p38MAPK在非对称性二甲基精氨酸所致内皮细胞骨架改变过程中的作用

目的：探讨非对称性二甲基精氨酸（asymmetricdimethylargine，ADMA）对人脐静脉内皮细胞（humanumbilicalveinendothelialcells，HUVECs）的细胞骨架改变的影响及p38丝裂原激活蛋白激酶（p38mitogen—activatedproteinkinase，p38MAPK）在该过程中的作用。方法：体外进行HUVEC培养，实验分为正常对照组、sB203580组、ADMA组（量效关系组、时效关系组）及sB203580＋ADMA组（SB203580＋ADMA量效关系组及SB203580＋ADMA时效关系组）。对各组细胞进行免疫荧光染色，利用激光扫描共聚焦显微镜观察肌动蛋白（F-actin）形态变化，图像分析软件行F-actin荧光灰度值分析，流式细胞仪行F．actin荧光定量分析。结果：ADMA可诱导HUVECs应力纤维形成，导致F．actin荧光灰度值、荧光定量增加；而SB203580可抑制ADMA的作用。结论：ADMA可呈时间及浓度依赖性地导致细胞骨架改变。p38MAPK特异性抑制剂SB203580可抑制ADMA对内皮细胞骨架的改变，提示p38MAPK参与了ADMA所导致的HUVECs内皮细胞的骨架改变。

Role of p38MAPK in asymmetric dimethylarginine induced cytoskeleton changes in endothelial cells

Objective： To explore the effect of asymmetric dimethylarginine （ADMA） on the cytoskeleton of human umbilical vein endothelial cells （HUVECs） and the role of p38 mitogen-activated protein kinase （p38MAPK） in this process. Methods： HUVECs were cultured in vitro and divided into four groups： a control group, a SB203580 group, a ADMA group （ including a subgroup of dose- relationship）, and a SB203580＋ADMA group （including a subgro time-effect relationship）. The treated cells were stained by immun effect relationship and a subgroup of time-effect up of dose-effect relationship and a subgroup of ofluorescence, then the conformational changes of F-actin of cells in all groups were observed under confocal microscope. The grey scale values of acquired imageswere analyzed by image analysis software and the fluorescence of F-actin was quantified by flow cytometry. Results： ADMA can increase the formation of stress fibers, then resulted in the increase of the grey scale values and fluorescent quantification of F-actin. SB203580 can inhibit the effect of ADMA on cytoskeleton. Conclusion： ADMA can induce the cytoskeleton changes in endothelial cells in a concentration or time-dependent manner, which is involved in p38MAPK pathway.