Expression Of VEGF In Rat Model Of Thromboangiitis Obliterans

Thromboangiitis Obliterans (TAO) is the most common vascular disease leading to young people's amputated extremity as well as a common peripheral arterial occlusive disease (PAOD).Thromboangiitis obliterans is defined as a non-arteriosclerosis and inflammatory disease, its lesion affecting small and medium-sized arteries and veins, with occasional thrombophlebitis in distal arterial occlusions. Early in 1908, Leo Buerger recognized thromboangiitis obliterans, a vessel inflammatory disease closely related to young people. But one century later, few are known about the etiopathogenic mechanisms of the vasculitis.With unknown etiological factor and complicated condition, it is very difficult to carry out the treatment of TAO. The surgical treatment, though relieves ischemic symptoms, fails still due to the lack of outflow track.At present, there are reports on the experimentation and clinic reports of curing cure chronic ischemia of low limb by applying VEGF, but reports regarding endogenous expression of VEGF are scarce. The objective of this study is to observe the relation between endogenous expression of VEGF and neovascularization of ischemic tissues, providing experimental and theoretic foundation for of ectogenesis gene therapy.The first step is to build rat model (50 male Wistar rats) of hindlimb ischemia byutilizing lauric acid. Then, divide them into two groups at random: 5 rats as control group while 45 rats as model group. Redivide model group, according to observing time limit, into 9 groups, namely, 12h group, Id group, 2d group, 3d group, lw group, 2w groupx 3w group, 4w group, 5w group. Finally, inject lauric acid Na solution into axifugal femoral artery under microscope.Putting rats to death and collecting specimen at different time: 12h> ld^ 2d, 3d, lw, 2w, 3w, 4w, 5w. Observing the expression of VEGF through immunohistochemistry staining and RT-PCR method; observing neovascularizative factor endothelial cell through immunohistochemistry staining of Von Willebrend, measuring MVD, and deducing ability of neovascularization of ischemic tissues.The rat model of hindlimb ischemia turns out to be successful. Through HE staining, the pathological characteristics of rat model of unilateral hindlimb ischemia is similar to that of TAO patients. Through immunohistochemistry staining and RT-PCR method, expression of VEGF can be detected in 12h group of rat model of unilateral hindlimb ischemia. The expression of VEGF reaches its peak in lw group, and fades away in the following groups. Through immunohistochemistry staining of Von Willebrend Factor, gradual enhance of MVD is detected after 2d group, and reaching its peak in 2w group, then gradually dropping in 3w group, 4w group and 5w group. There is a striking difference between lower limb of model and lower limb of correct in 2d group and the rest groups.Given the state of ischemia of low limb, the expression rule of VEGF is consistent with trend of MVD. This temporal rule of change offers theoreticalfoundation for genie treatment to patients of ischemia of low limb------when theexpression of VEGF begins to fall, the exterior put-in of VEGF should be considered.Our study first puts forward the idea of using microscope during the process of building animal model of unilateral hindlimb ischemia by applying lauric acid, thus improving the probability of building animal model. Our study discovers the expression rule of VEGF in the unilateral hindlimb ischemia of rat model, from the level of protein and molecular biology level. We have detected the expression of VEGF only 12 hours after the rat's hindlimb ischemia. By combining the expressionof VEGF and the measurement of MVD, we can detect the rats' ability to produce collateral circulation after the rat's hindlimb becomes ischemia.