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目的观察水平回转模拟微重力(modeled microgravity,MMG)后人骨肉瘤MG-63细胞受流体剪切应力(flow shear stress,FSS)作用后细胞骨架的改变,探讨模拟微重力情况下骨质丢失的细胞学机制。方法 MG-63成骨细胞采用回转器水平回转培养48 h模拟微重力效应,同时设静止培养为对照组。之后,细胞随机分为两组,一组利用流室系统实施FSS,FSS设为1.5 Pa,作用时间60 min,另一组无FSS作用。细胞经过免疫荧光染色后,激光共聚焦显微镜观察细胞骨架系统的变化。结果 FSS作用后,MG-63细胞微丝和微管的形态和分布均发生变化,微丝可见多束状排列,荧光强度增强,出现粗的应力纤维;微管可见其极性中心移向细胞质外缘,细胞外层荧光量加重,有少量束状结构出现。MMG后,MG-63细胞的细胞骨架发生解聚和重排,微丝向核周集聚;微管发生断裂,变短,弯曲等变化。MMG后,再给予FSS刺激,微丝未能形成应力纤维,仅见微丝、微管在应力方向有拉长。结论 1.5 Pa,60 min流体剪切应力未能在回转模拟微重力后成骨细胞内诱导形成应力纤维。
Objective To observe the changes of cytoskeleton in human osteosarcoma MG-63 cells under the action of fluid shear stress (FSS) after the horizontal gyration of simulated microgravity (MMG) Learning mechanism. Methods MG-63 osteoblasts were cultured in a gyrator with horizontal rotation for 48 h to simulate microgravity effects. At the same time, MG-63 osteoblasts were cultured as control group. After that, the cells were randomly divided into two groups. One group used flow chamber system to implement FSS. The FSS was set at 1.5 Pa and the action time was 60 min. The other group had no FSS effect. After immunofluorescence staining, the changes of the cytoskeletal system were observed by laser confocal microscopy. Results The morphology and distribution of microfilaments and microtubules of MG-63 cells changed after treated with FSS. Microfilaments were arranged in multi-bundles, the fluorescence intensity increased, and coarse stress fibers appeared. The polarity centers of microtubules shifted to the cytoplasm The outer edge, extracellular fluorescence increased, a small amount of bundles appear. After MMG, the cytoskeleton of MG-63 cells disaggregated and rearranged, and the microfilaments clustered into the perinuclear area. The microtubules ruptured, shortened and bent. MMG, then given FSS stimulation, micro-filaments failed to form stress fibers, only see the microfilaments, microtubules elongated in the direction of stress. Conclusions The fluid shear stress at 1.5 Pa and 60 min failed to induce stress fibers in osteoblasts after gyration simulating microgravity.