Pro-angiogenic Ginsenosides F1 and Rh1 Inhibit Vascular Leakage by Modulating NR4A1

Ji In Kang,Daeyoup Lee,Sun Chang Kim,Ho Min Kim,Chang-Hau Cui,Yoonjung Choi

doi:10.1038/s41598-019-41115-2

Abstract

Vascular endothelial growth factor (VEGF) plays a key role in angiogenesis, but VEGF-induced angiogenesis is often accompanied by a vascular permeability response. Ginsenosides are triterpenoid saponins from the well-known medicinal plant, ginseng, and have been considered a candidate for modulating angiogenesis. Here, we systemically investigated the effects of 10 different ginsenosides on human umbilical vein endothelial cells and newly identified that two PPT-type ginsenosides, F1 and Rh1 induce the migration and proliferation of endothelial cells. Interestingly, RNA transcriptome analysis showed that gene regulation induced by VEGF in endothelial cells is distinct from that of ginsenoside F1 and Rh1. In addition, F1 and Rh1 significantly inhibited vascular leakage both in vitro and in vivo, which are induced by vascular endothelial growth factor. Furthermore, comparative transcriptome analysis revealed that these effects of F1 and Rh1 on vascular leakage restoration are mainly caused by changes in VEGF-mediated TNFα signaling via NFκB, particularly by the suppression of expression and transcriptional activity of NR4A1 by F1 and Rh1, even in the presence of VEGF. These findings demonstrate that ginsenosides F1 and Rh1 can be a promising herbal remedy for vessel normalization in ischemic disease and cancer and that NR4A1 is the key target.

Highlights

Angiogenesis is the process of new blood vessel formation from pre-existing vessels, and is essential for development, reproduction and repair[1]
We systemically investigated the angiogenic effects of these variants using a tube formation assay with human umbilical vein endothelial cells (HUVECs)
We found that F1 and Rh1 promote tube formation of HUVECs, as well as human retinal microvascular endothelial cells (HRMECs), in a dose-dependent manner (Fig. 1B and Supplementary Fig. 2B)

Summary

Introduction

Angiogenesis is the process of new blood vessel formation from pre-existing vessels, and is essential for development, reproduction and repair[1]. Paracrine activation of endothelial cells by VEGF is a major initiating event in angiogenesis, deregulated expression of VEGF and subsequently induced vascular leakage can promote edema and extensive tissue injury in ischemic disease and tumor cell extravasation, and metastasis in cancer[13,14]. Unlike VEGF, several pro-angiogenic factors such as angiopoietin-1 (ANG1), fibroblast growth factor (FGF) and hepatocyte growth factor (HGF) enhance endothelial junctional integrity and vascular barrier function, and can even block VEGF-induced vascular leakage[17,18,19]. PPD-type ginsenosides, Rb1 and Rk1, have been reported to inhibit vascular permeability induced by VEGF, lipopolysaccharide, thrombin, or histamine[20,21], it is not yet clear whether pro-angiogenic ginsenosides can induce vascular leakage similar to VEGF, or act as a leakage inhibitor, similar to ANG1, FGF and HGF. We revealed that F1 and Rh1 do not induce vascular leakage and can even inhibit VEGF-induced vascular leakage in vitro and in vivo by suppressing NR4A1’s transcriptional activity as well as decreasing the gene expression of NR4A1, which mediates acute and chronic vascular hyperpermeability[22]

Methods

Results

Conclusion