| BackgroundWith the development of medical science,many prograss have been made in cardiovascular diseases.But the heart failure is still the difficult problem that menace the human health.Medicine,surgery and intervention operation can't substitute the putrescent tissue and prevent ventricular remodeling , Although they can improve the cardiac function.Studies show the transplate of stem cell can substitute the injured cardiac cell, increase the functional cardiac cell and reform new circulated vessel. Some studies show bone stem cells were "enlisted" to the circulation, automove to the injured cardiac zone and differentiate cardiac cell, endothelial cell and smooth muscle cell, so bone stem cells regenesis cardiac cell and improve the function of heart. But the effect of enlistment is feeble. G-CSF and SCF have the ability that canmobilize HSCs and MSCs. Aim①Combination of G-CSF and SCF mobilize BMSCs, observe the proliferation of BMSCs by giving different dose and medication;② observe the change of left heart function, heart ischemia realm and MI realm of different groups;③explore the possible mechanism of heart function improvement by H.E stain, immunofluorescence and double immunofluorescence. Methods and ResultsFirstly, 24 rats were divide into four groups.Control group(A group) was gived normal saline, other three groups were gived G-CSF 20μg/kg and SCF 200μg/kg one per day(B group), G-CSF 20μg/kg and SCF 200μg/kg twice per day(C group), G-CSF 40μg/kg and SCF 400μg/kg one per day(D group), all groups were gived for six days. PBMCs were isolated by Ficoll-Hypaque density gradient centrifugation, and quantity of BMSCs were detected by flow cytometry. Compare with the control group, positive rate of CD34 and CD44 in the B,C,D group were more than the rate in the control group(P<0.05); In the B,C,D groups, the effect of mobilization of B group was the best(P<0.05), D group is better than B group (p<0.05), but there wasn't multiple relation in the two group.Secondly, Myocardial ischemia of left ventricle was produced in male Wistar rats by ligation of left coronary artery. The rat models were randomly divided into 2 groups. Animals in mobilization groupreceived hypodermic injection of rhG-CSF 20ug/kg and rh-SCF 200ug/kg, twice per day, and continuously for 6 days. Those in Control group received hypodermic injection of an equivalent amount of normal saline for 6 days. After 28 days, heart function was examined and myocardial infarct size was determined by dual staining with triphenylterazolium chloride and Evan's blue. LVEDP of mobilization group was higher than that of control group, ±LVdp/dtmax of mobilization group were statistically higher than that of control group (P<0.05). Myocardial ischemic size and infarct size of mobilization group were statistically smaller than that of control group (p<0.05).Thirdly, myocardial infarction of left ventricle was produced in rats by ligation of left coronary artery.Bone marrow stem cell were mobilized by G-CSF and SCF. After 28 days, Heart were harvested for HEstain, immunofiuorescence and double immunofluorescence. Compare with the control group, fibrosis was less, myocardial tissue construction was more order state, and quantities of micrangium were obviously more in the mobilization group, blood vessal CD34+ positive rate in the mobilization group was more than the rate in the control group. In the mobilization group, there were some cell that CD44 and cardiac a-actin were positive.Conclusion1. Combinatively use of G-CSF and SCF can effectually mobilizate BMSCs, promote the generation of BMSCs.
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