细胞骨架完整性对流体剪切力诱导成骨细胞c-fos基因mRNA和蛋白表达的影响  

osteoblasts induced by fluid shear stress

作　　者：徐亚娟[1] 张艺平[1] 覃峰[1] 吴昌敬 李友瑞[1] 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]广州市中西医结合医院修复科

出　　处：《中华口腔医学杂志》2009年第11期681-685,共5页Chinese Journal of Stomatology

基　　金：基金项目：广东省科技计划（20088030301121）;广东省医学科研课题（A2008227）

摘　　要：目的研究细胞骨架完整性在流体剪切力诱导成骨细胞c-fos基因mRNA和蛋白表达中的作用，为进一步探索骨组织力传导机制提供依据。方法原代培养BALB／c小鼠成骨细胞，将原代培养的成骨细胞分为对照组和细胞松弛素D组，细胞松弛素D组使用细胞松弛素D破坏细胞骨架。每组一半细胞加载1．2Pa的流体剪切力，另一半不加载。分别用实时荧光定量聚合酶链反应（real-timePCR）和免疫荧光检测c—fos基因mRNA和蛋白、细胞骨架蛋白的表达水平，并对结果进行双因素方差分析和成组t检验。结果无论是对照组还是细胞松弛素D组，流体剪切力加载均可使成骨细胞c-fos基因mRNA相对水平（分别为0．1637±0．0303和0．0104±0．0070）和蛋白荧光强度（分别为177．14±9．37和150．95±6．17）升高，与未加载流体剪切力的对照组和细胞松弛素D组细胞（mRNA相对水平分别为0．0057±0．0021和0．0032±0．0014，蛋白荧光强度分别为117．96±4．11和119．77±5．19）相比，差异均有统计学意义（P〈0．05）。流体剪切力加载下，细胞松弛素D组细胞c—fos基因mRNA和蛋白的表达均低于对照组，差异均有统计学意义（P〈0．05）。结论细胞松弛素D对流体剪切力诱导成骨细胞c—fos基因mRNA和蛋白的表达起拮抗作用。保持细胞骨架的完整性是流体剪切力诱导成骨细胞c—fos基因表达过程中的重要条件。Objective To investigate the effect of cytoskeleton integrity on the expression of c-fos gene in osteoblasts induced by fluid shear stress. Methods BALB/c mouse primary osteoblasts were divided into four groups（ according to fluid shear stress loaded or not and cytochalasin D used or not）. The Tagman probe real-time PCR and immunofluorescence were performed to detect the expression levels of c-los mRNA, c-fos protein and cytoskeleton, respectively. The data were analysed using two-way ANOVA. Results In control group and cytochalasin D group, fluid shear stress could significantly increase the expression levels of c-fos mRNA （ 0. 1637 ± O. 0303 and 0. 0104 ± 0. 0070, respectively ） and protein （ 177. 14 ±9. 37 and 150. 95 ± 6. 17, respectively） in osteoblasts, compared with the unloaded osteoblasts of the control group and the eytochalasin D group（0. 0057±0. 0021 and 0. 0032±0. 0014, respectively for c- los mRNA, and 117. 96 ±4. 11 and 119. 77 ± 5.19, respectively for protein, P 〈 0.05）. Induced by the fluid shear stress, the expression levels of c-los mRNA and protein in cytochalasin D group were lower than control group, and the difference had statistical significance （ P 〈 0. 05 ）. Conclusions The cytoskeleton integrity in osteoblasts was essential to the expression of c-los gene induced by fluid shear stress.

关 键 词：细胞骨架 成骨细胞 基因 FOS 流体剪切力 

分 类 号：R686[医药卫生—骨科学]