改良的Pulsinelli四血管夹闭法建立大鼠脑缺血模型

刘小艳　严德萍　陈建鸣

【摘要】 目的：通過改良Pulsinelli四血管夹闭法制备大鼠脑缺血模型，为脑卒中的研究提供有效的动物模型。方法：实验选用健康成年雄性Wistar大鼠，随机分为对照组、假手术组和缺血模型组，自由饮食。术前禁食过夜，用水合氯醛麻醉（40 mg/100 g，腹腔注射），将大鼠固定于操作台上。沿第一颈椎内斜向上的神经来寻找翼突孔，用尖端细长的电烙铁电凝双侧翼小孔下方的椎动脉，缝合皮肤。颈部正中切口，分离双侧颈总动脉，穿线并缝合皮肤，夜间禁食。次日，用动脉夹夹闭清醒大鼠的双侧颈总动脉15 min，造成短暂前脑缺血。假手术组中除不电凝椎动脉和不夹闭颈总动脉外，其他操作与模型组相同。对照组不做任何处理。结果：大鼠造模成功43只，成功率为66.2%，模型成功后存活40只，存活率为93.0%。尼氏染色显示，对照组大鼠与假手术组大鼠海马CA1区神经元排列紧密整齐，结构正常，细胞核圆而规则。缺血模型组大鼠海马CA1区正常神经元数量显著减少，大量锥体神经元变性坏死，细胞核不规则并固缩深染，细胞间隙明显增大。结论：改良的Pulsinelli四血管夹闭法可以制备理想的大鼠脑缺血模型，成功率高，适合缺血性脑卒中的实验研究。

【关键词】 脑卒中； 动物模型； Pulsinelli四血管夹闭法； 大鼠

Establishment of Cerebral Ischemia Model in Rats by Modified Pulsinelli Four-vessel Occlusion Method/LIU Xiaoyan，YAN Deping，CHEN Jianming，et al.//Medical Innovation of China，2018，15（14）：029-032

【Abstract】 Objective：To establish a rat model of transient cerebral ischemiaby a modified Pulsinelli four-vessel occlusion method，which provided an effective animal model for the study of stroke.Method：Adult male Wistar rats were randomly divided into control group，sham operation group and ischemic group，free diet.Rats were fasted overnight and anesthetized with chloral hydrate（40 mg/100 g，i.p.），the rats were fixed on the console.Along the first cervical vertebra oblique nerve to find the wing protruding hole，bilateral vertebral artery below the pinhole were electro-coagulatedwith a sharp slender，and sutured the skin.The common carotid arterieswere separated，and rats were fasted at night.The next day，the common carotid artery was clamped for 15 min，resulting in transient forebrain ischemia.In the sham operation group，except the non-occlusion vertebral artery and the common carotid artery，the other operations were the same as the model group. The control group did not do any treatment.Result：Ischemic model were successfully established in 43 rats，with a success rate of 66.2% and 40 surviving after a successful model，the survival rate was 93.0%.Nissl staining showed that neurons in hippocampal CA1 region of rats in control group and sham operation group were arranged closely and neatly， with normal structure and regular nuclei.The number of normal neurons in hippocampal CA1 area of ischemic model group was significantly reduced.A large number of pyramidal neurons degenerated and necrotic，the nucleus was irregular and contracted deeply，and the intercellular space was significantly increased.Conclusion：The modified Pulsinelli four-vessel occlusion method can be used to establish an ideal model of cerebral ischemia in rats with high success rate and suitable for the experimental study of ischemic stroke.

【Key words】 Stroke； Animal model； Pulsinelli four-vessel occlusion； Rat

First-authors address：Shanxi Medical University，Taiyuan 030001，China

doi：10.3969/j.issn.1674-4985.2018.14.007

缺血性脑卒中是由于向脑组织提供血液的血管突然破裂或因血管阻塞导致血液不能流入大脑而引起脑组织损伤的一种疾病，颈内动脉和椎动脉闭塞可引起缺血性脑卒中[1]。脑卒中已成为我国第一位死亡原因，也是中国成年人残疾的首要原因，具有发病率高、死亡率高和致残率高的特点，严重危害人类健康和生活，至今没有很好的治疗方法[2-3]。……