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

    Abstract:

    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.

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王高频,杨雪佳,王继红,马海芳,刘国銮.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[J].中国动脉硬化杂志,2010,18(10):795~798.

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  • 收稿日期:2010-08-25
  • 最后修改日期:2010-10-16
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