Key Laboratory of Molecular Target and Clinical Pharmacology, School of Pharmaceutical Sciences and The Fifth Affiliated Hospital, Guangzhou, Guangdong 511436, P.R. China.
Department of Neonatology, Maternal and Children Hospital of Guangdong Province, Guangzhou, Guangdong 510260, P.R. China.
Mol Med Rep. 2022 Jan;25(1). doi: 10.3892/mmr.2021.12521. Epub 2021 Nov 5.
Modulating the biological status of endothelial progenitor cells (EPCs), such as function and survival, is essential for therapeutic angiogenesis in ischemic vascular disease environments. This study aimed to explore the role and molecular mechanisms underlying Netrin‑1 in the viability and angiogenic function of EPCs. EPCs were isolated from the bone barrow of adult C57/BL6 mice. The apoptosis and various functions of EPCs were analyzed by manipulating the expression of Netrin‑1. The TUNEL assay was performed to detect apoptotic EPCs. Cell migration and tube formation assays were performed to detect EPC function. Trypan blue staining was performed to detect cell viability. Western blot analysis was performed to detect the protein expression levels of Netrin‑1, CD146 and apoptotic factors. Quantitative PCR analysis was performed to detect the expression levels of Netrin‑1 receptors. The results demonstrated that treatment with exogenous Netrin‑1 promoted EPC migration and tube formation, whereas transfection with small interfering (si)RNA targeting Netrin‑1 exhibited the opposite effects. Exogenous Netrin‑1 protected EPCs from hypoxia‑induced apoptosis, whereas the interruption of endogenous Netrin‑1 enhancement under hypoxia by Netrin‑1‑siRNA exacerbated the apoptosis of EPCs. Furthermore, CD146, one of the immunoglobulin receptors activated by Netrin‑1, was screened for in the present study. Results demonstrated that CD146 did not participate in Netrin‑1‑promoted EPC function, but mediated the anti‑apoptotic effects of Netrin‑1 in EPCs. In conclusion, Netrin‑1 enhanced the angiogenic function of EPCs and alleviated hypoxia‑induced apoptosis, which was mediated by CD146. This biological function of Netrin‑1 may provide a potential therapeutic option to promote EPCs for the treatment of ischemic vascular diseases.
调节内皮祖细胞（EPC）的生物学状态，如功能和存活，对于缺血性血管疾病环境中的治疗性血管生成至关重要。本研究旨在探讨 Netrin-1 在 EPC 活力和血管生成功能中的作用和分子机制。EPC 从成年 C57/BL6 小鼠的骨髓中分离出来。通过操纵 Netrin-1 的表达来分析 EPC 的凋亡和各种功能。TUNEL 检测法用于检测凋亡的 EPC。细胞迁移和管形成试验用于检测 EPC 功能。台盼蓝染色用于检测细胞活力。Western blot 分析用于检测 Netrin-1、CD146 和凋亡因子的蛋白表达水平。实时定量 PCR 分析用于检测 Netrin-1 受体的表达水平。结果表明，外源性 Netrin-1 促进 EPC 迁移和管形成，而靶向 Netrin-1 的小干扰 (si)RNA 转染则表现出相反的效果。外源性 Netrin-1 可保护 EPC 免受缺氧诱导的凋亡，而 Netrin-1-siRNA 阻断缺氧下内源性 Netrin-1 增强作用则加剧了 EPC 的凋亡。此外，本研究还筛选了 Netrin-1 激活的免疫球蛋白受体之一 CD146。结果表明，CD146 不参与 Netrin-1 促进的 EPC 功能，但介导 Netrin-1 在 EPC 中的抗凋亡作用。综上所述，Netrin-1 增强了 EPC 的血管生成功能并减轻了缺氧诱导的凋亡，这是由 CD146 介导的。Netrin-1 的这种生物学功能可能为促进 EPC 治疗缺血性血管疾病提供一种潜在的治疗选择。
