应激性衰老导致血管内皮细胞能量代谢及线粒体功能异常的研究  

Research for disorder of energy metabolism and mitochondrial dysfunction in stress-induced endothelial senescence

作　　者：戴春梅 林桐 罗文威 李梓晴 刘培庆[1,2] 李卓明[1,2] DAI Chunmei;LIN Tong;LUO Wenwei;LI Ziqing;LIU Peiqing;LI Zhuoming(Laboratory of Pharmacology and Toxicology,School of Pharmaceutical Sciences,Sun Yat-sen University,Guangzhou 510006,China;National and Local United Engineering Lab of Druggability and New Drugs Evaluation,Guangzhou 510006,China)

机构地区：[1]中山大学药学院药理毒理实验室,广州510006 [2]中山大学新药成药性评估与评价国家地方联合工程实验室,广州510006

出　　处：《中国科技论文》2019年第8期867-873,共7页China Sciencepaper

基　　金：国家自然科学基金资助项目(81872860,81673433);广州市科技计划项目(201804010227)

摘　　要：为探究H2O2诱导的血管内皮应激性衰老中能量代谢和线粒体功能的变化,在H2O2诱导的人脐静脉内皮细胞衰老模型中,Western blotting检测细胞衰老指标p21、p53、γH2A.X的蛋白表达水平;检测衰老相关的β-半乳糖苷酶蓝染程度;荧光定量PCR检测端粒长度;利用安捷伦Seahorse XFe 96细胞能量代谢仪检测内皮细胞的糖酵解情况和氧耗情况;检测细胞活性氧(reactive oxygen species,ROS)、线粒体膜电位及ATP水平;检测细胞能量代谢关键酶的表达水平。与正常组相比,H 2O 2刺激的内皮细胞中p21、p53、γH2A.X的蛋白表达明显上调,衰老相关的β-半乳糖苷酶蓝染的细胞比例增加,端粒长度缩短,衰老内皮细胞糖酵解能力下降,线粒体呼吸功能明显降低,ROS水平增加,线粒体膜电位下降,ATP生成减少,但大部分能量代谢关键酶的表达水平不变。因此,内皮应激性衰老可引起内皮能量代谢紊乱及线粒体功能障碍,靶向内皮能量代谢及线粒体功能调节可能是干预内皮应激性衰老的重要策略。To investigate the changes of energy metabolism and mitochondrial function in senescent endothelial cells induced by H2O2,human umbilical vein endothelial cells were treated with H2O2 to induce senescence.The protein expressions of senescent markers p21,p53 and γH2A.X were detected by Western blotting.Senescence-associatedβ-galactosidase staining was performed.Telomere length was detected by real-time PCR.Glycolysis and oxygen consumption of the cells were measured by Seahorse XFe 96.Mitochondrial membrane potential,reactive oxygen species(ROS)level and ATP production were measured.The expressions of key metabolic enzymes were detected.Compared with the normal group,the expression of p21,p53 and γH2A.X protein in endothelial cells stimulated by H2O2 increased significantly.The proportion of aging-related beta-galactosidase blue-stained cells increased.Telomere length shortened.Glycolysis ability of aging endothelial cells decreased.Mitochondrial respiratory function decreased significantly.ROS level increased.Mitochondrial membrane potential increased.ATP production decreased,but the expression level of most key enzymes in energy metabolism remained unchanged.Therefore,endothelial senescence is coincided with disturbed energy metabolism and mitochondrial dysfunction.Strategies targeting energy metabolism and mitochondrial function might play a pivotal role in preventing endothelial senescence.

关 键 词：心血管药理学 应激性衰老 能量代谢 线粒体功能 

分 类 号：R966[医药卫生—药理学]