解放军医学杂志2017，Vol.42Issue(4)：314-319,6.DOI:10.11855/j.issn.0577-7402.2017.04.08
hHO-1联合GATA-4修饰大鼠骨髓间充质干细胞抗凋亡及向心肌分化的实验研究
hHO-1 combined with GATA-4 transduction promotes myocardial transdifferentiation and antiapoptosis of rat mesenchymal stem cells
摘要
Abstract
Objectives To explore if the rat bone marrow mesenchymal stem cells (BMSCs) modified by human heme oxygenase 1 (hHO-1) gene combined with GATA-4 gene may promote the ability of anti-apoptosis and myocardial transdifferentiation in vitro in hypoxia ischemic environment.Methods The rat BMSCs were isolated and cultured by whole bone marrow adherence and identified in vitro,and then were transfected with recombinant adenovirus;Western blotting was used to determinate the optimal time of gene expression;the genetically modified BMSCs were taken to hypoxia serum-free conditions simulating ischemia hypoxia microenvironment in vivo;CCK-8 kit and trypan blue staining were performed to detect the 12,24,48 and 72h survival rates in hypoxia ischemia respectively;flow cytometry was used to detect the apoptosis of BMSCs in hypoxia ischemia for 24h.The cardiomyocyte-specific cardiac troponin Ⅰ (cTnI) was detected by Western blotting and cellular immunofluorescence.Results The 12,24,48 and 72h survival rates were higher in hHO-1+GATA-4 group cultured in ischemia and hypoxia condition than in hHO-1 group (P＜0.05) and GATA-4 group (P＜0.05).After 24h cultivation in ischemia hypoxia condition,the apoptotic rates were lower in hHO-1+GATA-4 group than in hHO-1 group (P＜0.05) and GATA-4 group (P＜0.05).No significant difference existed in cTnI expressions between GATA-4 group and hHO-1+GATA-4 group.Conclusion Compared with transfection of hHO-1 or GATA-4 single gene,hHO-1 combined with GATA-4 transduction can significantly increase the survival rate of BMSCs in hypoxia ischemic condition,but myocardial transdifferentiation does not increase significantly.关键词
间质干细胞/基因,GATA4/血红素加氧酶-1/心肌分化Key words
mesenchymal stem cells/gene, GATA4/heme oxygenase-1/myocardial transdifferentiation分类
医药卫生引用本文复制引用
邓宁波,曾元清,韩腾龙,蒋知新..hHO-1联合GATA-4修饰大鼠骨髓间充质干细胞抗凋亡及向心肌分化的实验研究[J].解放军医学杂志,2017,42(4):314-319,6.基金项目
国家自然科学基金(30971235) （30971235）
军队“十二五”医学科研基金(2013XL010)This work was supported by the National Natural Science Foundation of China (30971235),and the "Twelfth Five-Year Plan" Medical Research Fund of PLA (2013XL010) （2013XL010）
