Construction of a rabbit model of abdominal aorta atherosclerosis by a modified balloon injury with a high⁃cholesterol diet
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摘要:
目的：评估改良球囊损伤联合高脂饮食建立兔腹主动脉粥样硬化模型方法的有效性，并检测早期斑块中免疫清除相关蛋白的表达。方法：采用20只新西兰大白兔，随机分为假手术组、经典手术组、改良手术组。假手术组予普通饮食，麻醉、切皮、分离股动脉加缝合，但不行球囊损伤。经典手术组和改良手术组予高脂饮食1周后，行腹主动脉球囊损伤术，其中经典手术组单向拉动球囊3次，改良手术组反复前后拉动球囊30~40次，术后继续高脂饮食4周。术后4周采用血管内超声检查腹主动脉内膜斑块情况，取腹主动脉损伤段行苏木素-伊红（hematoxylin-eosin，HE）染色和免疫组化检测。结果：术后4周，各组体重增加差异无统计学意义。血管内超声显示改良手术组斑块增生显著，管腔狭窄。HE染色显示经典手术组和改良手术组均有不同程度的内膜增生，造模成功率均为 100%。与经典手术组相比，改良手术组最大内膜厚度［（174.69±53.76）μm vs. （481.50±81.94）μm，P < 0.05］、平均内膜厚度［（77.49±18.02）μm vs.（262.63±53.04）μm，P < 0.05］、内膜中膜面积比［0.39±0.14 vs. 1.57±0.30，P < 0.05］和血管狭窄比例［（19.04±5.90）% vs.（52.13±11.31）%，P < 0.05］均显著增加，免疫组化提示改良手术组较经典手术组巨噬细胞浸润更为显著。相比假手术组和经典手术组，改良手术组增生内膜中抗吞噬蛋白CD47表达更为显著。结论：改良球囊损伤联合高脂饮食建立兔腹主动脉粥样硬化模型的方法进一步加速了斑块进展，造模更稳定，可靠性更高，且在早期斑块中即可观察到显著的坏死细胞清除缺陷。
Abstract:
Objective：To construct a rabbit model of abdominal aorta atherosclerosis by a modified balloon injury with a high-cholesterol diet，and evaluate the expression of immune clearance related proteins in early plaques. Methods：A total of 20 New Zealand rabbits were assigned randomly to 3 groups：a sham group，a classical intervention group and a modified intervention group. The high -cholesterol diet and normal diet were given to the intervention groups and the sham group，respectively for one week，after which the model was constructed through the femoral artery. The rabbits in the sham group were given a general diet，anesthetized，skin cut，isolated femoral artery and sutured，but without balloon injury. For the rabbits in the classic intervention group，a balloon was inserted into the abdominal aorta，inflated and unidirectionally pulled 3 times. For the rabbits in the modified intervention group，the balloon was inserted，inflated and a bidirectionally pushed and pulled for 30-40 times. Following the intervention，the two intervention groups received a high-cholesterol diet，while the sham group received a normal diet for another 4 weeks. The intravascular ultrasound was performed to examine the abdominal aorta via the femoral artery，and the injured segments of abdominal aorta were sampled for further hematoxylin-eosin（HE）and immunohistochemistry staining at the end of the study. Results：There was no significant difference in weight gain measured at 4 weeks post intervention among the 3 groups. The intravascular ultrasound demonstrated significant plaque hyperplasia and lumen stenosis in the modified intervention group. HE staining revealed varying degrees of intimal hyperplasia in all samples from both intervention groups. Compared with the classical intervention group，the modified intervention group exhibited significantly increased in maximum intimal thickness［（174.69±53.76）μm vs.（481.50±81.94）μm，P < 0.05］and mean intimal thickness［（77.49±18.02）μm vs.（262.63±53.04）μm，P < 0.05］，a higher intimal /media area ratio［0.39±0.14 vs. 1.57± 0.30，P < 0.05］and severer vascular stenosis［（19.04±5.90）% vs.（52.13±11.31）%，P < 0.05］. The immunohistochemistry staining revealed significantly enhanced macrophage infiltration in the modified intervention group，compared with the classical intervention group. Moreover，a higher expression of the anti-phagocytic protein CD47 in the proliferative intima was detected in the modified intervention group than the sham group and the classical intervention group. Conclusion：The novel rabbit model of abdominal aorta atherosclerosis was established by the modified balloon injury and the high-cholesterol diet，further accelerating early plaque development with an enhanced stability and reliability. A significant necrotic cell clearance impairment was detected in early plaque in this novel model.
