迷迭香酸拮抗H_2O_2诱导的人脐静脉内皮细胞凋亡及作用机制

目的研究迷迭香酸(Rosmarinic acid,RA)对H2O2诱导的人脐静脉内皮细胞株ECV304凋亡的抑制作用及其机制。方法以H2O2诱导ECV304细胞凋亡为模型,用MTT比色法检测RA对细胞活性的影响;流式细胞仪检测法和Tunel法检测RA对细胞凋亡率的影响;用Western-blotting检测凋亡相关基因Bcl-2、Bax蛋白的表达情况和第24hcaspase-3活性。结果用0.5mmol/LH2O2孵育细胞24h细胞凋亡明显。不同浓度的RA(1、3、10μmol/L)预处理细胞后,呈浓度依赖性的增加内皮细胞活性,降低细胞凋亡率,并且Bcl-2蛋白表达呈浓度依赖性的上调,而Bax蛋白的表达和caspase-3蛋白活性呈浓度依赖性下降。结论迷迭香酸可以拮抗H2O2诱导的内皮细胞凋亡,其抑制内皮细胞凋亡与激活Bcl-2活性有关。

Antiapoptotic Effect of Rosmarinic Acid on Human Umbilical Vein Endothelial Cell Apoptosis Induced by Hydrogen Peroxide

Objective To observe the antiapoptotic effect and its mechanism of rosmarinic acid on human umbilical vein endothelial cell apoptosis induced by hydrogen peroxide.Methods With the model of ECV304 cell apoptosis induced by H2O2,the cell viability was measured by the MTT assay.The cell apoptosis was determined by nuclear staining with AO,flow cytometric analysis and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling （Tunel）,respectively.Meanwhile,western-blotting was utilized to measure the expression of Bcl-2 and Bax,and the activity of caspase-3.Results After ECV304 cell was incubated with 0.5 mmol/L H2O2 for 24 hours,the apoptosis rate of cell was increased significantly.However,after pretreating different concentrations of RA （1 μmol/L,3 μmol/L,and 10 μmol/L）,the cell survival was promoted in a concentration-dependent manner,and the apoptotic rate of cell was decreased.Bcl-2 expression was increased in a dose-dependent way by rosmarinic acid,but the expression of Bax and caspase-3 was decreased in a dose-dependent way.Conclusions Rosmarinic acid can prevent H2O2 induced apoptosis of endothelial cells,and the inhibition of rosmarinic acid on apoptosis is correlated with an increase in Bcl-2 activity.