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Objective: general observation of 30 rabbit hind limb ischemia model, DSA count the number of collateral vessels, microvessel density and microvessel / muscle fiber beam ratios were measured to evaluate the effectiveness of different doses of rh-bFGF treatment of rabbit hindlimb ischemia screening rh -bFGF treatment rabbit hind limb ischemia optimal dose; liver and kidney function tests, angiography, and pathology observed to evaluate the safety of different doses of rh-bFGF treatment rabbit hindlimb ischemia; limb artery treatment for the clinical application of rh-bFGF stenosis or occlusive lesions provide a theoretical basis. Materials and Methods: 30 white rabbits, resection of the left femoral artery and ligation of its main branch limb ischemia model. Randomly divided into five groups (n = 6), A (control group), muscle local injection 5ml Tris buffer /; B, C, D, E group for the treatment group, B group, local injection of rh by muscle -bFGF 2.5μg 5ml Tris buffer / only; group C, muscle localized injection of rh-bFGF 5μg 5mlTris buffer / only; group D, 10 μg 5 ml of Tris buffer / muscle localized injection of rh-bFGF only; group E, by the muscle localized injection of rh-bFGF 20μg 5ml Tris buffer /. Each group 10 days after the modeling side limb vastus medialis muscles 5:00 by injection model were observed after 30 days killed. Each group prior to administration, 10 days after the administration, 20 days after administration of venous blood detection of liver and kidney function; animals in each group were sacrificed DSA observed limb collateral vessels, count the number of collateral vessels; killed animals taken after the vastus medialis muscle produced specimens, capillary density, microvessel / muscle fiber the beam ratio detection and pathological observation. The results: 1. General observation: no deaths in the group of animal experiments. Within 10 days after the making of the model animals left lower limb have varying degrees of lameness. Model-making after 10 days A, B group limp gradually increased; C, D, E group after treatment limp gradually ease. The better surgical wound healing leg ulcers gangrene. A group were killed after take pathological a healing incision abscess formation. Lower extremity arteriography: DSA images show the left femoral artery discontinuity group A small amount of delicate collateral vessels located in the the thigh middle of lack vascular area, B, C group by the deep iliac artery, femoral artery issued a number of support collateral vessels distal arterial see no development, D, E group the ischemic limbs more collateral vessels tortuous course, ischemic limb distal arterial through the development of collateral circulation, the filling velocity compared with the contralateral slightly slow; each group showed no tumor-like DSA and the formation of abnormal blood vessels. D E group, the number of collateral vessels were more than the control group and the B and C group (P <0.05), but the D and E groups the ratio of the number of collateral vessels was not statistically significant (P> 0.05); B, Group C and the number of collateral vessels in the control group and the B and C groups were not statistically significant (P> 0.05). Microvessel density and microvessel / muscle fiber bundle ratio: B, C group and the control group microvessel density and microvessel / muscle fiber bundle ratio difference was statistically significant (P> 0.05): B, C group microvessel density and microvessel / muscle fiber beam ratio D, E group ischemic limb microvascular density and microvascular / muscle ratio of the fiber bundles and B was not statistically significant (P> 0.05); increased D E microvessel density and microvessel / muscle fiber bundles ratio compared with the control group (P <0.05); C group difference was statistically significant (P <0.05); microvessel density and microvessel / muscle fiber bundles in groups D, E ratio was not statistically significant (P> 0.05). Pathology observed: A group of skeletal muscle fiber atrophy, degeneration, muscle fiber spacing increases, the stromal vascular scarce; B, C group skeletal muscle fiber atrophy, muscle fiber spacing increases the few interstitial capillary; D more Group E angiogenesis lumen visible red blood cells, no significant atrophy of muscle fibers; various-group skeletal muscle cells under the influence of rh-bFGF no abnormal proliferation or tumor-like growth performance. Blood liver and kidney function tests: each group before administration, 10 days after the administration 20 days after administration of the venous blood of serum ALT, AST, BUN, Cr detected value comparison of each group at different times of the indicators the difference was not statistically significant (P> 0.05). Conclusion: 1.rh-bFGF promote rabbit ischemic limb angiogenesis, collateral circulation: its efficacy with increasing dose of rh-bFGF enhanced to achieve the best therapeutic effect in 10μg, 20μg effect longer increase, 10μg rh-bFGF optimal dose for the treatment of limb ischemia. Application of experimental doses of rh-bFGF treatment of limb ischemia is safe, had no significant effect on liver and kidney function, will not cause the vascular smooth muscle, abnormal proliferation of skeletal muscle cells and tumor-like growth.
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