1日龄乳鼠心肌梗死模型制备及标准化评估  

Preparation and standardized assessment of a myocardial infarction model in 1⁃day⁃old suckling mice

作　　者：郝琰琰 布威麦热姆·热杰普 向晨莹 韩国玲 刘维静 刘俊 聂宇[3] 廉虹[3] 王玉瑶 HAO Yanyan;BUWEIMAIREMU·Rejiepu;XIANG Chenying;HAN Guoling;LIU Weijing;LIU Jun;NIE Yu;LIAN Hong;WANG Yuyao(Department of Biochemistry and Molecular Biology,School of Basic Medical Sciences,Shanxi Medical University,Taiyuan 030001,China;Department of Cardiac Surgery,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830011;State Key Laboratory of Cardiovascular Disease,Fuwai Hospital,National Center for Cardiovascular Diseases,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100037)

机构地区：[1]山西医科大学基础医学院,生物化学与分子生物学教研室,太原030001 [2]新疆医科大学第一附属医院,心脏外科乌鲁木齐830011 [3]北京协和医学院,中国医学科学院阜外医院,国家心血管病中心,心血管疾病国家重点实验室,北京100037

出　　处：《中国比较医学杂志》2023年第5期36-43,共8页Chinese Journal of Comparative Medicine

基　　金：国家自然科学基金项目(81900343);山西省重点研发计划项目(201903D321091)。

摘　　要：目的优化新生1 d乳鼠急性心肌梗死手术细节,建立稳定的模型并进行标准化评估。方法出生1日龄(postnatal 1 day,P1)CD1小鼠实施三组手术操作:第一组,冠状动脉左前降支(left anterior descending,LAD)结扎(左心耳缘下方1 mm,宽度为1 mm,结扎深度<0.5 mm)为标准深度结扎组(standard myocardial infarction,SMI);第二组,LAD结扎位置和宽度不变,结扎深度>0.5 mm为深度结扎组(deep MI,DMI);第三组,只开胸不进行LAD结扎组为假手术组(Sham)。通过TTC、伊文思蓝-TTC染色明确LAD结扎是否成功;术后3、7、14、21、28 d通过HE染色和Masson染色评估P1小鼠LAD结扎的心肌组织损伤、纤维化和再生程度;术后28 d通过超声心动图检测小鼠心脏结构和功能变化。结果首先详细描述了P1乳鼠心肌梗死模型构建过程,通过实现LAD暴露、结扎、术后护理等过程提供了一种稳定再生和高存活率的MI程序。术后1 d通过TTC、伊文思蓝-TTC染色确定了SMI模型结扎成功,术后28 d超声心动图发现与假手术组相比,LVEF、LVFS、LVIDd和LVIDs无统计学差异,说明心脏结构和功能基本恢复正常,术后3、7、14、21、28 d Masson染色发现,SMI组心脏组织纤维化面积分别为15.67%、3.34%、2.99%、2.73%、1.11%,说明术后28 d心肌梗死面积几乎完全恢复;DMI组与SMI组相比,小鼠存活率降低了35.71%(SMI为85.71%,DMI为50%),术后28 d超声结果显示,LVEF、LVFS分别降低了(17.25±6.03)%、(11.37±4.06)%,LVIDd、LVIDs分别升高(0.46±0.15)%、(0.69±0.20)%(P<0.05),纤维化面积DMI组为SMI组6倍,说明>0.5 mm的深度结扎,心脏无法实现术后28 d的完全再生修复。结论本研究详细描述了1日龄小鼠标准急性心肌梗死模型建立过程,通过TTC、伊文思蓝-TTC染色、心脏超声,Masson染色方法评估了术后不同时间点心脏梗死面积、心脏结构、功能以及纤维化程度;同时明确>0.5 mm的深度结扎,心脏无法实现术后28 d的完全修复;本研究为�Objective Optimization of the surgical details of acute myocardial infarction in neonatal 1⁃day⁃old mammary mice,and establishment standardized assessment of a stable model.Methods Postnatal day 1(P1)CD1 mice were subjected to three sets of surgical manipulations.Coronary artery left anterior descending(LAD)ligation(1 mm below the left auricular border,1 mm in width,and<0.5 mm ligation depth)was performed in the standard myocardial infarction(SMI)group.The LAD ligation position and width remained unchanged with a>0.5 mm ligation depth in the deep ligation group(Deep MI,DMI).Only the chest was opened without LAD ligation in the sham surgery group(Sham).The success of LAD ligation was verified by TTC and Evans blue⁃TTC staining.The extent of myocardial tissue damage,fibrosis,and regeneration of LAD ligation was assessed by HE and Masson staining at 3,7,14,21 and 28 days after surgery.Structural and functional changes of mouse hearts were assessed by echocardiography at 28 days after surgery.Results It describes the process of establishing a P1 neonatal mouse myocardial infarction model in detail and provides stable regeneration and a high survival rate of MI through realization of LAD exposure,ligation,postoperative care,and other processes.Successful ligation of the SMI model was determined by TTC and Evans blue⁃TTC staining at 1 day postoperatively.Echocardiography at 28 days postoperatively revealed no statistical difference in LVEF,LVFS,LVIDd,or LVIDs compared with the Sham group,indicating that the heart structure and function were restored to normal.Masson staining at 3,7,14,21 and 28 days postoperatively revealed that tissue fibrosis had occurred in 15.67%,3.34%,2.99%,2.73%and 1.11%of the heart area,respectively,in the SMI group,indicating almost complete recovery of the myocardial infarct area at 28 days postoperatively.The survival rate of the DMI group was reduced by 35.71%compared with the SMI group(85.71%for SMI and 50%for DMI).Ultrasound at 28 days postoperatively showed reduction in LVEF(17.25

关 键 词：乳鼠 心肌梗死 冠状动脉左前降支 模型评估 

分 类 号：R-33[医药卫生]