机械牵拉对培养人视网膜色素上皮细胞内钙离子的影响  

Effects of mechanical stress on i of human retinal pigment epithelial cells in vitro

作　　者：侯旭[1] 惠延年[1] 韩泉洪[1] 张晓光[1] 王建州[1] 郭长梅[1] 

机构地区：[1]第四军医大学西京医院眼科,中国陕西省西安市710032

出　　处：《国际眼科杂志》2005年第1期50-54,共5页International Eye Science

基　　金：中国国家自然科学基金资助项目(No.30200311)~~

摘　　要：目的:观察培养人视网膜色素上皮细胞(retinal pigment epithelial cells,RPEs)在受机械牵拉力时细胞内Ca^2+的变化。方法:将胶原包被的磁珠悬液加入培养人RPE细胞后孵育,附着磁珠的细胞放置在恒定磁场中,使用钙荧光指示剂fluo-3/AM和激光扫描共聚焦显微镜(LSCM)观察在机械牵拉作用的前后细胞内钙离子的变化情况。结果:RPE细胞荧光着色很强,遍布整个细胞,其中细胞核比细胞质更强。机械牵拉后荧光强度显著增强,加入氯化锰后迅速降低。EGTA预处理的细胞在受力后荧光强度增加不显著,加入氯化钙后明显增强。细胞松弛素D预处理的细胞在受力后荧光强度比正常组增高2倍以上。结论:机械牵拉能显著增加钙离子内流,并且增加细胞内的钙库释放。钙离子内流可能是RPE细胞损伤前的信号。AIM: To observe the calcium flux of human retinal pigment epithelial cells(RPEs)during mechanical stress in vitro.· METHODS: Ferric oxide microparticles, coated with collagen, were added into dishes containing substrate-attached RPEs. After incubation, the cell layer bounded with beads was laid in invariable magnetic field. Cultured human RPE cells were analyzed by using the fluorescence Ca2+ dye fluo-3/AM and laser scanning confocal microscope (LSCM) before and after force stimu- lation.· RESULTS: Normal fluorescence in RPE cells was strong and distributed throughout the cells. The nucleus appeared to be more fluorescent than the cytoplasm. After force stimulation, a rapid increase in fluorescence intensity was observed, followed by a rapid decrease by adding MnCl2. There was no obvious difference in increased curve in cells preincubated with EGTA. By adding CaCl2, the fluorescence intensity increased again. Moreover, after force stimulation, the fluorescence intensity in cells preincubated with cytochalasin D increased over twice than that in normal cells.· CONCLUSION: Mechanical stress may increase the calcium influx of RPEs and initiate the release of intracellular calcium store. The calcium influx may be a signal before RPEs injury.·

关 键 词：机械牵拉 RPE 人视网膜色素上皮细胞 胞内钙离子 钙离子内流 观察 氯化锰 培养 结论 LSCM 

分 类 号：R774[医药卫生—眼科] R781.05[医药卫生—临床医学]