体外原代培养肝细胞牵张损伤模型的建立及细胞骨架、 粘弹性变化的实验研究

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BioFlex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150kPa、250kPa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。

Establishment of stretch injury model of in vitro cultured primary liver cells and observation on changes of their cytoskeleton and viscoelasticity

Objective To establish the stretch injury model of cultured liver cells,observe the injury of liver cells and changes in their 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 arrangement and distribution of F-actin microfilament of the cytoskeleton in normal liver cells and those with stretch injury was conducted by using a laser con-focal microscope. Meanwhile, the viscoelasticity of the liver cells was determined.Results After the stretch injury, most of the liver cell microfilaments were broken. Furthermore, their network structure disappeared. The cells shrank to be shaped like flakes or there were only some big microfilament branches 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 mechanism of changes of dynamic features in the liver cells after stretch injury.