| Objective:Adipose-derived stem cell(ADSC) is a kind of therapeutic stem cell for cardiovascular diseases, especially for the treatment of myocardial infarction. However there has not been unified conclusion whether ADSC can effectively restore cardiac function. We want to study the therapeutic efficiency of autologous ADSC transplantation in myocardial infarction. And we try to find out a good way to improve the therapeutic efficiency by using the combination of gene therapy and cell therapy. Anti-apoptotic protein XIAP was selected to fight against the ischemic environment of myocardic infaction.Methods:1. Subcutaneous adipose tissue was removed from left side of groin,then culture ADSCs.2. ADSCs were induced to differentiate into adipocytes, osteoblasts, chondrocytes and cardiomyocyte which is used to prove ADSCs have multi-differentiation potential.3. ADSCs were analyzed by flow cytometry by use of a FACSCalibur flow cytometer and the CellQuest Pro software.4. XIAP experession plasmid was elertco-transduced to ADSC. The anti-apototic function of XIAP was tested by serum stavation induced apotosis.5. Western Blot was performed to find whether the ability of anti-apoptosis of ADSCs is better.6. Preparing the Myocardial infarction model, SD rats were randomly divided into AMI group(n=18), AMI with ADSC transplantation group(n=18), AMI with XIAP modified ADSCs transplantation group (n=18) and untreatment control group(n=7). ADSCs, XIAP modified ADSCs and normal saline were injected into the infarct zone and border zone.7. We measured cardiac function4weeks after ADSCs transplantation.8. Rats myocardial infarction area were measured by using special stain.9. Sections of other hearts were stained for tissue morphology (hematoxylin and eosin) plus fibrosis (Masson’s trichrome). 10. Factor VIII positive endothelial cells were identified immunohistochemically.11. Statistical analysis were performed.Results1. ADSCs were successful isolated from rat adipose tissue. That was confirmed by FACS assay. We found out that ADSC expressed both CD29、CD44、CD90and CD105. The expression ratio was greater than79%.2. ADSCs were induced to differentiate to adipocyte, chondrocyte and osteoplast in different medium. The successful differentiation was confirmed by specific staining. The ADSCs have multi-differentiation potential.3. Western blot analysis was performed for XIAP modified cells after serum starvation inducing apoptosis. Activated caspases7was used as indicators of apoptosis. Over expression of XIAP in ADSCs blocked serum starvation induced activation of caspase7. It strongly suggest that, XIAP over-expression block serum starvation induced apotosis and it may also block the apoptosis during cell-traplantation of ADSCs.4. We found out that LVDs and LVDd of ADSCs group were less than control group. LVEF was better in the ADSCs, comparison with control group. The reduction of LVEF of XIAP modified ADSCs group was significantly lower than the ADSCs.5. Left ventricular cavity and infarct size is reduced, compared with the control group (P<0.01). Infarction area in XIAP modified ADSCs group was smaller than ADSCs group(P<0.05) and the control group (P<0.01)6. The capillary count in infarct zone was significantly greater in the ADSC-treated animals than in the control group (P<0.05). The capillary count was the greatest in XIAP modified ADSCs group, compared with the control group(P<0.05)and ADSCs group(P<0.05).Conclusions:Autologus ADSC transplantation is an efficient therapeutic tool in myocardial infarction therapy. Over expression of XIAP can partly inhibit low serum induced apotosis of ADSC in vitro. Over expression XIAP of ADSC can improve the therapeutic efficiency compare to ADSC transplantation. |
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