Effects of a recombinant adenovirus expressing human hypoxia-inducible factor la double-mutant on the in vitro differentiation of bone marrow mesenchymal stem cells to cardiomyocytes
10.3760/cma.j.issn.0253-3758.2010.07.016
- VernacularTitle:人双突变型低氧诱导因子α促进骨髓间充质干细胞向心肌细胞分化的实验研究
- Author:
Jiao-Jie XUE
1
;
Ye-Song WANG
;
Hong MA
;
Yuan HU
;
Kang-Lin CHENG
Author Information
1. 中山大学附属第六医院
- Keywords:
Myocytes,cardiac;
Stem Cells;
Hypoxia-inducible factor 1,alpha subunint
- From:
Chinese Journal of Cardiology
2010;38(7):638-643
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the effects of mutant hypoxia-inducible factor-1α ( HIF-1α) adenovirus ( Adeno-HIF-1α-Ala402-Ala564 ) on cardiomyocytes ( CMCs ) differentiation from the mesenchymal stem cells ( MSCs) co-cultured with CMCs. Methods Following groups were studied: HIF-1α group ( MSCs + CMCs + Ad-HIF-1α), LacZ group ( MSCs + CMCs + Ad-LacZ) , Sham group (MSCs + CMCs + PBS) and MSC + HIF-la Group ( MSCs + Ad-HIF-1α). MSCs were co-cultured with myocardial cells in proportion of MSCs: CMCs 1:2, after 24 hours, cells were infect with virus (MOI = 100) or treated with PBS, cardiac troponin (cTnT) expression in MSCs was detected 7 days post infection by immunochemical analysis, mRNA expression of HIF-la, TGF-B,, Smad4, NKx2.5, GATA-4 was also detected by RT-PCR. Results HIF-la increased MSCs differentiation to myocardial cells (differentiation rate 32. 68%±6. 52% vs. 8. 28% ± 0.09% in the LacZ group and 10. 25 % ±2. 20% in the Sham group and 0.32% ±0.05% in the MSC group (all P< 0.05 vs. HIF-la group). mRNA expression of HIF, TGF-β1, Smad4, NKx2. 5 and GATA-4 was also significantly upregulated in HIF-1α group all P <0. 05 vs. Sham group). Conclusion HIF-1α promoted MSCs, co-cultured with myocardial cells, differentiating to cardiomyocytes via upregulating TGF-β1/Smad4 signaling pathway.
