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目的 :探讨成骨样细胞株UMR_10 6受机械力作用后细胞骨架微丝蛋白F_actin的变化。方法 :成骨样细胞株UMR_10 6体外扩增 ,实验组以 2 2 5×g相对离心力离心 10min后 ,分 7组分别继续培养 15、30min ,1、4、6、12、2 4h ,并设立相应对照组。所有样本经BODIPYFLPhallacidin处理后 ,在激光扫描共聚焦显微镜下观察、记录图片并测定单个细胞的平均荧光强度。结果 :对照组F_actin较粗 ,呈束状 ,排列较整齐 ;实验组除 2 4h外 ,其余各组的纤维明显较对照组纤细、短 ,散乱分布于胞浆内、无方向性 ;但两组间差异随时间延长逐渐减小。除 2 4h外 ,实验组的平均荧光强度较对照组明显降低 ,差异有显著性 ;随时间延长 ,实验组荧光强度逐渐回升 ,到 2 4h组时较对照组增强。结论 :成骨样细胞受力后细胞骨架微丝的结构明显变化 ,荧光染色变弱 ,表明在机械力作用下微丝结构趋于解聚。去除刺激后随时间延长 ,实验组逐渐恢复并比对照组增高 ,说明细胞骨架受力后的改变是可恢复和改建的
Objective: To investigate the changes of cytoskeleton fibroblasts F_actin after osteogenic cell line UMR_10 6 under mechanical force. Methods: The osteoblast-like cell line UMR_10 6 was expanded in vitro. The experimental group was centrifuged at 225 g for 10 minutes, and then cultured for 15, 30, 1, 4, 6, 12 and 24 hours respectively Corresponding control group. All samples were treated with BODIPYFLPhallacidin and observed under a laser scanning confocal microscope. The images were recorded and the average fluorescence intensity of individual cells was measured. Results: In the control group, F_actin was thicker and bundled and neatly arranged. In the experimental group except for 24 h, the fibers in other groups were slender, short and scattered in the cytoplasm without orientation; however, the two groups Differences between time gradually decreased with time. Except 2 4h, the average fluorescence intensity of the experimental group was significantly lower than that of the control group, the difference was significant; with time prolonged, the fluorescence intensity of the experimental group gradually rose to 24 hours group than the control group increased. CONCLUSION: The structure of cytoskeleton microfilaments changes obviously under the stress of osteoblast-like cells, and the fluorescence staining becomes weaker, which indicates that the microfilament structure tends to depolymerize under the mechanical force. After the stimulation was removed, the experimental group gradually recovered and increased compared with the control group, indicating that the cytoskeletal changes after stress were recoverable and remodeled