Study On Treatment Of Rat Acute Myocardial Infarction With Transplantation Of BMMSCs Co-inducted With Angiotensin Ⅱ And5-azacytidine

ObjectiveThe previous studies had confirmed that bone marrow mesenchymal stem cells(BMMSCs) could be induced by5-azacytidine (5-Aza) to differentiate intocardiomyocyte-like cells (CMCs) in vitro. But the differentiation ratio is very low. Forincreasing the differentiation ratio, BMMSCs were co-induced with angiotensin II(Ang II) and5-Aza in vitro. And the co-induced cells were transplantated into tissuesof rat acute myocardial infarction (AMI) to evaluate changes in cardiac function invivo.MethodBMMSCs were isolated from the bone marrow of Sprague-Dawley (SD) rats. Thefourth-passage cells were used for the succeeding researches. In vitro study: BMMSCswere divided into three groups: the un-induced group as control, the5-Aza group(10μmol/L), and the co-induced group (Ang II:0.1μmol/L,5-Aza:10μmol/L). Themorphological changes were observed. And the expression of β-MHC and CTn T were detected by immunofluorescence staining; the expression of GATA4and NKx2.5weredetected by PCR. In vivo study: Left anterior descending artery (LAD) was ligated toestablish a rat model of AMI on60SD rats. Then, the animals were randomly divided into4groups to receive variant treatments: Group A: rats injected the serum-free DMEM,Group B: rats injected un-induced BMMSCs. Group C: rats injected BMMSCs induced by5-Aza, and Group D: rats injected BMMSCs co-induced by Ang II and5-Aza. Beforetransplantation, all cells would be labeled with DAPI(10mg/L). Comparing the survival rateof transplanted cells and the changes in cardiac function in different groups.Results1) Comparing the5-Aza group with the co-induced group, we found the differentiationratio of BMMSCs between two groups had statistically significant differences.2) Compaired group D with B or C, the survival rate of DAPI-labeled transplanted cellswas increased, the collagen fibers in myocardium reduced, the VEGF expression increasedand the cardiac function improved in group D.ConclusionIt is confirmed that the co-induction of Ang II and5-aza can promote the differentiationof BMMSCs into CLCs, and the CLCs could improved the cardiac function of AMI rats.