Integrative network pharmacology, molecular dynamics simulation, and single-cell RNA sequencing strategies reveal the multi-target mechanisms of oridonin against cervical cancer.

Introduction: Cervical cancer remains a major threat to women's health worldwide, and effective therapeutic strategies with clear molecular mechanisms are still needed. Oridonin (Ori), a natural diterpenoid compound, has shown antitumor activity in multiple malignancies; however, its regulatory mechanisms in cervical cancer, particularly at the single-cell level, remain incompletely understood. Methods: In this study, an integrative strategy combining network pharmacology, bulk transcriptomic analyses, single-cell RNA sequencing (scRNA-seq), molecular docking, molecular dynamics simulations, and Results: Network pharmacology and transcriptomic analyses identified 25 core EMT-related targets of oridonin, with enrichment primarily in the PI3K/AKT signaling pathway. Immune infiltration analysis revealed that oridonin-associated targets were closely correlated with macrophages and T-cell subsets. Single-cell analysis demonstrated that PI3K/AKT and EMT-related pathways were predominantly enriched in epithelial tumor cells, suggesting cell-type-specific regulatory effects. Molecular docking and dynamics simulations indicated favorable binding potential between oridonin and AKT1. Conclusion: This study provides a systematic and multi-level characterization of the antitumor mechanisms of oridonin in cervical cancer. By integrating network pharmacology with single-cell transcriptomic analysis, our findings highlight the cell-type-specific modulation of the PI3K/AKT-EMT axis by oridonin, offering mechanistic insights and establishing a theoretical pharmacological basis that warrants future