显微操作条件下小鼠急性心肌梗死模型的建立与评估
|
摘要
目的建立小鼠急性心肌梗死模型,保证建模的稳定性,提高动物存活率,并使用超声心动图、心电图以及伊文氏蓝与TTC双染色等手段对模型的构建进行评估。方法 SPF级C57BL/6雄性小鼠16只,经口气管插管后开胸,显微镜直视条件下结扎冠状动脉左前降支。建模前后各检测一次心电图,术后1 d先用超声心动图检测心功能相关数据,随后经颈外静脉注射1%伊文氏蓝,开胸取心脏切片进行TTC染色。结果小鼠死亡率为6.25%(1/16),经心电图及超声心动图评估建模成功率为93.75%(15/16)。左心室射血分数由术前的63.97%±2.61%下降到23.04%±5.11%;左心室短轴缩短率由术前的34.24%±1.82%下降到10.56%±2.45%;左心室收缩末期内径由(2.39±0.15)mm上升到(3.52±0.18)mm;左心室收缩末期前壁厚度由(1.18±0.08)mm下降到(0.78±0.08)mm。心肌梗死后心电图显示T波高耸,ST段抬高。伊文氏蓝与TTC双染色可见明显心肌梗死区。结论经超声心动图、心电图及伊文氏蓝与TTC双染色评估后,显微操作条件下构建小鼠急性心肌梗死模型稳定可靠。
Objectives To establish a mice model of acute myocardial infarction,to ensure the stability of modeling,to improve the post surgery survival rate of animals and to evaluate the reconstruction of the models by ultrasound,electrocardiogram(ECG)and Evans blue/TTC double staining. Methods Sixteen SPF-class C57BL/6 male mice were thoracotomy and the left anterior descending coronary artery was ligated under microscope conditions. ECG was detected before and after modeling. Cardiac function data were assessed by ultrasound 1 day after surgery. 1% Evans blue was injected through the external jugular vein,and the heart was sliced for TTC staining. Results The mortality rate of mice was 6.25%(1/16). The success rate was 93.75%(15/16)which was confirmed by ECG and ultrasound. Left ventricular ejection fraction(LVEF)decreased from preoperative 63.97%±2.61% to 23.04%±5.11%;left ventricular short axis shortening rate(LVFS)decreased from preoperative 34.24%±1.82% to 10.56%±2.45%;left ventricular endsystolic diameter(LVESD)increased from(2.39±0.15)mm to(3.52±0.18)mm;left ventricular end-systolic anterior wall thickness(LVESAWT)dropped from(1.182±0.08)mm to(0.78±0.08)mm. ECG after myocardial infarction showed that the T-wave was high and peaked,and the ST-segment was elevated. Double staining of Evans blue/TTC showed obvious myocardial infarction. Conclusions Evaluated by the ultrasound,electrocardiogram and Evans blue/TTC double staining,the mice model of acute myocardial infarction was stable and reliable to build up under microscopic operating conditions.
Objectives To establish a mice model of acute myocardial infarction,to ensure the stability of modeling,to improve the post surgery survival rate of animals and to evaluate the reconstruction of the models by ultrasound,electrocardiogram(ECG)and Evans blue/TTC double staining. Methods Sixteen SPF-class C57BL/6 male mice were thoracotomy and the left anterior descending coronary artery was ligated under microscope conditions. ECG was detected before and after modeling. Cardiac function data were assessed by ultrasound 1 day after surgery. 1% Evans blue was injected through the external jugular vein,and the heart was sliced for TTC staining. Results The mortality rate of mice was 6.25%(1/16). The success rate was 93.75%(15/16)which was confirmed by ECG and ultrasound. Left ventricular ejection fraction(LVEF)decreased from preoperative 63.97%±2.61% to 23.04%±5.11%;left ventricular short axis shortening rate(LVFS)decreased from preoperative 34.24%±1.82% to 10.56%±2.45%;left ventricular endsystolic diameter(LVESD)increased from(2.39±0.15)mm to(3.52±0.18)mm;left ventricular end-systolic anterior wall thickness(LVESAWT)dropped from(1.182±0.08)mm to(0.78±0.08)mm. ECG after myocardial infarction showed that the T-wave was high and peaked,and the ST-segment was elevated. Double staining of Evans blue/TTC showed obvious myocardial infarction. Conclusions Evaluated by the ultrasound,electrocardiogram and Evans blue/TTC double staining,the mice model of acute myocardial infarction was stable and reliable to build up under microscopic operating conditions.
