| Because of the improving of living standard and increased elderly population, the morbality of coronary atherosclerotic heart disease increased significantly in recent years, so more and more patients need coronary artery bypass graft. The materials for coronary bypass graft include arteries and venous, but because the limited of arteries and more serious hurt to get arteries graft than venous, the great saphenous vein is widely used in clinical practice as the primary material of CABG, because the great saphenous vein is superficial and easily obtained, and has enough length to connect the aorta and the far coronary artery.The autogenous vein will be restenosis after vein grafts because of endometrial injury by operative, hemodynamic changes and a series of inflammatory factors release.That make the saphenous vein is restrictedin the clinical application.In recent years,a series of methods was used for preventing vein restenosis in clinical,It was reported that Ginsenoside Rbl has some positive effect on preventing vein restenosis.After established the animal models of vein grafting, we used a certain dose of Ginsenoside Rbl by intraperitoneal injection for the animal models. By observed the changes of vein past-grafting and endothelial progenitor cells proliferation of bone marrow, to investigate the influence of ginsenoside Rb 1 on vein graft restenosis in autogenous vein graft rabbits. Part one Expertment investigation on Ginsenoside Rbl mobilizing the endothelial progenitor cell proliferation of bone marrow in autogenous vein graft rabbitsBackground and ObjectiveEndothelial progenitor cells(Endothelial progenitor cells, EPCs) are a group of precursor cells which functions with migration and proliferation and differentiation of naive cells can not only migrate to the peripheral blood to differentiate into mature endothelial cells involved in angiogenesis during embryonic, postnatal angiogenesis growth, under certain conditions can also be transformed into smooth muscle cells, for the vein to prevent restenosis in the bridge provides a new way of thinking. Source of progenitor cells wide, and has easy to separate, proliferation ability, and many other features, has become an important vascular tissue engineering of seed cells. Under normal circumstances mature endothelial cells and endothelial damage and endothelial progenitor cells to repair in a dynamic balance. Pathological state, the body reserves the release of endothelial progenitor cells to endothelial damage to the peripheral parts of the directional migration, adhesion, assist in the repair of vascular endothelial cell injury, effective to reduce restenosis in vein grafts. Restenosis after coronary artery bypass grafting, mainly late atherosclerosis, endothelial injury is the initial factor, endothelial cell injury, macrophage adhesion, immersion and the excessive proliferation of smooth muscle cells led to stenosis or occlusion of target vessel. Promote vascular endothelial damage repair can effectively inhibit smooth muscle cell proliferation and neointimal formation, for the treatment of restenosis vein provides a new target.Previous studies found that ginsenoside Rbl can promote endothelial cell culture medium in the progenitor cell proliferation, this study based on animal vessel model based on the observation of animals, the femur bone marrow endothelial cell surface markers CD34+, CD 133+ and VEGFR2+ positive expression, to investigate the influence of ginsenoside Rbl on endothelial progenitor cells proliferation of bone marrow in autogenous vein graft rabbits.Materials and Methods45 New Zealand rabbits were randomly divided into experimental group, model group, control group 3 groups of 15 rats. Application of "no-touch" technology access after the external jugular vein, cut the carotid artery, using a continuous suture anastomosis external jugular vein and carotid artery, the establishment of external jugular vein and carotid artery bypass model. After 4 weeks, immunohistochemistry to detect bone marrow CD34+, CD133+ and VEGFR2+ positive cells in the situation, RT-PCR detection of bone marrow CD34+, CD133+ and VEGFR2+ positive mRNA expression.Results1. The degree of stenosis of grafted veinThe picture under-microscope showed that:After 4 weeks, the vascular intimal thickness of experimental group, model group and control group was(46.53±2.50)μm,(50.80±2.68)μm,(43.73±3.24)μm, the comparison among groups P<0.05, experimental group, model group and control group were significantly different among groups. The intimal/medial thickness ratio of three groups was (1.96±0.06), (2.11±0.07), (1.81±0.09), the comparison between groups P<0.05, the ratio of experimental group, model group and control group was significantly difference between groups.2. Immunohistochemistry of bone marrow CD34+, CD133+ and VEGFR2+ positive cells were observed Vein graft in each group shows CD34+, CD133+ and VEGFR2+.4 weeks after positive cells, positive cells in the experimental group was higher than the model group, model group, positive cells in the control group.3. bone marrow RT-PCRCD34+, CD133+ and VEGFR2+ gene expression observed The group transplanted veins visible CD34+, CD133+ and VEGFR2+ mRNA expression, CD34+ in the experimental group, model group and control group expressed by the relative coefficient is (2.32±0,08), (2.08±0.12), (1.88±0.11), the comparison between groups P<0.05, experimental group, model group, control group