目的研究Wnt7a基因过表达对大鼠骨髓间充质干细胞(mesenchymal stem cells,MSCs)增殖和向神经元样细胞分化的影响。方法构建Wnt7a腺病毒,利用病毒感染MSCs,MTT法检测Wnt7a基因腺病毒感染前后MSCs增殖情况,Western blot检测细胞质和细胞核中β-catenin及CyclinD1表达的变化,比较诱导后向神经元样细胞的分化率。结果与对照组细胞相比,Wnt7a腺病毒感染组细胞增殖能力明显增强(P〈0.05);细胞质和细胞核中β-catenin蛋白表达量均明显增加(P〈0.05);CyclinD1的蛋白表达量也明显增加(P〈0.05);诱导后向神经元样细胞分化率明显提高。结论 Wnt7a蛋白表达上调,可能通过提高β-catenin和CyclinD1表达来促进MSCs细胞增殖,Wnt7a蛋白同时促进MSCs向神经元样细胞的诱导分化。
Objective To explore the effect of up-regulation of Wnt7a on mesenchymal stem cells(MSCs) proliferation and differentiation into neuron-like cells.Methods An adenovirus vector expressing Wnt7a protein was constructed,and then MSCs were transfected with the recombinant vector.MSCs proliferation was detected by MTT assay,and the protein expression of cytoplasmic β-catenin,nuclear β-catenin and CyclinD1 was detected by Western blotting.The rates of MSCs differentiating into neuron-like cells were compared after transfection.Results Compared with the control MSCs,the MSCs transfected with Wnt7a recombinant adenovirus vector exhibited an increase in cell proliferation,and the protein expression of cytoplasmic β-catenin,nuclear β-catenin and CyclinD1 significantly increased(P0.05).The rate of neuron-like cells in the MSCs transfected with Wnt7a recombinant adenovirus vectors was significantly higher than that in the control group and the non-transfection group(P0.05).Conclusion Overexpression of Wnt7a protein promotes MSCs proliferation through up-regulating β-catenin and CyclinD1 expression,and stimulates MSCs differentiation into neuron-like cells,which provides further foundation for the therapy for spine cord injury.