引文
[1]SHARKEY S W,MARON B J,KLONER R A.The case for takotsubo cardiomyopathy(syndrome)as a variant of acute myocardial infarction[J].Circulation,2018,138(9):855-857.
[2]LOU L,TANG J,NISHI K,et al.Fabrication of synthetic mesenchymal stem cells for the treatment of acute myocardial infarction in mice[J].Circ Res,2017,120(11):1768-1775.
[3]成玲俐,吴素华,李志梁,等.大鼠心肌缺血再灌注损伤动物模型的建立与评估[J].中国实验诊断学,2010,14(8):1163-1165.
[4]刘建,范慧敏,汪进益,等.小鼠心梗模型的建立与无创评价[J].中国实验动物学报,2010,18(3):196-198.
[5]OH J G,WATANABE S,LEE A,et al.MiR-146a suppresses SUMO1 expression and induces cardiac dysfunction in maladaptive hypertrophy[J].Circ Res,2018,123(6):673-685.
[6]BHUSHAN S,KONDO K,POLHEMUS DJ,et al.Nitrite therapy improves left ventricular function during heart failure via restoration of nitric oxide-mediated cytoprotective signaling[J].Circ Res,2014,114(8):1281-1291.
[7]KANG E J,LEE K N,CHOI W J,et al.Left ventricular functional parameters and geometric patterns in korean adults on coronary ct angiography with a 320-detector-row CT scanner[J].Korean J Radiol.2017,18(4):664-673.
[8]LUGER D,LIPINSKI M J,WESTMAN P C,et al.Intravenously delivered mesenchymal stem cells:Systemic antiinflammatory effects improve left ventricular dysfunction in acute myocardial infarction and ischemic cardiomyopathy[J].Circ Res,2017,120(10):1598-1613.
[9]REBOLL M R,KORF-KLINGEBIEL M,KLEDE S,et al.EMC10(endoplasmic reticulum membrane protein complex subunit 10)is a bone marrow-derived angiogenic growth factor promoting tissue repair after myocardial infarction[J].Circulation,2017,136(19):1809-1823.
[10]DONG S,CHENG Y,YANG J,et al.MicroRNA expression signature and the role of microRNA-21 in the early phase of acute myocardial infarction[J].J Biol Chem,2009,284(43):29514-29525.
[11]CHENG Y,WANG X,YANG J,et al.A translational study of urine miRNAs in acute myocardial infarction[J].J Mol Cell Cardiol,2012,53(5):668-676.
[12]SHAN J,XIE W,BETZENHAUSER M,et al.Calcium leak through ryanodine receptors leads to atrial fibrillation in 3mouse models of catecholaminergic polymorphic ventricular tachycardia[J].Circ Res,2012,111(6):708-717.
[13]KIM H W,VAN ASSCHE L,JENNINGS R B,et al.Relationship of T2-weighted mri myocardial hyperintensity and the ischemic area-at-risk[J].Circ Res,2015,117(3):254-265.
[14]PUYMIRAT E,SIMON T,DANCHIN N,et al.Response by Puymirat et al to letter regarding article,“Acute myocardial infarction changes in patient characteristics,management,and6-month outcomes over a period of 20 years in the FAST-MIprogram(French Registry of Acute ST Elevation or Non-ST-Elevation Myocardial Infarction)1995 to 2015”[J].Circulation,2018,137(21):2307-2308.
[15]YANG F,CHEN Q,HE S,et al.MiR-22 is a novel mediator of vascular smooth muscle cell phenotypic modulation and neointima formation[J].Circulation,2018,137(17):1824-1841.
[16]SALTO-TELLEZ M,YUNG LIM S,EI OAKLEY RM,et al.Myocardial infarction in the C57BL/6J mouse.:a quantifiable and highly reproducible experimental model[J].Cardiovasc Pathol,2004,13(2):91-97.
