Change In C-kit And Nanog Gene Expressions In A Rat Model Of Chronic Heart Failure Induced By Adriamycin

Background It has draws great attention to prove the relations between c-kit and nanog and the disease in recent years as c-kit and nanog play an important role in the growth, proliferation and differentiation of stem cells.C-kit and nanog positive cells in the heart are cardiac stem cells. They can divide into cardiomyocytes. Our previous studies have shown that it is conducive to repair myocardial injury in the increase of nanog expression at the edge of the myocardial infarction.However,it isn’t still clear that cardiac stem cells are to increase or decrease, enhance or reduce myocardial regenerative capacity in the heart of heart failure.Objectives To explore the changes of c-kit and nanog gene expression in rat myocardial tissue after heart failure which was made by adriamycin, which has provided morphologi-cal basis for the further study of myocardial regeneration in the process of congestive heart failure.Methods1 There are one hundred and forty male SD rats.Fifteen of them were randomly divided into normal control groups and one huandred and twenty-five of them were randomly divided into heart failure model groups. Heart failure model:4 mg/kg intraperitoneal injection of adriamycin,once a week, a total of six weeks. Normal control groups were given equal amount of physiological saline. To detect the cardiac function of normal control groups and heart failur model groups during the eighth week to prove the heart failure.2 Gathering the rats’ hearts, Taking the atrioventricular organization border areas and making frozen sections;Using immunohistochemistry and immunofluorescence color technology to detect the changes of nanog and c-kit gene expression in myocardial tissue;3 Extract cardiac protein and mRNA of rats atrioventricular border areas;Using Western Blotting and RT-PCR to detect and quantitative myocardial tissues nanog and c-kit gene changes of the expression of quantity.Results1 immunohistochemistry shows that a small amount of nanog and c-kit positive cells all express in myocardial tissue of normal and heart failure..Nanog cells are less, small and round in normal myocardial tissue, which can be scattered or also be in groups in existence between the myocardial cells and blood vessels around.Most of the vascular endothelial cells express nanog, a small amount of myocardial cells also positive express., Three kinds of nanog positive cells were reduced, and only in myocardial intima and epicardial tissue was positive for distribution in heart failure rat myocardial tissues. C-kit positive cells in the small round or oval, small groups in normal and heart failure rat myocardial tissue, but did not see c-kit positive expression in myocardial cells and endothelial cells.Compared with the normal myocardium, c-kit positive cells decreased significantly in heart failure cardiac muscle.2 Immunofluorescence shows that nanog protein express more in normal control group myocardial, mainly expressed in part of the myocardial cells, endothelial and small round cells, strong positive cells uneven distribut. Nanog positive cells mainly distributed in epicardial and endocardial myocardium, the expression reduce significantly than normal myocardial and endocardial side express low compaired with epicardial side. C-kit protein is mainly expressed in the myocardium of small cells, the cells in myocardial tend to cluster together, the c-kit positive cells mainly gathered in epicardial myocardium. Compared with the normal myocardium, c-kit positive cells decrease significantly after heart failure. Not see myocardial cell and endothelial cell to express the c-kit fluorescence.In addition, you can observe a few cells express nanog and c-kit at the same time.3 Western blotting results show that c-kit and nanog expression quantity were 451846.3±7029.35 and 451846.3±3549.73 in the normal control groups; c-kit and nanog expression were 146428.7±8029.24 and 196803.3±3066.76 in heart failure model groups.The c-kit and nanog protein of normal control groups express more significantly than heart failure model groups (P< 0.05).4 PT-PCR results show that c-kit and nanog expression quantity were 115696.33 ± 8225.82 and 108401.7± 12090.2 in the normal control group; c-kit and nanog expression were 69273.7±16375 and 92935.67±14648.7 in heart failure model group.The c-kit and nanog mRNA of normal control group express more significantly than heart failure model groups (P<0.05).Conclusions Heart failure can induce the decrease of c-kit and nanog positive cells in myocardial and myocardial regeneration capacity decline.