Objective To investigate whether or not the endothelial progenitor cells (EPCs) participated in the repair process of aneurysm vascular injury in rabbits.Methods An aneurysm model was induced by elastase in 10 rabbits of clean grade.The rabbits were randomly allocated into in situ carotid EPCs transfusion group and ear vein EPCs transfusion group according to the different methods of transfusion (n = 5 in each group).After EPCs were cultured autologously, they were double-labeled by using Hoechst 33342 and carboxyfluorescein diacetate succinimidyl ester (CFSE).EPCs participating in the process of neointimal repair were analyzed by the methods of fluorescence staining, HE staining, and immunohistochemistry.Results ①The cultured EPCs were double-labeled via Hoechst 33342 and CFSE.1.45 ±0.09 × 106 and 1.47 ± 0.17 × 106 EPCs were transfused autologously in the in situ carotid EPCs transfusion group and the ear vein EPCs transfusion group respectively.②The double-labeled cell direct adhesion was observed both in the thrombi within the aneurysms and the neointimas in the in situ carotid EPCs transfusion group (5/5) , and no endothelial cell growth on the intimal surface was observed.③The double-labeled cells in part of the neointimas within the aneurysm walls rather than the intimal surface could be seen in the ear vein EPCs transfusion group ( 3/5 ).There was no endothelial cell growth.④The distribution of intimal epithelial-like cells of the aneurysm tissue was not observed with the Hematoxylin & Eosin (HE) staining in both groups.Immunohistochemistry showed that the yon Willebrand factor (vWF) staining was negative.Conclusion EPCs participated in the early intimal repair of aneurysm vascular injury, but they did not participate in the repair with the form of transducing to endothelial cells.%目的 探讨内皮祖细胞(EPCs)是否参与兔动脉瘤血管损伤的修复过程.方法 取清洁级兔10只,利用弹性酶诱导动脉瘤模型.按照EPCs回输方式不同,随机分为颈动脉原位回输组和耳缘静脉回输组,每组5只.自体培养EPCs后,用Hoechst 33342和羟基荧光素二醋酸盐琥珀酰亚胺脂(CFSE)对细胞进行双标记.采用荧光染色、HE染色、免疫组化等方法分析EPCs参与新生内膜修复的过程.结果 ①培养的EPCs经Hoechst 33342和CFSE双标记后,颈动脉原位回输组自体回输EPCs(1.45±0.09)×106个,耳缘静脉回输组自体回输EPCs(1.47±0.17)×106个.②颈动脉原位回输组的动脉瘤内血栓中及新生内膜内均可见双标记的细胞直接黏附(5/5),内膜表面未见内皮细胞生长.③静脉回输组的部分动脉瘤壁新生内膜内(非内膜表面)可见到双标记细胞(3/5),未见有内皮细胞生长.④两组动脉瘤组织的HE染色均未见内膜上皮样细胞分布,免疫组织化学均显示内膜Von Willebrand因子(vWF)染色阴性.结论 EPCs参与了动脉瘤损伤血管的早期内膜修复,但未以转化成内皮细胞形式参与修复.
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