蓝莓花色苷对过氧化氢所致血管内皮细胞凋亡的保护作用
Protective effects of blueberry anthocyanins on hydrogen peroxide- induced injury of vascular endothelial cells
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摘要: 目的:观察蓝莓花色苷对过氧化氢(H2O2)所致血管内皮细胞凋亡的保护作用及其机制。方法:人静脉血管内皮细胞EVC-304经不同浓度蓝莓花色苷(10、20、40μg·m L-1)预培养24 h后,加入100μmol/L H2O2再进行培养12 h。MTT法检测细胞存活率,流式细胞仪检测EVC-304细胞凋亡的变化,Western blot法检测细胞内激活型Caspase-3、细胞色素C(Cyto c)、Bax和Bcl-2蛋白表达变化。结果:同正常组相比,H2O2损伤组细胞存活率显著降低(p<0.01),细胞凋亡率显著升高(p<0.01),激活型Caspase-3、细胞质中Cyto c表达水平,Bax/Bcl-2比值升高(p<0.01)。与H2O2损伤组相比,20μg·m L-1和40μg·m L-1蓝莓花色苷处理组细胞存活率明显升高(p<0.01),凋亡细胞率明显下降(p<0.01),细胞内激活型Caspase-3,Cyto c表达,Bax/Bcl-2比值下降(p<0.01)。结论:蓝莓花色苷对H2O2所致血管内皮细胞凋亡具有保护作用,其机制可能与保护线粒体功能相关。Abstract: Objective: To investigate the protective effects of blueberry anthocyanins on hydrogen peroxide( H2O2)-induced apoptosis of vascular endothelial cells.Methods: The EVC-304 cells cultured in logarithmic growth phase were incubated with 100 μmol/L H2O2 for another 12 h after pretreatment with blueberry anthocyanins( 10,20,and40 μg·m L-1)for 24 h.Cell viability was determined by MTT assay,the change of cell apoptosis were measured with cytoanalyzer,and the expression levels of Cleaved Caspase-3,Cyto c,Bax,and Bcl-2 were detected by Western blotting.Results: Compared with the normal control group,the survival rate of H2O2-treated cells was decreased and the apoptosis ratio was increased significantly( p < 0.01),the proportion of cleaved Caspase-3,Cytoc and Bax/Bcl-2 was increased( p < 0.01). Compared with H2O2-treated group,the survival rate of cells in blueberry anthocyanins-pretreated groups was increased significantly( p < 0.01),the apoptosis rate was decreased significantly( p < 0.01).The expression of intracellular cleaved caspase-3,Cyto c and Bax/Bcl-2 was decreased( p < 0.01). Conclusion: Blueberry anthocyanin had protective effect on hydrogen peroxide-induced vascular endothelial cells apoptosis and the possible mechanism may be associated with protection the function of mitochondrion.