不同强度流体剪切力对MC3T3-E1成骨细胞内Piezo1蛋白表达的影响

Effect of different fluid shear stress intensities on expression of Piezo1 protein in MC3T3-E1 osteoblasts

目的:研究在不同强度的流体剪切力作用下,MC3T3-E1成骨细胞内Piezo1蛋白的表达差异。方法:对载玻片上贴附的MC3T3-E1成骨细胞加载不同强度(0、3、6、9、12、15、18 dyne/cm^2)流体剪切力45 min,通过免疫荧光和Western Blot分别观察MC3T3-E1成骨细胞内Pieoz1蛋白的表达。结果:免疫荧光及Western Blot结果显示,随着流体剪切力强度的增加,MC3T3-E1成骨细胞内Piezo1蛋白表达量逐渐递增,6 dyne/cm^2时达顶峰,此后逐渐下降,15、18 dyne/cm^2时与空白组水平相当,显微镜下可观察到细胞结构破坏,细胞被冲散。结论:流体剪切力可促进MC3T3-E1成骨细胞内Piezo1蛋白表达;加力6 dyne/cm^2时,Piezo1表达最高;较大的流体剪切力非但不能促进MC3T3-E1成骨细胞内Pizeo1蛋白的表达,反倒对细胞起破坏作用。

Objective To investigate the expression differences of Piezo1 protein in MC3T3-E1 osteoblasts under different fluid shear stress intensities.Methods The MC3T3-E1 osteoblasts attached to the slides were subjected to different fluid shear stress(0,3,6,9,12,15,18 dyne/cm2)for 45 minutes,and then the expression of Pieoz1 protein in MC3T3-E1 osteoblasts was respectively observed by immunofluorescence and Western Blot.Results The results from mmunofluorescence and Western Blot showed that with the increase of shear stress,the Piezo1 protein expression in MC3T3-E1 osteoblasts increased accordingly,but at 6 dyne/cm^2,the expression reached its highest and then decreased gradually,and at 15 or 18 dyne/cm^2,the expression kept the same with the blank group,in which the cells were found destroyed and dispersed.Conclusion The increase of fluid shear stress can improve the expression of Piezo1 protein,and when the stress is added to 6 dyne/cm^2,the expression reaches its highest point,but a stress greater than that cannot promote the expression of Pizeo1 protein in MC3T3-E1 osteoblasts any more,and instead it damages the cells.