切应力对与内皮细胞联合培养的血管平滑肌细胞粘附的影响及其机制  

Shear stress-induced regulation of vascular smooth muscle cells adhesion in a coculture system with endothelial cells

作　　者：王汉琴[1] 白玲[1] 王燕华[1] 姜宗来[1] 

机构地区：[1]上海交通大学力学生物学与医学工程实验室,上海200240

出　　处：《医用生物力学》2007年第1期4-7,20,共5页Journal of Medical Biomechanics

基　　金：国家自然科学基金(10572096;10132020)

摘　　要：目的细胞粘附是细胞迁移的关键性起始步骤。研究切应力和内皮细胞(endothelial cells,ECs)对血管平滑肌细胞(vascular smooth muscle cells,VSMCs)粘附的影响及其可能的信号通路,为探讨流体切应力诱导的血管壁细胞迁移行为的机制提供一些实验依据。方法应用VSMCs和ECs联合培养流动腔系统,对ECs面施加1．5Pa切应力,12h;以静止状态下,单独培养的VSMCs以及与ECs联合培养的VSMCs为对照,应用细胞粘附实验和Western Blot技术,观察切应力对与EC联合培养的VSMCs粘附的影响及蛋白激酶B(PKB/Akt)磷酸化水平表达变化。结果静态联合培养12h,VSMCs的粘附能力明显增强,同时磷酸化Akt的表达平行增高。切应力作用下,明显抑制了联合培养的VSMCs粘附,同时磷酸化Akt的表达平行降低。结论生理大小切应力明显抑制了ECs诱导的VSMCs粘附,其中Akt信号通路起了关键作用。Objective Cell adhesion is an essential process for cell migration. The influence of shear stress on the adhesion of VSMCs was elucidated in a co-culture system and to provide some experimental evident in molecular mechanisms of shear stress-induced call migration in vascular wall. Methods Using a parallel-plate co-culture flow chamber system, human VSMCs co-cultured with human ECs were exposed to static or laminar shear stress, 15dynes/ cm^2, conditions for 12 hours, with VSMCs cultured alone at static condition as a control. The adhesion of VSMCs was then assessed with call adhesion assays. The activation of Phosphatidylinositol-3 kinase （P13K/Akt） pathway in VSMCs was assessed by wastem blotting. Results It demonstrated that VSMCs co-cultured with ECs under static condition induced VSMC adhesion, whereas the shear stress applied to EC side for 12h significantly inhibited this process. Western blotting showed that there was a consistent correlation between the level of Akt phosphorylation and the efficacy of shear stress-inducad EC regulation VSMC adhesion. Conclusions Our findings indicate that shear stress protect against EC-inducad VSMC adhesion. Akt is a major downstream factor of P13K involved in the process.

关 键 词：细胞粘附 切应力 联合培养 血管平滑肌细胞 蛋白激酶B 

分 类 号：R318.01[医药卫生—生物医学工程]