Epimedium/Icariin promotes angiogenesis through HIF-1 signaling pathway

Abstract

Objective, With the help of network pharmacology and in vitro studies, this study aims to understand the mechanism by which Epimedium/ Icariin (EPI/ICA) promote angiogenesis through the HIF-1 signaling pathway, laying the groundwork for the treatment of bone discontinuity. Methods, 1) The TCMSP database provided the gene targets for Epimedium's effect in this investigation. Search the GeneCards database for gene targets associated with "bone discontinuity". GO, KEGG, and Reactome enrichment analysis is performed after identifying possible gene targets for EPI/ICA treatment of bone discontinuity. Molecular docking to confirm the HIF-1 pathway's essential proteins' capacity to bind to ICA. 2) ①. Human umbilical vein vascular endothelial cells (HUVECs) were divided into blank control groups and ICA groups with different concentrations (0.01, 0.1, 1, 10 mol/L), and CCK8 measured the proliferative activity of HUVECs in each group after ICA intervention. These in vitro experiments sought to identify the optimal intervention concentration of ICA. ②. A test for angiogenesis to determine whether it can encourage angiogenesis. ③. After ICA intervention, Western blot and Elisa measured the level of HIF-1 pathway-related protein activity in each group. Results, For the treatment of bone discontinuity, the bioinformatic analysis of EPI/ICA identified 164 potential therapeutic gene targets, including 45 important gene targets and 10 of these crucial gene targets (HIF1A, AKT1, TPT3, IL6, TNF, VEGFA, CASP3, MYC, EGFR, ESR1). The biological mechanisms examined in the GO of EPI/ICA for the treatment of osteonecrosis act primarily through the regulation of angiogenesis and the negative regulation of apoptotic processes, according to enrichment analysis of 45 important gene targets. The HIF-1 signaling pathway, TNF signaling pathway, IL-17 signaling pathway, and other signaling pathways were found to be primarily involved in the regulation of the EPI/ICA process in the therapy of bone discontinuity, according to KEGG analysis. Reactome enrichment analysis showed that the regulation of the EPI/ICA treatment of bone discontinuity mostly involves the regulation of inflammatory factors that are similar to interleukin and the body's autoimmune system. According to the results of molecular docking, ICA binds efficiently to the important target proteins of the HIF-1 signaling pathway, HIF-1A, VEGF, and IL-6. Further research revealed that ICA, the active ingredient in Epimedium, might increase the value-added differentiation of HUVECs, with the potential mechanism being angiogenesis via the HIF-1 signaling pathway. Conclusion: This research revealed that EPI/ICA controls HIF-1A, VEGF, and IL-6, which are important HIF-1 signaling pathway target proteins, increases the value-added differentiation of HUVECs and angiogenesis, and offers a new approach for the treatment of bone discontinuity using EPI/ICA.