| Objective:Application of vascular endothelial growth factor(VEGF) in autologous bone marrow to observe its mobilization of endothelial progenitor cells and its prevention of vascular stenosis after injury.Methods:21 Japanese big ear adult health rabbits(from the Experimental Animal Center in Henan Province) for the experimental study.Making iliac artery injury model through dragging balloon catheter in iliac artery.These rabbits were divided into three groups, randomly,7 rabbits in each group(n=7).Treatment group of injury:fter injury of rabbit iliac artery,through the injection of vascular endothelial growth factor(VEGF) to mobilize endothelial progenitor cells of in autologous bone marrow,once daily of a week.Control group of injury:after injury of rabbit iliac artery,injection of normal saline,once daily of a week.Sham-operated control group:only one side of iliac artery artery was ligated.Sources from the peripheral venous blood which was collected from ear vein of all rabbits on 1d,4d,7d will be examined the number of CD34~+ cell through using of flow cytometry.Each rabbit in these three groups will be measured routine of blood on 2d,4d,6d,8d.Killing all the rabbits on 21d and taking out all the rabbits' iliac artery as specimens to make histopathological slide.Every slide is carry out the HE(Hematoxylin and Eosin) staining and PCNA(proliferating cell nuclear antigen) immunohistochemical staining.Application of computer image analysis system to measue thickness of blood vessels and endangium,vascular neointima /media ratio of every slide.Observing PCNA immunohistochemistry which is the response effect of neointimal hyperplasiaSPSS13.0 was used to carry out statistical analysis.The experimental data was expressed with(?)±s.Results:The fourth and six days number of leukocytes and the percentage of mononuclear cells between all rabbits of intravenous injection VEGF treatment group and the injured artery control group and sham-operated control group were significantly different(P＜0.01);the control group injuries and control group sham-operated had no statistical significance between groups(P＞0.05).The rate of the CD34 positive cells detected by flow cytometry in peripheral circulation is 0.176 percent,injury control group1,4,7 days after the operation detected by flow cytometry of CD34 positive cell rate in the peripheral circulation was 1.735%,0.689%,0.307%, indicating that surgery and vascular endothelial injury of the rabbit has obvious effects in vivo endothelial progenitor cell mobilization,statistical analysis showed that the number of CD34 positive cells in the injury control group rabbit peripheral circulation after the operation of 1,4,7day were significantly higher than sham operation group(P＜0.01).VEGF treatment group after the operation of 1,4,7 days detected by flow cytometry of CD34 positive cell rate in the peripheral circulation was 2.736%,3.120%,2.358%,indicating that vascular endothelial growth factor can very significant mobilize EPCs on rabbits,the first 1 day,rabbit EPCs in the peripheral blood showed a significant increase in the trend in the administration of the first four days the number of CD34 positive cell peak,the increase trend has continued into the first 7 days.Statistical analysis showed that,VEGF treatment group in 1,4,7 days after administration of peripheral circulation the number of CD34 positive cell was significantly higher than sham-operated group and the control group,respectively compared with the above-mentioned two groups,both P＜0.01.HE staining method shows that the control group with simple balloon injured shows obvious neointimal proliferation and lumen stenosis after three weeks,while the VEGF treatment group is weaker compared to the control group.The average thickness of the vessel wall:the normal group is 127.36±10.39μm,the control group is 261.43±20.10μm,there are differences between the two groups(P＜0.01),the control group is thick than the normal group;the VEGF treatment group is 179.65±23.04μm,its vascular wall thickness is minor than the control group,there are differences between the two groups(P＜0.01).The average thickness of intima: sham-operated group is 29.07±8.21μm,the control group is 96.30±15.03μm,there are significant compared to normal group(P＜0.01);the VEGF treatment group is 54.82±12.03μm,its hyperplasia level is weakly than the control group,there is significant between the two groups.Endometrial neointima/media ratio:To calculate the medial ratio of the VEGF treatment group and the control group separately,the VEGF treatment group is 0.58±0.08,the control group is 1.01±0.05,there is difference between the two groups(P＜0.01).PCNA immunohistochemistry is used to calculate the PCNA- positive cells in intima and adventitial after the surgery was imposed for three weeks.It is found that very few cells for PCNA expression in normal group,while more PCNA positive cells in control group.The number of PCNA-positive cells in intima and adventitial of VEGF treatment group is less than the control group.It has statistical significant between the two groups(P＜0.01).Conclusions:1.Vascular endothelial growth factor can increase mobilization of CD34~+ cell of peripheral blood on bone marrow mononuclear cells in bone marrow,especially on vascular endothelial progenitor cells.2.Vascular endothelial growth factor can significantly reduce neointimal proliferation,to effectively reduce the intima after vascular injury in the extent of restenosis. |
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