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
[1] 虎喜成, 刘敬霞, 刘会贤, 等. 中药诱导骨髓间充质干细胞向神经样细胞分化研究进展[J]. 中国实验方剂学杂志, 2014, 20(3):219-224. HU Xicheng, LIU Jingxia, LIU Huixian, et al. Differentiation research progress of traditional Chinese medicine inducing bone marrow mesenchymal stem cells to neural cells [J]. Chinese Journal of Experimental Traditional Medical Formulae, 2014, 20(3):219-224.
[2] 卞琴, 黄建华,杨铸, 等. 三种补肾中药有效成分对皮质酮致骨质疏松大鼠骨髓间充质干细胞基因表达谱的作用[J]. 中西医结合学报, 2011, 9(2):179-185. BIAN Qin, HUANG Jianhua, YANG Zhu, et al. Effects of active ingredients in three kidney-tonifying Chinese herbal drugs on gene expression profile of bone marrow stromal cells from a rat model of corticosterone-induced osteoporosis [J]. Journal of Integrative Medicine, 2011, 9(2):179-185.
[3] Duryagina R, Thieme S, Anastassiadis K, et al. Overexpression of Jagged-1 and its intracellular domain in human mesenchymal stromal cells differentially affect the interaction with hematopoietic stem and progenitor cells [J]. Stem Cells Dev, 2013, 22(20):2736-2750.
[4] 付海燕, 杜红阳, 包翠芬. 地黄多糖诱导大鼠BMSCs向神经元样细胞分化及对Notch1和Jagged1蛋白表达的影响[J]. 解放军医学杂志, 2014, 39(6):448-453. FU Hanyan, DU Hongyang, BAO Cuifen. Effect of Rehmannia Glutinosa Polysaccharide on Notch1 and Jagged1 protein in Rat Bone Marrow Mesenchymal Stem Cells during Differentiation into Neuron-like Cells in vitro[J]. Medical Journal of Chinese People's Liberation Army, 2014, 39(6):448-453.
[5] 刘卫欣, 卢兖伟, 杜海涛, 等. 地黄及其活性成分药理作用研究进展[J]. 国际药学研究杂志, 2009, 36(4): 277-280. LIU Weixin, LU Yanwei, DU Haitao, et al. Pharmacological actions of Radix Rehmanniae and its active components research advances [J]. Journal of International Pharmaceutical Research, 2009, 36(4):277-280.
[6] Wang LJ, Zhang RP, Li JD. Transplantation of neurotrophin-3-expressing bone mesenchymal stem cells improves recovery in a rat model of spinal cord injury [J]. Acta Neurochir (Wien), 2014, 156(7):1409-1418.
[7] 杜红阳, 李东宁, 付海燕, 等. Notch1(NICD)过表达真核载体构建及对大鼠BMSCs增殖分化的影响[J]. 天津医药, 2014, 42(9): 883-888. DU Hongyang, LI Dongning, FU Haiyan, et al. Construction of Notch1(NICD) eukaryotic expression vector and its influence on the proliferation and differentiation of rat bone marrow mesenchymal stem cells in vitro [J]. Tianjin Medical Journal, 2014, 42(9):883-888.
[8] Kim M, Park YK, Kang TW, et al. Dynamic changes in DNA methylation and hydroxymethylation when hES cells undergo differentiation toward a neuronal lineage [J]. Hum Mol Genet, 2014, 23(3):657-667.
[9] Ille F, Atanasoski S, Falk S, et al. Wnt/BMP signal integration regulates the balance between proliferation and differentiation of neuroepithelial cells in the dorsal spinal cord[J]. Dev Biol, 2007, 304(1):394-408.
[10] Takeuchi H, Haltiwanger RS. Significance of glycosylation in Notch signaling[J]. Biochem Biophys Res Commun, 2014, 6. doi: 10.1016/j.bbrc.2014.05.115.
[11] LeBon L, Lee TV, Sprinzak D, et al. Fringe proteins modulate Notch-ligand cis and trans interactions to specify signaling states [J]. Elife, 2014, 25(3): 02950. doi: 10.7554/eLife.02950.
[12] Zhang Z, Meng Y, Guo Y, et al. Rehmannia glutinosa polysaccharide induces maturation of murine bone marrow derived Dendritic cells (BMDCs) [J]. Int J Biol Macromol, 2013, 54:136-143.
[13] Huang Y, Jiang C, Hu Y, et al. Immunoenhancement effect of rehmannia glutinosa polysaccharide on lymphocyte proliferation and dendritic cell [J]. Carbohydr Polym, 2013, 96(2):516-521.
[14] 杜红阳, 李东宁, 付海燕, 等. 地黄多糖诱导大鼠BMSCs向神经样细胞分化中对Notch信号通路的影响[J]. 山东大学学报: 医学版, 2013, 51(12): 1-6. DU Hongyang, LI Dongning, FU Haiyan, et al. Effect of rehmannia glutinosa polysaccharide on Notch signal pathway in rat bone marrow mesenchymal stem cells during differentiation into neuron-like cells in vitro[J]. Journal of Shandong University: Health Science, 2013, 51(12):1-6.
[15] El Khatib M, Bozko P, Palagani V, et al. Activation of Notch signaling is required for cholangiocarcinoma progression and is enhanced by inactivation of p53 in vivo [J]. PLoS One, 2013, 8(10):77433.
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