论文部分内容阅读
目的:体外观察碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)联合Sonic Hedgehog(Shh)的时序性基因传递对骨髓来源的间充质干细胞(bone marrow derived mesenchymal stem cells,BMSCs)中成骨相关因子的影响。方法:将前期实验中构建的重组病毒rAAV2-tet-off-bFGF及rAAV2-Shh感染原代培养的BMSCs,其中前者可被四环素调控系统调控bFGF的表达。real-time RT-PCR及Western blot检测各成骨相关因子的表达变化。结果:双病毒感染BMSCs后可被四环素衍生物强力霉素(doxycycline,Dox)调控。细胞内成骨相关因子的mRNA及蛋白水平较对照组都有不同程度的升高,变化具有统计学差异。结论:短期bFGF的表达联合Shh的基因传递能极大程度提高BMSCs的增殖能力,增加胞内成骨相关因子的表达水平,促进BMSCs成骨分化。
OBJECTIVE: To observe the effects of basic fibroblast growth factor (bFGF) combined with sequential gene delivery of Sonic Hedgehog (Shh) on bone marrow derived mesenchymal stem cells (BMSCs) Bone related factors. Methods: Primary cultured BMSCs were infected with recombinant viruses rAAV2-tet-off-bFGF and rAAV2-Shh constructed in the previous experiment. The former could be regulated by tetracycline regulatory system to regulate bFGF expression. Real-time RT-PCR and Western blot were used to detect the expression of osteogenesis related factors. RESULTS: BMSCs infected by double virus could be regulated by doxycycline (Dox), a tetracycline derivative. Compared with the control group, mRNA and protein levels of intracellular osteogenesis-related factors increased to some extent, with statistical significance. Conclusion: The expression of short-term bFGF combined with Shh gene transfer can greatly enhance the proliferation of BMSCs, increase the expression of intracellular osteogenesis-related factors and promote the osteogenic differentiation of BMSCs.