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Objective:The aim of this study was to test the protective effect of mesenchymal stem cells(MSCs) on cardiomyocytes in vitro and to investigate the anti-apoptotic signaling pathway.Methods:MSCs from Sprague-Dawley(SD) rats were separated and cultured.MSC medium was collected from MSCs cultured in serum-free Dulbecco’s modified eagle medium(DMEM) under hypoxia.Cultured cardiomyocytes from neonatal SD rats were exposed to hypoxia/reoxygenation(H/R) and treated with MSC medium.The apoptotic cardiomyocytes were stained with Annexin-V-fluorescein isothiocyanate(FITC),Hoechst 33342 and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL).The mitochondrial transmembrane potential of cardiomyocytes was assessed using a fluorescence microscope.The expression of Bcl-2,Bax,cytochrome C,apoptosis-induced factor(AIF),and caspase-3 was tested by Western blot analysis.Results:Our data demonstrated that MSC medium reduced H/R-induced cardiomyocyte apoptosis,increased the Bcl-2/Bax ratio,and reduced the release of cyto-chrome C and AIF from mitochondria into the cytosol.Conclusion:MSCs protected the cardiomyocytes from H/R-induced apoptosis through a mitochondrial pathway in a paracrine manner.
Objective: The aim of this study was to test the protective effect of mesenchymal stem cells (MSCs) on cardiomyocytes in vitro and to investigate the anti-apoptotic signaling pathway. Methods: MSCs from Sprague-Dawley (SD) rats were separated and cultured. MSC medium was collected from MSCs cultured in serum-free Dulbecco’s modified eagle medium (DMEM) under hypoxia. Cultured cardiomyocytes from neonatal SD rats were exposed to hypoxia / reoxygenation (H / R) and treated with MSC medium. Apoptotic cardiomyocytes were stained with Annexin-V-fluorescein isothiocyanate (FITC), Hoechst 33342 and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). The mitochondrial transmembrane potential of cardiomyocytes was assessed using a fluorescence microscope. The expression of Bcl-2, Bax, cytochrome C, apoptosis-induced factor (AIF), and caspase-3 was tested by Western blot analysis. Results: Our data demonstrated that MSC medium reduced H / R-induced cardiomyocyte apoptosis, increased the Bcl- Bax ratio, and reduced the release of cyto-chrome C and AIF from mitochondria into the cytosol. Confc: MSCs protected the cardiomyocytes from H / R-induced apoptosis through a mitochondrial pathway in a paracrine manner.