一种快速建立小鼠心肌缺血再灌注模型的方法  

A method for rapidly establishing myocardial ischemia-reperfusion model in mice

作　　者：臧光耀 李波[1] 耿田欣 严金川[1] ZANG Guang-yao;LI Bo;GENG Tian-xin;YAN Jin-chuan(Department of Cardiology, Affiliated Hospital of Jiangsu University, Zhenjiang Jiangsu 212001, China)

机构地区：[1]江苏大学附属医院心内科

出　　处：《江苏大学学报（医学版）》2019年第6期472-475,共4页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(81670405);江苏省自然科学基金资助项目(BK20161355);镇江市2018年度科技创新资金(SS2018008);镇江市社会发展项目(SH2017030)

摘　　要：目的:探讨一种快速高效且不需要借助呼吸机和开胸操作的方法,建立小鼠心肌缺血再灌注模型。方法:取40只雄性昆明小鼠(6~8周龄,体重18~25 g),随机分为假手术组、缺血15 min+再灌注24 h组、缺血30 min+再灌注24 h组和缺血45 min+再灌注24 h组,每组10只。给予2%异氟烷气体麻醉,分离胸大肌和胸小肌,于左侧第3、4肋间隙挤出心脏,迅速定位,假手术组仅穿线不结扎,手术组分别结扎冠状动脉左前降支(LAD)15、30和45 min后,松开结扎线24 h以建立心肌缺血再灌注模型。同时在结扎前、结扎后5 min及再灌注24 h后进行心电图检测。再灌注24 h后在结扎部位再次进行结扎,进行伊文思蓝-TTC染色,垂直于心脏长轴切片,检测心肌缺血区和梗死区面积。结果:至再灌注24 h后,小鼠生存率为90%,心肌缺血再灌注造模成功率86. 7%。心电图检测显示LAD结扎5 min后均出现T波高耸,ST段抬高,QRS波增宽。随着再灌注时间延长,ST段逐渐回落。结扎LAD 5 min后,肉眼可见结扎线以下心室壁呈现灰白色,再灌注24 h后可见与周围组织区分明显的白色梗死区,心脏搏动频率和幅度较结扎前明显减弱。心肌组织切片经伊文思蓝-TTC染色呈现界限明显的缺血区、梗死区和正常心肌组织区。缺血45 min+再灌注24 h组小鼠(切片缺血区面积+梗死区面积)/左室总面积比值为(63. 8±6. 36)%,梗死区面积/(梗死区+缺血区面积)比值为(46. 7±5. 77)%。结论:该方法建立小鼠心肌缺血再灌注模型,减少了呼吸机使用所需要的时间,操作快速便捷,造模成功率大于85%。Objective:We aim to develop a rapid and efficient method of myocardial ischemia reperfusion(MIR)without the requirement of ventilation and chest opening in mice.Methods:Forty male Kunming mice(6-8 weeks old and weighing 18-25 g)were randomly divided into sham operation group,ischemia 15 min+reperfusion 24 h group,ischemia 30 min+reperfusion 24 h group and ischemia 45 min+reperfusion 24 h group(10 mice in each group).Mice were anesthetized with 2%isoflurane gas,the heart were fully exposed through a small incision of the forth intercostal space on the left side.After rapidly locating and ligating the left anterior descending coronary artery(LAD)for 15,30 and 45 minutes,the myocardial ischemia-reperfusion model was established by loosening the ligature for 24 hours.At the same time,electrocardiogram was performed before,5 min and 24 h after surgery.The ligation site was re-ligated 24 h after surgery,and Evan′s Blue-TTC double staining was performed to detect myocardial ischemic area and infarct area.Results:By 24 hours after surgery,the survival rate of mice was 90%,and the success rate of MIR modeling was 86.7%.Electrocardiogram showed that the ST segment and the T wave was elevated after LAD ligation,and the QRS wave was broadened,followed by the gradually decrease of the ST segment.After ligation of LAD,the ventricular wall below the ligature line showed grayish white,and the pale infarct area was observed 24 h after operation.The frequency and amplitude of heart beat were significantly weaker than before ligation.Cardiac sections were stained with Evan′s Blued-TTC to present a clearly defined ischemic,infarcted,and normal myocardial tissue.In ischemia 45 min+reperfusion 24 h group,the ratio of the ischemia area+infarction area to the total myocardial area of the left ventricle was(63.8±6.36)%and the ratio of the area of the infarct area to the area of the ischemic area+infarct area was(46.7±5.77)%.Conclusion:This new rapid and convenient method of I/R in mice may be a more efficient model of MIR.It reduce

关 键 词：小鼠 冠脉结扎 缺血再灌注 动物模型 

分 类 号：R-332[医药卫生] R542.2