神经药理学报 ›› 2024, Vol. 14 ›› Issue (1): 11-.DOI: 10.3969/j.issn.2095-1396.2024.01.002

• 研究论文 • 上一篇    下一篇

Cyclin D1 在黄芪甲苷诱导BMSCs 向神经元样细胞分化中的表达

王晓寅,王会芳,罗强,张辉,张晓丽   

  1. 1 河北北方学院附属第二医院,张家口,075000,中国 

    2 河北北方学院生命科学研究中心,张家口,075000,中国

  • 出版日期:2024-02-26 发布日期:2024-05-24
  • 通讯作者: 张晓丽,E-mail:zhangxiaoli1972@163.com
  • 作者简介:王晓寅,副主任医师;研究方向:中药抗肿瘤及神经损伤与修复;E-mail: 289073528@ qq. com
  • 基金资助:
    河北省教育厅重点项目(No.ZD2014067)

Cyclin D1 Expression During Astragaloside A Induced Neuron- Like Cell Differentiation of BMSCs

WANG Xiao-yin, WANG Hui-fang, LUO Qiang, ZHANG Hui, ZHANG Xiao-li   

  1. 1. The Second Affiliated Hospital of HeBei North University, Zhangjiakou, 075000, China 

    2. Life Science Research Center of HeBei North University, Zhangjiakou, 075000, China

  • Online:2024-02-26 Published:2024-05-24
  • Contact: 张晓丽,E-mail:zhangxiaoli1972@163.com
  • About author:王晓寅,副主任医师;研究方向:中药抗肿瘤及神经损伤与修复;E-mail: 289073528@ qq. com

摘要:

目的:研究黄芪甲苷诱导骨髓间充质干细胞(bone marrow mesenchymal stem cells, BMSCs)分化的特 点和诱导过程中Cyclin D1 的表达特点。方法:采用100 mg·L-1 黄芪甲苷诱导BMSCs 分化,在1、12、24 h 倒 置显微镜观察细胞形态变化,免疫细胞化学方法观察巢蛋白(Nestin)、神经元特异性烯醇化酶(neuron specific enolase, NSE)、Cyclin D1 的表达, RT-PCR 检测Cyclin D1 mRNA 的表达。结果:收集获得的骨髓细胞采用原 代和传代培养,至第3 代BMSCs 细胞基本纯化。经100 mg·L-1 黄芪甲苷作用后,部分细胞自胞体长出突起,形 态似神经元样细胞且表达Nestin 和NSE 抗体,Nestin 的表达时相早于NSE 且表达强度高于NSE(P<0.05)。在 药物作用1 h 后,BMSCs 细胞Cyclin D1 不论在基因转录水平还是在蛋白表达水平均出现大幅度下调,此时 BMSCs 开始有Nestin 的表达;药物作用12、24 h,BMSCs 细胞Cyclin D1 表达与药物作用前表达水平无显著 性差异(P >0.05)。结论:黄芪甲苷可能首先通过调整细胞周期,延长了G1 期向S 期转变的时间,从而启动了 BMSCs 的分化程序,诱导BMSCs 向神经干细胞分化。在黄芪甲苷的营养作用下,细胞的突起进一步生长并向 神经元分化。

关键词: 黄芪甲苷, BMSCs, 分化, 神经元样细胞, Cyclin D1

Abstract:

Objective:To observe the differentiation of bone marrow mesenchymal stem cells (BMSCs) induced by astragaloside A and expression of Cyclin D1. Methods: BMSCs were induced by astragaloside A with 100 mg·L-1. At l, 12, 24 h after induetion, the morphological changes of BMSCs were observed with optical microscope and the expressions of Nestin, NSE, Cyclin D1 were identified by immunocytochemical techniques. Cyclin D1 mRNA was detected by RT-PCR.Results: Bone marrow cells were primarily cultured and subcultured. The third passage of BMSCs was purified. After treatment with 100 mg·L-1 astragaloside A, some cells presented processes, were similar to neuron-like cells in morphology, and expressed Nestin and neuron-specific enolase. Nestin expression occurred earlier (P<0.05) and at a higher level (P<0.05) than expression of neuron-specific enolase. At 1 hour after treatment with astragaloside A, cyclin D1 expression was substantially down-regulated in BMSCs at both gene and protein levels. Simultaneously, BMSCs began to express nestin. No significant differences in cyclin D1 expression in BMSCs were detected between 12 and 24 hours post-treatment and the pretreatment level (P>0.05). Conclusion: Our results indicated that astragaloside A regulated cell cycle, prolonged G1/S phase transition, triggered BMSC differentiation, and finally induced BMSC differentiation into neuronal stem cells. The trophism of astragaloside A accelerated further growth of cell processes and the differentiation of cells into neurons.

Key words: astragaloside A, BMSCs, differentiation, neuron-like cells, Cyclin D1