Li Yang, Li Xue, Han Shilong, Lian Weishuai, Cheng Jie, Xie Xiaoyun, Li Maoquan
Department of Interventional and Vascular surgery, Shanghai Tenth People's Hospital, Tongji University, Shanghai 200072, China.
Institute of Intervention Radiology and Vascular Surgery, Tongji University, Shanghai 200072, China.
J Interv Med. 2019 Apr 30;1(1):9-14. doi: 10.19779/j.cnki.2096-3602.2018.01.04. eCollection 2018 Feb.
To investigate the effects of exogenous basic fibroblast growth factor -2 (FGF-2) on the biological activity of endothelial progenitor cells (EPCs) exposed to high glucose conditions. 1) Bone marrow EPCs from C57BL/6 mice were isolated and cultured in vitro. EPC purity was identified by flow cytometry and immunofluorescence staining. 2) Apoptosis was detected by TUNEL assay. Migration and tube formation ability was detected by Transwell chamber and Matrigel assays, respectively. The expression and activation of β-catenin was detected by Western blot. 3) Doppler flowmetry was used to detect the effect of FGF2 on blood flow recovery in ischemic hind limbs of mice. 1) FGF-2 treatment reversed high glucose induced growth inhibition of EPCs. FGF-2 treatment also increased migration and tube formation ability of EPCs even in high glucose conditions. 2) Western blot analysis demonstrated that the percentage of activated β-catenin/total β-catenin in the high glucose group were significantly lower than that in the control group, while FGF-2 treatment reversed high glucose induced β-catenin inhibition. 3) In vivo experiments demonstrated that the blood flow recovery in ischemic hind limbs of mice was significantly improved after FGF-2 treatment. Exogenous FGF-2 could play a role in the functional repair of damaged EPC exposed to high glucose conditions, via the activation of the Wnt/β-catenin signaling pathway.
为研究外源性碱性成纤维细胞生长因子-2(FGF-2)对处于高糖环境下的内皮祖细胞(EPCs)生物学活性的影响。1)从C57BL/6小鼠中分离骨髓EPCs并进行体外培养。通过流式细胞术和免疫荧光染色鉴定EPC纯度。2)采用TUNEL法检测细胞凋亡。分别通过Transwell小室和基质胶实验检测细胞迁移和管腔形成能力。通过蛋白质免疫印迹法检测β-连环蛋白的表达和激活情况。3)使用多普勒血流仪检测FGF2对小鼠缺血后肢血流恢复的影响。1)FGF-2处理可逆转高糖诱导的EPCs生长抑制。即使在高糖条件下,FGF-2处理也能增强EPCs的迁移和管腔形成能力。2)蛋白质免疫印迹分析表明,高糖组中活化型β-连环蛋白/总β-连环蛋白的百分比显著低于对照组,而FGF-2处理可逆转高糖诱导的β-连环蛋白抑制。3)体内实验表明,FGF-2处理后小鼠缺血后肢的血流恢复情况得到显著改善。外源性FGF-2可通过激活Wnt/β-连环蛋白信号通路,对处于高糖环境下受损的EPCs发挥功能修复作用。