Elucidate the potential mechanism of Eucommiae Cortex against osteoporosis by network pharmacology and RNA-sequencing

Abstract

Abstract Purpose Eucommiae Cortex (Eucommia ulmoides Oliv., cortex) had possessed multiple curative effect since ancient time. Nevertheless, the mechanism of EC serves as anti-osteoporotic herb remains further investigated. Methods Cytotoxicity assay and osteogenesis assay were adopted to filtrate the TCMs and osteoporosis model rats of was utilized to verify the anti-osteoporosis ability of EC. Network pharmacology was used to investigate the potential mechanisms of the EC against osteoporosis. The database including TCMSP, BATMAN TCM and TCMID were utilized to obtain the active compounds of EC, and their potential targets were predicted by SwissTarget-Prediction. Osteoporosis related targets were found by OMIM, DisGeNET and Gene Cards databases. The target interaction network was analyzed by STRING, GO enrichment and KEGG pathway analysis were carried out by DAVID database. Results Results of in vitro and in vivo experiments illustrated that EC showed no cytotoxicity and exhibited anti osteoporosis effect. A total number of 19 active components and 124 osteoporosis related targets of the EC were selected. KEGG pathway enrichment from bioinformatics suggested that EC prevented osteoporosis through the HIF-1 signaling pathway and estrogen signaling pathway, while results of RNA- sequencing suggesting HIF-1 signaling pathway. Moreover, genes Akt1, MAPK3 and EGFR may serve as the critical targets regulated by EC. Conclusion Our results showed that HIF-1 signaling pathway was vital pathway in EC against osteoporosis, with the participation of gene AkT1, MAPK3 and EGFR. Estrogen and VEGF signaling pathway were synergetic pathway of anti-osteoporosis