Objective  To investigate the potential molecular mechanisms underlying Cassiae Semen in treating constipation by integrating network pharmacology with molecular docking and molecular dynamics (MD) simulations, and provide references for the modernization of traditional Chinese medicine (TCM) and drug development.

Methods  Chemical constituents of Cassiae Semen were initially screened from the TCMSP database. Potential bioactive compounds were selected based on oral bioavailability (OB)≥30% and drug-likeness (DL)≥0.18, ranked in descending order by OB values. The target proteins of these compounds were predicted using SwissTargetPrediction. Constipation-related disease targets were retrieved from the GeneCards and OMIM databases. Overlapping targets between the compound targets and constipation-associated targets were identified using Venny 2.1.0. A "compound-target-disease" network was constructed to analyze key nodes. Functional enrichment analysis was performed using GO function and KEGG pathway analysis for the core targets. Molecular docking was conducted using AutoDockVina, and a 100  ns molecular dynamics simulation was performed using GROMACS to validate the stability of protein-ligand complexes.

Results  A total of 14 bioactive compounds were identified, with aloe-emodin, stigmasterol, and obtusin as the core components. A total of 54 potential targets were predicted, among which 48 were associated with constipation. The core targets included prostaglandin-endoperoxide synthase 2 (PTGS2), tumor necrosis factor (TNF), and tumor protein P53 (TP53). Molecular docking reveals that aloe-emodin, stigmasterol, and obtusin bind stably to the core targets. Dynamic simulations validate that the complex maintains homeostasis under physiological conditions.

Conclusion  Cassiae Semen exerts their anti-constipation effects by targeting key pathways such as TNF and PTGS2 through active constituents including aloe-emodin, stigmasterol, and obtusin, thereby modulating inflammatory responses and neuromodulation.

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