Abstract
Formononetin (FMNT) is a major bioactive compound from Astragalus membranaceus (Fisch.) Bunge, and has been widely used to treat conditions related to vascular insufficiency. However, the molecular mechanism for the therapeutic effect has not been well defined. This study aimed to investigate the effect and mechanism of FMNT on endothelial function. The potential targets and signaling pathways of FMNT in the setting of ischemia were predicted using network pharmacology analysis. Human umbilical vein endothelial cells (HUVECs) were used for the in vitro studies and C57BL/6 mice were used for in vivo experiments. The results of the network pharmacology analysis showed that multiple signaling molecules including MAPK and PI3K-Akt pathways could be involved in the pharmacological actions of FMNT against ischemic diseases. The experimental validation data showed that FMNT significantly promoted the growth, proliferation, migration and tube formation of HUVECs in association with activation of endothelial nitric oxide synthase (eNOS) and promotion of intracellular nitric oxide (NO) production. FMNT also markedly activated Erk1/2 and Akt signaling in HUVECs. The enhanced endothelial function by FMNT was abolished when the cells were pre-treated with eNOS inhibitor. FMNT-induced eNOS/NO activation, endothelial function and angiogenesis was also effectively attenuated when Erk1/2 or Akt signaling pathway was inhibited. In addition, FMNT significantly promoted wound healing in C57BL/6 mice associated with activation of Erk1/2 and Akt signaling. Enhanced wound healing by FMNT in mice was prevented when eNOS-, Erk1/2, or Akt-medicated signaling was inhibited. Moreover, when Akt signaling was inhibited in HUVECs, FMNT was still able to activate Erk1/2 signaling without promotion of endothelial function. Similarly, FMNT could activate Akt signaling with no change in endothelial function when Erk1/2 signaling was attenuated in HUVECs. Conclusively, the present study demonstrated that FMNT significantly enhanced endothelial function and promoted angiogenesis in vitro and in vivo through activating Erk1/2- and Akt-mediated eNOS/NO signaling pathway. The data also suggested that simultaneous activation of Erk1/2 and Akt signaling was required for FMNT-induced promotion of endothelial function. Results from the present study might provide support and evidence for the application of FMNT during the clinical treatment of conditions related to vascular insufficiency.
Highlights
Endothelial cells play a vital role in the regulation of vascular function including angiogenesis (Carmeliet, 2003; Bauer et al, 2005; Falanga, 2005)
After analyzing the databases of TCMSP and PubChem (Hua et al, 2019; Zhu et al, 2019), a total of 54 potential pharmacological targets were identified for FMNT and the target network was visualized by Cytoscape 3.7.2 software (Gan et al, 2019; Zhu et al, 2019) (Figure 1A)
A group of 1096 genes associated with ischemia were identified through the CTD database (Gan et al, 2019) (Figure 1B). 27 potential targets of FMNT were associated with ischemia (Figure 1C)
Summary
Endothelial cells play a vital role in the regulation of vascular function including angiogenesis (Carmeliet, 2003; Bauer et al, 2005; Falanga, 2005). Erk1/2 signaling is critical to the regulation of eNOS phosphorylation and NO production, and endothelial function including survival, proliferation, migration, angiogenesis, and tube formation (Buckley et al, 1999; Meadows et al, 2001; Wang et al, 2013; Zhang et al, 2017). It is unknown whether FMNT could enhance endothelial function via PI3K/Akt- and/or Erk1/2mediated eNOS/NO signaling pathway
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
