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

发布时间：2018-05-06 05:23

本文选题：牵张成骨 + 骨髓间充质干细胞　；参考：《中国口腔颌面外科杂志》2015年03期

【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。
[Abstract]:Objective: To investigate the role and transduction mechanism of integrin signaling pathway in distraction osteogenesis (DO). Methods: by constructing focal adhesion kinase (FAK) specific Si RNA Lentivirus Expression Vector and infection of pre distraction bone marrow mesenchymal stem cells (bone marrow derived). An appropriate mechanical tensile stress was applied to the infected BMSCs with a self developed multi cell stretching and compression device. After the end of the stretch, the RNA and protein were collected after the time point 0,6,12 and 24 h. The RT-PCR and Western immunoblotting methods were used to detect the important signal molecules FAK, integrin beta 1 (integrin beta 1) and integrin kinase (integrin-li). The expression of nked kinase, ILK) and the expression of BMSCs cell proliferation and bone differentiation transcription factor Runx2, the expression of osteogenic factor alkaline phosphatase (alkaline phosphatase, ALP). The data were analyzed by single factor variance analysis using SPSS 20 software package. Results: FAK base was silent, FAK specificity Si slow virus group compared with the control group. The expression of bone differentiation factor Runx 2, ALP decreased, and the expression of integrin beta 1 and ILK of integrin important signal molecules also decreased significantly. Conclusion: after the inhibition of FAK gene, the osteogenic differentiation of BMSCs has a certain effect. The integrin signaling pathway participates in the transformation of the cell mechanical signal to the biochemical signal.

【作者单位】：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;
【基金】：国家自然科学基金(81100779)~~
【分类号】：R329.2

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