Department of Neurology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, P.R., China.
Cell Biol Int. 2013 Aug;37(8):812-9. doi: 10.1002/cbin.10103. Epub 2013 Apr 18.
To observe the effects of miR-125b on the differentiation of rat bone marrow mesenchymal stem cells (BMSCs) into neuron-like cells, rat BMSCs were isolated and transfected with Syn-rno-miR-125b* miScript miRNA Mimic (Mimic + BME group), with anti-rno-miR-125b* miScript miRNA Inhibitor (Inhibitor + BME group) and BME control was set up without transfection. Blank controls without transfection and induction by BME was also set up. BMSCs of three groups including Mimic + BME group, Inhibitor + BME group and BME group were induced to differentiate into neuron-like cells by β-mercaptoethanol (BME). Cells in the Blank group were not treated by BME. mRNA expression of miR-125b was determined with qRT-PCR in each group. mRNA and protein expressions of seven nerve cell markers, including β3 tubulin, neural microtubule-associated protein (MAP-2), neurofilament protein (NF), neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP), Nestin and Vimentin in the four groups were determined by RT-PCR and Western blots. GFAP and Nestin were detected by immunofluorescent and immunocytochemistry assays. Compared with BME group, mRNA and protein expression of β3 tubulin, MAP-2, NF, NSE, GFAP, Nestin were significantly increased in the Mimic + BME group (P < 0.01), but significantly decreased in the Inhibitor + BME group (P < 0.01). Cells in the Mimic + BME group were more like nerve cells in morphology than cells in the BME groups. Thus miR-125b can promote BME to induce rat BMSCs differentiation into neuron-like cells.
观察 miR-125b 对大鼠骨髓间充质干细胞（BMSCs）向神经元样细胞分化的影响，分离大鼠 BMSCs 并转染 Syn-rno-miR-125bmiScript miRNA Mimic（Mimic+BME 组），用 anti-rno-miR-125bmiScript miRNA Inhibitor（Inhibitor+BME 组）和 BME 对照组转染，同时设立空白对照组不转染和 BME 诱导。Mimic+BME 组、Inhibitor+BME 组和 BME 组的 BMSCs 均用β-巯基乙醇（BME）诱导分化为神经元样细胞，空白组不加 BME 处理。各组采用 qRT-PCR 检测 miR-125b 的 mRNA 表达水平，采用 RT-PCR 和 Western blot 检测 7 种神经细胞标志物，包括β3 微管蛋白、神经微管相关蛋白（MAP-2）、神经丝蛋白（NF）、神经元特异性烯醇化酶（NSE）、胶质纤维酸性蛋白（GFAP）、巢蛋白（Nestin）和波形蛋白（Vimentin）的 mRNA 和蛋白表达水平，采用免疫荧光和免疫细胞化学检测 GFAP 和 Nestin。与 BME 组比较，Mimic+BME 组的β3 微管蛋白、MAP-2、NF、NSE、GFAP、Nestin 的 mRNA 和蛋白表达明显增高（P<0.01），而 Inhibitor+BME 组明显降低（P<0.01）。Mimic+BME 组细胞形态更接近神经细胞，而 BME 组细胞则不同。miR-125b 可促进 BME 诱导大鼠 BMSCs 分化为神经元样细胞。
