【摘要】 目的探讨转录抑制因子-锌指蛋白和同源框3(zinc fingers and homeoboxes 3,ZHX3)基因过表达对BMSCs体外成骨能力的影响。方法采用ZHX3过表达慢病毒载体转染大鼠BMSCs,同时设空载病毒转染BMSCs作为阴性对照及不做任何处理的BMSCs做空白对照。采用荧光显微镜计数细胞转染率,并采用Western blot检测ZHX3蛋白表达状况。于体外培养,经定向成骨诱导21d后采用碱性磷酸酶和茜素红染色观察各组细胞成骨能力。结果 (1)贴壁法获得的细胞具有BMSCs表型。(2)转染后,ZHX3过表达组和阴性对照组均表达绿色荧光,空白对照组不表达荧光,且ZHX3过表达组ZHX3基因表达显著高于阴性对照组。(3)ZHX3过表达、阴性对照组和空白对照组均可见碱性磷酸酶阳性表达的细胞。Kaplow评分显示前者阳性表达显著高于后两者(P<0.05)。ZHX3过表达组、阴性对照组和空白对照组均可见明显的矿化结节。前者矿化结节的数量和体积均大于后两者。结论 ZHX3基因过表达可促进BMSCs体外成骨能力。更多还原

【Abstract】 Objective To investigate the influence of zinc fingers and homeoboxes 3 gene(ZHX3) transfected BMSCs and osteogenesis induced in vitro. Method Transfect the rat BMSCs with lentivirus overexpressing of ZHX3 genet. At the same time, BMSCs transfected with no-load virus serves as the negative control group and BMSCs without any treatment as the blank control group. Fluorescent microscope were used to count cell number and calculate the rate of transfection. Western blot was used to detect the protein expression of ZHX3 gene in the transected cells. The cells received In-vitro inducement of for 21d, after which alkaline phosphates staining and alizarin red staining were performed to observe osteogenic ability. Results(1) Cells express BMSCs phenotype could be obtained by adherent method.(2) After transfection, the ZHX3 gene overexpression group and the negative control group both expressed green fluorescence, blank control group do not express, and ZHX3 gene expression in overexpression group is higher than the negative control group.(3) cells in each group were found to be positive with expression of alkaline phosphatase. Kaplow show that the score of overexpression group is higher than the other two groups( P≤0.05). And mineralized nodules were visible among all groups, But the number and volume of mineralized nodules in overexpression group was larger than the others. Conclusion ZHX3 gene may promote BMSCs in vitro osteogenesis.更多还原