[Effects of a recombinant adenovirus expressing human hypoxia-inducible factor 1α double-mutant on the in vitro differentiation of bone marrow mesenchymal stem cells to cardiomyocytes].
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-1α 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-1α, TGF-β(1), Smad4, NKx2.5, GATA-4 was also detected by RT-PCR.


RESULTS
HIF-1α 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-1α group). mRNA expression of HIF, TGF-β(1), Smad4, NKx2.5 and GATA-4 was also significantly upregulated in HIF-1α group all P < 0.05 vs. Sham group).


CONCLUSIONS
HIF-1α promoted MSCs, co-cultured with myocardial cells, differentiating to cardiomyocytes via upregulating TGF-β(1)/Smad4 signaling pathway.
Department of Cardiology, Sixth Affiliated Hospital of Sun Yat-sen University, Guangzhou 510655, China.
Jiao-jie
JJ
Xue
Ye-song
YS
Wang
Hong
H
Ma
Yuan
Y
Hu
Kang-lin
KL
Cheng
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics; D002478:Cells, Cultured; D005822:Genetic Vectors; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction; D051901:Smad4 Protein / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000378:metabolism
C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D051901:Smad4 Protein; C490811:Smad4 protein, rat; C508962:Tgfb1 protein, rat; D053773:Transforming Growth Factor beta1