Akt/NF-κB信号转导通路在AngⅡ诱导的血管内皮细胞衰老中的表达及意义  

Effect of Akt/NF-κB signaling transduction pathway in vascular endothelial cell senescence induced by Angiotensin Ⅱ

作　　者：单海燕[1] 张四洋[2] 祁慧萌[1] 李婉[1] 赵心[1] 白小涓[1] 陈香美[3] 

机构地区：[1]中国医科大学附属第一医院老年病科 [2]中国医科大学实验技术中心三部,辽宁沈阳110001 [3]中国医科大学中国人民解放军总院肾病中心,北京100853

出　　处：《中国现代医学杂志》2013年第32期1-6,共6页China Journal of Modern Medicine

基　　金：国家自然科学基金资助(No:81200245);高等学校博士学科点专项科研基金联合资助课题(No:20122104120003);辽宁省科学技术研究项目(No:2012225091;201202263);沈阳市科学技术计划项目(No:F10-205-1-44)

摘　　要：目的研究血管紧张素Ⅱ（AngⅡ）对人脐静脉内皮细胞（HUVECs）衰老的影响及其相关蛋白激酶B（PKB/Akt）、核转录因子-KB（NF-KB）表达的变化，阐明血管内皮细胞衰老对诊治动脉粥样硬化的重要意义。方法制备血管紧张素ⅡP．PMI1640培养液（10^-6mol/L）培养HUVECs，通过β-半乳糖苷酶（β-gal）染色、流式细胞仪检测细胞衰老，Western blotting测定Akt、磷酸化Akt、NF-KB蛋白水平。结果AngII诱导组β-gal阳性细胞数与对照组相比为（80．10±6．81）％；流式细胞仪检测细胞周期停滞于G0～G，证实细胞衰老；与对照组相比，16、32和48hAkt磷酸化蛋白表达水平明显增高（P〈0．05），Akt磷酸化水平于32h达到最高峰（P〈0．01）；磷酸化Akt表达呈持续增加时，NF-KB呈显著上升趋势。结论血管紧张素Ⅱ诱导内皮细胞的衰老可能与Akt/NF-KB信号转导通路有关。[Objective] To investigate the activation of Akt/NF-KB signaling transduction pathway involved in the senescence of vascular endothelial cell senescence induced by Ang Ⅱ, and the underlying mechanism of cell senescence in the pathogenesis of atherosclerosis. [Methods] Human umbilical vein endothelial cells （HUVECs） were cultured in RPMI 1640 supplemented with Ang Ⅱ （10^-6 mol/L）. β-gal staining was applied for the analysis of cell cycle progression. Flow cytometry was used for analyzing the cell cycle changes. By means of Western blotting, the levels of Akt, p-Akt and NF-KB proteins were examined at different time points. [Results] The number of positive cells which were identified by β-gal staining was significantly high er in the Ang Ⅱ group （80.10± 6.81）% than that in the control group; the cell cycle was at G0-G1 （91.36 ± 6.45）% in Ang Ⅱ group. Compared with control cells, the phosphorylation of p-Akt was constantly increased at 16, 32 and 48 h （P〈0.05）, and reached its peak at 32 h （P〈0.01）. The significant activation of NF-KB was also observed. [Conclusion] It is presumed that the activated Akt/NF-KB signaling pathway is involvedin the process of pathologic and physiologic reaction in the senescence of vascular endothelial cells induced by Ang Ⅱ, which is closely correlated with atherosclerosis.

关 键 词：血管紧张素Ⅱ 内皮细胞 血管衰老 蛋白激酶B 核转录因子-KB 

分 类 号：Q255[生物学—细胞生物学]