华西口腔医学杂志

• 基础研究 • 上一篇    下一篇

辐射对体外培养MC3T3-E1成骨前体细胞增殖和分化的影响

李玉梅1  赵铱民2  查年保1  舒震3  张松4   

  1. 1.襄阳市中心医院口腔科,襄阳 441021;2.第四军医大学口腔医院修复科;
    3.第四军医大学药学系生物技术中心,西安 710032;4.第四军医大学唐都医院药剂科,西安 710038
  • 出版日期:2014-08-01 发布日期:2014-08-01
  • 通讯作者: 赵铱民,主任医师,博士, E-mail:zhaoym@fmmu.edu.cn
  • 作者简介:李玉梅,主治医师,博士,E-mail:lymzxf@163.com

Effects of irradiation on proliferation and differentiation of MC3T3-E1 osteoblastic cells in vitro

 Li Yumei1, Zhao Yimin2, Zha Nianbao1, Shu Zhen3, Zhang Song4   

  1. 1. Dept. of Stomatology, Xiangyang Central Hospital, Xiangyang 441021, China; 2. Dept. of Prosthodontics, School of Stomatology, The Fourth Military Medical University, Xi’an 710032, China; 3. Biotech-nology Center, School of Pharmacy, The Fourth Military Medical University, Xi’an 710032, China; 4. Dept. of Pharmacy, Tangdu Hospital, The Fourth Military Medical University, Xi’an 710038, China
  • Online:2014-08-01 Published:2014-08-01

摘要:

目的  观察60Co γ射线对MC3T3-E1成骨前体细胞增殖和分化能力的影响。方法  MC3T3-E1细胞接种24 h后进行60Co γ射线照射,单次照射剂量分别为0、4、8 Gy。辐射后第 1、3、5、7天,采用噻唑蓝比色法检测细胞的增殖能力;辐射后第12天,用黏胶纤维红染色法检测细胞的胶原分泌情况;第16天,用实时荧光定量聚合酶链反应检测细胞成骨相关基因mRNA的表达;第28天,采用茜素红染色及定量分析检测细胞基质的矿化能力。结果  与对照组(0 Gy)相比,4、8 Gy剂量的射线可以明显降低MC3T3-E1细胞的增殖,并下调其Osterix和骨钙素基因的表达;8 Gy剂量的射线可以明显抑制细胞的胶原分泌和基质矿化能力。结论  60Co γ射线可以降低MC3T3-E1细胞的增殖、胶原分泌及基质矿化能力,并下调其成骨相关基因表达,且随辐射剂量增加,其作用增强。

关键词: 成骨细胞, &gamma, 辐射, 细胞分化, 基因表达

Abstract:

 Objective  The aim of this study is to investigate the effects of irradiation on the proliferation and differentiation of MC3T3-E1 osteoblastic cells. Methods  MC3T3-E1 cells were irradiated 24 h after initial seeding. Gamma-radiation was administered at 0, 4, and 8 Gy as single doses by using a 60Co source. Cell proliferation was assessed at days 1, 3, 5, and 7 post-irradiation by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylte-trazolium bromide assay. The collagen secretion of the cells was measured through sirius red staining at day 12 post-irradiation. The expressions of osteogenesis-related genes were assessed throughreal time fluorescence quantitative polymerase chain reaction at day 16 post-irradiation. The matrixminera-lization caused by cells was evaluated through alizarin red staining at day 28 post-irradiation. Results  The cells exposed to 4 Gy or 8 Gy demonstrated significantly lower proliferation rates compared with the non-irradiated group. Doses of 4 Gy or more significantly inhibited the expressions of osteogenesis-related genes(Osterix and osteocalcin). Collagen secretion and cell mineralization were significantly reduced by the 8 Gy dose. Conclusion  60Co γ-rays dose-dependently suppress the proliferation, collagen secretion, and mineralization of MC3T3-E1 cells. Furthermore, radiation seems to dose-dependently inhibit the expressions of osteogenesis-related genes of the cells.

Key words: osteoblast, gammairradiation, celldifferentiation, geneexpression