The Mechanism of Ermiao Powder In Treating Prostatitis: A Study Based on Network Pharmacology and Experimental Verification.

This protocol presents a combined network pharmacology and experimental validation approach to investigate the mechanism of action of Ermiao Powder (EMP) in the treatment of prostatitis. The workflow follows a stepwise design: (i) database-driven screening of active compounds and targets, (ii) construction and analysis of interaction networks, and (iii) in vitro experimental validation. First, active ingredients of EMP and their corresponding targets were retrieved from the TCMSP database, while prostatitis-related targets were obtained from the GeneCards database. The intersection of EMP targets and disease targets was used to identify key therapeutic targets. Second, a "Drug-Active Ingredient-Target-Disease" network was constructed using Cytoscape. Protein-protein interaction (PPI) analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment were performed on the key targets, revealing that inflammatory factors such as IL6 and TNF served as core targets, and pathways including PI3K/AKT and MAPK were significantly enriched. Finally, in vitro validation was conducted using an LPS-induced inflammatory model of prostate epithelial cells. Treatment with EMP significantly reduced the release of pro-inflammatory factors IL-6 and TNF-α compared to the LPS-treated group (p < 0.05), confirming the anti-inflammatory efficacy of EMP. This integrative approach provides a reproducible framework that can be adapted to investigate other multi-component therapeutic systems. In conclusion, this study preliminarily elucidates that EMP exerts its anti-prostatitis effects through a multi-component, multi-target, and multi-pathway mechanism, closely associated with the suppression of inflammatory responses, thereby offering a scientific basis for its clinical application.