尼莫地平对H2O2诱导的PC12细胞凋亡的保护作用

摘要

目的 探讨尼莫地平对过氧化氢(H2O2)诱导的PC12细胞凋亡的保护作用.方法 PC12细胞随机分为正常组,模型组(200 μmol/L H2O2),尼莫地平低、中、高浓度组(1、10、100 μmol/L尼莫地平+200 μmol/L H2O2).MTT法检测各组细胞活力,Hoechst染色各组细胞凋亡情况,比色法检测天冬氨酸蛋白水解酶(caspase 3),caspase 9及超氧化物歧化酶(SOD)活性与丙二醛(MDA)含量,western blot分析B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)及p53蛋白.结果 与正常组比较,模型组中细胞活力下降,细胞凋亡率提高,caspase 3及caspase 9提高,SOD活性降低,MDA含量降低,Bcl-2表达量下降,Bax及p53表达量上升,差异均具有统计学意义(P<0.01);与模型组比较,尼莫地平低、中、高浓度组细胞活力提高,细胞凋亡率降低,caspase 3及caspase 9活性降低,SOD活性提高,MDA含量提高,Bcl-2蛋白表达量提高,Bax及p53表达量降低,差异均具有统计学意义(P<0.01).结论 尼莫地平可通过调控细胞凋亡相关蛋白表达,从而抑制H2O2诱导的PC12细胞凋亡.%Obejective To explore the protective effect of nimodipine on PC12 cell apoptosis induced by hydrogen peroxide (H2O2).Methods The cells were randomized into five groups: normal group,model group (200 μmol/L H2O2),nimodipine low-,medium-and high-dose groups (1,10,100 μmol/L nimodipne+200 μmol/L H2O2).Cell viability was measured by MTT assay.Cell apoptosis was assessed by Hoechst staining.The activity of cysteinyl aspartate specific proteinase 3 (caspase 3),caspase 9 and superoxide dismutase (SOD),the level of malonic aldehyde (MDA) were measured by colorimetry.The expression of B-cell lymphoma-2 (Bcl-2),Bcl-2 associated X protein (Bax) and p53 were detected by western blot.Results Compared with the normal group,the cell apoptosis rate,the activity of caspase 3 and caspase 9,the MDA level,and the expression of Bax and p53 were significantly increased,the cell viability,SOD activity,and expression of Bcl-2 were decreased in the model group (P<0.01).Compared with the model group,the cell apoptosis rate,the activity of caspase 3 and caspase 9,the level of MDA,and the expression of Bax were decreased;and the cell viability,SOD activity,and the expression of Bcl-2 were increased in the nimodipine low-,medium-and high-dose groups (P<0.01).Conclusions Nimodipine suppresses cell apoptosis induced by H2O2 in PC12 cells,which may be related to regulation of expression of cell apoptosis-related proteins.