| BackgroundFollow with the develop of industry and traffic, each factor lead to the limbs mutilation, crush injury, avulsed wound, and the situation is very common. In the event if the operation is not in a timely manner or postoperative improper handling often cause limb ischemia reperfusion (LIR) injury. Influence the function of local or systemic organ. If the situation becomes worse, injury limb will be amputated even threat to life. For this disease, oxygen radicals theories, calcium overload theories and lipid peroxidation theories have been put forward, but the precise mechanism is not yet folly understand. So clarify the mechanism of this disease, explore prevention and control measures, reduce the damage, reduce the amputation rate and mortality is particularly important.ObjectiveTo observe the dynamic expression of caveolin-3and skeletal muscle cell apoptosis in a rabbit limb ischemia reperfusion injury model. To investigate the rule and correlation of cell apoptosis and expression of caveolin-3.Methods1. Choose the Chinese white rabbit as the research object. Establish the rabbit model by Shao Heng’s methods. Open the abdomen and find the bifurcation of iliac artery. Then free out the left iliac artery. Establish the ischemia model by blocking the left iliac artery use the sterile vascular clamp. Sew up the incision step by step. Establish the ischemia reperfusion model by releasing the vascular clamp after ischemia5h. Take gastrocnemius muscle for sample when ischemia5h and reperfusion4h and8h. Then start the next experiment.2. Grab15Chinese white rabbits randomly. Establish the ischemia reperfusion model by the the first part method. A total of30lower limb divided into two groups. The right lower limb marked as blank group A which without ischemia reperfusion. The left lower limb marked as experimental group B which ischemia reperfusion. Add the rabbit when it dead. Take two gastrocnemius muscles for sample when ischemia5h and reperfusion4h and8h. One sample fixed with10%formaldehyde24h, decalcified, dehydration, transparent with xylene, paraffin embedding, serial section4μm. Detect the sample with TUNEL POD kit, then observe by fluorescence microscope. Another samples were used to detect caveolin-3by fluorescence quantitative PCR. In the end, we analysed the outcomes of this experiments with SPSS19.0. Vaue of P>0.05were considered statistically significance.Results1. Through the anatomical observation, the left gastrocnemius muscle of the chinese white rabbit were pale when ischemia5h and restore ruddy gradually.2. The change of cell apoptosis:No positive apoptotic cells was observed in the control group. Lots of apoptotic cells were seen in the ischemia group. There is a trend of increase after decreases first for ischemia5hours, reperfusion4hours, reperfusion8hours3. The change of expression of caveolin-3:the expression quantity of Caveolin group are higher than the control group. There is a trend of increase after decreases first for ischemia5hours, reperfusion4hours, reperfusion8hours.Conclusions1. Block the left iliac artery by the sterile vascular clamp can create successful animal model of rabbit limb ischemia reperfusion.2. The change of caveolin-3may be related to apoptosis during limb ischemia reperfusion injury. 3. Caveolin-3participates in regulating the skeletal muscle apoptosis during limb ischemia-reperfusion injury. |
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