Network pharmacology and molecular dynamics simulation elucidate the potential mechanism of Batatasin-III in Bletilla...

Batatasin-III, a phenanthrene compound isolated from Bletilla striata, has demonstrated potential anti-inflammatory and immunomodulatory effects, yet its precise molecular mechanism against ulcerative colitis (UC) remains largely unexplored. This study integrates network pharmacology, molecular docking, ADMET profiling, and molecular dynamics (MD) simulations to systematically elucidate the multitarget therapeutic potential of Batatasin-III in UC treatment. Batatasin-III-related targets were retrieved from SwissTargetPrediction, while UC-associated genes were collected from GeneCards and OMIM databases. A total of 101 intersecting genes were identified and subjected to PPI network construction using STRING and topological analysis in Cytoscape. GO and KEGG enrichment analyses revealed significant involvement in key biological processes and pathways such as MAPK signaling, PI3K-Akt signaling, protein phosphorylation, and cytokine-mediated inflammation. Molecular docking showed strong binding affinities between Batatasin-III and core targets ALB (- 8.4 kcal/mol), MAPK3 (- 8.2 kcal/mol), ESR1 (- 7.7 kcal/mol), and HSP90AA1 (- 5.7 kcal/mol). ADMET evaluation via ADMETlab 3.0 predicted favorable drug-likeness, bioavailability, and low toxicity for Batatasin-III. Subsequent 100-ns MD simulations demonstrated high conformational stability (RMSD < 3.7 Å), sustained hydrogen bonding, and compact binding dynamics, particularly in ESR1-Batatasin-III and MAPK3-Batatasin-III complexes. MM/PBSA binding free energy analysis supported strong binding thermodynamics, with ALB-Batatasin-III exhibiting the most favorable ΔG_bind (- 29.65 kcal/mol). Residue energy decomposition highlighted critical contributions from TYR411, MET125, HIS524, and ASN171, among others. To validate these computational predictions, in vitro assays were conducted. A CCK-8 assay confirmed Batatasin-III was non-cytotoxic to RAW 264.7 macrophages. In an LPS-stimulated model, Batatasin-III significantly and dose-dependently inhibited the mRNA expression of key pro-inflammatory mediators, including TNF-α, IL-6, IL-1β, and NOS2. Overall, Batatasin-III may exert therapeutic effects against UC through multitarget modulation of inflammation, kinase regulation, and epithelial repair, primarily via the MAPK and PI3K-Akt pathways. This study provides a validated mechanistic foundation for Batatasin-III as a potential bioactive compound for UC intervention and supports further in vivo validation.