CD146 mediates the anti-apoptotic role of Netrin-1 in endothelial progenitor cells under hypoxic conditions

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 in vitro 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.