between the two groups were significantly different; CD133+ in the experimental group, model group, control the expression of the relative coefficient is (2.46±0.12), (2.24±0.09) and (1.97＝0.15), the comparison between groups P<0.05, experimental group, model group, control group between the two groups were significantly different; VEGFR2+ in the experimental group, model group, control the expression of the relative coefficient is (1.92±0.14), (1.79±0.15) and (1.55±0.09), the comparison between groups P<0.05, experimental group, model group and control group were significantly different between groups.Conclusion1.The expression of CD34+, CD133+ and VEGFR2+ increased significantly in autogenous graft vein rabbits.2. Ginsenoside Rbl can promote the endothelial progenitor cells proliferation of bone marrow.Part two Expertment investigation on Ginsenoside Rbl to prevent the restenosis of autogenous vein graft in rabbitsBackground and ObjectiveCABG (coronary artery bypass grafting, CABG) is one of the important treatments for coronary heart disease.the great saphenous vein is widely used in clinical practice as the primary material of CABG, because the the great saphenous vein is superficial and easily obtained, and has enough length to connect the aorta and the far coronary artery.The autogenous vein will be restenosis after vein grafts because of endometrial injury by operative, hemodynamic changes and a series of inflammatory factors release.That make the saphenous vein is restrictedin the clinical application.In recent years,a series of methods was used for preventing vein restenosis in clinical, such as drug treatment, gene therapy, extravascular stent, the improve expansion fluid and endothelial progenitor cells, but the effect was so poor. It was reported that:Ginsenoside Rbl has some positive effect on preventing vein restenosis. After established the animal models of vein grafting, we used a certain dose of Ginsenoside Rbl by intraperitoneal injection for the animal models.By observed the changes of vein past-grafting, proliferating cell nuclear antigen (PCNA), transforming growth factor (Transforming growth factor, TGF-P) and nitric oxide synthase(Endothelial nitric oxide synthase, eNOS)in endometrial cells,we found that the ginsenoside Rbl has some positive effect on the vein graft restenosis inhibition. This was a basis theoretical for future clinical application.Materials and Methods45 New Zealand rabbits were randomly divided into three groups:the experimental group, the model group and the control group,15 rabbits in each group, "no-touch" technology was used for getting the external jugular vein. Establish the animal model with using a continuous way anastomosis external jugular vein to ipsilateral carotid artery by end-to-end fashion.4 weeks later, grafting vein was obtained as sample, the vein intimal thickness,and the intima/media thickness ratio, and the expression of TGF-βand eNOS gene was examined by RT-PCR.Results1. The degree of stenosis of grafted veinThe picture under-microscope showed that:After 4 weeks, the vascular intimal thickness of experimental group, model group and control group was(46.53±2.50) um,(50.80±2.68)μm,(43.73±3.24)μm, the comparison among groups P<0.05, experimental group, model group and control group were significantly different among groups. The intimal/medial thickness ratio of three groups was (1.96±0.06), (2.11±0.07), (1.81±0.09), the comparison between groups P<0.05, the ratio of experimental group, model group and control group was significantly difference between groups.2.The results observed by immunohistochemical stainingPCNA, eNOS, TGF-P positive cells can be seen in each group grafted vein. After 4 weeks.the experimental group, model group, the control group expression of positive PCNA index were (40.63±2.30),(60.45±3.38),(19.89±2.56), The expression of TGF-p were (18.15±4.31), (29.28±6.37), (12.63±3.72) respectively, The expression of eNOS were (25.64±5.03),(17.91±3.75), (10.25±2.31)respectively, compared among the groups P<0.05. the expression of experimental group, model group and control group were significantly different among groups.3 The results observed by RT-PCR of grafted veinThe expression of eNOS, TGF-βgene can be seen in each group grafted vein. the expression of eNOS relative coefficients in experimental group, model group, control group were (2.12±0.05),(1.98±0.09),(1.72±0.09), the expression of eNOS relative coefficient in the experimental group is higher than the model group and control group, compared among groups P<0.05, the expression of experimental group, model group and control group were significantly different among groups. The expression of TGF-βrelative coefficients in experimental group, model group, control group were (1.95±0.08),(2.10±0.01),(1.82±0.01), the expression of TGF-βrelative coefficient in the model group is higher than in the experimental group and control group, compared among groups P<0.05, the expression of experimental group, model group and control group were significantly different between groups.Conclusion1. Grafted vein intimal injury is the initial factor of restenosis.The remodeling of vascular is the most important reason of restenosis.2. Ginsenoside Rbl can prevent restenosis of vein graft effectively.3. Ginsenoside Rbl can promote NOS gene expression in grafted vein, inhibited TGF-βgene expression in grafted vein. |
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