丝切蛋白在成骨细胞细胞骨架改建中的作用探讨  

Effects of cofilin phosphorylation on the actin cytoskeleton reorganization induced by shear stress

作　　者：刘艳辉[1] 李友瑞[1] 邵敏锋[1] 张晓娟[2] 付强[1] 

机构地区：[1]中山大学光华口腔医学院·附属口腔医院修复科·中山大学口腔医学研究所,广州510055 [2]广州中医药大学第一附属医院口腔科

出　　处：《中华口腔医学杂志》2010年第12期763-766,共4页Chinese Journal of Stomatology

基　　金：广东省医学科研项目（A2008227）;广东省科学事业费科技计划（20088030301121）;广东省产业技术研究与开发资金计划（20098050700027）

摘　　要：目的 采用流体剪切力加载成骨细胞,通过观察细胞骨架改建和测定丝切蛋白及磷酸化丝切蛋白的含量,探讨丝切蛋白在流体剪切力引起成骨细胞细胞骨架改建中的作用.方法 采用1.2 Pa的流体剪切力作用于成骨细胞0（空白对照组）、15、30、45、60、120 min后,蛋白质印迹法测定细胞内丝切蛋白及磷酸化丝切蛋白的含量,采用异硫氰酸荧光素标记的鬼笔环肽染色观察纤维型肌动蛋白的变化.结果 成骨细胞受流体剪切力刺激后,随加载时间延长,细胞内磷酸化丝切蛋白含量持续升高.流体剪切力作用60 min内丝切蛋白随加载时间延长有所下降,但加载120 min时细胞内丝切蛋白含量（0.254±0.026）为加载60 min（0.162±0.004）的1.5倍.随流体剪切力作用时间延长,各组细胞纤维型肌动蛋白染色逐渐增强,纤维增粗;加载120 min时的平均荧光强度（42.93±6.41）为空白对照组（15.41±3.60）的2.8倍（P＜0.05）.结论 流体剪切力刺激可通过丝切蛋白磷酸化,使成骨细胞细胞骨架发生改建.Objective To explore the effects of cofilin on the actin eytoskeleton reorganization in osteoblasts induced by fluid shear stress. Methods Fluid shear stress （ 1.2 Pa） was applied to osteoblasts for 0（control group）, 15, 30, 45, 60, 120 min in vitro. Cells were stained with fluorescein isothiocyanate （FITC）-phalloidin for fiber-actin, and confocal laser scanning microscope（ CLSM ） was used to observe the fluorescence of fiber-actin. Western blotting was used to detect the expression of the cofilin and the phosphocofilin. Results Actin filaments became organized into stress fibers that were thicker and more abundant than those in non-flowed cells. The fluorescence intensity （ 38. 00 ± 6. 88 ） of fiber-actin after 120 min （42.93 ±6.41） loading it was 2.8 times as much as that in control group（15.41 ±3.60, P 〈0.05）.Additionally, the level of phospho-cofilin protein was dramatically elevated after loading Fluid shear stress induced an initial decrease of cofilin at 60 min. However, at 120 min cofilin（0. 254 ±0. 026） increased to 1.5 times as much as that at 60 min （0. 162 ± 0. 004 ）. Conclusions The results indicate that cofilin phosphorylation mediates fiber-actin reorganization in the osteoblasts induc ed by fluid shear stress.

关 键 词：成骨细胞 细胞骨架 肌动蛋白解聚因子类 流体剪切力 

分 类 号：R686[医药卫生—骨科学]