体外原代培养肝细胞牵张损伤模型的建立及细胞骨架、 rn粘弹性变化的实验研究

摘要

目的　建立培养肝细胞牵张损伤模 型， 探讨肝细胞损伤、细胞骨架和粘弹性的改变及其发生机制。方法　用BioFl ex培养皿培养原代肝细胞贴壁生长后，通过小型生物撞击机分别以驱动压力150 kPa、250kP a牵张损伤细胞。然后对正常肝细胞及牵张损伤后肝细胞骨架Factin微丝排列及分布变化在 激光共聚焦显微镜下进行形态学观察，同时测定肝细胞的粘弹性。结果　 牵张损伤后肝细胞微丝大部分断裂，网状结构消失，收缩呈絮状，或遗留粗大的微丝束，粘 弹性显著降低。结论　肝细胞粘弹性显著降低与细胞骨架的损伤断裂，可 能是细胞损伤力学特性变化的分子生物学机制。%Objective　To establish the stretch injury mode l of cultured liver cells,observe the injury of liver cells and changes in th eir cytoskeleton and viscoelasticity,and explore the mechanisms.Methods 　After the primary liver cells were cultured in BioFlex culturing plate , they were impacted with a small biological impacting machine with pressures of 150 kPa and 250 kPa respectively. The morphological observation on changes of a rrangement and distribution of F-actin microfilament of the cytoskeleton in norm al liver cells and those with stretch injury was conducted by using a laser con- focal microscope. Meanwhile, the viscoelasticity of the liver cells was determin ed.Results　After the stretch injury, most of the liver cell mic rofilaments were broken. Furthermore, their network structure disappeared. The c ells shrank to be shaped like flakes or there were only some big microfilament b ranches left. The viscoelasticity of the liver cells was significantly decreased .Conclusion　The remarkable decrease of the viscoelasticity and breaking of cytoskeleton in the liver cells might be the molecular biological me chanism of changes of dynamic features in the liver cells after stretch injury.