RESUMEN
Arnebin-1, a naphthoquinone derivative, plays a crucial role in the wound healing properties of Zicao (a traditional wound healing herbal medicine). It has been noted that Arnebin-1, in conjunction with vascular endothelial growth factor (VEGF), exerts a synergistic pro-angiogenic effect on human umbilical vein endothelial cells (HUVECs) and accelerates the healing process of diabetic wounds. However, the mechanisms responsible for the pro-angiogenic effect of arnebin1 on HUVECs and its healing effect on diabetic wounds have not yet been fully elucidated. In this study, in an aim to elucidate these mechanisms of action of arnebin1, we investigated the effects of arnebin1 on the VEGF receptor 2 (VEGFR2) and the phosphoinositide 3-kinase (PI3K)dependent signaling pathways in HUVECs treated with VEGF by western blot analysis. The proangiogenic effects of arnebin1 on HUVECs, including its effects on proliferation and migration, were evaluated by MTT assay, Transwell assay and tube formation assay in vitro. The expression levels of hypoxia-inducible factor (HIF)1α, endothelial nitric oxide synthase (eNOS) and VEGF were determined by western blot analysis in the HUVECs and wound tissues obtained from nondiabetic and diabetic rats. CD31 expression in the rat wounds was evaluated by immunofluorescence staining. We found that the activation of the VEGFR2 signaling pathway induced by VEGF was enhanced by arnebin1. Arnebin1 promoted endothelial cell proliferation, migration and tube formation through the PI3Kdependent pathway. Moreover, Arnebin1 significantly increased the eNOS, VEGF and HIF1α expression levels in the HUVECs and accelerated the healing of diabetic wounds through the PI3Kdependent signaling pathway. CD31 expression was markedly enhanced in the wounds of diabetic rats treated with arnebin1 compared to the wounds of untreated diabetic rats. Therefore, the findings of the present study indicate that arnebin-1 promotes the wound healing process in diabetic rats by eliciting a pro-angiogenic response.