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背景:国外小鼠局灶性脑缺血模型技术已经成熟,由于条件限制国内现多以大鼠制作脑缺血模型。目的:建立小鼠局灶性脑缺血再灌注模型,以适应在基因水平进行脑缺血和再灌注损伤分子机制的研究。设计:随机对照实验。单位:承德医学院基础医学研究所。材料:实验于2002-09／2003-05在承德医学院基础研究所完成。选择 18只昆明小鼠2-3月龄,体质量20-25 g,雌雄兼用。分为假手术缎、缺血3 h再灌18 h和24 h组,每组6只。干预:通过神经病学评分,红四氮唑染色及病理形态学的观察,对造模的可靠性进行评价。选用尼龙钓丝(φ0.128mm)经过液体石蜡浸泡后, 从颈外动脉向颈内动脉插入,阻断小鼠右侧大脑中动脉3 h后,拔出线栓再灌注18 h和24 h。主要观察指标:小鼠脑梗死体积及神经学评分。结果:实验共18只小鼠均进入结果分析。①假手术组未出现神经缺损症状。②手术组小鼠神经症状表现为提尾悬空时头向左歪,左前肢内收、肩内旋,左前肢肌张力降低,不能完全伸直,自主活动时,以左后腿为圆心向左旋转,左前肢压在腹下与右前肢交叉成剪刀状,左侧肢体瘫痪更为明显,尤其是前肢,神经症状随再灌注时间延长而加重。③红四氨唑染色可清晰地显示出脑梗死范围,缺血3 h再灌18 h梗死体积为(13+2.3)mm3,缺血3 h再灌注24 h梗死体积为(16+4.5)mm3,并相对稳定,光学显微镜下可见脑缺血后的病理改变。阻断3 h再灌18,24 h 后病理损伤性改变随时间延长加重。结论:插线法制作小鼠脑缺血再灌注模型无需开颅,创伤小,缺血的部位较恒定、可以准确控制缺血和再灌注时间,此模型可做为观察脑血管疾病的病理生理改变、预后及药物治疗作用的理想实验动物模型。
BACKGROUND: The technology of focal cerebral ischemia model in foreign mice has matured. Due to the limitation of conditions, most rats in China are now making cerebral ischemia models. OBJECTIVE: To establish a mouse model of focal cerebral ischemia-reperfusion in order to adapt to the molecular mechanism of cerebral ischemia and reperfusion injury at the genetic level. Design: Randomized controlled experiment. Unit: Institute of Basic Medicine, Chengde Medical College. Materials: The experiment was performed at the Institute of Basic Medicine, Chengde Medical College from September 2002 to May 2003. Select 18 Kunming mice 2 - 3 months old, body weight 20-25 g, both male and female. Divided into sham surgery, ischemia 3 h and then 18 h and 24 h group, 6 in each group. Intervention: The reliability of modeling was evaluated by neurological score, red tetrazolium staining and histopathology. After the nylon line (φ0.128mm) was soaked in liquid paraffin, the carotid artery was inserted from the external carotid artery to the internal carotid artery, and the right middle cerebral artery was blocked for 3 hours. Then the reperfusion was withdrawn for 18 hours and 24 hours. MAIN OUTCOME MEASURES: Volume and neurological score of cerebral infarction in mice. Results: A total of 18 mice were involved in the result analysis. ① There was no neurological deficit in sham operation group. ② The neurological symptoms in the operation group were as follows: the left head left crooked, the left forelimb adduction, the internal rotation of the left shoulder and the left forelimb muscle tension decreased, which could not be completely straightened. When left uninvolved, Turn, left forelimb pressure in the abdomen and right forelimb cross into scissors, left limb paralysis more obvious, especially forelimb, neurological symptoms with the reperfusion time to aggravate. (3) Red tetrazolium staining can clearly show the range of cerebral infarction. The volume of infarction at 18 h after ischemia was (13 + 2.3) mm3 at 3 h and the volume of infarction at 24 h after ischemia at 3 h was (16 + 4.5) mm3 , And relatively stable, visible under optical microscope pathological changes after cerebral ischemia. Blocking 3 h reperfusion 18,24 h after injury pathological changes with the extension of time. CONCLUSION: The model of cerebral ischemia-reperfusion in mice produced by patch-line method does not require craniotomy and trauma. The site of ischemia is relatively constant and the time of ischemia and reperfusion can be accurately controlled. This model can be used to observe the pathophysiology of cerebrovascular disease Change, prognosis and drug treatment of the ideal experimental animal model.