The Promoting Angiogenesis And Mechanisms Of Caveolin-1on Limb Ischemia Region Of Diabetic Rats

Objective：After the establishment of the limb ischemia animal model of diabetic rats, caveolin-1plasmid was transfected into ischemic muscular tissue by injection of the vein, toinvestigate the effect of caveolin-1on limb ischemia region of diabetic rats and therelationship with AKT signal pathway by observation the angiogenesis.Method:The healthy、male、SD rats were obtained from the Animal Center of the NanhuaUniversity of South China. To structure the diabetic rat model, it was adoptedintraperitoneal injection of streptozotocin (STZ). To establish animal model of acute lowerextremity ischemia, it was adopted amputation of the left femoral artery and its branches.According to the experimental design, they were randomly divided into3groups (n=10):the sham operation group、the ischemia group and the diabetic group. After14days,ischemic limb gastrocnemius muscle tissues were collected, which were used to measureCaveolin-1expression by Western-blot. The same according to the experimental design,using liposome transfection of Caveolin-1plasmid, they were randomly divided into4groups (n=20): the sham operation group、the ischemia group、the transfected group andthe empty transfected group. After14days, ischemic limb gastrocnemius muscle tissueswere collected from each group of ten, which were used to measure Caveolin-1and AKTexpression by Western-blot. After28days of operation, ischemic limb gastrocnemiusmuscle tissues were collected from remaining rats, which were used to evaluate vasculardensity by CD34immunohistochemical staining and HE staining.Result:After detected the groups expression of caveolin-1by Western-Blot, compared with thesham operation group, the ischemia group and the diabetic group expression of caveolin-1was significantly increased (P <0.05); the diabetic group expression of caveolin-1was increased lower than the ischemia group (P <0.05);compared with the sham operationgroup、the ischemia group and the empty transfected group, the transfected groupexpression of caveolin-1and AKT was significantly increased (P <0.05), which indicatessuccessful transfection. After detected the ratio of micro-vessel/muscle fiber beam and themicro-vessel density (MVD) by HE staining and CD34immunohistochemical staining,compared with the sham operation group, the MVD of the ischemia group and the emptytransfected group was significantly increased, which indicates the significantly difference(P <0.05), but it was lower than the transfected group(P <0.05). This shows thatcaveolin-1would obviously promote angiogenesis in the lower limb ischemic region ofdiabetic rats after over-expression and that it would may be related to the activation ofAKT signal pathway.Conclusion:1. The over-expression of caveolin-1can promoto angiogenesis on limb ischemicregion of diabetic rats.2. The promoting angiogenesis of caveolin-1may be related to the activation of AKTsignal pathway on limb ischemic region of diabetic rats.