ABSTRACT
AIM:To investigate the effects of integrin-linked kinase (ILK) overexpression on survival and proliferation of cardiac c-Kit + cells,and the role of ILK-overexpressing c-Kit + cell transplantation in cardiac function in a rat myocardial infarction (MI) model.METHODS:Cardiac c-Kit + cells were isolated from the hearts of neonatal Sprague-Dawley (SD) rats and cultured to prepare the ILK-c-Kit+ cells by infected with recombinant adenoviral vector harboring human wild-type ILK cDNA.The survival and proliferation of cardiac c-Kit + cells were detected by cell counting and CCK-8 assay at 48 h after infection,respectively.The protein levels of cyclin D1 and proliferating cell nuclear antigen (PCNA) in the cardiac c-Kit+ cells were examined by Western blot.MI was induced by coronary artery ligation in 40 adult rats.After 15 min,ILK-c-Kit + cells were transplanted into the hearts by myocardial injection at 3 different sites in the infracted zone and border zone.All rats were randomly divided into 4 groups:sham group,MI plus saline injection group (MI group),MI plus null vector-infected cardiac c-Kit+ cell injection group (Ad-null-c-Kit+ cell group),and MI plus ILK-overexpressing cardiac c-Kit + cells injection group (ILK-c-Kit+ cell group),with 10 rats in each group.At 2 weeks after MI,the protein levels of c-Kit in MI hearts were investigated by immunohistochemical assay.At 4 weeks,left ventricular function was examined by hemodynamic measurement.RESULTS:The survival and proliferation of cardiac c-Kit + cells and the protein levels of cyclin D1 and PCNA were enhanced by ILK overexpression compared with Ad-null group.In MI rat model,the number of c-Kit + cells was increased by ILK-c-Kit + cell injection compared with Ad-null-c-Kit + cell group at 2 weeks after MI.Cardiac function was significantly improved in ILK-c-Kit + cell-transplanted rats.CONCLUSION:ILK overexpression improves survival and proliferation of cardiac c-Kit + cells by increasing the protein levels of cyclin D1 and PCNA.ILK-c-Kit + cell transplantation enhances the therapeutic efficiency of cardiac c-Kit + cells in the postMI hearts of rats.
ABSTRACT
AIM:To investigate the effects of integrin-linked kinase (ILK) overexpression on survival and proliferation of cardiac c-Kit + cells,and the role of ILK-overexpressing c-Kit + cell transplantation in cardiac function in a rat myocardial infarction (MI) model.METHODS:Cardiac c-Kit + cells were isolated from the hearts of neonatal Sprague-Dawley (SD) rats and cultured to prepare the ILK-c-Kit+ cells by infected with recombinant adenoviral vector harboring human wild-type ILK cDNA.The survival and proliferation of cardiac c-Kit + cells were detected by cell counting and CCK-8 assay at 48 h after infection,respectively.The protein levels of cyclin D1 and proliferating cell nuclear antigen (PCNA) in the cardiac c-Kit+ cells were examined by Western blot.MI was induced by coronary artery ligation in 40 adult rats.After 15 min,ILK-c-Kit + cells were transplanted into the hearts by myocardial injection at 3 different sites in the infracted zone and border zone.All rats were randomly divided into 4 groups:sham group,MI plus saline injection group (MI group),MI plus null vector-infected cardiac c-Kit+ cell injection group (Ad-null-c-Kit+ cell group),and MI plus ILK-overexpressing cardiac c-Kit + cells injection group (ILK-c-Kit+ cell group),with 10 rats in each group.At 2 weeks after MI,the protein levels of c-Kit in MI hearts were investigated by immunohistochemical assay.At 4 weeks,left ventricular function was examined by hemodynamic measurement.RESULTS:The survival and proliferation of cardiac c-Kit + cells and the protein levels of cyclin D1 and PCNA were enhanced by ILK overexpression compared with Ad-null group.In MI rat model,the number of c-Kit + cells was increased by ILK-c-Kit + cell injection compared with Ad-null-c-Kit + cell group at 2 weeks after MI.Cardiac function was significantly improved in ILK-c-Kit + cell-transplanted rats.CONCLUSION:ILK overexpression improves survival and proliferation of cardiac c-Kit + cells by increasing the protein levels of cyclin D1 and PCNA.ILK-c-Kit + cell transplantation enhances the therapeutic efficiency of cardiac c-Kit + cells in the postMI hearts of rats.