JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES) ›› 2015, Vol. 53 ›› Issue (1): 34-40.doi: 10.6040/j.issn.1671-7554.0.2014.552

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Effect of rehmannia glutinosa polysaccharide on the differentiation and proliferation of over-expressed Notch1(NICD) gene in rat bone marrow mesenchymal stem cells in vitro

FU Haiyan1, HU Zhansheng1, DU Hongyang2, LI Chao3, BAO Cuifen4   

  1. 1. Department of ICU, First Affiliated Hospital of Liaoning Medical University, Jinzhou 121001, Liaoning, China;
    2. Department of Dermatology, First Affiliated Hospital of Liaoning Medical University, Jinzhou 121001, Liaoning, China;
    3. Department of Pathology, First Affiliated Hospital of Liaoning Medical University, Jinzhou 121001, Liaoning, China;
    4. Department of Anatomy, Liaoning Medical University, Jinzhou 121001, Liaoning, China
  • Received:2014-08-26 Revised:2014-12-09 Published:2015-01-10

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Abstract: Objective To explore the effect of rehmannia glutinosa polysaccharide (RGP) on the differentiation and proliferation of over-expressed Notch1(NICD) gene in rat bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods The P3 to P5 of BMSCs were randomly divided into the control group (CON group), empty vector group (VEC group) and transfection group (TRA group). The NICD eukaryotic expression vectors were transfected into BM-SCs which were then induced with RGP (concentration 0.2 mg/mL). The general morphology was observed. Protein expression of NICD, neurone specific enolase (NSE) and glial gibrillary acidic protein (GFAP) were detected with immunocytochemetry assay and Western blotting assay. Cell survival rate was assessed with CCK-8 assay. Results The BMSCs in TRA group were glial-like cells after transfection, and the induced cells were neuron-like, especially in TRA group. Immunofluorescence assay showed that the positive cell percentage of Notch1 and GFAP protein expressions in TRA group were significantly higher than those in CON group and VEC group (P<0.05). However, NSE in the three groups had no statistical difference after transfection (P>0.05). The positive cell percentage of NSE in TRA group was remarkably higher than that in CON group and VEC group after induction (P<0.05). The positive rate of Notch1 expression in TRA, CON and VEC groups decreased (P<0.05). The positive rate of NSE expression in TRA, CON and VEC group markedly increased (P<0.01). The positive rate of GFAP expression in TRA group decreased significantly (P<0.01). The CCK-8 assay showed that cell survival rate decreased with time in each group after RGP induction. Conclusion The transfection of NICD over eukaryotic expression vector may induce rat BMSCs into glial-like cells. RGP inhibits the expression of Notch1 protein significantly and induces BMSCs into neuron like cells.

Key words: Notch signal pathway, Transfection, Cell differentiation, Neurone specific enolase, Induction, Mesenchymal stem cells

CLC Number: 

  • R285.5
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