解剖学报

解剖学报 ›› 2019, Vol. 50 ›› Issue (4): 459-464.doi: 10.16098/j.issn.0529-1356.2019.04.009

• 细胞和分子生物学 • 上一篇    下一篇

槲寄生多糖促进小鼠颅骨前骨细胞系MC3T3-E1的增殖及抑制凋亡的机制

赵磊1 田守进1 葛建飞1 黄群1 朱宏岩2 董启榕3*   

  1. 1.张家港市第一人民医院骨科，江苏 张家港 215600; 2.张家港市第一人民医院转化医学中心，江苏 张家港 215600; 3.苏州大学附属第二医院骨科，江苏 苏州 215004
  • 收稿日期:2018-06-14 修回日期:2018-07-05 出版日期:2019-08-06 发布日期:2019-08-06
  • 通讯作者: 董启榕 E-mail:zhaolei5492@163.com
  • 基金资助:
    基金项目：张家港市科技局科技支撑项目（编号：ZKS1741）。

Effect of mistletoe polysaccharose on mouse osteoblast cell line MC3T3-E1 promoting proliferation and  inhibiting apoptosis and its possible mechanism

ZHAO Lei1 TIAN Shou-jin1 GE Jian-fei1 HUANG Qun1 ZHU Hong-yan2 DONG Qi-rong 3*   

  1. 1.Department of Orthopaedics, the First People’s Hospital of Zhangjiagang, Jiangsu Zhangjiagang 215600, China;   2.Transformation Medical Center, the First People’s Hospital of Zhangjiagang, Jiangsu Zhangjiagang 215600, China;  3.Department of Orthopaedics, the Second Affiliated Hospital of Soochow University, Jiangsu Suzhou 215004, China
  • Received:2018-06-14 Revised:2018-07-05 Online:2019-08-06 Published:2019-08-06
  • Contact: DONG Qi-rong E-mail:zhaolei5492@163.com

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摘要：

目的 检测槲寄生多糖对小鼠颅骨前骨细胞（MC3T3-E1）增殖、凋亡的影响并探讨其机制。 方法 提取槲寄生多糖，分别配置成不同质量浓度（0.625 g/L，1.25 g/L，2.5 g/L，5 g/L）作用MC3T3-E1小鼠颅骨前骨细胞及对照组。流式细胞术检测细胞周期及凋亡情况，用BrdU和碱性磷酸酶（ALP）试剂盒分别检测细胞增殖及细胞外碱性磷酸酶分泌变化，通过Real-time PCR检测成骨细胞相关基因mRNA的表达水平。 结果 与对照组相比，药物处理组随槲寄生多糖浓度的上升，细胞周期S期和G2/M的占比增多，在Annexin Ⅴ+/PI-象中的细胞百分数明显下降，细胞数量明显递增，细胞外ALP的活性呈明显递减，Runt相关转录因子2（Runtx2）、Ⅰ型胶原α1（COL1A1）链转录水平升高，且递增、递减效果都在2.5 g/L浓度之后趋于平稳。而骨钙素（OC）mRNA表达水平逐渐升高，在1.25 g/L之后平稳。 结论 槲寄生多糖能促进成骨细胞MC3T3-E1的增殖，抑制其凋亡，其机制可能与抑制碱性磷酸酶分泌以及促进骨钙素、Runt相关转录因子2、Ⅰ型胶原α1链的mRNA表达水平有关。

关键词: 槲寄生多糖, 小鼠颅骨前骨细胞, 细胞周期, 流式细胞术, 小鼠

Abstract:

Objective To observe the effect of mistletoe polysaccharose on proliferation and apoptosis of osteoblast and to explore the possible mechanism. Methods The mistletoe polysaccharose was extracted and made different concentrations (0.625 g/L, 1.25 g/L, 2.5 g/L, 5 g/L) to treat mouse skull bone cell line MC3T3-E1. Cell cycle and apoptosis were detected by flow cytometry. Cell proliferation and extracellular alkaline phosphatase secretion were detected by BrdU and alkaline phosphatase ELISA kits. The mRNA expression level of osteoblast related gene was detected by Real-time PCR. Results Compared with the control group, the result of treatment groups with increasing mistletoe polysaccharose concentrations were as follows: the proportions of S phase and G2/M phase in cell cycle increased and the cell count Annexin Ⅴ+/PI-significantly decreased, the number of cells increased, extracellular alkaline phosphatase activity obviously decreased. The increasing transcription levels of Runtrelated transcription factor 2(Runtx2) and collagen typeⅠ alpha 1(COL1A) were stabled over 2.5 g/L and the upregulation of mRNA expression of osteocalcin (OC) was steady at 1.25 g/L. Conclusion Mistletoe polysaccharose can promote the proliferation and inhibit the apoptosis of MC3T3-E1 cells, and its possible mechanism might be related to down-regulating of extracellular alkaline phosphatase activity and the grow exhanced expression of OC, Runtx2 and COL1A.

Key words: Mistletoe polysaccharose, MC3T3-E1 cell, Cell cycle, Flow cytometry, Mouse