Systematic Pharmacology-Based Strategy to Investigate the Mechanism of Vladimiriae Radix in Benign Prostatic Hyperplasia

Objective: The present study aimed to elucidate the mechanism of action of the dried roots of Vladimiriae Radix against benign prostatic hyperplasia (BPH) using network pharmacology, molecular docking, and cell-level experimental verification technologies, thereby providing experimental evidence for basic research, clinical application, and the modernization research of Tibetan-Chinese medicine integrated medication. Methods: Firstly, the active components of Vladimiriae Radix were screened using the TCM Systems Pharmacology Database and Analysis Platform (TCMSP) and PubChem, with the criteria of oral bioavailability (OB) ≥30% and drug-likeness (DL) ≥0.18. Subsequently, the SwissTargetPrediction database was used to identify potential targets for the components, and an overlap analysis was conducted on the BPH-related targets from GeneCards, Online Mendelian Inheritance in Man (OMIM), and the Therapeutic Target Database (TTD) to identify the common targets. Then, STRING and Cytoscape 3.10.3 analyses were used to construct the protein-protein interaction (PPI) network and the “Chinese medicine-component-target-disease” network for screening core targets. Gene ontology (GO)/kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses were performed using the Database for Annotation, Visualization and Integrated Discovery (DAVID) database and bioinformatics platforms. Discovery Studio 2019 was used to verify the binding between components and targets, and AutoDockTools1-2 was employed to calculate the binding energy. Cell-level experiments (CCK-8 assay and RT-PCR) were conducted using BPH-1 cells to validate the effect of the representative component oleanolic acid. Results: A total of 235 common targets were identified between Vladimiriae Radix and BPH, and 6 core targets, including AR, CYP17A1, CYP19A1, ACHE, F2, and HMGCR, were further screened. These core targets are mainly involved in biological functions such as steroid hormone response, cellular response to nutrient levels, and regulation of membrane potential, and are enriched in BPH-related pathways including lipid and atherosclerosis, cholinergic synapse, and AGE-RAGE. Molecular docking verification found that the active components form stable bindings with the core targets. Cell experiments found that oleanolic acid significantly inhibits BPH-1 cell proliferation and regulates the mRNA expression of the six core targets at concentration of 10 μM, 20 μM, 40 μM (significantly downregulated the mRNA expression of AR and HMGCR (p < 0.05), significantly upregulated the mRNA expression of CYP17A1, CYP19A1, and ACHE (p < 0.05), and had no significant effect on F2). Conclusion: Costunolide, dehydrocostus lactone, luteolin, quercetin, taraxasterol and oleanolic acid are the main bioactive ingredients in Vladimiriae Radix. Among them, oleanolic acid exhibited the highest binding energy with 6 core targets and exhibits anti-BPH properties. The present study fills the research gap in the anti-BPH mechanism of Vladimiriae Radix, validates the efficacy of the active components in Vladimiriae Radix at the cellular level, and provides clear targets and theoretical support for subsequent pharmacological verification, active component development, and clinical translation.