横向主动脉缩窄术致兔心力衰竭模型的建立

doi:10.12300/j.issn.1674-5817.2022.020

实验动物与比较医学 ›› 2022, Vol. 42 ›› Issue (5): 401-408.DOI: 10.12300/j.issn.1674-5817.2022.020

横向主动脉缩窄术致兔心力衰竭模型的建立

罗庆祎¹^,²^,³(), 章体玲²^,³^,⁴, 丁云川¹^,²^,³, 陈剑¹^,²^,³, 赵丽¹^,²^,³, 王庆慧¹^,²^,³()()

^1.昆明市延安医院超声医学科, 昆明 650200
^2.云南省心血管疾病重点实验室, 昆明 650200
^3.云南省心脏疾病临床医学中心, 昆明 650200
^4.昆明市延安医院心内科, 昆明 650200

收稿日期:2022-02-24 修回日期:2022-08-04 出版日期:2022-10-25 发布日期:2022-11-04
通讯作者: 王庆慧（1977—），女，主任医师，研究方向：心血管疾病超声诊断。E-mail： wqh962099@163.com。ORCID：0000-0001-6360-9508
作者简介:罗庆祎（1981—），女，副主任医师，博士，研究方向：心血管疾病超声诊断及基础研究。E-mail：1481215034@qq.com
基金资助:
云南省科技厅科技计划项目“围术期多模态超声心动图在冠状动脉旁路移植术中的应用研究”(2019FE001-266)

Establishment of a Rabbit Heart-failure Model Using Transverse Aortic Constriction

Qingyi LUO¹^,²^,³(), Tiling ZHANG²^,³^,⁴, Yunchuan DING¹^,²^,³, Jian CHEN¹^,²^,³, Li ZHAO¹^,²^,³, Qinghui WANG¹^,²^,³()()

^1.Department of Ultrasound Medicine, Yan'an Hospital Affiliated to Kunming Medical University, Kunming 650200, China
^2.Key Laboratory of Cardiovascular Diseases of Yunnan Province, Kunming 650200, China
^3.Medical Center of Heart Disease of Yunnan Province, Kunming 650200, China
^4.Department of Cardiology Medicine, Yan'an Hospital Affiliated to Kunming Medical University, Kunming 650200, China

Received:2022-02-24 Revised:2022-08-04 Published:2022-10-25 Online:2022-11-04
Contact: WANG Qinghui (ORCID: 0000-0001-6360-9508), E-mail: wqh962099@163.com

摘要/Abstract

摘要：

目的探讨应用横向主动脉缩窄（transverse aortic constriction，TAC）术建立兔心力衰竭模型的方法及可行性。方法 25只普通级健康雄性日本大耳兔，随机分为手术组（n=15）和假手术组（n=10）。手术组动物行TAC术，建立兔心力衰竭模型；假手术组行开胸手术但不进行缩窄操作。术后4周和8周超声检查心脏功能，术后8周检测降主动脉流速；结合分子生物学指标及病理学观察，评估TAC术建立兔心力衰竭模型的可行性和稳定性。结果 TAC术后8周，手术组兔存活13只，假手术组兔存活10只，手术组主动脉弓缩窄处血流速度均大于2.20 m/s。与假手术组比较，手术组心率、升主动脉内径、左房舒张末期内径、左室舒张末期内径和左室收缩末期内径均明显增加（P＜0.05），左室射血分数明显降低（P＜0.05）；与假手术组相比，手术组左心室组织中NT-proBNP和β-MHC含量明显升高（P＜0.05）。病理切片显示，TAC术后8周，手术组心肌细胞肥大且排列紊乱，心肌纤维束增宽。TAC术后4周，与假手术组相比，手术组左室舒张末期室间隔厚度和左室后壁舒张末期厚度均显著增加（P＜0.05）。结论应用TAC术建立实验兔心力衰竭模型是有效且可行的。心脏超声是评估主动脉弓缩窄程度及心力衰竭建模成功与否的有效手段。手术组实验兔在TAC术后8周时心肌出现失代偿反应，形成不可逆的心力衰竭。

