| Objective:Establishment of the limb ischemia animal model of diabetic rats,Caveolin-1 recombinant adenovirus was injected through the tail vein of rats, to investigate the effect of overexpression of Caveolin-1 on lower limb ischemia in diabetic rats by up regulation of VEGF expression and its mechanism. Methods:80 male SD rats(200 + 20 g), which were established to the model of diabetic rats. Randomly selected 56 diabetic rats were divided into four groups, each group was divided into 14 groups and 21 days groups( 7 rats for each group): normal group(sham group), operation group, operation untransfected group, operation transfection group.The lower limb ischemia model was established by ligation and cutting off the femoral artery in rats. On the first day of operation,the Caveolin-1 gene was injected into the tail vein by adenovirus vector. On the 14 day and 21 day after surgery, the gastrocnemius muscle of the lower limb was taken from the rats, specimens with formaldehyde fixed paraffin buried, hematoxylin eosin staining was used to observe the morphology of muscle cells and the infiltration of inflammatory cells, The expression of Caveolin-1 and vascular endothelial growth factor(VEGF) and the distribution of vascular endothelial growth factor were detected by immunohistochemistry. Analysis of effects of the overexpression of Caveolin-1 in diabetic rats with diabetic lower limb ischemia. Result:14 days after operation in rats:Operated lower extremity Piwimbi the rest of the group of high, good healing of the wound scab, the body does not appear necrosis and only acrocyanosis, normal walking, 21 days after surgery in rats: Acrocyanosis improved freely, toenail normal, thigh palpable arterial pulsation. HE staining: 14 days after operation, the arrangement of the gastrocnemius muscle cells was slightly in order, the morphology of the muscle cells was normal. There was a small amount of blood vessel formation in the muscle fibers, and there was a significant inflammatory cell infiltration, 21 days after operation, the arrangement of the gastrocnemius muscle cells was normal, the distribution of the blood vessels were more, and the inflammatory cells were decreased. The expression of Caveolin-1 and VEGF was detected by immunohistochemical method, the expression of Caveolin-1 in rat gastrocnemius muscle was significantly higher than that in operation group, empty transfection group and sham operation group(P<0.05) after 14 days of transfection, the expression of 21 days transfection group was higher than that of 14 days group; In 14 and 21 days, the VEGF expression in the gastrocnemius muscle of diabetic lower limb was significantly higher than that in operation group and empty transfection group(P<0.05). Microvessel density(MVD) determination: The expression of CD-34 was detected by immunohistochemical method, and the expression of MVD was found to be increased in operation group and empty transfection group(P<0.01), but the expression of CD-34 was significantly higher in the surgical side of the diabetic rats than in the operation group and the empty transfection group(P<0.05), and in the 21 days after transfection, the MVD was basically close to the sham operation group. Conclusion:Over expression of Caveolin-1 can induce the expression of VEGF, promote angiogenesis, improve the condition of ischemia limbs. |
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