ABSTRACT
BACKGROUND: Loss of cardiomyocytes in the myocardial infarction leads to regional contractile dysfunction, and necrotized cardiomyocytes in infracted ventricular tissues are progressively replaced by fibroblasts forming scar tissue. Although cardiomyoplasty, or implantation of ventricular assist device or artificial heart was tried in refractory heart failure, the cardiac transplantation was the only therapeutic modality because these other therapeutic strategies were not permanent. Cell transplantation is tried instead of cardiac transplantation, especially bone marrow is the most popular donated organ. But because bone marrow aspiration procedure is invasive and painful, and it had the fewer amounts of cellular population, the adipose tissue is recommended for harvesting of mesenchymal stem cells. MATERIAL AND METHOD: After adipose tissues were extracted from abdominal subcutaneous adipose tissue and intra-abdominal adipose tissue individually, the cellular components were obtained by same method. These cellular components were tried to transformation with the various titers of 5-azacytidine to descript the appropriate concentration of 5-azacytidine and possibility of transformation ability of adipose tissue. Group 1 is abdominal subcutaneous adipose tissue and Group 2 is intra-abdominal adipose tissue-retroperitoneal adipose tissue and omentum. Cellular components were extracted by collagenase and NH4Cl et al, and these components were cultured by non-induction media-DMEM media containing 10% FBS and inducted by none, 3 micromol/L, 6 micromol/L, and 9 micromol/L 5-azacytidine after the 1st and 2nd subculture. After 4 weeks incubation, the cell blocks were made, immunostaining was done with the antibodies of CD34, heavy myosin chain, troponin T, and SMA. RESULT: Immunostaining of the transformed cells for troponin T was positive in the 6 micromol/L & 9 micromol/L 5-azacytidine of Group 1 & 2, but CD34 and heavy myosin chain antibodies were negative and SMA antibody was positive in the 3 micromol/L & 6 micromol/L 5-azacytidne of Group 2. CONCLUSION: These observations confirm that adult mesenchymal stem cells isolated from the abdominal subcutaneous adipose tissues and intra-abdominal adipose tissues can be chemically transformed into cardiomyocytes. This can potentially be a source of autologous cells for myocardial repair.
Subject(s)
Adult , Animals , Humans , Rats , Adipose Tissue , Antibodies , Azacitidine , Bone Marrow , Cardiomyoplasty , Cell Transplantation , Cicatrix , Collagenases , Fibroblasts , Heart Failure , Heart Transplantation , Heart, Artificial , Heart-Assist Devices , Intra-Abdominal Fat , Mesenchymal Stem Cells , Myocardial Infarction , Myocytes, Cardiac , Myosins , Omentum , Subcutaneous Fat, Abdominal , Transplants , Troponin TABSTRACT
Objective To investigate the protective effects of Astragaloside Ⅳ(ASⅣ)on cardiomyocytes against hydrogen peroxide(H2O2)-induced injury.Methods Cultured neonatal rat cardiomyocytes were divided into control group;0.1%DMSO solven group;H2O2 group,in which cells were treated with H2O2( 0.2 mmol?L-1) for 24 h;ASⅣ+H2O2 groups,in which cells were pretreated with ASⅣ(with final concentration of 3,10,30 mg?L-1) for 30 min before H2O2 treatment;Astragaloside injection group(150 g?L-1).Morphological changes of myocardial cells damaged for 24 h by 0.2 mmol?L-1 H2O2 under the inverted microscope were observed.The cardiomyocyte viability was analysed by MTT assay;lactate dehydrogenase(LDH) activity,superoxide dismutase(SOD) activity,malonaldehyde(MDA) content and nitric oxide(NO) content were detected in culture media.Results Most of cells in H2O2 group were crenulated,the nucleus were dim and the pseudopodia were disappeared.The degrees of lesion in ASⅣ(3,10,30 mg?L-1) groups were lessened obviously,the nucleus were bright and the the pseudopodia were thinned slightly.Cardiomyocyte viabilities in ASⅣ(3,10,30 mg?L-1) groups were higher than that in H2O2 group(P