摘要
目的改良小鼠心肌缺血再灌注损伤(MIRI)造模方法,并对其系统性评价。方法将70只SPF级雄性C57BL/6小鼠随机分为假手术组(Sham)、经典组(CM)、单圈活结组(SM)和改良活结组(MM)。通过结扎冠状动脉左前降支30 min后再灌注24 h建立MIRI模型,记录并统计各组小鼠建模时间、模型存活率及实时心电图的变化。采用2,3,5-氯化三苯基四氮唑(TTC)、HE、Masson和TUNEL染色,比较MIRI心肌组织病理改变。生化仪测定血清谷草转氨酶(AST)、肌酸激酶(CK)、肌酸激酶同工酶MB(CK-MB)和乳酸脱氢酶(LDH)水平。ELISA法分析肿瘤坏死因子α(TNF-α)、白细胞介素2(IL-2)、白细胞介素6(IL-6)和单核细胞趋化因子1(MCP-1)水平。结果MM组小鼠心电图ST段明显抬高;MM组建模时间(36.2 min)显著少于CM组(66.2 min,P<0.0001),且MM组相较于CM组存活率显著提升(P<0.05)。MM组小鼠心肌组织梗死区细胞肿胀变形,心肌纤维化程度高于其他组并明显发生凋亡。CM组和MM组小鼠心肌酶AST、CK、CK-MB、LDH水平较Sham组均显著升高,其中MM组的CK和CK-MB水平较CM组明显增加(P<0.05)。CM组和MM组小鼠炎性因子TNF-α(P<0.01)、IL-6(P<0.05)较Sham组显著升高,CM组IL-2水平显著高于Sham组和MM组(P<0.05),MCP-1水平各组间差异无统计学意义。结论与经典和单圈活结造模方式相比,利用改良活结法建立的小鼠MIRI模型,造模时间缩短,模型更稳定,小鼠存活率显著提高。
Objective To improve and systematically evaluate the modeling methods of myocardial ischemia and reperfusion injury(MIRI)in mice.Methods 70 SPF-level male C57BL/6 mice were randomly divided into sham operation group(Sham),conventional modeling group(CM),single-loop slip knot group(SM),and modified slip knot group(MM).The MIRI model was established by ligating the left anterior descending branch of the coronary artery(LAD)for 30 min followed by reperfusion for 24 h.The modeling time,model survival rate,and changes in real-time electrocardiogram were recorded and analyzed for each group.The histopathological changes in MIRI myocardial tissue were compared using 2,3,5-triphenyltetrazolium chloride(TTC),HE,Masson,and TUNEL staining methods.The levels of myocardial enzyme spectrum glutamic-pyruvic transaminase(AST),creatine kinase(CK),creatine kinase isoenzyme MB(CK-MB),and lactate dehydrogenase(LDH)were analyzed by biochemical analyzer,and the levels of inflammatory factors tumor necrosis factor-alpha(TNF-α),interleukin-2(IL-2),interleukin-6(IL-6),and monocyte chemotactic protein-1(MCP-1)were detected by ELISA.Results The ST segment of the electrocardiogram in the MM group mice was significantly elevated.The modeling time in the MM group(36.2 min)was significantly shorter than that in the CM group(66.2 min,P<0.0001),and the survival rate in the MM group was significantly increased(P<0.05).The pathological examination of myocardial tissue showed that the myocardial infarction area in the MM group mice had significantly more cell swelling and deformation,and the degree of myocardial fibrosis was significantly higher than that in the other groups,with significant apoptosis.The serum AST,CK,CK-MB,and LDH levels in the CM and MM groups were significantly higher than those in the Sham group,and the serum CK and CK-MB levels in the MM group were significantly higher than those in the CM group(P<0.05).TNF-α(P<0.01)and IL-6(P<0.05)were significantly higher in the CM and MM groups than in the Sham group,and IL-2 was signific
作者
宋远
王一卉
高瑭
董念国
谢明星
Song Yuan;Wang Yihui;Gao Tang;Dong Nianguo;Xie Mingxing(Department of Ultrasound Medicine,Union Hospital,Huazhong University of Science and Technology,Wuhan 430022,China;Clinical Research Center for Medical Imaging in Hubei Province,Wuhan 430022,China;Hubei Province Key Laboratory of Molecular Imaging,Wuhan 430022,China;Department of Cardiovascular Surgery,Union Hospital,Huazhong University of Science and Technology,Wuhan 430022,China)
出处
《中华胸心血管外科杂志》
CSCD
2024年第4期214-221,共8页
Chinese Journal of Thoracic and Cardiovascular Surgery
基金
国家自然科学基金资助(82230066,82202234)
中央高校基本科研业务费专项资金资助(YCJJ20230212)。
关键词
心肌缺血再灌注损伤
结扎
动物模型
小鼠
Myocardial ischemia reperfusion injury
Ligation
Animal model
Mice
