CAJ | 학술논문

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BM－SCs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。
목적：체외관찰감성성섬유세포생장인자（basic fibroblast growth factor，bFGF）연합Sonic Hedgehog（Shh）적시서성기인전체대골수래원적간충질간세포（bone marrow derived mesenchymal stem cells，BMSCs）중성골상관인자적영향。방법：장전기실험중구건적중조병독rAAV2－tet－off－bFGF급rAAV2－Shh감염원대배양적BMSCs，기중전자가피사배소조공계통조공bFGF적표체。real－time RT－PCR급Western blot검측각성골상관인자적표체변화。결과：쌍병독감염BMSCs후가피사배소연생물강력매소（doxycycline，Dox）조공。세포내성골상관인자적mRNA급단백수평교대조조도유불동정도적승고，변화구유통계학차이。결론：단기bFGF적표체연합Shh적기인전체능겁대정도제고BM－SCs적증식능력，증가포내성골상관인자적표체수평，촉진BMSCs성골분화。
Objective:The aim of this study was to delivery basic fibroblast growth factor(bFGF) and sonic hedgehog (Shh) to bone marrow derived mesenchymal stem cells (BMSCs),and then to detect the effect for osteogenic marker genes. Method:bFGF and Shh were constructed to the recombinant adeno-associated virus,respectively(rAAV2-tet-off-bFGF and rAAV2-Shh). The previous viral vector allowed for regulation of the bFGF expression by the addition of doxycycline,a te-tracycline analogue. Several osteogenic markers were detected by quantitative real-time reverse transcriptase polymerase chain reaction and western blot. Result:BMSCs were constructed successfully by two recombinant virus that were regulated by Dox. The expression of osteogenic marker mRNA and protein had an increased tendency after two genes transduction(P<0.05). Conclusion:Sequential delivery of angiogenic and osteogenic factors likely has a synergistic effect,which could not only enhance the cell proliferation,but also improve the expression of osteogenic marker of BMSCs.