Papain combined with'Left renal artery stenosis'for establishing an abdominal aortic aneurysm model
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目的 建立一种更加符合腹主动脉瘤(AAA)演变过程的动脉瘤模型.方法 选择体重为2 kg的16只新西兰兔,对其进行编号,并采取随机数字表法将16只兔分为实验组和对照组,每组各8只.实验组采用"左侧肾动脉狭窄"联合木瓜蛋白酶局部湿敷的方法建模,对照组采用局部生理盐水湿敷.术后分别在第2周和第4周对两组新西兰兔进行彩色多普勒超声检查,测量湿敷段主动脉直径的变化.处死新西兰兔后,对第2周和第4周的主动脉标本进行苏木精-伊红(HE)、Verhoeff's Van Gieson(EVG)染色、Masson、转位蛋白(TSPO)和波形蛋白(Vimentin)染色,以及TSPO、Vimentin、α-平滑肌动蛋白(α-SMA)/成纤维细胞活化蛋白(FAP)的免疫荧光染色.利用SPSS和Image J软件对所得数据进行统计学分析.计数资料以频数(％)表示,计量资料以均数±标准差表示,以P＜0.05为差异有统计学意义.结果 超声检查显示,实验组的腹主动脉直径随造模进程持续增加,与对照组比较,差异有统计学意义[(0.54±0.05)cm比(0.28±0.02)cm,Z=-2.6,P＜0.01].此外,第4周实验组的动脉直径明显高于第2周实验组[(0.54±0.05)cm比(0.41± 0.03)cm,Z=-2.9,P＜0.01].免疫组织化学结果显示,实验组的动脉瘤壁比较对照组,具有弹力纤维断裂、中膜增厚及外膜胶原纤维沉积等表型.第4周实验组的动脉壁与第2周实验组比较,病变程度加重,相反动脉壁活化巨噬细胞浸润程度下降,且FAP+/α-SMA+的肌成纤维细胞表达明显增高,反映了在动脉瘤进展过程中血管平滑肌细胞的表型转化现象.结论 采用"左侧肾动脉狭窄"联合木瓜蛋白酶局部湿敷的方法可以成功构建更符合人AAA演变的模型.
Objective To establish an aneurysm model that closely mimics the progression of ab-dominal aortic aneurysms(AAA).Methods Totally,16 New Zealand rabbits weighing 2 kg.Using the random number table method,the 16 rabbits were divided into experimental and control groups,with 8 rab-bits in each group.The experimental group underwent modeling using two-kidney,one clip combined with local application of papain,while the control group received local application of physiological saline.Color Doppler ultrasound was performed on both groups of rabbits at the 2nd and 4th weeks post-surgery to meas-ure changes in the diameter of the treated aortic arterial segment.After euthanasia of the rabbits,arterial specimens obtained at the 2nd and 4th weeks were subjected to hematoxylin-eosin(HE),Verhoeff's Van Gieson(EVG),Masson's trichrome,translocator protein(TSPO),and Vimentin staining.Additionally,fluorescence staining was performed for TSPO,Vimentin,and α-smooth muscle actin(α-SMA)/fibroblast activation protein(FAP).SPSS and Image J software was used for statistical analysis of the collected data.Categorical variables were presented as ratios(％)and continuous variables were presented by mean± standard deviation.P＜0.05 was considered to be statistically significant difference.Results Color Doppler ultrasound revealed continuous expansion of the abdominal aortic diameter in the experimental group,and statistically significant differences were observed compared to the control group at 4th week(0.54±0.05 cm vs.0.28±0.02 cm,Z=-2.6,P＜0.01).Moreover,the arterial diameter at 4th week in the experimental group further increased compared to that at the 2nd week[(0.54±0.05)cm vs.(0.41±0.03)cm,Z=-2.9,P＜0.01].Pathological findings showed that the aneurysm wall of the ex-perimental group exhibited characteristics such as elastic fiber rupture,medial thickening,and deposition of outer membrane collagen fiber,distinct from the arterial wall of the control group.The degree of arterial wall pathology was more severe in the 4th week experimental group than the 2nd week experimental group.Additionally,infiltration of activated macrophages in the arterial wall decreased,and the expression of FAP+/α-SMA+ myofibroblasts significantly increased in the 4th week experimental group compared to the 2nd week experimental group,reflecting the phenotypic transition of vascular smooth muscle cells(SMC)during AAA progression.Conclusion The approach of"two-kidney,one clip"combined with local appli-cation of papain successfully induced AAA in rabbits,which can more accurately mimics the progression process of human AAA.
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