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  

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

作　　者：张常勤[1] 张东亮[2] 刘文虎[2] 

机构地区：[1]首都医科大学附属北京天坛医院肾内科,北京100050 [2]首都医科大学附属北京友谊医院肾内科,北京100054

出　　处：《临床与病理杂志》2014年第1期34-40,共7页Journal of Clinical and Pathological Research

摘　　要：目的:探讨非对称性二甲基精氨酸(asymmetric dimethylargine,ADMA)对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)的细胞骨架改变的影响及p38丝裂原激活蛋白激酶(p38 mitogen activated protein kinase,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。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.

关 键 词：非对称性二甲基精氨酸 肌动蛋白 p388丝裂原激活蛋白激酶 细胞骨架 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]