| Background: As the Chinese people's living standards and changes in the structure of diet make vascular disease risk factors increase,as well as the development of the ageing of society,lower limb ischemia causing limb gangrene disease has become one of the major causes of lower limb amputation.Lower limb ischemia of the lower extremity arterial diseases includes atherosclerosis, thrombosis obliterans, and diabetic foot.It often can lead to lower extremity vascular outflow tract obstructed, secondary ischemic necrosis of lower limb amputations result.Lower extremity arterial blood flow in patients with the redevelopment is to improve the quality of life, reduce amputation rate.Conventional treatment has many limitations in the current clinical application,and long-term treatment is not precise.Because of poor conditions for lower extremity arterial not receiving arterial bypass surgery, or ineffective drug therapy,or the patient was unwilling to accept the lower extremity arterial bypass, or frail elderly patients with lower extremity arterial have no tolerance bypass surgery,autologous bone marrow stem cell transplantation (Autologous Bone-marrow Cells Transplantation, ABMCT )in the treatment of lower extremity ischemia can be significantly improved symptoms , which introduces a new line of thought for the treatment of lower extremity ischemia.However,the differentiation of bone marrow stem cells and control mechanism are still not clear, and lower limb ischemia patients were elderly.The quality of bone marrow is poor, stem cells may be less content,so how to induce vascular endothelial cell differentiation orientation,and promote angiogenesis growth factor secreted to enhance its function to promote angiogenesis, needs further study.This is the aim for the design of this experiment.Objective:The experiment is designed to study the expression trend of VEGF,Flk-1 and tie-2 in hind limb ischemia of the rat and observe the effect of autologous bone marrow stem cell transplantation VEGF, Flk-1,tie-2 expression using panax notoginseng saponins(PNS) to discuss the role and mechanisms of ischemic tissue angiogenesis after stem cell transplantation. Methods:80 SD rats were randomly divided into normal group, ischemic group (4h ischemic group, 3d ischemic group, one week ischemic group,two weeks ischemic group, four weeks ischemic group),and treatment group(2 weeks stem cells group,4 weeks stem cell group,2 weeks stem cell + medicine group,4 weeks stem cell + medicine group),cut,and ligated on the left hind limb femoral artery and its branches,established rat model of hindlimb ischemia.Ischemia group were executed in batches at 4h, 3d,1 week,2 weeks,4 weeks,used western blot method to detect VEGF,Flk-1, tie-2 expression of ischemic tissue in rats;get bone marrow mononuclear cells from the donor rat;the treatment group two weeks later after modeling success,had ischemic hind limb stem cell injection, at the same time the medicine group began to inject Xuesaitong (PNS),and the only stem cells group was injected saline in the same proportion,then was executed at two weeks and four weeks respectively,used western blot method to detect VEGF, Flk-1, tie-2 expression in the gastrocnemius muscle of the ischemic tissue.Results:The results of this study show that:①Ischemia after 4h VEGF protein expression was increased , peaked at one week, then declined lower than the normal level at four weeks.②Flk-1:after 4h ischemic Flk-1 protein of the expression hind limb began to increase, one week at the peak, and then slowly declined, slightly lower than the normal level also at 4 weeks.③Tie-2: after 4h ischemia, tie -2 protein expression was slightly lower, then slowly increased, reached a peak at the second weeks, and then slowly declined. Ischemia time of the Tie-2 expression were lower than normal.④stem cell + medicine four weeks group was significantly higher than that of VEGF expression in the normal group and two weeks stem cells group; two weeks stem cell +medicine group expressed VEGF significantly higher than the normal group and two weeks stem cell group.⑤Flk-1 expression in four weeks stem cell +medicine group was higher than the other four groups.⑥tie-2 expression in four weeks stem cell+ medicine group was significantly higher than the normal group, two weeks stem cells group and four weeks stem cell group; tie-2 expression in two weeks stem cell +medicine group was significantly higher than the normal group and two weeks stem cell group; tie-2 expression in two and four weeks stem cell groups were higher than ischemia after 4h group; tie-2 expression in the normal group was higher than ischemia after 4h group.Conclusions: 1.Hypoxia-ischemia can cause VEGF,Flk-1 high expression of the reaction within one week ischemia,subsequently the impact reduced.2.Stem cell transplantation can promote VEGF,Flk-1 and Tie-2 high expression in the ischemic tissue.3.PNS can increase VEGF,Flk-1 and Tie-2 expression in the ischemic tissue after bone marrow mononuclear cell transplantation,and promote EPC differentiation,proliferation,further promote the proliferation of endothelial cells and angiogenesis. |
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