| Objective:To establishment a stable reproduction animal model of diabetic food ulcer and investigation the VEGF expression on allogeneic bone marrow-derived mesenchymal stem cell (MSCs) transplantation in the treatment of rats with diabetic foot ulcers in the systemic and local.Method:Wistar rats were randomly divided into two groups, control group (n=10) and experiment group (n=40). Rats in experiment group were injected Steptozotocin(STZ) after high lipids food feeding two months while those in control group were injected normal buffer instead. During the experimental period, variety of body weight, water drinking amount, urine, fasting plasma glucose and femoral artery Doppler in each group were observed and measured.The diabetic foot rats were then used for the development of the ulcer model. by remove a layer of rectangle skin in full thickness. MSCs were isolated from the bone marrow, purified and cultured in vitro. Wistar rats(n=90) were randomly divided into three groups, normal control group (A group) and MSCs treatment group (B group) and diabetic control group(C group). Establish diabetic foot ulcer model and obseve three groups'injury area, nuclear tracer, VEGF expression in peripheral flood and local by ELISA and western-blotting methods and histopathology. Result:1. Compared with control group, experiment group had significant syndrome of diabetic foot, remarkably decreased body weight, increased fasting plasma glucose, drinking water amount and urine. Doppler detection of the experiment group appeared ischemic changes after three weeks STZ injection.2.The woud healing of experiment group(12.51±1.21天)is more difficult compared with control group (9.67±0.87天)(P< 0.01).3. MSCs cultured primarily were oval. short spindle. shaped. long Spindle shaped or polygonal by adhesion separation method. After purification and proliferation, they were uniformly long spindle shaped form. The growth behavior of MSCs from passage 2 to passage 4 was quite stable, exhibiting a large proliferative potential. MSCs were passaged every 9 days.4. The MSCs was found gathered around the fascia layer of ulcer observed by fluorescence method after injecting transplantation 4th and 8th day.5. The average healing area (4d, A 20.16±3.68d, B 23.62±2.26, C 25.50±6.79, p=0.002; 8d, A 3.64±0.92, B 7.56±1.91, C 13.20±1.74, p=0.000) are different. Group B is faster than C but slower than A..6. VEGF on peripheral blood (4d 1.58±0.06,1.53±0.08,1.04±0.05, p=0.000; 81.70±0.04,1.50±0.04,1.14±0.07, p=0.000) and local VEGF (1d, A 1.04±0.05, B 2.44±0.25, C 0.21±0.0.02; 4d, A1.81±0.06, B2.86±0.11, C0.24±0.02; 8, A1.64±0.06, B0.85±0.06, C 0.19±0.02, p=0.000). are have significant statistically difference.7. B group is stronger in MVD standing for CD34 expression,but weaker than A.8.HE staining pathology found that epidermis of B group covered faster than C group but slower than A. Conclusion:1. This rat model of diabetic foot ulcer is easy to establish, well reproducible and area easily controled which provides a stable model.2. Allogeneic bone marrow-derived mesenchymal stem cell transplantation can promote diabetic foot ucler healing.3. The mechanism may be increase the expression of VEGF in systemic especially local.
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