阿司匹林抗高糖诱导内皮细胞衰老过程中对一氧化氮-一氧化氮合酶系统的影响  

The effect of aspirin on NO-NOS system invovled in the endothelial cell senescence induced by 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,沈阳110031 [2]中国医科大学附属第一医院内分泌科

出　　处：《中国糖尿病杂志》2014年第9期841-845,共5页Chinese Journal of Diabetes

基　　金：辽宁省博士启动基金(20121101)

摘　　要：目的 探讨阿司匹林是否通过影响一氧化氮（NO）-一氧化氮合酶（NOS）系统发挥抗高糖诱导的内皮细胞衰老的作用. 方法 人脐静脉血管内皮细胞（HUVECs）分别于正常糖浓度培养液（5.5 mmol/L）、高糖培养液（33 mmol/L）、含阿司匹林（0.01～1mmol/L）培养液及含L-NAME（300 μmol/L）培养液中培养48 h.采用β-半乳糖苷酶（SA-β-gal）染色鉴定衰老细胞,流式细胞仪检测细胞内活性氧簇（ROS）水平.Griess法检测细胞总NO水平,荧光酶标仪检测细胞内NOS活性.Westernblot分析eNOS蛋白及eNOS人丝氨酸（Ser-1177）蛋白表达. 结果 高糖作用内皮细胞48 h后与正常糖浓度相比,SA-β-gal阳性细胞数明显增多.阿司匹林剂量依赖性地减少SA-β-gal阳性细胞数,降低ROS的水平,升高NO的水平、总NOS活性和eNOS Ser-1177蛋白表达（P＜0.05）.L-NAME（NOS的抑制剂）能完全抑制高糖环境下阿司匹林的上述作用（P＜0.05）.各组间eNOS总蛋白表达差异无统计学意义（P＞0.05）. 结论 高糖环境下阿司匹林通过提高细胞NOS活性而升高NO水平,降低氧化应激反应而发挥抗衰老作用.Objective To investigate whether aspirin exerts its effect on the endothelial cell senescence induced by high glucose through influencing NO-NOS system. Methods The human umbilical vein endothelial cells(HUVECs)were cultured for 48 hours in different culture medium including the normal glucose(5.5mmol/L),high glucose(33mmol/L),aspirin(0.01~1mmol/L)and LNAME(300μmol/L)culture medium.SA-β-gal staining was used to identify the senile cells.The level of intracellular reactive oxygen species(ROS)was monitored by flow cytometry.Griess reaction was used to measure the total nitric oxide(NO)production and NO synthase(NOS)activity was measured using a fluorescence microplate reader.Endothelial NOS(eNOS)protein and eNOS Ser-1177 protein expressions were analyzed by western blotting. Results After exposure to high glucose for 48 hours,the number of SA-β-gal positive cells was significantly increased compared with the normal glucose culture medium.Aspirin dose-dependently reduced the number of SA-β-gal positive cells,decreased the level of ROS,and elevated the level of NO,the total NOS activity and the eNOS Ser-1177 protein expression(P<0.05).LNAME(NOS inhibitor)could completely inhibit those effects of aspirin in high glucose condition(P<0.05).However,the difference of total eNOS protein expression between groups was not statistically significant(P>0.05). Conclusion Aspirin exerted its anti-senescent effects by increasing NO production and decreasing oxidative stress reaction via up-regulation of NOS activity in high glucose condition.

关 键 词：阿司匹林 高糖 内皮细胞衰老 活性氧簇 一氧化氮 

分 类 号：R587.2[医药卫生—内分泌]