ABSTRACT

The endothelialization of endothelial progenitor cells (EPCs) was proven to facilitate the vascular repair of aneurysm. MiR-17-5p regulated angiogenesis in various cancers. This research focused on exploring the effect of miR-17-5p on EPCs and the vascular repair of aneurysm. In vivo study: the aneurysm rat model was established and treated with AgomiR-17-5p; the histopathology of aneurysm tissues was examined by hematoxylin-eosin staining; and the level of EPCs in the aneurysm tissues and peripheral blood of rats were evaluated by immunofluorescence and flow cytometry, respectively. In vitro study: EPCs were cultured and identified using flow cytometry; the target of miR-17-5p was proven by dual-luciferase reporter assay; after transfection, the viability, migration, and tube formation of the EPCs were detected by MTT, wound healing, and tube formation assays, respectively; the expressions of VEGFA and factors related to PTEN-mediated PI3K/AKT pathway were detected by ELISA, qPCR, or Western blot as needed. In vivo study: miR-17-5p overexpression promoted the vascular repair in aneurysm rats and increased the level of EPCs in the aneurysm tissues and peripheral blood of the rats. In vitro study: miR-17-5p overexpression promoted the viability, migration, and tube formation of EPCs, up-regulated the expressions of VEGFA, p-PI3K, and p-AKT, and down-regulated the PTEN expression in EPCs; miR-17-5p silencing did the opposite; PTEN was targeted by miR-17-3p and further abrogated the effects of miR-17-5p overexpression on EPCs. MiR-17-5p promoted the endothelialization of EPCs to facilitate the vascular repair of aneurysm by regulating PTEN-mediated PI3K/AKT/VEGFA pathway.

摘要

内皮祖细胞 (EPC) 的内皮化已被证明有助于动脉瘤的血管修复。MiR-17-5p 调节各种癌症中的血管生成。本研究重点探讨miR-17-5p对EPCs和动脉瘤血管修复的影响。体内研究：动脉瘤大鼠模型的建立和AgomiR-17-5p治疗；通过苏木精-伊红染色检查动脉瘤组织的组织病理学；分别采用免疫荧光和流式细胞术检测大鼠动脉瘤组织和外周血EPCs水平。体外研究：使用流式细胞术培养和鉴定EPC；双荧光素酶报告基因检测证实了miR-17-5p的靶点；转染后，分别通过MTT、伤口愈合和管形成试验检测EPCs的活力、迁移和管形成；根据需要采用ELISA、qPCR或Western blot检测VEGFA及PTEN介导的PI3K/AKT通路相关因子的表达。在体内研究：的miR-17-5p过表达促进了血管修复动脉瘤大鼠和在动脉瘤组织和大鼠外周血增加内皮祖细胞的水平。在体外研究：miR-17-5p过表达促进EPCs的活力、迁移和管形成，上调VEGFA、p-PI3K和p-AKT的表达，下调EPCs中PTEN的表达；miR-17-5p 沉默则相反；PTEN 被 miR-17-3p 靶向并进一步消除了 miR-17-5p 过表达对 EPC 的影响。MiR-17-5p 通过调节 PTEN 介导的 PI3K/AKT/VEGFA 通路促进 EPCs 的内皮化以促进动脉瘤的血管修复。