毛蕊异黄酮抑制氧糖剥夺/复氧复糖PC12细胞凋亡研究

Inhibitory effect of calycosin on apoptosis of PC12 cells under oxygen and glucose deprivation/reoxygenation

目的研究毛蕊异黄酮对氧糖剥夺/复氧复糖PC12细胞凋亡的影响。方法将对数生长期PC12细胞随机分为4组:正常对照组(Control)、模型组(Model)、毛蕊异黄酮组(Calycosin,0.07μmol·L^-1)和尼莫地平组(Nimodipine,5.00μmol·L^-1,阳性对照药组)。用CCK-8检测细胞存活率;流式细胞术和TUNEL染色检测细胞凋亡率和凋亡指数;免疫荧光染色检测细胞Bax/Bcl-2比值;Western blot检测细胞凋亡蛋白caspase-3表达。结果与Control组相比,Model组细胞存活率明显降低(P<0.05),细胞凋亡率和凋亡指数明显升高(P<0.05),Bax/Bcl-2比值和caspase-3蛋白表达明显升高(P<0.05);与Model组相比,Calycosin组和Nimodipine组细胞存活率均明显升高(P<0.05),细胞凋亡率和凋亡指数均明显降低(P<0.05),Bax/Bcl-2比值和caspase-3蛋白表达均明显降低(P<0.05),差异有统计学意义。结论毛蕊异黄酮可明显提高氧糖剥夺/复氧复糖PC12细胞存活率,抑制细胞凋亡,其作用机制与毛蕊异黄酮调控凋亡蛋白Bax、Bcl-2、caspase-3表达有关。

Aim To study the effect of calycosin on apoptosis of PC12 cells under oxygen and glucose deprivation/reoxygenation.Methods PC12 cells in logarithmic phase were randomly divided into four groups:normal control group,model group,calycosin group(0.07μmol·L^-1)and nimodipine group(5.00 micromol L-1,positive control group).CCK-8 assay was used to detect cell survival rate;flow cytometry and TUNEL staining were used to detect apoptotic rate and apoptotic index;immunofluorescence staining was used to detect Bax/Bcl-2 ratio;Western blot was used to detect the expression of caspase-3 apoptotic protein.Results Compared with control group,the cell survival rate significantly declined(P<0.05),the apoptotic rate and apoptotic index significantly rose(P<0.05),and Bax/Bcl-2 ratio and caspase-3 protein expression were significantly up-regulated(P<0.05);compared with model group,the cell survival rate significantly increased in calycosin group and nimodipine group(P<0.05),the mortality and apoptotic index significantly decreased(P<0.05),the Bax/Bcl-2 ratio and caspase-3 protein expression significantly decreased(P<0.05),and the difference was statistically significant.Conclusions Calycosin can significantly improve the survival rate of oxygen-glucose deprivation/reoxygenation PC12 cells and inhibit cell apoptosis.Its mechanism is related to the regulation of expression of apoptotic proteins Bax,Bcl-2 and caspase-3 by calycosin.