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
| 作者: |
1张常勤,
2张东亮,
2刘文虎
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 |
| 通讯: |
张常勤
Email: zhangchangqin138@163.com |
| DOI: | 10.3978/j.issn.2095-6959.2014.01.005 |
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。
关键词：
非对称性二甲基精氨酸；肌动蛋白；p38丝裂原激活蛋白激酶；细胞骨架
Role of p38MAPK in asymmetric dimethylarginine induced cytoskeleton changes in endothelial cells
CorrespondingAuthor: ZHANG Changqin Email: zhangchangqin138@163.com
DOI: 10.3978/j.issn.2095-6959.2014.01.005
Abstract
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-effect relationship and a subgroup of time-effect relationship), and a SB203580+ADMA group (including a subgroup of dose-effect relationship and a subgroup of time-effect relationship). The treated cells were stained by immunofluorescence, then the conformational changes of F-actin of cells in all groups were observed under confocal microscope. The grey scale values of acquired images were 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.