关键词: 超声, 横向主动脉缩窄术, 心力衰竭, 兔

Abstract:

Objective To investigate the method and feasibility of establishing a rabbit heart-failure model using transverse aortic constriction (TAC). Methods Twenty-five healthy male Japanese white rabbits were randomly divided into an operation group (n=15) and a sham-operation group (n=10). The animals in the operation group underwent TAC to establish the rabbit model of heart failure. The sham-operation group underwent thoracotomy but no narrowing operation. Cardiac function was evaluated using ultrasound at 4 and 8 weeks after operation. Eight weeks after operation, the descending aortic flow velocity was detected using cardiac ultrasound, combined with molecular biological indexes and pathological observations to verify the feasibility and stability of TAC in establishing the rabbit heart-failure model. Results Eight weeks after operation, 13 rabbits in the operation group and 10 in the sham-operation group survived. The blood flow velocities at the constriction in the operation group were >2.20 m/s. Compared with the sham-operation group, the following outcomes in the operation group increased significantly (P<0.05): heart rate, ascending aortic diameter, left atrial end-diastolic diameter, left ventricular end-diastolic diameter, and left ventricular end-systolic diameter. Left ventricular ejection fraction decreased significantly (P<0.05). Compared with the sham-operation group, the contents of left ventricular N-terminal pro B type natriuretic peptide (NT-proBNP) and β-myosin heavy chain (β-MHC) significantly increased in the operation group (P<0.05). Pathological sections showed that 8 weeks after TAC, cardiomyocytes in the operation group were hypertrophic and disordered, and myocardial fiber bundles widened. Interventricular septal end-diastolic thickness and left ventricular posterior wall end-diastolic thickness significantly increased in the operation group (P<0.05), compared to those in the sham-operation group at 4 weeks. Conclusion It is effective and feasible to establish an experimental rabbit heart-failure model using TAC. Cardiac ultrasound is an effective method to evaluate and determine the degree of aortic arch constriction and the success of heart-failure modeling. Rabbits in the operation group showed myocardial decompensation at 8 weeks after TAC, resulting in irreversible heart failure.

Key words: Ultrasound, Transverse aortic constriction, Heart failure, Rabbits

中图分类号:

Q95-33

罗庆祎, 章体玲, 丁云川, 陈剑, 赵丽, 王庆慧. 横向主动脉缩窄术致兔心力衰竭模型的建立[J]. 实验动物与比较医学, 2022, 42(5): 401-408.

Qingyi LUO, Tiling ZHANG, Yunchuan DING, Jian CHEN, Li ZHAO, Qinghui WANG. Establishment of a Rabbit Heart-failure Model Using Transverse Aortic Constriction[J]. Laboratory Animal and Comparative Medicine, 2022, 42(5): 401-408.

图/表 6

图1 实验兔横向主动脉缩窄术结扎部位图注：A，TAC术主动脉缩窄部位示意图，黑色线圈所示为结扎部位。B，TAC术中操作图，箭头所示为结扎部位。

Figure 1 Sites of surgical ligation of the transverse aortic constriction (TAC) in rabbitsNote：A, Schematic diagram of the site of coarctation of the aorta in TAC; the black circle indicates the ligation site. B, Intraoperative opration diagram of TAC; the arrow shows the ligation site.

图2 多普勒超声检测实验兔降主动脉流速注：A，手术组兔术后8周时，于胸骨上窝切面测量降主动脉缩窄处流速为2.25 m/s（n=13）。B，假手术组兔术后8周时，于胸骨上窝切面测量降主动脉流速为0.22 m/s（n=10）。C，两组实验兔降主动脉流速对比柱形图，*P＜0.05。

Figure 2 Flow velocity of the descending aorta in rabbitsNote： A, The flow velocity at the coarctation of the descending aorta measured in the suprasternal fossa was 2.25 m/s at 8 weeks after transverse aortic constriction operation (n=13); B, The velocity of the descending aorta measured in the suprasternal fossa section was 0.22 m/s (n=10) at 8 weeks after operation in rabbits in the sham-operated group; C, Bar graph comparing the descending aortic flow velocity between the two groups, *P<0.05.

