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

发布时间：2018-03-02 02:36

本文关键词： 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-　出处：《中国动脉硬化杂志》2016年05期 　论文类型：期刊论文

【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(P0.05)。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。
[Abstract]:Objective to observe the effect of aspirin on the expression of Caveolin-1 protein and the system of DDAH- asymmetric dimethylarginine hydrolase (DDAH- ADMA) in endothelial cells induced by high glucose. Methods Human umbilical vein endothelial cells (HUVECs) were cultured in normal glucose concentration medium (5.5 mmol / L), high glucose medium (33 mmol / L), high glucose aspirin (0.01 mmol / L) and L-NAMEE (300 渭 mol / L). 尾 -galactosidase (尾 -galase) staining was used to identify aging cells after 48 h culture. Flow cytometry was used to detect the level of reactive oxygen species (Ros) in the cells. Western blot was used to detect the expression of Caveolin-1 protein. The content of ADMA in the supernatant of the cells and the activity of DDAH were measured by HPLC. Results the endothelial cells were treated with high glucose for 48 h. The number of 尾 -gal staining positive cells increased significantly and the expression of Caveolin-1 protein increased. DDAH activity decreased. 0.01 mmol/L aspirin decreased the Ros and ADMA levels and the expression of Caveolin-1 protein in a dose-dependent manner. At the same time, the inhibition of DDAH activity was inhibited completely by increasing the activity of DDAH. L-NAME-NOS). Conclusion Aspirin (0.01 mmol / L) can inhibit the expression of Caveolin-1 and improve the activity of DDAH-ADMA system in high glucose environment, thus further exerting antioxidant damage. The antiaging effect of cells.
【作者单位】：辽宁中医药大学附属医院内分泌科;中国医科大学附属第一医院内分泌科;
【基金】：辽宁省博士启动基金资助项目(20121101)
【分类号】：R587.2

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