Arnebin-1 promotes angiogenesis by inducing eNOS, VEGF and HIF-1α expression through the PI3K-dependent pathway

Abstract

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 arnebin-1 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 arnebin-1, we investigated the effects of arnebin-1 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 pro-angiogenic effects of arnebin-1 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 non-diabetic 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 arnebin-1. Arnebin-1 promoted endothelial cell proliferation, migration and tube formation through the PI3K-dependent pathway. Moreover, Arnebin-1 significantly increased the eNOS, VEGF and HIF-1α expression levels in the HUVECs and accelerated the healing of diabetic wounds through the PI3K-dependent signaling pathway. CD31 expression was markedly enhanced in the wounds of diabetic rats treated with arnebin-1 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.

Endometrial stem cells repair injured endometrium and induce angiogenesis via AKT and ERK pathways

“…AKT and MAPK signal pathways play a vital role in angiogenesis , Zeng et al 2015, Lv et al 2016. In this study, we also found that phosphorylated level of AKT and ERK in HUVECs increased notably after treatment with CM-48h ( Fig.…”

“…Several studies had found that stem cells could paracrine a panel of cytokines and growth factors, contributing to repair of injured tissues (Kinnaird et al 2004a, Boomsma & Geenen 2012, Rahbarghazi et al 2013. AKT and MAPK signal pathway had been proved to play a vital role in angiogenesis , Zeng et al 2015, Lv et al 2016. Interestingly, we found that AKT and ERK could be activated by MenSCs-CM and the transcriptional expression level of eNOS, VEGFA, VEGFR1, VEGFR2 and TIE2 in HUVECs, which are the downstream targets of AKT and ERK pathway, increased after MenSCs-CM treatment.…”

Bufalin enhances anti-angiogenic effect of sorafenib via AKT/VEGF signaling

“…Serine-threonine protein kinase AKT is a major downstream target of PI3K for regulating tumor growth and angiogenesis (37,38). For instance, PI3K/AKT may regulate angiogenesis by several downstream targets such as mTOR/p70S6K1, FOXO, NOS, and GSK-3β (39)(40)(41).A study has reported that Arnebin-1 promotes angiogenesis by inducing eNOS, VEGF and HIF-1 expression through the PI3K-dependent pathway (42).…”

INPP4B overexpression suppresses migration, invasion and angiogenesis of human prostate cancer cells

“…In this study, we found that PC3 and DU145 cells transfected with the Flag‐INPP4B plasmid showed significantly decreased protein levels of p‐Akt, but this had no significant influence on the total protein levels of Akt. It was confirmed that PI3K/Akt pathway functions upstream of VEGF . Therefore, we assumed that INPP4B decreased VEGF secretion through inhibiting the activation of the PI3K/Akt signalling pathway.…”