大鼠全脑缺血再灌注模型方法的改进及其在热休克蛋白70方面的应用

目的　改进大鼠全脑缺血再灌注模型的方法。方法　采用 42只成年雄性SD大鼠 ,四动脉阻断法制作全脑缺血再灌注模型 ,术中乙醚麻醉 ,41/ 2号注射器针头破坏椎动脉 ,动脉夹夹闭颈总动脉 ,苏木素 伊红 (HE)染色观察海马区病理改变 ,免疫组织化学法检测海马CA1区热休克蛋白 70 (HSP70 )表达 ,苏木素在体染色大体观察脑血供的阻断情况。结果　全脑缺血模型成功率为 77.8% ,四动脉阻断的动物海马区神经元大量坏死 ,HSP70表达 ,苏木素在体染色阴性 ,单纯颈总动脉阻断组和假手术组未见上述表现。结论　改进后的四动脉阻断方法简便 ,效果可靠 ,利于推广。

The modified method of global cerebral ischemia-reperfusion model in rat and the application of HSP70 in the confirmation

Objective To modify the method of global cerebral ischemia-reperfusion model in rats.Methods In 42 male Sprague-Dauley rats,the global cerebral ischemia-reperfusion model was established by the occlusion of the 4 vessels.During the anesthesia,the rats' vertebra arteries were destroyed by pinhead of 4-1/2 injector through the alar foramina of the first cervical vertebra and reversible claps placed loosely around the common carotid arteries.At different time points of reperfusion,the rats were killed and the pathological changes in the hippocampal were assessed by HE staining.The expression of heat-shock protein 70 (HSP70),as a sensitive marker of cerebral injury,in hippocampal CA1 region was detected and hematoxylin solution was perfused through the aorta to investigate the occlusion of the Willi's circle.Results The successful rate was 77.8% in the modified method of global cerebral ischemia-reperfusion model in rats.After 4 vessels were occluded,great many necrotic neurons were observed in hippocampal region. HSP70 staining was positive and Willi's circle staining was negative.The above changes were not observed in single common carotid occluded rats and sham-operation group.Conclusion The modified method of global cerebral ischemia-reperfusion model in rats is easily performed and has sure effects.