盐酸法舒地尔对血管紧张素Ⅱ诱导的人脐静脉内皮细胞骨架损伤的保护作用  

Protective Effect of Fasudil Hydrochloride on Cytoskeleton of Human Umbilical Vein Endothelial Cells Injury Induced by AngiotensinⅡ in Vitro

作　　者：王高频[1] 杨雪佳[1] 王继红[1] 马海芳[1] 刘国銮[1] 

机构地区：[1]辽宁医学院附属第一医院心血管内科,辽宁省锦州市121001

出　　处：《中国动脉硬化杂志》2010年第10期795-798,共4页Chinese Journal of Arteriosclerosis

摘　　要：目的探讨盐酸法舒地尔对血管紧张素Ⅱ诱导的人脐静脉内皮细胞Rho激酶和肌球蛋白轻链磷酸化表达的影响。方法体外培养人脐静脉内皮细胞,并将其分为对照组、血管紧张素Ⅱ组、特异性阻断剂组(Y-27632)和盐酸法舒地尔组,MTT法观察血管紧张素Ⅱ对细胞活力的影响;免疫荧光法观察各组细胞骨架肌动蛋白F-actin形态及分布的变化,Western Blotting法测定内皮细胞Rho激酶和磷酸化肌球蛋白轻链的表达。结果与对照组相比,血管紧张素Ⅱ对内皮细胞活力有明显抑制作用(P<0.01),Rho激酶特异性阻断剂和盐酸法舒地尔能明显减弱血管紧张素Ⅱ对内皮细胞活力的抑制作用(P<0.01)。与对照组相比,血管紧张素Ⅱ组内皮细胞F-actin形态学发生明显的改变,细胞间裂隙增大。与血管紧张素Ⅱ组比较,Rho激酶阻断剂和盐酸法舒地尔组均可使Rho激酶蛋白表达降低(P<0.01),但仍高于对照组(P<0.01)。结论盐酸法舒地尔对血管紧张素Ⅱ诱导的人脐静脉内皮细胞的保护作用是通过抑制Rho/Rho激酶信号通路中Rho激酶和磷酸化肌球蛋白轻链蛋白的表达,以减弱血管紧张素Ⅱ对内皮细胞骨架的损伤。Aim To investigate the effects of fasudil hydrochloride on the injury of human umbilical vein endothelial cells（HUVEC）induced by angiotensinⅡ（AngⅡ）which influence on the expression of Rho associated kinase（ROCK）and phospho-myosin light chain（P-MLC）.Methods HUVEC were incubated in vitro and treated with AngⅡ,AngⅡ＋specific blocker（Y-27632）,AngⅡ＋fasudil hydrochloride for different time.Cell activities were detected by MTT method.Morphous and distribution changes of F-actin were observed by immunofluorescence,expression of ROCK and P-MLC were measured by Western Blotting.Results Compared with control group,AngⅡ had significant inhibitory effect on endothelial cells vigor（P〈0.01）,furthermore,these effects were dose-dependent（P〈0.01）in the other three groups.Morphous and distribution changes of F-actin by imunofluorescence,compared with control group,the rupture of stree fibers constructed by F-actin,deformation and digitation of HUVEC were exhibited.Compared with AngⅡ group,expression of ROCK and P-MLC in the specific blocker group and the drug group were remarkably restrained（P〈0.01）,but higher than the control group（P〈0.01）.Conclusion Fasudil hydrochloride can protect the cytoskeletal of HUVEC which were induced by AngⅡ,through obviously controlling the expression of ROCK and P-MLC in the Rho/Rho kinase signaling pathways,thus weakened the damage in cytoskeletal.

关 键 词：动脉粥样硬化 血管紧张素Ⅱ 人脐静脉内皮细胞 细胞骨架 RHO激酶 

分 类 号：R363[医药卫生—病理学]