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“锚定结”法小鼠颈部异位心脏移植模型构建及其稳定性测试

Establishment and stability assessment of mouse cervical heterotopic heart transplantation model with“Anchoring Node”
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摘要 目的在传统Cuff法(“袖套”法)血管吻合技术基础上进行改良,优化小鼠颈部异位心脏移植操作步骤、过程,以建立更稳定的小鼠颈部异位心脏移植模型。方法将6~8周龄雄性C57BL/6小鼠(体质量20~24 g)分为实验组(通过“锚定结”法进行小鼠颈部异位心脏移植)和对照组(用传统Cuff法进行小鼠颈部异位心脏移植)。记录各项操作步骤时间、血管外翻次数、血管修剪次数、实验失败原因等。术后每天观察、触摸判断小鼠颈部供心存活情况,存活>48 h为移植成功。术后第7、14天HE染色观察实验组小鼠吻合血管处组织;第7天心肌肌钙蛋白T(cardiac troponin T,cTnT)染色观察2组小鼠供心。结果①实验组共进行25只移植,失败2只,供心存活率92%;对照组共进行25只移植,失败8只,供心存活率68%。实验组动脉平均外翻1.16次,修剪0.04次;静脉平均外翻1.16次,修剪0.04次。对照组动脉平均外翻2.00次,修剪0.21次;静脉平均外翻2.28次,修剪0.46次。②实验组、对照组的颈部心脏移植总时间[(77.22±3.82)vs(87.49±8.01)min]、分离血管+套管时间[(30.06±2.68)vs(36.50±6.67)min]、颈部血管吻合时间[(7.31±1.08)vs(12.34±2.58)min]差异有统计学意义(P<0.05)。③术后第7、14天实验组吻合血管处组织HE染色显示血管通畅。④术后第7天供心cTnT染色情况提示对照组、实验组均无大面积坏死。结论“锚定结”法进行小鼠颈部异位心脏移植保证了单人显微操作的稳定、高效,术后供心存活时间长、吻合血管通畅率高、心功能状态好、术后并发症少。 Objective To optimize the operational steps and processes in mouse cervical heterotopic heart transplantation by modifying the conventional cuff technique for vascular anastomosis and consequently establish a more stable cervical heterotopic heart transplantation model in mice.Methods C57BL/6 male mice(6~8 weeks old,weighing 20~24 g)were categorized into control(conventional cuff technique)and experimental groups(our“Anchoring Node”technique).Time for each surgical step,frequencies of vascular everting and vascular trimming,and the reasons for failure were recorded and compared between the 2 groups.Postoperative survival of heart allograft was determined by daily observation and touching,and the mice with the survival time>48 h were considered as successful model establishment.On the 7th and 14th days after surgery,HE staining was used to observe the pathologic changes in the vascular tissues at anastomosis.The expression of troponin T(cTnT)in the heart on the 7th day was detected with immunofluorescence assay.Results①In the 25 hearts from each group,2 hearts from the experimental group and 8 from the control group failed,and the survival rate of heart allografts was 92%and 68%,respectively.In the experimental group,arterial and venous everting occurred at an average of 1.16 times,with an average frequency of trimming of 0.04 times,while in the control group,arterial and venous everting was 2.00 and 2.28 times,respectively,with an average frequency of trimming of 0.21 and 0.46 times,respectively.②Significant differences were observed in the overall duration for cervical heterotopic heart transplantation(77.22±3.82 vs 87.49±8.01 min),vascular separation plus cannulation(30.06±2.68 vs 36.50±6.67 min),and cervical anastomosis(7.31±1.08 vs 12.34±2.58 min)between the experimental and control groups(all P<0.05).③HE staining displayed that vascular patency was observed in the experimental group on the 7th and 14th days after surgery.④cTnT staining indicated no massive myocardial necrosis was seen in bo
作者 李子炫 方易冰 程伟 LI Zixuan;FANG Yibing;CHENG Wei(Department of Cardiac Surgery,First Affiliated Hospital,Army Medical University(Third Military Medical University),Chongqing,400038,China)
出处 《陆军军医大学学报》 CAS CSCD 北大核心 2024年第8期822-828,共7页 Journal of Army Medical University
关键词 心脏移植 袖套法 小鼠 模型 heart transplantation cuff technique mice model
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