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.