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成纤维细胞生长因子1型受体-显性负性作用对骨髓基质干细胞成骨诱导培养后碱性磷酸酶活性的影响 被引量：5

Effect of fibroblast growth factor receptors 1-dominant negative strategy on alkaline phosphatase activity of bone marrow stromal cells after osteogenic induction
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摘要 目的探讨成纤维细胞生长因子1型受体-显性负性作用（FGFR1-DN）对骨髓基质干细胞（BMSCs）成骨诱导培养后碱性磷酸酶（ALP）活性的影响。方法真核表达质粒pcDNA3．1（＋）．DNFGFR1、pcDNA3．1（＋）．FGFR1转染BMSCs，实验分为FGFR1-DN转染组、FGFR1转染组、空载体转染组和未转染组。细胞对数生长期进行成骨诱导培养后检测ALP的活性，定性检测采用免疫组化方法，定量检测采用细胞内ALP（cALP）试剂盒。比较4组细胞诱导培养7、14d的ALP活性。结果与诱导培养7d比较，4组14d后ALP活性评分均明显增高，但FGFRl．DN转染组评分增高最明显，差异有统计学意义（P〈0．05）。7、14d FGFR1-DN转染组ALP活性评分最高，FGFR1转染组最低，差异均有统计学意义（P〈0．05）。结论FGFR1-DN可促进BMSCs成骨期ALP的活性，为实现局部基因治疗与组织工程的联合应用及构建生物相容性更理想的组织工程骨提供了实验理论依据。 Objective To explore the effect of fibroblast growth factor receptors 1-dominant negative strategy （FGFR1-DN） on alkaline phosphatase （ALP） activity of bone marrow stromal stem cells （BMSCs） after osteogenic induction. Methods BMSCs were transfected with eukaryotic expression plasmid pcDNA 3.1 （ ＋ ） -DN FGFR1 and pcDNA3. 1 （ ＋ ） -FGFR1. The experiment was conducted in 4 groups： FGFR1-DN transfection group, FGFR1 transfection group, pcDNA3.1（ ＋ ） empty vector transfection group and non-transfection group. The ALP activity of BMSCs was detected in logarithmic growth phase after osteogenic culture. The qualitative detection of ALP activity was carried out immunohistochemically while the quantitative detection by cALP kit. The ALP activity was compared between the 4 groups at 7 and 14 days after osteogenic induction. Results Compared with 7 days, the ALP activity at 14 days was significantly increased in the 4 groups, and the increase in FGFR1-DN transfection group was significantly higher than in the other 3 groups （ P 〈 0. 05）. At both 7 and 14 days, the ALP activity in FGFR1-DN transfection group was the highest while that in FGFR1 transfection group was the lowest （ P 〈 0.05）. Conclusions FGFR1-DN can promote the ALP activity of BMSCs during osteogenesis. This may provide an experimental basis for the joint application of local gene therapy and tissue engineering and for construction of tissue engineered bone with better biocompatibility.
作者 李霞 徐文漭 简洪 徐永清 李福兵
机构地区 成都军区昆明总医院全军创伤骨科研究所 成都军区昆明总医院病理科
出处 《中华创伤骨科杂志》 CAS CSCD 北大核心 2016年第6期526-531,共6页 Chinese Journal of Orthopaedic Trauma
基金 国家自然科学基金（81171734） 云南省创新团队基金（2009C1008） 国家临床重点专科军队建设项目 国家科技支撑计划项目（2012BA117801） 中国博士后科学基金（20100481529）
关键词 骨髓细胞 成纤维细胞生长因子 成纤维细胞生长因子受体1 显性负性作用 Bone marrow ceils Fibroblast growth factors Fibroblast growth factor receptor 1 Dominant negative effect
分类号 R68 [医药卫生—骨科学]
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参考文献18

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二级参考文献21

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中华创伤骨科杂志
2016年 第6期

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