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

中国口腔颌面外科杂志 ›› 2015, Vol. 13 ›› Issue (3): 193-199.

• 基础研究 • 下一篇

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

姜喜亮, 朱晓文, 胡静, 宋健, 叶斌

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

收稿日期:2014-09-03 修回日期:2014-11-11 出版日期:2015-05-20 发布日期:2015-06-18
通讯作者: 叶斌,Tel：028-85502334,E-mail:yb6560@126.com
作者简介:姜喜亮（1989-）,男,博士,E-mail:jiangxl0221@foxmail.com
基金资助:
国家自然科学基金（81100779）

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

JIANG Xi-liang, ZHU Xiao-wen, HU Jing, SONG Jian, YE Bing

Department of Orthognathic and Joint Surgery, West China Hospital of Stomatology, Sichuan University; State Key Laboratory of Oral Disease. Chengdu 610041, Sichuan Province, China

Received:2014-09-03 Revised:2014-11-11 Online:2015-05-20 Published:2015-06-18
Supported by:
Supported by National Natural Science Foundation of China (81100779)

摘要/Abstract

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siRNA 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siRNA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。

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

Abstract: 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 siRNA 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 integrinβ1,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. SPSS 20.0 software package was used for one way analysis of variance. RESULTS: The expression of FAK specific siRNA lentiviral expression vector-bone differentiation factors Runx-2, ALP decreased compared to control group. And the expression of important signal molecules integrinβ1 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 conversion of cellar mechanical signals to biological chemical signals.

Key words: Distraction osteogenesis, BMSCs, Integrin signaling pathway, FAK

中图分类号:

Q257

姜喜亮, 朱晓文, 胡静, 宋健, 叶斌. 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[J]. 中国口腔颌面外科杂志, 2015, 13(3): 193-199.

JIANG Xi-liang, ZHU Xiao-wen, HU Jing, SONG Jian, YE Bing. The function and mechanism of integrin signal pathway in the stretched rat BMSCs[J]. China J Oral Maxillofac Surg, 2015, 13(3): 193-199.

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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

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