摘要
目的研究培养人视网膜色素上皮细胞(RPE)受机械牵拉力后细胞骨架的变化。方法将胶原包被的磁珠悬液加入培养的人RPE细胞后孵育,洗去未结合的磁珠。在磁场垂直方向力作用下0、4、8、12和24h,以及加入细胞松弛素D(0.05mmol/L,25min)预处理后,用免疫荧光双标染色法着染细胞中的肌动蛋白和波形蛋白,并用激光共聚焦显微镜观察。结果机械牵拉作用4h后,人RPE细胞的形态和微丝的极性发生改变。肌动蛋白微丝的排列与受力方向一致。细胞骨架微丝表达主要在细胞核周及磁珠聚集处周围。而加入细胞松弛素D预处理的细胞,在8h时发生以上变化。结论机械牵拉可引起人RPE细胞骨架分布的改变,使其表现出肌细胞的某些生物学特征,这对细胞的移行和增生可能有促进作用。
Objective To observe changes of the cytoskeleton of cultured human retinal pigment epithelium （hRPE） cells in a mechanical stress model in vitro. Methods Ferric oxide beads, coated with collagen, were added to dishes containing substrate-attached hRPE cells. After incubation, the cells were washed to remove unbound beads. The cells were pretreated by actin polymerization inhibitor cytochalasin D （0.05 mmol/L, 25 min）. Before and 4, 8, 12 and 24 h after vertical magnetic force application the cells were stained for actin and vimentin using immunity fluorescence double labelled technique and analyzed with confocal microscopy. Results Four hours after force application, the polarity of hRPE cells was changed, and actin was polymerized into filaments in array of them along with the direction of the force. Cytoskeleton filaments were located mainly around the nucleus and the ferric oxide beads. Pretreatment with cytochalasin D, the same changes as described above occurred 8 h after magnetic force application. Conclusions Mechanical force could induce changes of distribution of cytoskeleton in bRPE cells, which are some biological characteristics of muscle cells, and this may facilitate the cellular migration and proliferation.
出处
《中华眼科杂志》
CAS
CSCD
北大核心
2006年第2期121-126,共6页
Chinese Journal of Ophthalmology
基金
国家自然科学基金资助项目(30200311)
