Hypoxia promotes proliferation of nucleus pulposus-derived mesenchymal stem cells in rats
10.3969/j.issn.1001-6325.2017.11.010
- VernacularTitle:低氧培养促进大鼠髓核间质干细胞增殖
- Author:
Liang ZHANG
1
,
2
;
北京大学第三医院骨科
;
jun Zhong LIU
;
cheng Jing WANG
;
nan Ze HUANG
;
Tao CHEN
;
min Xin FENG
;
ping Yu TAO
Author Information
1. 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
2. 北京大学第三医院骨科
- Keywords:
nucleus pulposus mesenchymal stem cells;
hypoxia;
proliferation;
HIF-1α;
silent information regulator protein
- From:
Basic & Clinical Medicine
2017;37(11):1546-1551
- CountryChina
- Language:Chinese
-
Abstract:
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 .