[17]ZHANG Y,JIAO L,SUN L,et al.LncRNA ZFAS1 as a SERCA2a inhibitor to cause intracellular Ca2+overload and contractile dysfunction in a mouse model of myocardial infarction[J].Circ Res,2018,122(10):1354-1368.
[2]LOU L,TANG J,NISHI K,et al.Fabrication of synthetic mesenchymal stem cells for the treatment of acute myocardial infarction in mice[J].Circ Res,2017,120(11):1768-1775.
[3]成玲俐,吴素华,李志梁,等.大鼠心肌缺血再灌注损伤动物模型的建立与评估[J].中国实验诊断学,2010,14(8):1163-1165.
[4]刘建,范慧敏,汪进益,等.小鼠心梗模型的建立与无创评价[J].中国实验动物学报,2010,18(3):196-198.
[5]OH J G,WATANABE S,LEE A,et al.MiR-146a suppresses SUMO1 expression and induces cardiac dysfunction in maladaptive hypertrophy[J].Circ Res,2018,123(6):673-685.
[6]BHUSHAN S,KONDO K,POLHEMUS DJ,et al.Nitrite therapy improves left ventricular function during heart failure via restoration of nitric oxide-mediated cytoprotective signaling[J].Circ Res,2014,114(8):1281-1291.
[7]KANG E J,LEE K N,CHOI W J,et al.Left ventricular functional parameters and geometric patterns in korean adults on coronary ct angiography with a 320-detector-row CT scanner[J].Korean J Radiol.2017,18(4):664-673.
[8]LUGER D,LIPINSKI M J,WESTMAN P C,et al.Intravenously delivered mesenchymal stem cells:Systemic antiinflammatory effects improve left ventricular dysfunction in acute myocardial infarction and ischemic cardiomyopathy[J].Circ Res,2017,120(10):1598-1613.
[9]REBOLL M R,KORF-KLINGEBIEL M,KLEDE S,et al.EMC10(endoplasmic reticulum membrane protein complex subunit 10)is a bone marrow-derived angiogenic growth factor promoting tissue repair after myocardial infarction[J].Circulation,2017,136(19):1809-1823.
[10]DONG S,CHENG Y,YANG J,et al.MicroRNA expression signature and the role of microRNA-21 in the early phase of acute myocardial infarction[J].J Biol Chem,2009,284(43):29514-29525.
[11]CHENG Y,WANG X,YANG J,et al.A translational study of urine miRNAs in acute myocardial infarction[J].J Mol Cell Cardiol,2012,53(5):668-676.
[12]SHAN J,XIE W,BETZENHAUSER M,et al.Calcium leak through ryanodine receptors leads to atrial fibrillation in 3mouse models of catecholaminergic polymorphic ventricular tachycardia[J].Circ Res,2012,111(6):708-717.
[13]KIM H W,VAN ASSCHE L,JENNINGS R B,et al.Relationship of T2-weighted mri myocardial hyperintensity and the ischemic area-at-risk[J].Circ Res,2015,117(3):254-265.
[14]PUYMIRAT E,SIMON T,DANCHIN N,et al.Response by Puymirat et al to letter regarding article,“Acute myocardial infarction changes in patient characteristics,management,and6-month outcomes over a period of 20 years in the FAST-MIprogram(French Registry of Acute ST Elevation or Non-ST-Elevation Myocardial Infarction)1995 to 2015”[J].Circulation,2018,137(21):2307-2308.
[15]YANG F,CHEN Q,HE S,et al.MiR-22 is a novel mediator of vascular smooth muscle cell phenotypic modulation and neointima formation[J].Circulation,2018,137(17):1824-1841.
[16]SALTO-TELLEZ M,YUNG LIM S,EI OAKLEY RM,et al.Myocardial infarction in the C57BL/6J mouse.:a quantifiable and highly reproducible experimental model[J].Cardiovasc Pathol,2004,13(2):91-97.
[17]ZHANG Y,JIAO L,SUN L,et al.LncRNA ZFAS1 as a SERCA2a inhibitor to cause intracellular Ca2+overload and contractile dysfunction in a mouse model of myocardial infarction[J].Circ Res,2018,122(10):1354-1368.