阿司匹林抗高糖诱导的内皮细胞衰老过程中对DDAH-ADMA系统及Caveolin-1蛋白的影响  

Effect of Aspirin on ADMA-DDAH System and Caveolin-1 Protein Expression in Endothelial Senescence Exposed to High Glucose

作　　者：伊桐凝[1] 张锦[2] 于世家[1] 

机构地区：[1]辽宁中医药大学附属医院内分泌科,辽宁省沈阳市110031 [2]中国医科大学附属第一医院内分泌科,辽宁省沈阳市110001

出　　处：《中国动脉硬化杂志》2016年第5期469-473,共5页Chinese Journal of Arteriosclerosis

基　　金：辽宁省博士启动基金资助项目(20121101)

摘　　要：目的观察阿司匹对高糖诱导的内皮细胞衰老模型二甲基精氨酸二甲胺水解酶（DDAH）-非对称性二甲基精氨酸（ADMA）系统和Caveolin-1蛋白表达的影响,探讨阿司匹林抗高糖诱导内皮细胞衰老的机制。方法人脐静脉内皮细胞（HUVEC）分别培养于正常糖浓度培养液（5.5 mmol/L）、高糖培养液（33 mmol/L）、高糖＋阿司匹林（0.01~1 mmol/L）培养液以及含L-NAME（300μmol/L）培养液,48 h后采用β-半乳糖苷酶（β-gal）染色鉴定衰老细胞,流式细胞仪检测细胞内活性氧（ROS）水平,Western blot分析Caveolin-1蛋白表达,液质联用仪检测细胞上清液中ADMA的含量并计算DDAH的活性。结果高糖作用内皮细胞48 h后,细胞β-gal染色阳性细胞数明显增多,ROS、ADMA水平以及Caveolin-1蛋白表达升高,DDAH活性降低。0.01~1 mmol/L阿司匹林剂量依赖性地降低β-gal染色阳性细胞数、ROS和ADMA水平以及Caveolin-1蛋白表达,同时升高细胞DDAH活性（P均〈0.05）。L-NAME（NOS的抑制剂）能完全抑制阿司匹林的上述作用（P〈0.05）。结论高糖环境下阿司匹林（0.01~1mmol/L）能抑制Caveolin-1表达,改善DDAH-ADMA系统的活性而进一步发挥抗氧化损伤、抗细胞衰老的作用。Aim To observe the anti-senescence effects of aspirin on activity of dimethylarginine dimethylaminohydrolase（ DDAH）-asymmetric dimethylarginine（ ADMA） system and caveolin-1 protein expression in human umbilical vein endothelial cells（ HUVEC） exposed to high glucose condition and explore the mechanism of aspirin of antisenescence. Methods The human umbilical vein endothelial cells（ HUVEC） were cultured in Dulbecco＇s modified Eagle＇s medium（ DMEM） containing 5. 5 mmol / L glucose as normal level,33 mmol / L glucose as high glucose,aspirin（ 0. 01~1 mmol/L） with high glucose and 300 μmol/L L-NAME was added to the culture medium when needed for 48 hours. The activity of DDAH were reflected by ADMA concentration determined by high-performance liquid chromatography. The level of intracellular reactive oxygen species（ ROS） was monitored by flow cytometry. Caveolin-1 protein expressions were analyzed by Western blot. Results After the endothelial cells were treated with high glucose concentration for 48 hours,the number of SA-β-gal positive cells,the level of ROS,ADMA and caveolin-1 protein were increased significantly. While,the activity of DDAH was decreased dramatically（ P〈 0. 05）. All these changes were reversed by aspirin（ 0.01 ~ 1 mmol / L） remarkably in a dose-dependent manner（ P〈 0.05）. However,all the effects of aspirin on senescence were completely inhibited by L-NAME,the NOS inhibitor（ P〈 0.05）. Conclusion The anti-senescent effects of aspirin were fulfilled by inhibiting caveolin-1 protein expression and regulating the activity of DDAH-ADMA system.

关 键 词：阿司匹林 细胞衰老 活性氧 二甲基精氨酸二甲胺水解酶 非对称性二甲基精氨酸 Caveolin-1 

分 类 号：R5[医药卫生—内科学]