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目的:探讨锌指蛋白521(Zfp521)在大鼠骨髓间充质干细胞分化为神经元过程中的表达变化及意义。方法:体外培养大鼠MSCs,实验分为未转染组、转染组(转染Rn-Zfp521-siRNA)和阴性对照组(转染nega-tive control siRNA),采用β-巯基乙醇诱导骨髓间充质干细胞分化为神经元。倒置荧光显微镜下观察MSCs转染后荧光表达情况。采用免疫细胞化学法、RT-PCR法及Western blotting法检测诱导后神经元特异性烯醇化酶(NSE)和微管相关蛋白2(MAP-2)的表达情况及诱导前、后Zfp521的表达变化。结果:(1)siRNA转染72 h荧光表达最强,转染率可达84.1%±2.3%,转染组骨髓间充质干细胞的Zfp521 mRNA表达下降(P<0.05);(2)β-巯基乙醇可以诱导骨髓间充质干细胞分化为神经元,以转染组诱导效果最佳,NSE和MAP-2表达显著高于其它各组(P<0.05);(3)Zfp521在各组细胞中均有表达,诱导后Zfp521表达明显低于诱导前(P<0.01)。结论:Zfp521在大鼠骨髓间充质干细胞神经分化中表达下降,抑制Zfp521表达可促进神经元的分化,提示Zfp521在骨髓间充质干细胞的神经分化中可能发挥重要调控作用。
OBJECTIVE: To investigate the changes and significance of zinc finger protein 521 (Zfp521) in the differentiation of rat bone marrow mesenchymal stem cells into neurons. Methods: Rat MSCs were cultured in vitro. The experiment was divided into untransfected group, transfected group (transfected with Rn-Zfp521-siRNA) and negative control group (transfected with nega-tive control siRNA), and β-mercaptoethanol MSCs differentiate into neurons. Fluorescence microscopy was used to observe the expression of MSCs after transfection. The expression of neuron-specific enolase (NSE) and microtubule-associated protein 2 (MAP-2) and the expression of Zfp521 before and after induction were detected by immunocytochemistry, RT-PCR and Western blotting . Results: (1) The expression of Zfp521 mRNA in transfected group was significantly lower than that in transfected group (P <0.05) at 72 h after transfection. The transfection rate was 84.1% ± 2.3% Mercaptoethanol induced the differentiation of BMSCs into neurons. The transfection group had the best induction effect and the expression of NSE and MAP-2 was significantly higher than that of other groups (P <0.05). (3) The expression of Zfp521 in each group of cells The expression of Zfp521 was significantly lower than that before induction (P <0.01). Conclusion: The expression of Zfp521 is decreased in neural differentiation of rat bone marrow mesenchymal stem cells. Inhibition of Zfp521 expression can promote the differentiation of neurons, suggesting that Zfp521 may play an important regulatory role in the differentiation of bone marrow mesenchymal stem cells.