高糖诱导小鼠脑缺血后出血转化的研究

High glucose-induced hemorrhagic transformation in mice after cerebral ischemia

目的 探讨稳定的高糖诱导小鼠脑缺血后出血转化模型的实验条件。方法 采用线栓法制备小鼠大脑中动脉闭塞模型,将小鼠按照葡萄糖浓度(10%,20%,30%,40%,50%)与缺血时间(30 min, 1、2、3 h)进行组合,每个浓度4个时间点,共分为20组合,每组6只。术前15 min腹腔注射不同浓度葡萄糖,分别在脑缺血术后30 min, 1、2、3 h拔出线栓恢复再灌注,诱导小鼠脑缺血后出血转化模型。以术后24 h内死亡率以及术后7 d内脑出血大体观察,脑皮质和白质出血量,出血发生率及死亡率为指标。结果 大体观察显示,随着缺血时间延长或葡萄糖浓度升高,脑组织出血发生率呈逐渐上升趋势。苏木精-伊红染色镜下观察显示,大部分模型组合小鼠随着缺血时间延长或葡萄糖浓度升高,大脑白质及皮质内出血量呈逐渐上升趋势。术后7 d内,葡萄糖浓度升高或缺血时间延长均增加模型小鼠出血发生率,差异有统计学意义(F=15.282,F=6.632,P<0.01)。术后24 h和术后7 d内,葡萄糖浓度升高或缺血时间延长均显著增加模型小鼠死亡率,差异有统计学意义(P<0.05,P<0.01)。结论 当葡萄糖浓度为30%,缺血时间为30 min时,高糖诱导可制备死亡率低,出血转化发生率高的脑梗死后出血转化模型。

Objective To explore the stable experimental conditions of high glucose-induced hemorrhage transformation model in mice after cerebral ischemia.Methods The mouse model of middle cerebral artery occlusion was established by thread occlusion.Different concentrations of glucose(10%,20%,30%,40%,50%)were injected intraperitoneally 15 min before cerebral ischemia.In 30 min and 1,2 and 3 h after cerebral ischemia,the suture was removed to restore blood reperfusion to induce the model of hemorrhage transformation.Thus,the mice were divided into 20 groups with 6 mice each group.Death within 24 h after operation,gross view of cerebral hemorrhage 7 d after operation,amount of hemorrhage in cerebral cortex and white matter,incidence of hemorrhage and mortality were used to evaluate.Results Gross observation showed that the incidence of cerebral hemorrhage was increased gradually with the duration of ischemia time or increment of glucose concentration.HE staining displayed that the hemorrhage amount in the white matter and cortex was elevated with the prolongation of ischemia time or the increase of glucose concentration in most model mice.Within 7 d after operation,increase of glucose concentration or prolongation of ischemia time increased the incidence of hemorrhage in model mice(F=15.282,F=6.632,P<0.01).Elevated glucose level and prolonged ischemic duration significantly raised the mortality rates in 24 h and 7 d post-operatively(P<0.05,P<0.01).Conclusion Mouse model of hemorrhage transformation can be induced with 30%glucose concentration and 30 min ischemic duration,for higher incidence of transformation and low mortality.