超声评价法优化三氯化铁诱导大鼠颈总动脉血栓模型的研究

Optimization of a rat model of ferric chloride-induced carotid arterial thrombosis by ultrasonic continuous evaluation

目的探讨采用连续超声评价法优化不同三氯化铁浓度梯度和作用时间诱导大鼠颈总动脉血栓模型,为溶栓及取栓研究提供合适的在体血栓模型。方法选取20只(Sprague–Dawley)SD大鼠,每组5只,分别采用20%、30%、40%、50%四种三氯化铁浓度外敷大鼠单侧颈总动脉,使用14 L高频超声探头,分别在外敷10min、15 min、20 min三个时间点观察血管二维超声图像及彩色多普勒血流情况。选取血管闭塞最佳组大鼠,于干预120 min后观察血管自发再通率及静脉注射尿激酶后的血管再通率。实验结束后,取出干预血管甲醛固定,石蜡包埋,切片,进行HE染色病理分析。结果三氯化铁外敷大鼠颈总动脉20 min、50%浓度组血管闭塞率100%,40%、30%、20%浓度组外敷20 min血管闭塞率分别20%、0%、0%(P<0.001)。外敷120 min后,50%浓度组组血管自发再通率0%、40%、30%、20%组血管自发再通率100%(P<0.001),50%三氯化铁浓度造模组尾静脉注射尿激酶后血管部分再通率为40%。50%浓度组镜下病理显示血栓结构致密,尿激酶干预后管腔部分再通,血栓结构松散。结论采用50%三氯化铁浓度外敷大鼠颈总动脉20 min,可能是在体血栓模型形成的有效条件,该动物模型可能适合溶栓研究。

Objective The aim of this study was to establish a reasonable protocol for a rat model of ferric chloride-induced carotid arterial thrombosis by ultrasonic continuous evaluation. Methods Twenty SD rats were ran- domly divided into four groups and then treated with 20%, 30%, 40% or 50% concentrations of FeCl3, respectively. The vascular conditions were evaluated by 14L ultrasonic probe, at 10 mins, 15 mins, and 20 rains. We then selected the best group through the examination of the rate of spontaneous reperfusion of blood vessels and the rate of reperfu- sion after intravenous injection of urokinase. At the end of the experiment, vessels were fixed in 10% formalin solution and stained with HE. Results After external application of FeCl3 on rat common carotid artery for 20 mins, the artery occlusion rate was 100%, 20%, 0% and 0% in animals receiving 50%, 40%, 30% and 20% FeCl3, respectively. Af- ter external application of FeCl3 on rat common carotid artery for 120 mins, the spontaneous revascularization rate was 0% in 50% concentration group whereas were 100% in rest other groups （P〈 0.001）. In 50% concentration group, the partial recanalization rate was 40% after intravenous injection of urokinase. HE staining revealed that the thrombuswas dense and the lumen was partially recanalized after the urokinase intervention in 50% concentration group. Con- clusion By use of uuhrasonic continuous evaluation of ferric chloride-induced thrombosis of rat common carotid artery, we have demonstrated that external application of 50% ferric chloride solution for 20 rains is effective for the formation of thrombus model, which may be suitable for the study of thrombolysis.