整合素信号通路在大鼠骨髓间充质干细胞牵张中的作用和转导机制  

The function and mechanism of integrin signal pathway in the stretched rat BMSCs

作　　者：姜喜亮 朱晓文[1] 胡静[1] 宋健[1] 叶斌[1] 

机构地区：[1]四川大学华西口腔医院正颌与关节外科,口腔疾病研究国家重点实验室,四川成都610041

出　　处：《中国口腔颌面外科杂志》2015年第3期193-199,共7页China Journal of Oral and Maxillofacial Surgery

基　　金：国家自然科学基金(81100779)~~

摘　　要：目的:探讨整合素信号通路在牵张成骨(distraction osteogenesis,DO)中的作用及转导机制。方法:通过构建粘着斑激酶(focal adhesion kinase,FAK)特异性si RNA慢病毒表达载体,感染牵张前骨髓间充质干细胞(bone marrow derived mesenchymal stem cells,BMSCs),应用自行研制的多单元细胞拉伸与压缩装置,对感染后的BMSCs施加适当的机械张应力。在牵张结束后,收取时间点0、6、12和24 h后的RNA及蛋白,采用RT-PCR与Western免疫印迹法,检测重要信号分子FAK、整合素β1(integrinβ1)和整合素连接激酶(integrin-linked kinase,ILK)的表达,同时检测BMSCs细胞增殖及骨向分化转录因子Runx2、成骨因子碱性磷酸酶(alkaline phosphatase,ALP)的表达变化情况。采用SPSS 20.0软件包对数据进行单因素方差分析。结果:FAK基因沉默后,与对照组相比,FAK特异性si RNA慢病毒组骨向分化因子Runx 2、ALP表达减弱,整合素重要信号分子integrinβ1和ILK表达也显著减弱。结论:FAK基因被抑制后,对BMSCs的骨向分化有一定影响。整合素信号通路参与细胞力学信号向生物化学信号的转化。PURPOSE： To detect the role and mechanism of integrin signal pathway acting on distraction osteogenesis（DO）. METHODS： A distraction model was established on the basis of an uniaxial static stretching device, and si RNA technology was used to silence the focal adhesion kinase（FAK） of bone marrow derived mesenchymal stem cells（BMCs）,an important signal transduction molecule in integrin signaling pathway. After distraction, at 0, 6, 12 and 24 h, RNA and protein of important signal transduction molecules such as FAK, ILK and integrin β1,were detected by RT-PCR and Western blot. And the cell proliferation,expression of the osteogenic genes Cbfa 1/Runx 2 and ALP were detected. SPSS20.0 software package was used for one way analysis of variance. RESULTS： The expression of FAK specific si RNA lentiviral expression vector-bone differentiation factors Runx-2, ALP decreased compared to control group. And the ex-pression of important signal molecules integrin β1 and ILK in integrin pathway decreased dramatically. CONCLUSION：BMSCs bone differentiation is inhibited after FAK gene silence and the integrin signal pathway takes part in the conver-sion of cellar mechanical signals to biological chemical signals.

关 键 词：牵张成骨 骨髓间充质干细胞 整合素信号通路 黏着斑激酶 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]