力学刺激对软骨细胞细胞骨架的影响

摘要

Objective To Explore the Changes of Chondrocyte Cytoskeleton by Dynamic and Static Mechanical Stimuli in Vitro.Methods Chondrocytes derived from healthy New Zealand rabbits,then the protocols (1Hz,500με1000με 1500με and 0Hz,500με 1000με 1500με) were applied 6 hours every day for 3 days,Immunofluorescence was employed to directly observe the organization of the cytoskeleton.Morphological remodeling of the cytoskeleton was visualized by fluorescence microscope.Fluorescence intensity of chondrocytes were recorded by densitometry.The dynamic and static remodeling of during the mechanical protocols was observed morphologically and mechanically by fluorescence microscope and Fluorescence analysis respectively.Results The variations in both the morphology and the mechanical property actin were highly synchronous to vimentin.It's fluorescence became stronger after dynamic mechanical stimuli at first and became weakness again,but it's continuing weaken under static mechanical stimuli.The fibers of actin and vimentin became thin,sparse,and disordered,and the cell stiffness decreased concomitantly.The variations in both the morphology and the mechanical property both were highly synchronous to the mechanical stimuli in a time-dependent manner.Conclusion Mechanical stimuli on chondrocytes has been discovered and defined in this study.The chondrocyte cytoskeleton changed by mechanical stimuli.Meanwhile it's related the time and different forces.%目的 探讨不同应力刺激下软骨细胞细胞骨架的荧光强度和形态分布的变化.方法 取新西兰兔关节软骨细胞,分离传代增殖然后将第3代软骨细胞分别给以3组不同的动态(1Hz,500με 1000με 1500με)和静态(0Hz,500με 1000με1500με)力学刺激分组培养,然后行细胞骨架肌动蛋白(actin)、中间丝波形蛋白(vimentin)、微管(tubulin)的免疫荧光染色,吸光法对标本进行测定,分析荧光强度和细胞骨架分布的变化.结果 软骨细胞在未受到应力刺激时其分布相对分散并显示比较均匀,在力学刺激后,细胞骨架发生的排列和强度均发生了变化.vimentin在动态的力学刺激下荧光强度逐渐增强,Actin先减弱后逐步增强,而Tublin是先增强在减弱再增强.vimentin、actin和tublin的荧光强度在静态的力学刺激下有明显的一致性的变化,随压力的增加其强度明显的下降,其排列疏松变化明显.结论 在力学刺激下,细胞骨架发生的明显的变化,而在各种变化中,其变化和力学刺激的大小和时间和刺激的方式有关系.