剪切力对视网膜色素上皮细胞syndecan-1,血管细胞黏附分子-1,E-选择素蛋白表达和肌动蛋白的影响

Effects of shear stress on the protein expressions of syndecan-1,vascular cell adhesion molecule-1,E-selectin and actin in retinal pigment epithelial cells

目的:研究剪切力对视网膜色素上皮(RPE)细胞synde-can-1,血管细胞黏附分子(VCAM)-1和E-选择素(se-lectin)表达和细胞骨架的影响。方法:体外培养人RPE细胞,应用2dyns/cm2大小的剪切力干预0,0.25,0.5,0.75,1,1.5,3h后,用免疫荧光染色法观察RPE细胞syndecan-1,VCAM-1和E-selectin表达和肌动蛋白的变化。结果:静态下的RPE细胞呈多角形和不规则形。2dyns/cm2剪切力作用0.75h后,细胞间隙加大,部分细胞形状变圆、变小;作用后1.5h,细胞极性改至与剪切力作用方向一致。静态条件下,肌动蛋白呈黄绿色丝状分布于中央细胞内,排列松散不规则且微丝较细;剪切力作用3h后,肌动蛋白增多增粗,且沿切力方向排列于中央和远周边细胞。剪切力作用后,RPE细胞syndecan-1的表达在0.5h时最弱,1h时最强;VCAM-1的表达在0.5h时最弱,1.5h时最强;E-selectin的表达在0.75h时最低,3h时最高。结论:剪切力可引起RPE细胞syndecan-1,VCAM-1和E-selectin及细胞骨架蛋白发生变化,它们之间的相互协作对RPE细胞的形态、黏附和力学信号转导起重要作用。

AIM： To observe the effects of shear stress on the expressions of syndecan-1, vascular cell adhesion molecule （VCAM） -1, E-selectin and actin in cultured retinal pigment epithelial （RPE） cells. METHODS： The cultured RPE cells were treated with 2 dyns/cm2 shear stress for 0, 0.25, 0.5, 0.75, 1, 1.5 and 3 hours. Immunofluorescence staining was used to detect the expression of syndecan-1, VGAM-1, E-selectin and actin in RPE cells. RESULTS： RPE cells under static conditions were in multi-angle or atactic shapes. After treated with 2 dyns/cm2 for 0.75 hour, the cellular spacing was widened, and some cells became rounded and smaller; after the treatment for 1.5 hour, the cell polarity changed to accord with the direction of the force. Under static conditions, actin was located in the cellular center with yellow-green filiform, and the cytoskeleton filament was loose, irregular and thin. Actin increased obviously after treatment for 3 hours, and arranged in the central and far peripheral cell with the array of the force. With shear stress, syndecan-1 was the weakest in 0.5 hour, and the strongest in 1 hour; VCAM-1 was lowest in 0.5 hour, and highest in 1.5 hour; E-selectin decreased to the weakest in 0. 75 hour, and increased to the strongest in 3 hours.CONCLUSION： Shear stress can induce changes of syndecan-1, VCAM-1, E-selectin and actin in RPE cells. Their interaction may play important roles in the morphology, adhesion and mechanical force signal transduction, which may participate in the occurrence and development of some eye diseases.