表 1 术后4周及8周实验兔心脏彩超指标变化 (xˉ± s, n=5)

Table 1 Changes in cardiac ultrasound parameters at 4 and 8 weeks after operation

项目 Items	假手术组 Sham-operation group		TAC手术组 TAC operation group
项目 Items	4周 4 weeks	8周 8 weeks	4周 4 weeks		8周 8 weeks
心率HR/(次·min^-1)	218.00±18.00	225.00±15.00	251.00±15.00	300.00±20.00^*
升主动脉内径AAOd/mm	4.31±0.74	4.52±0.91	4.72±0.77	7.57±0.88^*
左房舒张末期内径LADd/mm	3.56±0.84	3.75±0.55	3.84±0.92	4.94±0.42^*
左室舒张末期内径LVEDd/mm	11.04±1.70	12.15±1.51	12.55±1.84	15.45±1.66^*
左室收缩末期内径LVEDs/mm	4.64±0.75	4.89±0.68	5.14±0.92	8.83±0.72^*
左室舒张末期室间隔厚度IVSd/mm	1.73±0.34	1.87±0.26	2.95±0.41^*	1.51±0.32
左室后壁舒张末期厚度LVPWd/mm	1.55±0.32	1.76±0.24	2.65±0.56^*	1.23±0.41
左室射血分数LVEF/%	57.33±7.24	60.16±11.97	58.31±8.74	40.29±7.62^*

图3 心脏彩超显示各组实验兔心尖四腔心切面图注：A，假手术组术后4周时心尖四腔切面图；B，假手术组术后8周时心尖四腔切面图；C，手术组4周时心尖四腔切面图；D，手术组8周时心尖四腔切面图。

Figure 3 Doppler echocardiography shows the apical four-chamber view of each groupNote： A, Apical four-chamber view of the sham-operation group 4 weeks after operation; B, Apical four-chamber view of the sham-operation group 8 weeks after operation; C, Apical four-chamber view of the operation group 4 weeks after transverse aortic constriction operation; D, Apical four-chamber view of the operation group 8 weeks after transverse aortic constriction operation.

图4 横向主动脉缩窄（TAC）术后8周实时荧光定量PCR测定实验兔左心室心衰标志物水平变化注：A为术后8周NT-proBNP含量对比；B为术后8周β-MHC含量对比。与假手术组相比，*P＜0.05；n=3。

Figure 4 Changes in the contents of left ventricular heart failure markets 8 weeks after TAC detected by real-time quantitative PCRNote：A, NT-proBNP content at 8 weeks after operation; B， β-MHC content at 8 weeks after operation. Compared with the sham-operation，*P<0.05； n=3.

图5 手术组及假手术组实验兔心肌组织的HE染色与Masson染色注：A，假手术组和横向主动脉缩窄（TAC）手术组术后8周的HE染色切片，上下图分别放大20倍和40倍；B，假手术组和手术组术后8周的Masson染色切片，上下图分别放大40倍和100倍。

Figure 5 HE staining and Masson staining of experimental rabbit myocardial tissues in the operation and sham-operation groupsNote： A, HE stained sections of the sham-operation group and the transverse aortic constriction (TAC) operation group at 8 weeks after operation (upper, ×20; lower, ×40); B, Masson’s trichrome stained sections of the sham-operation group and the TAC operation group at 8 weeks after operation (upper, ×40; lower, ×100).

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横向主动脉缩窄术致兔心力衰竭模型的建立

Establishment of a Rabbit Heart-failure Model Using Transverse Aortic Constriction

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