丹参多酚酸盐对过氧化氢诱导人内皮细胞EA.hy926氧化应激损伤的保护作用及机制  

Salvianolate attenuates oxidative stress injury of human endothelial EA.hy926 cells induced by hydrogen peroxide

作　　者：吕林林 安姿旖 梁家健 刘革修 LU Lin-lin;AN Zi-yi;LIANG Jia-jian;LIU Ge-xiu(Institute of Hematology,Basic Medical College of Jinan University,Guangzhou 510632.China)

机构地区：[1]暨南大学基础医学院血液研究所,广东广州510632

出　　处：《中国病理生理杂志》2019年第5期865-872,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81270568)

摘　　要：目的:探讨丹参多酚酸盐(salvianolate)对过氧化氢诱导人脐静脉内皮细胞EA.hy926氧化损伤的保护作用及机制。方法:体外培养EA.hy926细胞,随机分为:正常对照组、损伤组和抗损伤(丹参多酚酸盐+过氧化氢)组。采用CCK-8法检测细胞活性,Transwell实验检测内皮细胞迁移能力;一氧化氮(nitric oxide,NO)试剂盒和ELISA法分别检测细胞培养液中NO和血管内皮生长因子(vascular endothelial growth factor,VEGF)的水平;流式细胞术检测细胞内超氧化物阴离子水平、线粒体跨膜电位和细胞凋亡情况;采用Western blot法检测caspase-3、cleaved-caspase3、Bcl-2、Bax、NF-κB和p53蛋白表达水平。结果:与损伤组比较,不同剂量的丹参多酚酸盐预处理可提高EA.hy926细胞的活性、降低内皮细胞的凋亡,并促进细胞的迁移(P<0.05);细胞上清液中VEGF和NO含量及细胞内线粒体膜电位水平显著升高(P<0.05);细胞内ROS水平显著下降;NF-κB、p53、Bax和cleaved caspase-3蛋白水平显著降低(P<0.05);Bcl-2蛋白水平显著升高(P<0.05)。结论:丹参多酚酸盐能减轻过氧化氢对EA.hy926细胞氧化应激的损伤作用,其机制可能与NF-κB通路的阻滞相关。AIM:To investigate the effect of salvianolate on oxidative damage induced by hydrogen peroxide in human endothelial EA.hy926 cells.METHODS:EA.hy926 cells were cultured in vitro and divided into the following groups:control group,damage group,and anti-damage groups(salvianolate+damage groups).The cell viability was measured by CCK-8 assay.The migration ability of the EA.hy926 cells was detected by Transwell assay.The content of nitric oxide(NO)in the culture supernatant of the EA.hy926 cells was examined.The levels of vascular endothelial growth factor(VEGF)were detected by ELISA.The apoptosis,mitochondrial membrane potential and intracellular superoxide anion content of the EA.hy926 cells were analyzed by flow cytometry.The protein levels of caspase-3,cleaved caspase-3,Bcl-2,Bax,NF-κB and p53 were determined by Western blot.RESULTS:Compared with damage group,the viability of EA.hy926 cells pretreated with salvianolate at different concentrations was significantly increased(P<0.05).The apoptotic rate was significantly decreased(P<0.05).Savianolate enhanced the migration ability of the cells.The levels of VEGF,NO and mitochondrial transmembrane potential were increased(P<0.05),and the intracellular ROS level was significantly decreased(P<0.05).The protein levels of NF-κB,p53,Bax and cleaved caspase-3 were significantly decreased,and the protein level of Bcl-2 was markedly increased(P<0.05).CONCLUSION:Savianolate reduces the damage of EA.hy926 cells by hydrogen peroxide exposure,and its mechanism may be related to the blocking of NF-κB signaling pathway.

关 键 词：丹参多酚酸盐 血管内皮细胞 细胞凋亡 氧化损伤 NF-ΚB信号通路 

分 类 号：R363.2[医药卫生—病理学] R329.25[医药卫生—基础医学]