GW25-e0245 Effects of allogenic bone marrow mesenchymal stem cells modified by bcl-2 gene transplantation on cardiac dysfunction rabbits after acute myocardial infarction

  • Shuren L
  • Man W
  • Qing G
  • et al.
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Objectives: Ventricular remodeling of MI has seriously affected the quality of life in patients. Modified stem cells have a better angiogenesis ability, anti-apoptotic, antiinflammatory and improve cardiac dysfunction.Bcl-2 modified BMSCs to be used as a means to improve the survival rate of transplanted cells. The purpose of this study is to observe the effects of transplantation of allogeneic BMSCs modified by Bcl-2 on cardiac dysfunction in rabbits after myocardial infarction. Methods: We transferred the pShuttle-CMV-EGFP-Bcl-2 to the pAdeno carrier. Got pAdeno-Bcl-2 virus plasmid and transfected to 293 cells, collected virus. Used the adenovirus of passage one to infection to 293 cells, collected the virus which repeatedly amplified three generations. We cultured bone marrow mesenchymal stem cells by density gradient centrifugation and adherent until to the ninth generation. BMSCs were transfected by empty adenovirus vector or by adenovirus vector labeled with enhanced green fluorescent protein and modified Bcl-2 gene. Cardiac dysfunction eabbits models were made by ligating the left anterior descending artery. Two weeks later the successful rabbit models were selected according to echocardiography, randomly divided them into Ad-EGFP-Bcl-2-BMSCs group (n=6), Ad-EGFPBMSCs group (n= 6), DMEM group (n=6). Thoracotomy for cell transplantation, transplanted Ad-EGFP-Bcl-2-BMSCs, Ad-EGFP-BMSCs, DMEM. The cardiac function was detected by echocardiography at four weeks after cells transplantation. Four weeks after BMSCs transplantation, the mRNA of Bcl-2 in the infarct border zone of myocardial was determined by real time quantitative PCR. We analyzed the correlation between hearts function and Bcl-2 expression. Myocardial pathological changes were assessed by hematoxylin-eosin staining. The survival of labeled BMSCs was observed by fluorescence microscope. Results: 1.Titer of Ad-Bcl2-GFP adenovirus was 1010PFU/ml; 2.This experiment used the ninth passage of BMSCs, the best MOI was 500, the expression of EGFP was observed after infection about 36h, the transfect efficiency of adenovirus was 100%; 3.Ad-EGFP-Bcl-2-BMSCs group and Ad-EGFP-BMSCs group were better than DMEM group; the Ad-EGFP-Bcl-2-BMSCs group was the best. EF[(68.222 +/- 3.763) %, (57.056 +/- 3.555) %, (47.389 +/- 5.567) %,P=0.000],EDD[( 9.556 +/- 0.981) mm, (11.611 +/- 0.828) mm, (13.389 +/- 1.569) mm, P=0.000],ESD[(5.722 +/- 0.647) mm, (7.889 +/- 0.544) mm, (9.889 +/- 0.749) mm,P=0.000], FS[(39.167 +/- 3.817) %, (31.833 +/- 2.492) %, ( 25.778 +/- 3.851) %,P=0.000]. Bcl-2 mRNA was higher in Bcl-2 group (1.015 +/- 0.115, 0.358 +/- 0.057, 0.090 +/- 0.065, P=0.000). 4. EF and Bcl-2 expression are positive correlation, the correlation coefficient r was 0.940, the coefficient of determination ( R2 ) was 0.883, regression equation, the regression equation was Y=46.664+22.329X; 5.The survival rate of transplanted stem cells was improved, in the fluorescence microscope, the number of BMSCs in Ad-EGFP-Bcl-2 group was 1.5 times of Ad-EGFP group; 6.Ad-EGFP-Bcl-2 group and Ad-EGFP group are compared with DMEM group, the pathological changes were lighter, better effect in Ad-EGFP-Bcl-2 group. Conclusions: Transplantation the Bcl-2 modified BMSCs can significantly improve cardiac dysfunction in cardiac dysfunction rabbits models. The possible mechanisms is to inhibit cells apoptosis and improve the survival rate of transplanted stem cells.




Shuren, L., Man, W., Qing, G., & Shuren, L. (2014). GW25-e0245 Effects of allogenic bone marrow mesenchymal stem cells modified by bcl-2 gene transplantation on cardiac dysfunction rabbits after acute myocardial infarction. Journal of the American College of Cardiology, 64(16), C34.

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