小鼠全脑缺血再灌注损伤模型的研究

    Research on the creation of global cerebral ischemia-reperfusion injury model in mice

    • 摘要: 目的: 探讨结扎双侧颈总动脉、颈部软组织加压人工通气等方法,制作小白鼠脑缺血再灌注损伤动物模型的成功率和可靠性。方法: 结扎双侧颈总动脉,并用粗棉线在颈部血管、神经和气管下方绕颈后结扎加压,以脑电波变平作为全脑完全缺血的标志,造成全脑缺血30 min,然后解除结扎、恢复血液灌流60 min;使用微通气量小动物呼吸机进行人工通气;测大脑皮质乳酸脱氢酶(LDH)、肌酸磷酸激酶(CPK)的活性;观察大脑皮质和海马的形态学变化。结果: 缺血后数秒钟内脑电波变为一条直线,缺血30 min和再灌60 min内未见恢复。LDH和CPK活性,缺血组和再灌组显著低于对照组(P<0.01);再灌组CPK活性明显低于缺血组(P<0.01)。病理检查:缺血30 min,大脑皮质和海马锥体细胞固缩,少数神经元变性,轻度充血、水肿;再灌60 min,上述损伤进一步加重。结论: 本文建立的改进模型方法简便易行,缺血和再灌注损伤显著,模型成功率高,动物死亡率低。

       

      Abstract: Objective: To investigate the success rate and reliability of creating global cerebral ischemia reperfusion injury model by obstructing the bilateral common carolid arteries and pressing the cervix and ventilating the mice in the artificial way and so on.Methods: Artificial respiration was performed using rodent micro-ventilator during the whole experiment course.ECG and EEG were detected carefully.EEG turnning to a straight line was used as the criterion of full cerebral ischemia.Pathological examination was performed in the cerebral cortex and hippocampe.Lactate dehydrogenase (LDH) and creatine phosphokinase(CPK) were determined in the cerebral cortex.Results: EEG turned to a straight line after a few seconds of ischemia and didn't recover within 30 minutes of ishemic and 60 minutes of reperfusion.The LDH and CPK activity of ischemia and reperfusion group was significantly lower than that of control group(P<0.01).CPK activity of reperfusion group was significantly lower than that of ischemia group(P<0.01).Pathologic examination showed that after 30 minutes of ischemia,the cells of cerebral cortex and hippocampe pyramid pyknosed,a small number of neurons degenerated and swelled.The above mentioned damage worsened after 60 minutes of reperfusion.Conclusions: This improved operative method is simple and practical.The global cerebral ischemia-reperfusion injury is obvious in the model,with higher success rate and lower mortality.

       

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