Molecular mechanism study on the regulation of osteoporosis by Congrongyishen Granules based on network pharmacology and its preliminary validation

Objective:To explore the effective targets and signaling pathways of Congrongyishen Granules in the treatment of osteoporosis and investigate its potential molecular mechanism through network pharmacology.Methods:First, cellular assay was used to verify whether Congrongyishen Granules could inhibit osteoclast dif-ferentiation in vitro.Then, the TCMSP database was used to screen the active ingredients and targets of Congron-gyishen Granules, and the GeneCards database was used to obtain the relevant targets for osteoporosis, with the target information normalized in the Uniprot database.After that, the intersection of the both was taken to obtain the intersection genes of Congrongyishen Granules for the treatment of osteoporosis.Finally, protein-protein in-teraction (PPI) analysis was performed using STRING 11.0 database for drug and disease target genes to con-struct network diagrams, which were uploaded to Cytoscape 3.9.1 software to produce PPI network diagrams and whose potential protein functional modules were analyzed using MCODE.GO enrichment analysis and KEGG pathway analysis were also performed on the core target genes using Metascape database.Results:The results of cellular assay showed that Congrongyishen Granules could inhibit osteoclastogenesis in vitro.There were 69 ac-tive ingredients of Congrongyishen Granules, corre-sponding to 220 targets, and there were 1, 167 osteo-porosis-related disease targets.Among them, the main active ingredients of Congrongyishen Granules for osteoporosis were quercetin, β-sitosterol, kaempferol, isorham-netin and sesamin, etc.The core targets were NCOA2, PTGS2, PTGS1, HSP90AB1 and PIK3CG, etc.The KEGG pathway involved cancer signaling pathway, AGE-RAGE signaling pathway, NF-κB signaling pathway, VEGF signaling pathway, etc.Conclusion:The multi-component, multi-target, and multi-pathway features of Congrongyishen Granules are revealed by network pharmacology, providing a foundation for future study on the mechanism of Congrongyishen Granules in the treatment of osteoporosis.