| Title | 低氧培养促进大鼠髓核间质干细胞增殖 |
| Other Titles | Hypoxia promotes proliferation of nucleus pulposus-derived mesenchymal stem cells in rats |
| Authors | 张亮 刘忠军 王静成 黄泽楠 陈涛 冯新民 陶玉平 |
| Affiliation | 江苏省苏北人民医院骨科/江苏省苏北人民医院骨科研究所江苏扬州225001 开云app体育 第三医院骨科,北京100191 开云app体育 第三医院骨科,北京,100191 江苏省苏北人民医院骨科/江苏省苏北人民医院骨科研究所江苏扬州225001 |
| Keywords | 髓核间质干细胞 低氧 增殖 缺氧诱导因子-1α 沉默信息调节蛋白 nucleus pulposus mesenchymal stem cells hypoxia proliferation HIF-1α silent information regulator protein |
| Issue Date | 2017 |
| Publisher | 基础医学与临床 |
| Citation | 基础医学与临床. 2017, 37(11), 1546-1551. |
| Abstract | 目的 研究低氧培养对大鼠髓核间质干细胞(NPMSCs)增殖的影响.方法 分离大鼠尾椎髓核获得NPMSCs并进行体外扩增培养,观察细胞形态,流式细胞仪鉴定细胞免疫表型.取第3代NPMSCs分为常氧组和2%低氧组,分别培养7 d及14 d,观察细胞形态;MTT法检测细胞增殖;流式细胞仪检测细胞活力和RT-qPCR检测低氧诱导因子1α(HIF-1α)、葡萄糖转运蛋白1(GLUT-1)、血管内皮生长因子(VEGF)、沉默信息调节蛋白1(SIRT1)及SIRT6的mRNA表达情况.结果 原代NPMSCs细胞早期可形成葵花样细胞集落,传代后细胞增殖明显加快,细胞形态以纺锤形为主.第3代细胞高表达干细胞相关阳性表面抗原分子,低表达干细胞相关阴性表面抗原分子.低氧组细胞形态以多角形为主,细胞更容易聚集成团块,且细胞增殖速度明显加快(P<0.05),细胞凋亡率明显下降(P<0.05).低氧组HIF-1α、VEGF、GLUT-1、SIRT1及SIRT6表达量明显高于常氧组(P<0.05).结论 2%O2的低氧环境促进髓核间质干细胞增殖,其机制可能与SIRT1及SIRT6介导的HIF-1α信号通路有关. Objective To investigate the effect of hypoxia on the proliferation of nucleus pulposus-derived mesen-chymal stem cells ( NPMSCs) in vitro and explore its possible mechanism .Methods NPMSCs were isolated from nucleus pulposus of Sprague-Dawley rats.Cellular morphology was observed and expression of CD 11b, CD45, CD73, CD90 and CD105 was detected using flow cytometry .The third generation NPMSCs were cultured under nor-moxia (20%O2) and hypoxia (2%O2) for 14 days.Cell viability was determined by the annexin-V-FITC/propidi-um iodide doublestaining assay and cell proliferation was measured by MTT assay .The expressions of hypoxia-in-ducible factor-1α( HIF-1α) , glucose transporter 1( GLUT-1) , vascular endothelial growth factor ( VEGF) , silent information regulator protein 1( SIRT1) and silent information regulator protein 6 ( SIRT6) at the mRNA level were examined by semi-quantitative reverse transcription polymerase chain reaction ( RT-qPCR ).Results NPMSCs formed sunflower-like colony and the third passage NPMSCs became homogeneous and exhibited spindle -like mor-phology .Meanwhile , high expression level of stem cell-related positive antigen molecules and low expression levels of negative antigen molecules .Hypoxia promoted proliferation of NPMSCs and promoted gene expression of HIF-1α, GLUT-1, VEGF, SIRT1 and SIRT6 significantly(P<0.05).Conclusions Hypoxia has a significant impact on the proliferation of NPMSCs and SIRT 1, SIRT6 mediated HIF-1αpathway is potentially involved in the mechanism . |
| URI | http://hdl.handle.net/20.500.11897/498192 |
| ISSN | 1001-6325 |
| DOI | 10.3969/j.issn.1001-6325.2017.11.010 |
| Indexed | 中国科学引文数据库(CSCD) |
| Appears in Collections: | 第三医院 |
