Apelin-13对ADMA所致内皮细胞骨架损害的影响

Effect of Apelin-13 on the cytoskeleton of endotheliocyte injuried by asymmetrical dimethyl arginine

目的:探讨Apelin-13对非对称性二甲基精氨酸(asymmetrical dimethyl arginine,ADMA)所致的人脐静脉内皮细胞(human umbilical endothelial cell,HUVEC)细胞骨架损害是否有保护作用及其可能的机制。方法:体外培养HUVEC,根据培养液中加入不同的药物分为正常对照组、ADMA 100μmol/L组、Apelin-13 10-6 mol/L组、Apelin-13 10-7 mol/L组、Apelin-13 10-8 mol/L组、ADMA(100μmol/L)+Apelin-13(10-6 mol/L)组、ADMA(100μmol/L)+Apelin-13(10-7 mol/L)组和ADMA(100μmol/L)+Apelin-13(10-8 mol/L)组,孵育24 h后,测量细胞骨架成分肌动蛋白(F-actin)、黏着斑蛋白(vinculin)含量,并测定细胞上清液一氧化氮(nitric oxide,NO)的含量。结果:1)ADMA 100μmol/L单独作用可诱导人脐静脉内皮细胞F-actin和vinculin形成,细胞上清液的NO含量显著减少(P<0.01)。2)联合给予Apelin-13可以减少ADMA诱导的F-actin和vinculin的增加,增加细胞上清液中的NO含量(P<0.01)。3)相关性分析显示上清液中的NO含量分别与F-actin和vinculin含量呈负相关(r=-0.809,P<0.05;r=-0.781,P<0.05)。结论:Apelin-13能够减弱ADMA诱导的人脐静脉内皮细胞骨架损害,这一作用可能与NO生成增加有关。

Objective To examine whether Apelin-13 could protect cytoskeleton of endothelial cell against asymmetrical dimethyl arginine（ADMA） induced injury and the underlying mechanisms. Methods Human umbilical vein endothelial cell（HUVEC） were divided into 8 groups as follow： 1） a control（ECM）group;2） an ADMA 100 μmol/L group;3） an Apelin-13 10-6 mol/L group;4） an Apelin-13 10^-7 mol/L group;5）an Apelin-13 10^-8 mol/L group;6） an ADMA（100 μmol/L） and Apelin-13（10-6 mol/L）group;7）an ADMA（100 μmol/L） and Apelin-13（10^-7 mol/L）group;8） an ADMA（100 μmol/L） and Apelin-13（10^-8 mol/L）group.After incubated for 24 h,the protein content of stress fibre（F-actin）,focal adhesion（vinculin） and the content of nitric oxide（NO） in the culture medium were measured. Results 1） ADMA at concertration of 100 μmol/L could induce the formation of F-actin and vinculin in HUVECs and cause a decrease in NO content in the culture medium（P〈0.01）.2） Co-administration of Apelin could inhibite the ADMA-induced formation of F-actin and vinculin and increase the NO production in the cell culture medium（P〈0.01）.3） Correlation analysis showed that the content of NO in the cell culture medium was negatively related with the content of F-actin and vinculin respectively（r=-0.809,P〈0.05;r=-0.781,P〈0.05）. Conclusion Apelin-13 can inhibite the ADMA-induced cytoskeleton damage and increase NO production,which might be the underlying mechanism.