Expression and secretion of neuregulin-1 in cardiac microvascular endothelial cells treated with angiogenic factors.

Abstract

Neuregulin-1 (NRG-1) is a positive regulator of angiogenesis, which suggests there may be an association between NRG-1 and angiogenic factors. The aim of the present study was to investigate the effect of treating human cardiac microvascular endothelial cells (HCMECs) with angiogenic factors on NRG-1 expression and secretion. HCMECs were cultured and stimulated with vascular endothelial growth factor (VEGF; 100 ng/ml), angiopoietin (Ang)-1 (100 ng/ml) or Ang-2 (100 ng/ml) under normal or hypoxia/serum deprivation (Hypo/SD) conditions for 24 h. The expression of ErbB receptors and NRG-1 in HCMECs was measured by western blot analysis and the secretion of NRG-1 in HCMECs was determined by ELISA. The results demonstrated that ErbB2, ErbB3 and ErbB4 were expressed in HCMECs and that ErbB2 expression levels were notably higher than those of ErbB3 and ErbB4. Under normal culture conditions the expression and secretion of NRG-1 was significantly increased in HCMECs treated with VEGF or Ang-1 (P<0.05), however levels significantly decreased in HCMECs treated with Ang-2 (P<0.05). Under Hypo/SD conditions the expression and secretion of NRG-1 significantly increased (P<0.05) and VEGF or Ang-1 treatment significantly increased these effects further (P<0.05). Conversely Ang-2 treatment significantly decreased these effects (P<0.05). The expression and release of NRG-1 were significantly increased in HCMECs with VEGF or Ang-1 treatment (P<0.05), which suggests that VEGF and Ang-1 may regulate myocardial angiogenesis and survival via the NRG-1/ErbB signaling pathway.