Experimental study on the facilitative effects of miR-125b on the differentiation of rat bone marrow mesenchymal stem cells into neuron-like cells

Authors

  • Rui Wu,

    1. Department of Neurology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, P.R., China
    2. Department of Neurological Rehabilitation, the Second Affiliated Hospital of Zhengzhou University, Zhengzhou, P.R., China
    Search for more papers by this author
  • Na Wang,

    1. Department of Microbiology and Immunology, College of Basic Medical Sciences, Zhengzhou University, Zhengzhou, P.R., China
    Search for more papers by this author
  • Min Li,

    1. Department of Microbiology and Immunology, College of Basic Medical Sciences, Zhengzhou University, Zhengzhou, P.R., China
    Search for more papers by this author
  • Wenqiao Zang,

    1. Department of Microbiology and Immunology, College of Basic Medical Sciences, Zhengzhou University, Zhengzhou, P.R., China
    Search for more papers by this author
  • Yuming Xu

    Corresponding author
    • Department of Neurology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, P.R., China
    Search for more papers by this author

Corresponding author: e-mail: xuyumingpaper64@yahoo.cn

Abstract

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.

Ancillary