大鼠在体心肌缺血再灌注模型的建立

Establishment of an in Vivo Myocardial Ischemia-Reperfusion Injury Model in Rat

目的探讨在体心肌缺血再灌注模型的建立的新方法。方法 SD大鼠,随机分为3组(A组、B组、C组),每组10只(n=10)。麻醉后连续记录心电图,气管切开呼吸机通气,开胸结扎冠状动脉前降支,各组缺血/再灌注时间为:A组15/120min;B组30/120min;C组30/60min。再灌注结束后经左心房注入0.2%的伊文思蓝2mL。迅速取出心脏,去除非心脏组织,洗净残血,置于-20℃冷冻10min。然后将其切成1.5mm厚的薄片,拍照后浸入1%氯化三苯基四氮唑溶液液中,37℃孵育20min。10%福尔马林固定24h,拍照,通过图像分析软件计算心肌梗死面积。结果 B组与A组心律发生率差异有统计学意义(P=0.020),两组的发生率分别为90%和30%;三组心肌梗死面积分别为13.6±2.5、87.4±7.5、39.7±4.5,差异有统计学意义(P=0.000)。结论缺血30min,再灌注120min是建立心肌缺血再灌注模型的最佳时间。

Objective To investigate a new method of establishing an in vivo myocardial ischemia-reperfusion injury model in rats. Methods SD rats were randomly divided into three groups （A, B, C; n = 10）. Electrocardiograms were recorded continuously after rats were anesthetized. Rats were mechanically ventilated following tracheotomy. Ischemia-reperfusion was achieved by the ligation and deligation of left anterior descending artery. Ischemia and reperfusion duration were 15/120 rain in Group A, 30/120 min in Group B, and 30/60 min in Group C. 2 mL Evans blue at concentration of 0.2% was injected into left atrium after reperfusion. The heart was cut and rinsed with cold normal saline, then put into fridge at -20 %. After 10 min, the heart was sliced into 1.5 mm thickness and photographed, incubated in 1% 2, 3, 5-Triphenyltetrazolium chloride at 37 ℃ for 20 min, fixed in 10% formalin for 24 h, and photographed again. The photographs were analyzed with software to determine the infarction size. Results The incidence of arythmia in Group B and Group A was statistically different （90% and 30% respectively, P = 0.020）; the infarction size between three groups were statistically different （13.6 ± 2.51 87.4 ± 7.5, 39.7 ± 4.5 respectively, P = 0.000）. Conclusions Ischemia for 30 min and reperfusion for 120 min can make the best ischemia-reperfusion injury model.