofluorescence method was used to detect Bax/Bcl-2 ratio,Western blotting was used to detect the expression of key proteins cytochrome C(Cyt-C),apoptotic protease activating factor-1(Apaf-1)and Caspase-3 in mitochondrial apoptotic pathway.Results Compared with the control group,the survival rate of cells in model group decreased significantly(P<0.05),and the apoptotic rate increased significantly(P<0.05),the ratio of Bax/Bcl-2 was significantly increased(P<0.05),and the expression of key proteins Cyt-C,Apaf-1 and Caspase-3 in mitochondrial apoptotic pathway were significantly increased(P<0.05).Compared with the model group,the cell survival rates of calycosin group and nimodipine group increased significantly(P<0.05),apoptotic rate decreased significantly(P<0.05),the ratio of Bax/Bcl-2 was significantly decreased(P<0.05),and the expression of key proteins of mitochondrial apoptotic pathway,Cyt-C,Apaf-1 and Caspase-3 were significantly decreased(P<0.05).The difference has statistical significance.Conclusion Calycosin can significantly improve the survival rate of oxygen-glucose deprivation/reoxygenation PC12 cells and inhibit cell apoptosis.Its mechanism is closely related to the inhibition of the expression of key proteins Cyt-C,Apaf-1 and Caspase-3 in mitochondrial apoptotic pathway by calycosin.
作者
张彐宁
靳晓飞
周晓红
高维娟
ZHANG Xue-ning;JIN Xiao-fei;ZHOU Xiao-hong;GAO Wei-juan(Hebei University of Chinese Medicine,Hebei Key Laboratory of Chinese Medicine Research on Cardio-cerebrovascular Disease,Shijiazhuang 050091,China)
出处
《解剖学报》
CAS
CSCD
北大核心
2020年第3期332-337,共6页
Acta Anatomica Sinica
基金
国家自然科学基金(81873180)
河北省教育厅青年基金(QN2018059)
河北省中医药管理局科研计划项目(2019093)
河北省研究生创新项目(CXZZSS2019066)
河北中医学院优秀青年教师基础研究计划项目(YQ2018005)
河北中医学院2019年省属高校基本科研业务费项目(YXZ2019012)。
